Fragment Coupling Approach To C₁₉- AND C₂₀-Diterpenoid Alkaloids: Total Synthesis of (–)-Talatisamine, (–)-Liljestrandisine, and (–)-Liljestrandinine Citation Fastuca, Nicholas James (2022) Fragment Coupling Approach To C₁₉- AND C₂₀-Diterpenoid Alkaloids: Total Synthesis of (–)-Talatisamine, (–)-Liljestrandisine, and (–)-Liljestrandinine. Dissertation (Ph.D.), California Institute of Technology. doi:10.7907/vf11-jy04. https://resolver.caltech.edu/CaltechTHESIS:05292022-214045444 Abstract A unified, convergent fragment coupling approach to the C₁₉- and C₂₀-diterpenoid alkaloid natural products is presented. The highly-caged aconitine, denudatine, and napelline cores are disconnected through the central B-ring cyclohexane to an A/E/F-ring fragment common to the structures all three subfamilies. 1,2-addition of an appropriate organometallic C/D-bicycle to an A/F-ring hydrindane epoxy ketone fragment followed by a Lewis acid-catalyzed semipinacol reaction couples the two fragments together and sets a key all-carbon quaternary center at C11. This strategy is realized in the synthesis of the C₁₉-aconitine core by using a [3.2.1]-bicyclooctene C/D-fragment as the nucleophile in the 1,2-addition. This C/D-fragment is prepared using a meta -photocycloaddition; this represents an alternative approach to the commonly employed biomimetic Wagner-Meerwein rearrangement of a [2.2.2]-bicyclooctane. To complete the aconitine core, a radical cyclization cascade to form the E-ring piperidine and B-ring cyclohexane in a single step is investigated. N -centered radicals were evaluated to initiate the cascade via a 6- exo -trig cyclization. A neutral aminyl radical gave rise to an unexpected Hoffman-Löffler-Freytag type product resulting from 1,5-hydrogen atom transfer. Employing Lewis-acidic single electron reducing metal catalyst led to formation of the E-ring cyclized product, however the second cyclization to close the B-ring did not occur. As an alternative approach, the E-ring was closed via an intramolecular aziridination. Treatment of this aziridine with acetyl bromide results in an aziridine-opened alkyl bromide product. This alkyl bromide is used as a functional group handle to form the final ring of the aconitine core. From there, the total synteheses of the C₁₉-diterpenoid alkaloids (–)-talatisamine, (–)-liljestrandisine, and (–)-liljestrandinine were completed in short order. These synthetic efforts led to revision of the proposed structure of (–)-liljestrandisine. Item Type: Thesis (Dissertation (Ph.D.)) Subject Keywords: diterpenoid; alkaloid; natural product; total synthesis; fragment-coupling Degree Grantor: California Institute of Technology Division: Chemistry and Chemical Engineering Major Option: Chemistry Thesis Availability: Public (worldwide access) Research Advisor(s): Reisman, Sarah E. Thesis Committee: Stoltz, Brian M. (chair) Fu, Gregory C. Peters, Jonas C. Reisman, Sarah E. Defense Date: 26 April 2022 Funders: Funding Agency Grant Number NIH T32GM007616 Record Number: CaltechTHESIS:05292022-214045444 Persistent URL: https://resolver.caltech.edu/CaltechTHESIS:05292022-214045444 DOI: 10.7907/vf11-jy04 Related URLs: URL URL Type Description https://doi.org/10.1021/acscentsci.1c00540 DOI Article adapted for portions of Chapters 2–4. https://doi.org/10.15227/orgsyn.097.0327 DOI Article adapted for a portion of Chapter 2. ORCID: Author ORCID Fastuca, Nicholas James 0000-0003-4081-6031 Default Usage Policy: No commercial reproduction, distribution, display or performance rights in this work are provided. ID Code: 14650 Collection: CaltechTHESIS Deposited By: Nicholas Fastuca Deposited On: 07 Jun 2022 15:35 Last Modified: 08 Nov 2023 00:42 Thesis Files PDF - Final Version See Usage Policy. 18MB Repository Staff Only: item control page

FRAGMENT COUPLING APPROACH TO C19- AND C20-DITERPENOID ALKALOIDS: TOTAL SYNTHESIS OF (–)-TALATISAMINE, (–)LILJESTRANDISINE, AND (–)-LILJESTRANDININE Thesis by Nicholas James Fastuca In Partial Fulfillment of the Requirements for the Degree of Doctor of Philosophy CALIFORNIA INSTITUTE OF TECHNOLOGY Pasadena, California 2022 (Defended April 26th, 2022) ii © 2022 Nicholas James Fastuca All Rights Reserved ORCID: 0000-0003-4081-6031 iii ACKNOWLEDGEMENTS I am so grateful for all of the wonderful people in my life who have encouraged and supported me over the years and made my life so fulfilling. I consider myself extremely lucky to be surrounded by such caring people. I would first like to thank my family for all of their love and support. As a young person, it is easy to take for granted how lucky I am to have a family who are always there for me, without judgement, whenever I need their help. Over these past years, I’ve realized how fortunate I am to know, without a second thought, that they would drop everything and help me whenever I need it. To my parents, Debra and Stephen, thank you for all you have done for us. You have always encouraged us to follow our own paths in life, giving us a sense of freedom to do what we feel is best and not worry what you will think of it. Thank you for all of the support. To my sisters, Jessica and Ally, thank you for being such caring people and inspiring me to better myself. I love you all, and I’m looking forward to getting to spend more time with you all when I move back to Philadelphia. I am very fortunate to have a wonderful (and large) extended family as well. Growing up with 16 aunts and uncles, 14 cousins and 4 loving grandparents was always a lot of fun. Going over to see Lou, Joe, and Anthony on the weekends, seeing you all attend my sporting events as a kid, holiday dinners full of laughs are treasured memories I will have forever. I’m so thankful to my grandparents Mary and Salvatore Fastuca and Katherine and James Knowles for creating such a close-knit family environment for all of us. We are all very different people, but we come together so well. iv Thank you to my Aunt Monette and Uncle Robert for being like a second set of parents. Growing up around the corner from you all and spending so much time with Lou, Joe, and Anthony as kids was great. Thank you to my Aunt Carol Fastuca for her guidance and support for navigating graduate school as a young adult. I always feel better about the future after we talk! Uncle Jimmy, thank you for always being there for us and for all of the memories as a kid. I’ll never forget my first Phillies game at Citizens Bank Park with you! Thank you to my Aunt Nancy and Uncle Donald for all of the fun times we’ve spent together and for always believing in me. I’d next like to thank all of the people who took the time and made a conscious effort to mentor me over my career in chemistry. To my high school chemistry teacher Josh Gellner, thanks for going above and beyond to help me find a love for chemistry. To my academic advisor at Penn State, Dr. Joseph Keiser, thank you for all of the confidence you inspired in me. I always looked forward to our meetings in your office—I could tell you really cared about me and all of your students as people and wanted to see us succeed. Thanks for helping to make the undergraduate chemistry program at Penn State such an exciting environment. You and your colleagues have built a great community environment for students; we always felt supported—even after class was over. I have great memories from my time in Whitmore Lab. Thank you to Dr. Katherine Masters for inspiring a love for organic chemistry in me! You made the material really exciting and gave us confidence to tackle hard problems. Your mechanisms and spectroscopy classes were some of my favorite ever! Thank you to Professor Alex Radosevich for believing in me and giving me the opportunity to start learning to do research. I was extremely v nervous coming into your lab, and you helped me develop a lot of confidence. Thanks for all of the time you set aside to teach me and challenge me to think and grow. As for my time at Caltech, I thank Professor Sarah Reisman for giving me the opportunity to work on such exciting projects. Again, I was very nervous starting work on total synthesis, so thank you for giving me space to learn and grow. Thank you for advising me, pushing me to get the most out of my work, and for putting so much effort into teaching. I learned so much from the synthesis class and really enjoyed it. Thank you for your support during tough times as well. Thanks to my thesis committee chairman Professor Brian Stoltz for helping me navigate through the degree requirements. This has been really difficult, and your help and support has meant a lot. Thank you for your passion for teaching as well; starting classes at Caltech with your 242a where there was “only one stupid question” helped me feel a lot less intimidated by total synthesis. It was also great to learn from you during our group meetings—the relationship between your lab and Sarah’s made for a great environment to work and grow. Thank you to the other two members of my committee, Professor Gregory Fu and Professor Jonas Peters, for all of your helpful feedback during our committee meetings and proposals exam. Thank you to Dr. Scott Virgil for all your advice and assistance on projects in the lab. The way you solve any problem we present you with inspires me to work to take on new challenges outside of my comfort zone. Thank you for all you do for the department. Thank you to Dr. David VanderVelde for always taking time to help us solve our structures by proposing experiments and helping us to execute them and interpret their results. Your dedication to the department and to maintaining a world-class NMR facility vi is greatly appreciated. In addition, thank you for the opportunity to work with you as a GLA and to learn more about NMR. Thank you to Lynne Martinez, Veronica Triay, and Beth Marshall for all of their help and for always being there to talk. Thank you to Alison Ross for keeping the department running smoothly and helping graduate students to have a wonderful experience at Caltech. For my mentors in lab, thank you to Dr. Denise Grünenfelder for being my first mentor on the acutumine project. Thanks for being patient with me as I learned and for teaching me how to do things the right way in lab! Thanks for all of the fun times outside of the lab as well; it was great to have a friend after moving across the country. I’d like to thank my project partner and mentor Dr. Alice Wong: you encouraged me to believe in myself and my ideas. I really loved talking about the chemistry with you, going over data and potential routes. Thanks for all of your help getting me up to speed on all of the literature and work that had been done before I joined the project. You put your heart and soul into your project, so I appreciate having the opportunity to work on it with you. As my last mentor, thank you to Dr. Yasuaki Nakayama for going out of your way to help me develop skills for running tough reactions. The effort and process you put into your project was really inspiring and helped me learn a lot. One thing I will never forget is how the structures in your notebook overlap perfectly from page to page! I want to thank Dr. Victor Mak, Professor Susan Stevenson, and Jeff Kerkovius for being great project partners. It’s been great to learn from all of you and work on such a fun project! vii I want to thank all of the people I’ve had the opportunity to work with in our lab. I want to thank Caitlin Lacker and Skyler Mendoza for being the best cohort I could have asked for. You are some of the kindest, most selfless people I have ever met. I’m so lucky to have had you two to lean on during grad school—I don’t know if I could have done it without you. Thanks for always being there for me when times were tough. I have so many wonderful memories with you that have been the highlight of my time in grad school. Our trip to Yosemite was one of the most fun trips I’ve ever been on—and half of the fun times were just from the car ride! I loved goofing around with you guys in lab and making up handshakes. Caitlin, I loved when you would come by my hood while waiting for the glovebox or rotovap and chat. Skyler, I loved sitting next to you in the office and feeding off of each other’s hyper energy at times. I also loved when you would bake something you saw on GBB and let us eat it. I hope we can have more tea times together soon! Alice Wong, thanks for being such a great friend. Thanks for taking me under your wing and helping me feel at home in Pasadena. I had a hard time moving here and not knowing anyone, but once we became friends, it all changed for the better. Hanging out at 533 eating Oakland hippie food, making pizza, or baking an impromptu cake were some of the best times I had in grad school. Thanks for always making time to talk to me and for just being a great friend. To Ray Turro, thanks for always being there for me when times were tough. I’m so glad I was able to bully you into being my friend. When we first met when you were a G0 working in the hood next to me, I knew right away you were a great person. Thanks for introducing me to cats, beer brewing, and board games—I always have so much fun viii hanging out with you. I also loved talking about chemistry with you—I have learned so much from you! Yasu, thanks for being such a great friend. When you baked me those green tea cookies after my candidacy, I was touched. I loved all of the fun stuff we got to do together exploring California! Thanks for always going to Dodgers and Angels games with me—seeing Ohtani pitch in Anaheim was so awesome! Our late-night In-N-Out trips were so fun, too. I really hope I get to see you soon! Thank you to Emily Chen for all of her support over the past year. Thanks for always making time when I need help and for supporting me through some of my most stressful times. I want to thank all of the people I shared a bay with in lab. You all made being in lab a lot of fun. Thanks to Suzie for being the best lab DJ, resident beer expert, and legendary cat photographer. I am so glad I got to know you and for all the times you made me laugh. Thanks to Dr. Dave Charboneau for being a great labmate to talk chemistry with and for always making us laugh. I’ll keep listening to the mid-2000s playlist on Fridays! Thanks to Karli Holman and Travis DeLano who I just started sharing a bay with for the last year of grad school—it’s been a ton of fun! Thanks to Dr. Justin Su for always being encouraging and great to talk to in lab. Thanks to Yujia Tao for being my desk mate and one of the funniest people I know. Thanks to the ping-pong players: Dr. Marco Brandstätter, Dr. Chris Lavoie, Dr. Mike Maser, Dr. Mike Rombola, Dr. Tyler Fulton, Nick Hafeman, Dr. Eric Alexy, Dr. Eric Welin, Dr. Veronica Hubble, and Farbod Moghadam. ix I want to thank the members of the Stoltz lab for being such great friends and people to work with. The 2021 Roundbottoms softball season was one of the most fun times I had throughout all of grad school. Thanks to all of my other friends from Caltech. Thanks to Dr. Carson Matier for his help when talking chemistry and for being a great friend. Thanks to Dr. Jaika Dörfler for looking out for all of us when we were young grad students and making us feel at home. Thanks to Alexia Kim for being a great friend and for all of the fun times going to the movies and seeing cute dogs. Thanks to Veronica Hubble for all of her support over the past year and for always making me smile. Thanks to the Caltech Wood Bat Baseball Club for getting me back to playing baseball for the first time in 10 years. I had forgotten how happy it makes me to play, and getting back out there has been a blast. Thanks to Brian Stoltz for being our faculty mentor and hitting instructor, Mike Maser for starting the club, and Veronica Hubble, Nathan Harper, Alex Shimozono, and Chris Kenseth for being great members. Thanks to the Southern California Amateur Baseball League and my team Los Bandolitos for making Sunday’s so fun. I want to thank all of the people in my life from outside of grad school for always being there for me. I am so blessed to have been surrounded by so many wonderful people throughout all of the stages of my life. Words cannot express how much you all mean to me. Thank you to Jordyn Richman for being the most caring person I know. I cannot overstate how much you have influenced me. You are always working to help those in x need, and you inspire me to do more for others. Thank you for always being there for me, no matter what. I am so lucky to have you in my life! Thank you to Shaan Prabhakar, Dom Braccia, Ray Gerhart, and Jason Conaway. I am so lucky to have met such an awesome group of friends. I feel so at home when we get to spend time together, and I’m looking forward to seeing more of you all soon! Thank you to Jared Tinari for being such a great friend and listener. I’m so glad our friendship has grown for the past 20 years. Thanks to Sanil Shah for being so supportive and willing to talk whenever I need. Thank you to Ameya Naik for being a great friend and always making time to keep our friendship going across the country. Thanks to Evan Harkins, Kyle Hoch, and Joe Palladino for being great friends. Thanks to Lia Siegelman for being a great friend and making time to hang out on the weekends—I always loved going to the farmers’ market! Thanks to Mattia Poinelli for being a great roommate and friend, and for teaching me how to surf! Lastly, I’d like to thank my cat Pepita for all of the joy she has brought into my life. She has been through a lot in terms of health, but has never lost her love for people. xi ABSTRACT A unified, convergent fragment coupling approach to the C19- and C20-diterpenoid alkaloid natural products is presented. The highly-caged aconitine, denudatine, and napelline cores are disconnected through the central B-ring cyclohexane to an A/E/F-ring fragment common to the structures all three subfamilies. 1,2-addition of an appropriate organometallic C/D-bicycle to an A/F-ring hydrindane epoxy ketone fragment followed by a Lewis acid-catalyzed semipinacol reaction couples the two fragments together and sets a key all-carbon quaternary center at C11. This strategy is realized in the synthesis of the C19-aconitine core by using a [3.2.1]-bicyclooctene C/D-fragment as the nucleophile in the 1,2-addition. This C/D-fragment is prepared using a meta-photocycloaddition; this represents an alternative approach to the commonly employed biomimetic WagnerMeerwein rearrangement of a [2.2.2]-bicyclooctane. To complete the aconitine core, a radical cyclization cascade to form the E-ring piperidine and B-ring cyclohexane in a single step is investigated. N-centered radicals were evaluated to initiate the cascade via a 6-exo-trig cyclization. A neutral aminyl radical gave rise to an unexpected Hoffman-Löffler-Freytag type product resulting from 1,5-hydrogen atom transfer. Employing Lewis-acidic single electron reducing metal catalyst led to formation of the E-ring cyclized product, however the second cyclization to close the B-ring did not occur. As an alternative approach, the E-ring was closed via an intramolecular aziridination. Treatment of this aziridine with acetyl bromide results in an aziridineopened alkyl bromide product. This alkyl bromide is used as a functional group handle to form the final ring of the aconitine core. From there, the total synteheses of the C19- xii diterpenoid alkaloids (–)-talatisamine, (–)-liljestrandisine, and (–)-liljestrandinine were completed in short order. These synthetic efforts led to revision of the proposed structure of (–)-liljestrandisine. xiii PUBLISHED CONTENT AND CONTRIBUTIONS Portions of the work described herein were disclosed in the following publications: Wong, A. R.; Fastuca, N. J.; Mak, V. W.; Kerkovius, J. K.; Stevenson, S. M.; Reisman, S. E. ACS Cent. Sci. 2021, 7, 1311–1316. DOI: 10.1021/acscentsci.1c00540 Fastuca, N. J.; Wong, A. R.; Mak, V. W.; Reisman, S. E. Org. Synth. 2020, 97, 327-338. DOI: 10.15227/orgsyn.097.0327 N. J. F. conducted experiments towards (–)-talatisamine, (–)-liljestrandisine, and (–)-liljestrandinine and contributed to the preparation of the manuscripts and supporting information. xiv TABLE OF CONTENTS CHAPTER 1 1 C19-Diterpenoid Alkaloids: Structure and Synthesis 1.1 INTRODUCTION ............................................................................................. 1 1.2 STRUCTURAL ANALYSIS AND BIOSYNTHESIS OF C19-DITERPENOID ALKALOIDS ........................................................................................................... 2 1.3 TOTAL SYNTHESES OF C19-DITERPENOID ALKALOIDS ................................. 4 1.3.1 Biomimetic Wagner-Meerwein Approaches to Aconitine Core ............ 4 1.3.2 Gin’s Synthesis of Neofinaconitine ...................................................... 8 1.4 CONCLUDING REMARKS ............................................................................... 9 1.5 NOTES AND REFERENCES ............................................................................. 11 CHAPTER 2 12 Convergent Approach to C19-Diterpenoid Alkaloids via 1,2-Addition/Semipinacol Sequence 2.1 INTRODUCTION ........................................................................................... 12 2.2 RETROSYNTHETIC ANALYSIS OF C19-DITERPENOID ALKALOIDS ............... 13 2.3 SYNTHESIS OF FRAGMENT COUPLING PARTNERS ..................................... 15 2.3.1 Synthesis of [3.2.1]-Bicyclooctene C/D-Bicycle ................................. 15 xv 2.3.2 Enantioselective Synthesis of A/F-Bicycle Epoxy Ketone 71 ............... 18 2.4 1,2-ADDITION/SEMIPINACOL REARRANGEMENT FRAGMENT COUPLING SEQUENCE ....................................................................................... 19 2.5 CONCLUDING REMARKS ............................................................................. 20 2.6 EXPERIMENTAL SECTION .............................................................................. 21 2.7 NOTES AND REFERENCES ............................................................................. 49 CHAPTER 3 50 Towards C19-Diterpenoid Alkaloids: Approaches to the C4 Chiral Quaternary Center 3.1 INTRODUCTION ........................................................................................... 50 3.2 SEQUENTIAL REDUCTION OF C18 AND C19 ESTERS.................................. 51 3.3 SELECTIVE AMINOLYSIS OF C19 LACTONE ................................................. 53 3.4 TOWARDS A C18/C19 FUNCTIONALIZED EPOXY KETONE FRAGMENT .... 54 3.4.1 Selective Functionalization of a C18/C19 Diol .................................. 55 3.4.2 Setting C4 Quaternary Center by Michael Addition of a Prochiral Nucleophile ............................................................................................... 58 3.5 CONCLUDING REMARKS ............................................................................. 60 3.6 EXPERIMENTAL SECTION .............................................................................. 61 3.7 NOTES AND REFERENCES ........................................................................... 117 xvi CHAPTER 4 118 Completion of Aconitine Core and Synthesis of (–)-Talatisamine, (–)-Liljestrandisine and (–)-Liljestrandinine 4.1 INTRODUCTION ......................................................................................... 118 4.2 RADICAL CYCLIZATION CASCADE ............................................................ 118 4.2.1 Reaction of Neutral Aminyl Radical ................................................ 106 4.2.2 Reactivity of Aminyl Radical Cations ............................................... 119 4.3 SYNTHESIS OF C19-DITERPENOID ALKALOIDS (–)-TALATISAMINE, (–)-LILJESTRANDISINE AND (–)-LILJESTRANDININE ............................... 126 4.3.1 Completion of the Aconitine Core................................................... 126 4.3.2 Reduction/Oxidation of D-ring for Divergent Syntheses of (–)Talatisamine, (–)-Liljestrandisine and (–)-Liljestrandinine .......................... 129 4.4 CONCLUDING REMARKS ........................................................................... 131 4.5 EXPERIMENTAL SECTIONS .......................................................................... 133 4.6 NOTES AND REFERENCES ........................................................................... 192 ABOUT THE AUTHOR 193 APPENDIX 1 194 Spectra Relevant to Chapter 2 xvii APPENDIX 2 230 Spectra Relevant to Chapter 3 APPENDIX 3 Spectra Relevant to Chapter 4 327 xviii LIST OF ABBREVIATIONS [α]D angle of optical rotation of plane-polarized light Å angstrom(s) ABNO 9-azabicyclo[3.3.1]nonane N-oxyl p-ABSA para-acetamidobenzenesulfonyl azide Ac acetyl acac acetylacetonate AIBN azobisisobutyronitrile aq aqueous Ar aryl group atm atmosphere(s) BINOL 1,1'-bi-2,2'-naphthol BINAP 2,2′-bis(diphenylphosphino)-1,1′-binaphthyl Bipy/bpy 2,2'-bipyridine Bn benzyl Boc tert-butoxycarbonyl bp boiling point br broad Bu butyl i-Bu iso-butyl n-Bu butyl or norm-butyl t-Bu tert-butyl xix BQ 1,4-benzoquinone BRSM yield based on recovered starting material Bz benzoyl c concentration of sample for measurement of optical rotation 13 C carbon-13 isotope /C supported on activated carbon charcoal °C degrees Celsius calc’d calculated CAN ceric ammonium nitrate cat. catalyst Cbz benzyloxycarbonyl cf. consult or compare to (Latin: confer) cis (zusammen) on the same side cm–1 wavenumber(s) CoA Coenzyme A conc. concentrated conv. conversion Cp cyclopentadienyl CSA camphor sulfonic acid Cy cyclohexyl Δ heat or difference δ chemical shift in ppm d doublet xx d deutero or dextrorotatory D deuterium dba dibenzylideneacetone DBU 1,8-diazabicyclo[5.4.0]undec-7-ene DCE 1,2-dichloroethane DDQ 2,3-dichloro-5,6-dicyano-1,4-benzoquinone de novo starting from the beginning; anew DIPEA N,N-diisopropylethylamine DHQ dihydroquinine DHQD dihydroquinidine DIAD diisopropyl azodicarboxylate DIBAL diisobutylaluminum hydride DMAP 4-(dimethylamino)pyridine DMBA 1,3-dimethylbarbituric acid DME 1,2-dimethoxyethane DMEDA N,N'-dimethylethylenediamine DMF N,N-dimethylformamide DMP Dess-Martin Periodinane DMPU 1,3-dimethyl-3,4,5,6-tetrahydro-2(1H)-pyrimidinone DMSO dimethylsulfoxide DTBMP 2,6-di-tert-butyl-4-methylpyridine dppe 1,2-bis(diphenylphosphino)ethane dppf 1,1'-bis(diphenylphosphino)ferrocene xxi dr diastereomeric ratio ee enantiomeric excess E methyl carboxylate (CO2CH3) E+ electrophile E trans (entgegen) olefin geometry EDCI N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride e.g. for example (Latin: exempli gratia) EI electron impact ent enantiomer of epi epimeric equiv equivalent(s) ESI electrospray ionization Et ethyl et al. and others (Latin: et alii) FAB fast atom bombardment FTIR fourier transform infrared spectroscopy g gram(s) h hour(s) 1H proton [H] reduction HDA hetero-Diels–Alder HFIP hexafluoroisopropanol HMBC heteronuclear multiple-bond correlation spectroscopy xxii HMDS hexamethyldisilazide HMPA hexamethylphosphoramide hν irradiation with light HPLC high performance liquid chromatography HRMS high resolution mass spectrometry Hz hertz IC50 half maximal inhibitory concentration (50%) i.e. that is (Latin: id est) iso isomeric in situ in the reaction mixture J coupling constant in Hz k rate constant kcal kilocalorie(s) kg kilogram(s) L liter or neutral ligand l levorotatory LA Lewis acid LC/MS liquid chromatography–mass spectrometry LDA lithium diisopropylamide m multiplet or meter(s) M molar or molecular ion m meta µ micro xxiii m-CPBA meta-chloroperbenzoic acid Me methyl MeO bpy 4,4’-dimethoxy-2,2’-bipyridine mg milligram(s) MHz megahertz MIC minimum inhibitory concentration min minute(s) mL milliliter(s) MM mixed method mol mole(s) MOM methoxymethyl Ms methanesulfonyl (mesyl) MS molecular sieves MTT 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide m/z mass-to-charge ratio NBS N-bromosuccinimide NCS N-chlorosuccinimide nd not determined NHC N-heterocyclic carbene nm nanometer(s) nM nanomolar NMI N-methylimidazole NMO N-methylmorpholine N-oxide xxiv NMR nuclear magnetic resonance NOE nuclear Overhauser effect NOESY nuclear Overhauser enhancement spectroscopy NPh naphthyl Nu nucleophile o ortho [O] oxidation P peak p para PCC pyridinium chlorochromate PDC pyridinium dichromate Ph phenyl pH hydrogen ion concentration in aqueous solution PHAL 1,4-phthalazinediyl diether PIFA [bis(trifluoroacetoxy)iodo]benzene Pin pinacol PivOH pivalic acid pKa acid dissociation constant pm picometer(s) PMB para-methoxybenzyl ppm parts per million PPTS pyridinium para-toluenesulfonate Pr propyl xxv i-Pr isopropyl n-Pr propyl or norm-propyl psi pounds per square inch py pyridine PYR 2,5-diphenyl-4,6-pyrimidinediyl diether q quartet QD Quinidine QN Quinine quant. quantitative R generic (alkyl) group RL large group R rectus RCM ring-closing metathesis recry. recrystallization ref reference Rf retention factor rgt. reagent rt room temperature s singlet or seconds sat. saturated t triplet TBAF tetra-n-butylammonium fluoride TBAI tetra-n-butylammonium iodide xxvi TBME tert-butyl methyl ether TBS tert-butyldimethylsilyl TC thiophene-2-carboxylate TCA trichloroacetic acid temp temperature Tf trifluoromethanesulfonyl TFA trifluoroacetic acid THF tetrahydrofuran TIPS triisopropylsilyl TLC thin layer chromatography TMS trimethylsilyl TOF time-of-flight tol tolyl TPAP tetrapropylammonium perruthenate trans on the opposite side Ts para-toluenesulfonyl (tosyl) UV ultraviolet vide infra see below w/v weight per volume X anionic ligand or halide xs excess Z cis (zusammen) olefin geometry Chapter 1 – C19-Diterpenoid Alkaloids: Structure and Synthesis 1 Chapter 1 C19-Diterpenoid Alkaloids: Structure and Synthesis 1.1 INTRODUCTION The extracts of the Aconitum and Delphinium genera of plants have long been used in traditional medicine as local anesthetics and as poisons for hunting and battle.1 Common names for these flowering plants alluding to their toxicity include “wolf’s bane,” “the devil’s helmet,” and “the queen of poisons.” Diterpenoid alkaloids are a broad family of natural products associated with the bioactivity of these flowering plants,2–4 and many of these compounds exhibit analgesic,5,6 anti-inflamatory, antihypertensive, and antiarrhythmic7 properties. Talatisamine (1) selectively blocks inwardly rectifying K+ ion channels over Na+ and Ca2+ ion channels in rat neurons.8 1 has also been shown to attenuate neurocytotoxicity induced by β-amyloid oligomers.9 The structural complexity of these highly caged natural products have drawn significant attention from the synthetic community for over 50 years.10–13 Chapter 1 – C19-Diterpenoid Alkaloids: Structure and Synthesis 2 Figure 1.1 Varying carbon frameworks of diterpenoid alkaloid subfamilies. (a) C20-diterpenoid alkaloids. HO OH OH Me Me N OH O N Et veatchine (4) napelline (5) veatchine type OH napelline type O HO OH Me Me N H H N Et O OH OH Me Me Ac H N Me H O N OAc OH O atisine (6) denudatine (7) delcarduchol (8) guan-fu base A (9) atisine type denudatine type hetidine type hetisine types (b) C19- and C18-diterpenoid alkaloids. OMe NEt MeO OMe OMe NEt MeO OMe NEt MeO NHAc O O OH OMe NEt MeO N Et OMe HO HO OMe HO talatisamine (1) H HO OH HO HO HO liljestrandisine (2) HO liljestrandinine (3) (proposed) MeO HO OMe lappaconitine (10) C18 lappaconitine type C19 aconitine type 1.2 STRUCTURAL ANALYSIS AND BIOSYNTHESIS OF C19-DITERPENOID ALKALOIDS Figure 1.2 Structural analysis of C19-diterpenoid alkaloid (–)-talatisamine (1). 3 1 18 4 A 19 NEt E 5 F 11 17 10 B 6 12 7 15 9 8 C D 13 14 16 Me azaOMe NEt [3.3.1] MeO 2 •Unique polybridged hexacyclic framework 11 Me 11 A 4 E N Et C19 diterpenoid alkaloid core C B F D H [3.2.1] HO [3.2.1] HO OMe •12 stereocenters, including 2 all-carbon quaternary centers •Highly oxygenated (–)-talatisamine (1) Diterpenoid alkaloids are a broad class of natural products with a variety of carbon skeletons with over 1200 natural products isolated to date.3,14 They are classified by the Chapter 1 – C19-Diterpenoid Alkaloids: Structure and Synthesis 3 number of carbons in their backbones (C18, C19, and C20). The highly-caged polycyclic core of these compounds, such as (–)-talatisamine (1, Figure 1.2), present a significant synthetic challenge. The C19-aconitine core is comprised of an array of bridged bicycles, including a synthetically challenging [3.2.1]-bicyclooctane. Biosynthetically, diterpenoid alkaloids are derived from the structurally similar kaurane and atisane diterpenoid families by incorporation of serine as a source of nitrogen.15 These complex skeletons are believed to arise from a series of cationic polyene cyclizations and skeletal rearrangements (Figure 1.3).16 It is believed that C19-diterpenoid alkaloids are derived from Wagner-Meerwein rearrangement of the C20denudatine core, wherein the [2.2.2]-bicyclooctane is converted to a [3.2.1]-bicyclooctane (Figure 1.4). The cationic rearrangements of the carbon skeletons which are crucial the biosynthesis of these natural products have also played an important role in many of the total syntheses of C18- and C19-diterpenoid alkaloids. Figure 1.3 Biosynthesis of diterpenoid alkaloid cores. Me Me Me Me geranylgeranyl diphosphate (11) copalyl Me diphosphate Me synthase Me 14 Me Me 18 Me 13 Me 16 Me Me 13 16 L-serine 13 Me H Me 16 16 Me 16 Me 13 14 15 16 Me L-serine N Et N Et Me Me Me H Me Me 14 Me Me 17 12 Me Me Me 14 14 13 12 13 Me Me OPP Me 13 Me Me Me ent-copallyldiphosphate (ent-CPP) (12) OPP Me Me ent-atiserene Me synthase Me Me Me 13 14 atisane C20 atisine-type C20 vetachine-type kaurane (19) (20) (22) (21) 16 Me Chapter 1 – C19-Diterpenoid Alkaloids: Structure and Synthesis 4 Figure 1.4 Wagner-Meerwein rearrangement of denudatine core to aconitine core. OH OH Me 10 Me oxidation N Et N 17 Et C7-C17 bond formation C20 atisine-type HO E 7 8 9 C O B D E C HO C19 aconitine-type O 24 Oxidation WagnerMeerwein + (-OH) OH NEt NEt Me 10 H B OH OPP OH A HO F F 10 OPP (23) OH N Et 8 17 C20 denudatine-type (22) A 7 NEt Me 9 9 8 H D HO OH HO 25 (26) 1.3 TOTAL SYNTHESES OF C19-DITERPENOID ALKALOIDS 1.3.1 Biomimetic Wagner-Meerwein Approaches to Aconitine Core The Wagner-Meerwein rearrangement of a [2.2.2]-bicyclooctane which comprises the C and D rings of the C20 atisine and denudatine scaffolds to the [3.2.1]-bicyclooctane characteristic of the C18- and C19-diterpenoid alkaloids has inspired biomimetic approaches in many of the total syntheses reported to date. Targeting a [2.2.2]-bicyclooctane as an intermediate in the synthesis of C18- and C19-diterpenoid alkaloids has two main strategic advantages. Firstly, it allows for the divergent synthesis of two families of natural products via a late stage diversification of the carbon skeleton. Secondly, the [2.2.2]-bicyclooctane structural motif is a classic keying element for retrosynthetic disconnection via a Diels-Alder cycloaddition, one of the most well studied transformations in organic chemistry. Wiesner’s synthesis of talatisamine (1) employing such an approach represents the first total synthesis of a C19-diterpenoid alkaloid. Chapter 1 – C19-Diterpenoid Alkaloids: Structure and Synthesis 5 Scheme 1.1 Wiesner’s synthesis of the atisine core. OMe NMs MeO 1. Na/NH3 2. Ac2O OMe NAc MeO 3. HCl, MeOH 27 OMe NAc MeO OMe 28 • 1. HO(CH2)2OH 2. OsO4, NaIO4 hυ 3. NaBH4 O OMe NAc MeO 7 steps OMe NAc MeO O 29 OMe NAc MeO TsO 33 OMe NAc MeO HCl X = ethylene ketal X OMe X = ethylene ketal O 32 O OH O 31 30 X OH atisine core Wiesner’s route to talatisamine is summarized here (Schemes 1.1 and 1.2). Beginning from tetracyclic arene 27, prepared in 21 steps, Birch reduction followed by N-acetylation affords enone 28 upon treatment with acid. [2 + 2] photocycloaddition with ketene affords cyclobutane 29. Ketalization of 29 with ethylene glycol followed by oxidative-cleavage of the cyclobutane exo-methylene and subsequent ketone reduction gives cyclobutanol 30. Treatment with acid effects retro-aldol/aldol cascade to give the less strained cyclohexane isomer 32, forming the atisine core. In seven further steps, they access a [2.2.2]-bicyclooctane with a tosylate leaving group suitable for the desired rearrangement. Treatment with tetramethylguanidine in DMSO at high temperature forms the rearranged product as a mixture of alkenes after deprotonation of the resulting tertiary carbocation. To complete the core, the C7–C17 bond was formed by an aza-Prins reaction upon oxidation of the amine. Chapter 1 – C19-Diterpenoid Alkaloids: Structure and Synthesis 6 Scheme 1.2 Wiesner’s synthesis of talatisamine (1). NH OMe NAc MeO Me2N xx OMe NAc MeO OMe NAc MeO OMe NAc MeO NMe2 + DMSO 180 °C TsO X OMe O X = ethylene ketal O 33 atisine core O 37 OMe 36 OMe NEt MeO talatisamine (1) OMe 38 40% yield 17 3 steps 7 H 2O HO OMe O OMe NEt MeO Hg(OAc)2 HO OMe 40% yield OMe NEt MeO O O 40% yield HO OMe 40 HO OMe 39 In a later synthesis of the C19-diterpenoid alkaloid 13-desoxydelphonine (39), Wiesner and coworkers sought to rearrange the denudatine core, where the C7–C17 bond is already in place (Scheme 1.3).19 Starting from ortho-quinone mono-ketal 34, Diels-Alder cycloaddition with benzyl vinyl ether followed by global reduction with Zn and acetic acid affords the denudatine core in 35. This can be elaborated in 12 steps to the substrate for the WagnerMeerwein rearrangement, which procedes smoothly to deliver the aconitine core in excellent yield. This product can be advanced in 4 steps to 13-desoxydelphonine. Chapter 1 – C19-Diterpenoid Alkaloids: Structure and Synthesis 7 Scheme 1.3 Wiesner’s synthesis of 13-desoxydelphonine (39). OMe O NMe MeO Diels-Alder 1. MeO R O 2. Zn, AcOH 12 steps MeO Br denudatine core OMe NMe OMe O NMe MeO HO HO OMe 13-desoxydelphonine (39) N 38 4 steps MeO X OMe X = ethylene ketal 36 35 34 R = H, Br (2:1) MeO MeO OBn O O OMe O NMe MeO OBn O O OMe O NMe MeO N DMSO/o-xylene 180 °C MeO O O OMe 89% yield 37 The Sarpong lab has utilized this strategy for the synthesis of a variety of diterpenoid alkaloid carbon skeletons, including the aconitine-type C19-diterpenoid alkaloids (Scheme 1.4).20,21 In their elegant 2015 synthesis of liljestrandinine (3), they employ an intramolecular Diels-Alder cycloaddition of an ortho-quinone monoketal with a pendant olefin to build the [2.2.2]-bridged C/D-bicycle 42. This intramolecular approach also provides an efficient way to form the central B-ring. In short order, they elaborate 42 to an alkyl triflate, which serves as the substrate for the Wagner-Meerwein rearrangement. Similar approaches have been used by the Fukuyama and Inoue labs in their syntheses of C19-diterpenoid alkaloids.22,23 Chapter 1 – C19-Diterpenoid Alkaloids: Structure and Synthesis 8 Scheme 1.4 Sarpong’s synthesis of liljestrandinine (3). OMe NCO2Me MeO 1. 2N HCl MeO MeO O 2. PIDA MeO MOMO 98% yield (2 steps) 40 OMe NEt MeO OMe NCO2Me MeO NCO2Me 150 °C 4 Steps MeO O OMe denudatine framework 42 77% yield 41 OMe NCO2Me MeO OMe NCO2Me MeO OMe MeO OMe MeO NCO2Me DBU 4 steps DMSO 120 °C HO HO liljestrandinine (3) MOMO 1.3.2 HO MOMO 45 H 2O 44 OMOM OTf Wagner–Meerwein rearrangement 43 Gin’s Synthesis of Neofinaconitine (60) Scheme 1.5 Gin’s approach to aconitine core. Et3N TBSOTf + 47 TIPSO [6 steps] MeO 46 8 steps O 11 11 O 17 NEt 17 3 steps + CO2Me 50 MeO 49 Et N Et N O CO2Me Br Tf2NH H 11 Br OH Diels–Alder O Br OTIPS 48 1.6 : 1 rr MeO2C N 17 Br OMe HO Et Et N 0 °C CO2Me O Br TBSO Diels–Alder TBSO O Br O CO2Me H H CH2Cl2 O Mannich Cyclization MeO 54 O 75% yield O MeO O MeO 53 52 O MeO 51 Gin and coworkers used a different approach to the C/D-bicycle in their synthesis of the C18-diterpenoid alkaloid neofinaconitine (60, Schemes 1.5 and 1.6).24 They assemble the [3.2.1]-bicycle directly via a Diels-Alder reaction between a cyclopentadiene derived from 46 and cyclopropane dienophile (47). Prior to the work presented in subsequent chapters, this is Chapter 1 – C19-Diterpenoid Alkaloids: Structure and Synthesis 9 the only example of a total synthesis of a C18- or C19-diterpenoid alkaloid which does not involve a Wagner-Meerwein rearrangement to form the C/D-bicycle. Upon elaboration of 48 to diene 49, they perform a fragment coupling Diels-Alder with dienophile 50 to form the Aring of the natural product. Intramolecular Mannich reaction forms the A/E/F-tricycle of the natural product and afforded the unexpected product 54 after intramolecular oxy-Michael. Scheme 1.6 Completion of C18-diterpenoid alkaloid neofinaconitine (60). CO2Me O Br N Et CAN O MeO NH2 O O Br MsCl, Et3N O O MeO2C neofinaconitine (60) H O 57 O O MeO2C NEt NEt Bu3SnH, AIBN H H HO OMe O MeO 56 11 steps MeO N Et 66% yield (2 steps) O OMe NEt Br CH2Cl2, 50 °C O MeO CO2Me O N Et MeCN/H2O 54 O CO2Me OH O MeO O 59 aconitine core [21 steps] Br PhH, 80 °C 99% yield O MeO 58 To break the unnecessary C–O bond of the enol ether 54, allylic oxidation and mesylate formation enabled elimination to bis-enone 59 (Scheme 1.6). The final bond of the natural product core is forged using a Giese addition of a C7-radical to the D-ring enone to give 59. This intermediate is elaborated to the C18-diterpenoid alkaloid neofinaconitine (60) in 11 further steps. 1.4 CONCLUDING REMARKS The complex carbon frameworks of the diterpenoid alkaloids have provoked a number of creative synthetic approaches. The early structural elucidation work on diterpenoid alkaloids Chapter 1 – C19-Diterpenoid Alkaloids: Structure and Synthesis 10 provides insight into the probable biosynthetic relationship between different subfamilies of diterpenoid alkaloids. This has informed many of the total synthetic efforts, as multiple groups have reported approaches which convert the carbon framework of one natural product subfamily to another. Despite the success of synthetic chemists in accessing these natural products over the past 50 years, they still remain attractive targets for total synthesis and inspire development of creative synthetic strategies. Chapter 1 – C19-Diterpenoid Alkaloids: Structure and Synthesis 11 1.5 NOTES AND REFERENCES (1) (2) (3) (4) (5) (6) (7) (8) (9) (10) (11) (12) (13) (14) (15) (16) (17) (18) (19) (20) (21) (22) (23) (24) Sung, Y. Sun, E.-T. Z., Sun, S.-C., Translators; University Park: Pennsylvania State University, 1966. Wang, F.-P.; Liang, X.-T. In The Alkaloids; Elsevier Science: New York, 2002; Vol. 59, pp 1–280. Wang, F.-P.; Chen, Q.-H.; Liu, X.-Y. Nat. Prod. Rep. 2010, 27 (4), 529. Wada, K. In Studies in Natural Products Chemistry; Elsevier, 2012; Vol. 38, pp 191– 223. Friese, J.; Gleitz, J.; Gutser, U. T.; Wilffert, B.; Selve, N.; Heubah, J. F.; Matthiesen, T. Eur. J. Pharmacol. 1997, 337, 165–174. Gutser, U. T.; Friese, J.; Heubach, J. F.; Matthiesen, T.; Selve, N.; Wilffert, B.; Gleitz, J. Naunyn. Schmiedebergs Arch. Pharmacol. 1997, 357 (1), 39–48. Vakhitova, Yu. V.; Farafontova, E. I.; Khisamutdinova, R. Yu.; Yunusov, V. M.; Tsypysheva, I. P.; Yunusov, M. S. Russ. J. Bioorganic Chem. 2013, 39 (1), 92–101. Song, M.-K.; Liu, H.; Jiang, H.-L.; Yue, J.-M.; Hu, G.-Y.; Chen, H.-Z. Neurosci. 2008, 155 (2), 469–475. Wang, Y.; Song, M.; Hou, L.; Yu, Z.; Chen, H. Neurosci. Lett. 2012, 518 (2), 122– 127. Wiesner, K. Tetrahedron 1985, 41 (3), 485–497. Liu, X.-Y.; Qin, Y. Asian J. Org. Chem. 2015, 4 (10), 1010–1019. Dank, C.; Sanichar, R.; Choo, K.-L.; Olsen, M.; Lautens, M. Synthesis 2019, 51, 3915–3946. Liu, X.-Y.; Wang, F.-P.; Qin, Y. Acc. Chem. Res. 2021, 54 (1), 22–34. Wang, F.-P.; Chen, Q.-H. In The Alkaloids: Chemistry and Biology; Elsevier, 2010; Vol. 69, pp 1–577. Zhao, P.-J.; Gao, S.; Fan, L.-M.; Nie, J.-L.; He, H.-P.; Zeng, Y.; Shen, Y.-M.; Hao, X.-J. J. Nat. Prod. 2009, 72 (4), 645–649. Hong, Y. J.; Tantillo, D. J. J. Am. Chem. Soc. 2010, 132 (15), 5375–5386. Wiesner, K.; Tsai, T. Y. R.; Huber, K.; Bolton, S. E.; Vlahov, R. J. Am. Chem. Soc. 1974, 96 (15), 4990–4992. Wiesner, K. Pure Appl. Chem. 1975, 41, 93–112. Wiesner, K.; Tsai, T. Y. R.; Nambiar, K. P. Can. J. Chem. 1978, 56 (10), 1451–1454. Marth, C. J.; Gallego, G. M.; Lee, J. C.; Lebold, T. P.; Kulyk, S.; Kou, K. G. M.; Qin, J.; Lilien, R.; Sarpong, R. Nature 2015, 528 (7583), 493–498. Kou, K. G. M.; Kulyk, S.; Marth, C. J.; Lee, J. C.; Doering, N. A.; Li, B. X.; Gallego, G. M.; Lebold, T. P.; Sarpong, R. J. Am. Chem. Soc. 2017, 139 (39), 13882–13896. Nishiyama, Y.; Yokoshima, S.; Fukuyama, T. Org. Lett. 2016, 18 (10), 2359–2362. Kamakura, D.; Todoroki, H.; Urabe, D.; Hagiwara, K.; Inoue, M. Angew. Chem. Int. Ed. 2020, 59 (1), 479–486. Shi, Y.; Wilmot, J. T.; Nordstrøm, L. U.; Tan, D. S.; Gin, D. Y. J. Am. Chem. Soc. 2013, 135 (38), 14313–14320. Chapter 2 – Convergent Approach to C19-Diterpenoid Alkaloids 12 Chapter 2 Convergent Approach to C19-Diterpenoid Alkaloids via 1,2Addition/Semipinacol Sequence 2.1 INTRODUCTION This chapter details our fragment coupling approach to the C19-diterpenoid alkaloids as an alternative to the commonly employed biomimetic skeletal rearrangement of the denudatine scaffold. Our retrosynthetic analysis of (–)-talatisamine (1) is presented. The asymmetric syntheses of our two coupling partners are described, which includes a direct synthesis of the [3.2.1]-bicyclooctene C/D-ring bicyclic fragment via an arene-alkene meta- photocycloaddition. Lastly, our two-step fragment coupling sequence of 1,2-addition of an alkenyl lithium C/D-bicyclic fragment to an epoxy ketone A/F-bicyclic fragment followed by Lewis acid-catalyzed semipinacol rearrangement is presented. Chapter 2 – Convergent Approach to C19-Diterpenoid Alkaloids 13 Figure 2.1 Unified approach to C19 and C20 diterpenoid alkaloids: disconnection through the central B-ring. HO OH Me OH NEt Me OH B B N Et napelline (5) H HO OH B N Et H HO lepenine (62) cardiopetaline (61) (C20 denudatine type) HO (C20 napelline type) OH Me (C19 aconitine type) Fragment Coupling 64 HO HO OH H R 2N A 63 Me HO OH HO F H 65 O O 66 2.2 RETROSYNTHETIC ANALYSIS OF C19-DITERPENOID ALKALOIDS The structural similarities of various diterpenoid alkaloid subfamilies inspired us to devise a unified approach to access natural products with a variety of carbon skeletons (Figure 2.1). The C19-aconitine, C20-napelline and C20-denudatine type diterpenoid alkaloids share a common A/E/F-tricycle connected by two bonds of the central B-ring to a C/D-bicycle characteristic of each subfamily. As such, we envisioned a retrosynthetic disconnection through the two bonds of the B-ring would allow us to access each subfamily through a fragment coupling of the proper C/D-bicycle with a common A/F-bicycle. We sought to apply this strategy first to the synthesis of C19-diterpenoid alkaloids. Our retrosynthetic analysis of (–)-talatisamine (1) is shown in Figure 2.2. Disconnection of the C19-N bond via lactam formation and reduction gives secondary amine Chapter 2 – Convergent Approach to C19-Diterpenoid Alkaloids 14 67, which we envisioned installing through a late stage reductive amination of a C17 ketone (68). Disconnection of the C7–C8 bond via enolate functionalization opens the central B-ring. Key to our fragment coupling strategy, we envisioned engaging epoxy-alcohol 70 in a semipinacol rearrangement, using the strain release of the epoxide opening as the enthalpic driving force to form the hindered C11 quaternary center. To form the C10–C17 bond of 70, we envisioned a 1,2-addition of an organolithium C/D-bicyclic fragment (72) to an A/F-epoxy ketone fragment (71). The [3.2.1]-bicycle was envisioned to arise from an arene-alkene metaphotocycloaddition. Epoxy ketone 71 was disconnected to 2-cyclopenten-1-one (73), dimethyl malonate (74) and 2-(2-bromoethyl)-1,3-dioxolane (75). Figure 2.2 Retrosynthetic analysis of (–)-talatisamine (1). OMe A MeO 19 lactam formation/ reduction NEt E 17 F MeO2C 4 OMe CO2Me NHEt reductive amination 17 OMe CO2Me O MeO2C 17 B B Michael reaction MeO2C 11 OMe CO2Me O 17 7 C HO HO OMe HO (–)-talatisamine (1) Br O 75 HO 67 (C19 aconitine type) O HO OMe MeO2C 74 O CO2Me A MeO2C MeO2C H 69 O F O 1,2-addition 71 MeO2C MeO2C H 11 O Li OH 17 10 70 O C 77 76 O Si t-Bu O 72 t-Bu O Si D t-Bu HO semipinacol rearrangement + metaphotocycloaddition O OR 68 73 OEt HO OMe t-Bu Chapter 2 – Convergent Approach to C19-Diterpenoid Alkaloids 15 2.3 SYNTHESIS OF FRAGMENT COUPLING PARTNERS Synthesis of [3.2.1]-Bicyclooctene C/D-Bicycle 90 2.3.1 As mentioned above, we envisioned synthesis of the [3.2.1]-bicyclooctane prior to fragment coupling. When considering methods to form these bridged bicycles,1,2 we were intrigued by a report from Sugimura and coworkers for a diastereoselective intramolecular arene-alkene meta-photocycloaddition (Scheme 2.1).3,4 Following the meta- photocycloaddition, Sugimura and coworkers hydrogenate the olefin; we, however, envision leveraging this alkene to incorporate the oxidation pattern of the natural product. Epoxidation of this olefin, followed by Grob-fragmentation would lead to formation of C/D-bicycle 82 which is oxidized at C16. Scheme 2.1 Sugimura’s diastereoselective intramolecular meta-photocycloadition for the synthesis of [3.2.1]-bicyclooctanes. Sugimura 2004 meta-photocycloaddition 6 7 O hν (254 nm) O Me R S Me 78 70% yield Me Me H 6 7 1. H2, Pd/C O 2. Hg(OAc)2, H2O then NaBH4 O H 79 [2 steps from phenol] H xx Me Me OH O 7 6 xx O 45–50% yield (2 steps) 80 Proposed Fragment Synthesis 6 H H 7 O epoxidation O Me H 16 H 79 O Me O H Me H O Me H 81 Me Grob Fragmentation Me OH O 82 O 16 OH To begin, chiral diol 84 was synthesized by Noyori hydrogenation of acetylacetone (83, Scheme 2.2).5 Mitsunobu reaction of 84 with phenol (85) followed by transetherfication of ethyl vinyl ether (86) affords meta-photocycloaddition substrate 78. Irradiation of 78 with UV Chapter 2 – Convergent Approach to C19-Diterpenoid Alkaloids 16 light results in formation of Sugimura’s product 79 in yields comparable to those reported in the original communication. These intramolecular photochemical reactions must be run at low concentrations (~0.01 M) to prevent intermolecular reactions, limiting the largest viable scale of this reaction to 1.5 grams per batch. With 79 in hand, we set out to test the proposed epoxidation and Grob-fragmentation to access 82. Epoxidation with mCPBA followed by treatment with aqueous hydrochloric acid furnishes the desired C/D-bicycle 82. Luche reduction of 82 yields the axial C14 alcohol 83. The C14/C16-1,3-diol is protected as the siliconide 84. Having functionalized the D-ring, the chiral auxiliary needed to be removed. Oxidation of alcohol 84 using Stahl’s conditions produces β-alkoxyketone 85 which is treated directly with basic methanol to cleave the auxiliary via E1cB elimination to give alcohol 86 as a single enantiomer. This alcohol is then oxidized to give ketone 87. Scheme 2.2 Synthesis of [3.2.1]-bicyclic ketone 87 enabled by Sugimura’s metaphotocycloaddition 85 (1.1 equiv) 1. O Ru[R-BINAP]Cl2 (0.1 mol %) H2 (1100 psi) O Me OH EtOH 30 °C Me Me R OH DIAD, PPh3 THF, 20 °C OH Me OH MeOH, -78 °C 83 OH Cu(MeCN)4OTf (5 mol %) MeOBpy (5 mol %) O 84 O Si O t-Bu OH 89% yield Me Me OH t-Bu Me R pentane 13 S Me 67% avg. yield ( 5 x 1.5 g batches) 78 Me NaBH4 CeCl3•7H2O O 93% yield Me hν (254 nm) O 56% yield (2 steps) Me CH2Cl2, –78 °C O OEt 86 reflux 85% yield 45 g scale t-Bu2Si(OTf)2 2,6-lutidine 14 2. Hg(OAc)2 (20 mol%) R Me 84 $235/g from Sigma 83 9 12 10 ABNO (5 mol %) NMI (10 mol %) MeCN, air, 20 °C Me OH O 10 12 9 82 14 O C then HCl (aq.) 19 °C D 13 Cu(MeCN)4OTf (5 mol %) MeOBpy (5 mol %) HO O then K2CO3 85 MeOH O Si O t-Bu t-Bu not isolated 93% yield O Si O t-Bu t-Bu 86 14 O Me 82% yield OH 13 10 Me O H 12 m-CPBA CH2Cl2, 0 to 19 °C; ABNO (5 mol %) NMI (10 mol %) MeCN, air, 20 °C 91% yield 9 H O H 79 Me O O Si O t-Bu t-Bu 87 Chapter 2 – Convergent Approach to C19-Diterpenoid Alkaloids 17 To access a precursor to an organometallic C/D-bicycle fragment, C10-ketone 87 was converted to an alkenyl halide (Scheme 2.3). The ketone was first converted to alkenyl triflate 88 using KHMDS and Comins’ reagent. A Stille coupling of 88 and hexamethylditin forms the corresponding alkenyl stannane which is converted to alkenyl iodide 89 upon treatment with N-iodosuccinimide. While this sequence was effective, it requires the use of stoichiometric quantities of the expensive and highly toxic reagent hexamethylditin. Recently, our lab reported a convenient nickel-catalyzed method to achieve the same transformation.6 The conditions were optimized for substrate 88, screening nickel (II) catalysts, catalyst loading and additives (Table 2.1). 5 mol % loading of Ni(OAc)2•4H2O with NMI as the additive were the optimal conditions, delivering alkenyl bromide 90 in 87% yield. The main limitation was scalability of the reaction—scaling the reaction past 2.3 mmol (1.0 grams) of substrate results in incomplete conversion. This is likely due to the heterogeneity of the reaction; vigorous stirring was key to achieving complete conversion of starting material. In all, this represents a 10-step synthesis of the C/D-bicycle coupling partner 90. Scheme 2.3 Synthesis alkenyl halide coupling partner 89 via Stille coupling. I TfO O 10 Me6Sn2, Pd(PPh3)4 LiCl, THF, 70 °C KHMDS, THF, –78 °C then NIS 0 °C then Comins’ reagent O Si O t-Bu t-Bu 87 96% yield O Si O t-Bu t-Bu 88 80% yield O Si O t-Bu t-Bu 89 Chapter 2 – Convergent Approach to C19-Diterpenoid Alkaloids 18 Table 2.1 Optimization of Ni-catalyzed conversion of enol triflate 88 to alkenyl bromide 90. Br TfO [Ni] (x mol %) Zn (2x mol %) additive (2x mol %) O Si O t-Bu t-Bu 88 LiBr (1.5 equiv.) 3:1 THF:DMA O Si O t-Bu t-Bu 90 Entry Scale (mmol) [Ni] [Ni] loading (x) Additive Temp yield 1 0.1 NiBr2 10 mol % DMAP 21 °C 30%a 2 0.1 NiBr2•dme 10 mol % ‘’ 21 °C 60%a 3 0.1 Ni(OAc)2 •4H2O 2.5 mol % “ 26 °C 3:1 pdt:SM 4 0.1 “ 5 mol % ‘’ 26 °C 1.4:1 pdt:SM 5 0.1 " 7.5 mol % ‘’ 21 °C 74%a 6 0.1 “ 10 mol % ‘’ 26 °C 69% 7 0.1 “ 10 mol % NMI 25 °C 66% 8 0.3 “ 7.5 mol % “ 21 °C 78% 9 0.3 “ 5 mol % “ 21 °C 81% 10 1.0 “ 5 mol % “ 21 °C 87% 11 2.3 Ni(OAc)2 • 4H2O 5 mol % NMI 21 °C 87% aNMR yield using dimethylfumarate as internal standard 2.3.2 Enantioselective Synthesis of A/F-Bicycle Epoxy Ketone 71 Our synthesis of epoxy ketone fragment 71 began with an asymmetric Michael addition of dimethyl malonate (74) to 2-cyclopenten-1-one (73) using Shibasaki’s heterobimetallic catalyst 97 (Scheme 2.4).7 We developed a highly scalable protocol for this reaction, providing 91 in 88% yield and 91% enantiomeric excess (ee) on 31 g scale.8 The ketone of 91 was converted to ketal 92 by treating with ethylene glycol and catalytic acid in benzene at reflux using a Dean-Stark reaction setup. Alkylation of the malonate with three-carbon fragment 75 forms the C4 quaternary center in 93. Acid-catalyzed double dioxolane deprotection and concomitant aldol condensation closes the A-ring to give enone 94. A two-step sequence was employed to introduce the epoxide on the concave face: treatment of 94 with the strongly Chapter 2 – Convergent Approach to C19-Diterpenoid Alkaloids 19 electrophilic bromonium ion source N-bromosacharrin (95) and water furnishes bromohydrin 96, isolated as a single diastereomer in 63% yield. The crude diastereomeric ratio (d.r.) for this reaction was ~3:1. Elimination of HBr to yield the epoxide 71 was easily achieved by treating 96 with triethylamine at ambient temperature. 71 was recrystallized to enantiopurity, giving access to this A/F-fragment in six steps from commercial starting materials. Scheme 2.4 Enantioselective synthesis of epoxy ketone 71. MeO2C + MeO2 C 74 GaNa-(S)-BINOL (10 mol%) NaOt-Bu (7 mol%) O MeO2C THF/Et2O, 21 °C 73 ethylene glycol p-TsOH•H2O MeO2C O H 91 88% yield, 91% ee [31 g scale] MeO2C O MeO2C benzene, reflux 85% yield O S O O O 95 MeCN/H2O, 21 °C 63% yield 75 O N Br O H 92 HCl (aq) MeO2C MeO2C H O acetone 70 °C 94 74% yield 2 MeO2C 4 MeO2C H 93 O Br NaH, n-Bu4NI O O O DMF, 0 to 21 °C 91% yield OH MeO2C MeO2C H 96 Et3N Br O CH2Cl2, 21 °C 99% yield MeO2C MeO2C H Na O Ga O O O O A F O 71 GaNa-(S)-BINOL 97 recrystallized to 99% ee 2.4 1,2-ADDITION/SEMIPINACOL REARRANGEMENT FRAGMENT COUPLING SEQUENCE Having synthesized our key fragments for the synthesis of 1, we set out to test our fragment coupling sequence (Scheme 2.5). Alkenyl bromide 90 was converted to the alkenyl lithium 72 upon treatment with tert-butyllithium at low temperature. This alkenyl lithium was added slowly by cannula to a solution of the epoxy ketone 71 at -94 °C. The 1,2-addition was quenched with TMSCl to give epoxy silyl ether 98 in good yield on 3.8 gram scale. Treatment of the 1,2-addition product 98 with a catalytic amount of the Lewis acid TMSNTf2 and a pyridine base affords semipinacol rearrangement product 99 in near quantitative yield on 4.3 Chapter 2 – Convergent Approach to C19-Diterpenoid Alkaloids 20 gram scale. This remarkable two-step sequence brings together all of the carbon atoms needed to access the aconitine core and forms the key C11 quaternary center, demonstrating the power of 1,2-addition/semipinacol rearrangement as a fragment coupling tactic. Scheme 2.5 1,2-Addition/semipinacol rearrangement fragment coupling to prepare 99. Br Li MeO2C MeO2C H t-BuLi C C D F O 71 MeO2C MeO2C H O 11 OTMS O Si O t-Bu t-Bu 72 THF, –94 °C; then TMSCl, –78 to 23 °C 77% yield [3.8 g scale] MeO2C 17 10 D THF, -78 °C O Si O t-Bu t-Bu 90 O A 11 TMSNTf2 (10 mol %) 10 OTMS CO2Me O 17 2,6-t-Bu2-4-MePy CH2Cl2, –78 °C 98 O Si O t-Bu t-Bu 97% yield [4.8 g scale] O Si O 99 t-Bu t-Bu 2.5 CONCLUDING REMARKS A 1,2-addition/semipinacol rearrangement fragment coupling strategy to access C19 and C20 diterpenoid alkaloids has been described. Retrosynthetic analysis of the C19 diterpenoid alkaloid (–)-talatisamine (1) identifies [3.2.1]-bicyclooctene 91 and epoxy ketone 71 as practical coupling partners. Synthesis of a single enantiomer of 91 is achieved through diastereoselective intramolecular meta-photocycloaddition of an arene tethered to an alkene by a cleavable chiral auxiliary. An enantioselective Michael addition enables the enantioselective synthesis of epoxy ketone 71. The aforementioned fragment coupling between 91 and 71 forms the key C11 quaternary center of the natural product in 99. Chapter 2 – Convergent Approach to C19-Diterpenoid Alkaloids 21 2.6 EXPERIMENTAL SECTION Preparation of GaNa-(S)-BINOL solution (0.05 M in 9:1 THF:Et2O):1,2 OH OH Na NaOtBu O GaCl3 O THF/Et2O, 19 °C (S)-BINOL ((S)-100) Ga O O Ga-Na-(S)-BINOL ((S)-97) In a flame-dried 500 mL round-bottom flask, NaOtBu (10.92 g, 0.114 mol, 4.0 equiv) was dissolved in THF (200 mL) under N2 atmosphere. In a flame-dried 1 L round-bottom flask, (S)-(–)-1,1’-bi(2-naphthol) ((S)-BINOL, (S)-100) (16.26 g, 56.8 mmol, 2.0 equiv) was dissolved in THF (172 mL) under N2 atmosphere. The solution of NaOtBu was transferred via cannula to the solution of (S)-100 over ~20 minutes at ambient temperature (19 °C). The mixture was stirred for an additional 30 minutes. In a flame-dried 1 L round-bottom flask, gallium (III) chloride (5.00 g, 28.4 mmol, 1.0 equiv) was dissolved in a mixture of THF (142 mL) and Et2O (56 mL) under N2 atmosphere. The solution of (S)-S1 and NaOtBu was transferred to the solution of GaCl3 via cannula over ~45 minutes at ambient temperature (19 °C). The mixture was stirred for an additional 2 hours before the heterogeneous mixture was allowed to settle, unperturbed for 18 hours. Enantioselective synthesis of (S)-(–)-14: O MeO2C CO2Me 74 73 O (S)-97 (10 mol %) NaOtBu (7 mol %) THF/Et2O 19-20°C, 40h MeO2C CO2Me 88% yield, 91% ee (S)-91 A flame-dried 1 L, 3-necked round-bottom flask with 24/40 joints and a 1” Teflon coated egg-shaped magnetic stir bar was charged with NaOtBu (0.97 g, 10.1 mmol, 0.07 equiv). The flask was fitted with an oven-dried 250 mL graduated addition funnel and two rubber Chapter 2 – Convergent Approach to C19-Diterpenoid Alkaloids 22 septa. The flask was evacuated and backfilled with nitrogen three times (5 minutes under vacuum per cycle) before anhydrous THF (22.5 mL) was added by syringe. The supernatant solution of (S)-100 (288 mL, 14.4 mmol, 0.10 equiv) was measured by transferring to the addition funnel via cannula and then added to the suspension of NaOtBu. Dimethyl malonate (74) (16.5 mL, 144 mmol, 1.0 equiv) was then added by syringe, followed by 2-cyclopenten1-one (73) (12.0 mL, 144 mmol, 1.0 equiv). The mixture was stirred at room temperature (1920 °C) for 40 hours. The reaction was transferred to a 2L Erlenmeyer flask and 900 mL of 1M HCl (aq.) was added slowly while the mixture stirred. The mixture was transferred to a 4L separatory funnel and extracted with EtOAc (3 x 600 mL). The combined organic phases were washed with saturated aqueous NaHCO3 (400 mL). The aqueous NaHCO3 wash was back-extracted with EtOAc (200 mL). The combined organic phases were washed with brine (400 mL), dried over Na2SO4 (~300 g) filtered through cotton and concentrated in vacuo. The crude product was transferred to a tared 100 mL round-bottom flask with a 0.25” stir bar and dried on highvacuum while heating to 75 °C with stirring for 1h. The crude product was obtained as a yellow oil (38.5 g). The flask of crude product was fitted with a short-path distillation head. The product was purified by distillation under reduced pressure (180 °C oil bath, vacuum line at 210-214 mTorr. Four fractions were collected in the following quantities: 0.48 g (40-108 °C), 19.40 g (108-112 °C), 5.46 g (112-114 °C), 3.05 g (108-120 °C). The first and fourth fractions were combined in a 25 mL round-bottom flask with a 14/20 joint and a 0.25” egg-shaped stir bar and redistilled in the same manor to give 2.16 g (108-112 °C), which was combined with the second and third fractions of the initial distillation to give (S)-91 as a colorless oil (27.0 g, 126 Chapter 2 – Convergent Approach to C19-Diterpenoid Alkaloids 23 mmol, 88% yield, 91% ee). Enantiomeric excess was measured using chiral supercritical fluid chromatography (SFC) using a Mettler SFC supercritical CO2 analytical chromatography system (CO2 = 1450 psi, column temperature = 40 °C) with a Chiralcel AD-H column (4.6 mm x 25 cm) eluting with 5 % isopropanol at a flow rate of 2.5 mL/min over 12 minutes. A racemic standard of (±)-14 was prepared using previously reported methods.3 1 H NMR (600 MHz, CDCl3): δ 3.77 (s, 3H), 3.75 (s, 3H), 3.38 (d, J = 9.4 Hz, 1H), 2.90-2.83 (m, 1H), 2.51 (dd, J = 18.4, 7.6 Hz, 1H), 2.34 (dd, J = 17.5, 8.2 Hz, 1H), 2.30 – 2.16 (m, 2H), 2.01 (dd, J = 18.5, 11.0 Hz, 1H), 1.69 – 1.61 (m, 1H). 13 C NMR (101 MHz, CDCl3): δ 217.1, 168.6, 168.56, 56.2, 56.2, 52.8, 43.0, 38.3, 36.5, 27.6. FTIR (NaCl, thin film): 2956, 1738, 1439, 1322, 1236, 1211, 1172, 1154, 1017, 787 cm-1. HRMS: (ESI-TOF) calc’d for C10H14O5 [M + H]+ 215.0914, found 215.0915 (0.46 ppm difference). [𝜶]𝟐𝟓 𝐃 = –84.8° (c = 1.22, CHCl3). TLC (2:1 Hexanes:EtOAc), Rf: 0.2 (purple in p-anisaldehyde) SFC Data for racemic 91: Chapter 2 – Convergent Approach to C19-Diterpenoid Alkaloids 24 SFC Data for enantioenriched (–)-91: Preparation of ketal 92: MeO2C MeO2C ethylene glycol p-TsOH O H 91 PhMe, reflux 85% yield MeO2C MeO2C O H O 92 In a flame dried 1 L, round-bottom flask equipped with a reflux condenser and Dean– Stark trap, ketone 91 (38.6 g, 180 mmol, 1.0 equiv) was dissolved in toluene (600 mL). To this solution was added ethylene glycol (20.2 mL, 360 mmol, 2.0 equiv) and p-TsOH•H2O (3.42 g, 18.0 mmol, 0.10 equiv), and the reaction was heated to reflux for 11 h. The reaction was then Chapter 2 – Convergent Approach to C19-Diterpenoid Alkaloids 25 cooled to room temperature and diluted with Et2O (1200 mL). The solution was washed with saturated aqueous NaHCO3 (2 x 400 mL), and the combined aqueous phase was extracted with Et2O (750 mL). The combined organic phase was washed with brine (500 mL), dried over Na2SO4, filtered, and concentrated in vacuo. Purification by silica gel chromatography (25% EtOAc in hexanes with 2% NEt3) provided ketal 92 (39.6 g, 153 mmol, 85% yield) as a colorless oil. 1 H NMR (400 MHz, CDCl3): δ 3.93 – 3.84 (m, 4H), 3.72 (m, 6H), 3.31 (d, J = 10.2 Hz, 1H), 2.75 – 2.62 (m, 1H), 2.08 (ddd, J = 13.7, 8.0, 1.2 Hz, 1H), 1.99 – 1.86 (m, 2H), 1.85 – 1.75 (m, 1H), 1.56 (dd, J = 13.6, 9.5 Hz, 1H), 1.48 – 1.36 (m, 1H). 13 C NMR (101 MHz, CDCl3): δ 169.0, 168.9, 117.0, 64.3, 64.2, 56.7, 52.4, 52.4, 40.6, 36.8, 35.5, 28.2. FTIR (NaCl, thin film): 2956, 2885, 1754, 1734, 1435, 1316, 1216, 1151, 1079, 1017, 947 cm-1. HRMS: (PMM) calc’d for C12H19O6 [M + H]+ 259.1176, found 259.1169. [𝜶]𝟐𝟓 𝐃 = +2.2° (c = 1.4, CHCl3). TLC (2:1 Hexanes:EtOAc), Rf: 0.3 (purple in p-anisaldehyde) Preparation of bis-ketal 93: 75 Br MeO2C MeO2C O O O O 91% yield 92 O NaH, n-Bu4NI DMF, 0°C to rt H O MeO2C MeO2C H O O 93 A flame dried 1 L round-bottom flask was charged with diester 92 (39.9 g, 155 mmol, 1.0 equiv) and DMF (386 mL). The solution was cooled to 0 °C before NaH (60% dispersion Chapter 2 – Convergent Approach to C19-Diterpenoid Alkaloids 26 in mineral oil, 7.42 g, 185 mmol, 1.2 equiv) was added in portions. n-Bu4NI (11.39 g, 30.8 mmol, 0.20 equiv) and 2-(2-bromoethyl)-1,3-dioxolane (75, 27.2 mL, 232 mmol, 1.5 equiv) were added sequentially, and the reaction was allowed to warm to room temperature and stirred for 72 hours. The reaction was then quenched by addition of sat. NH4Cl (600 mL). The mixture was extracted with EtOAc (3 x 1 L), and the combined organic extracts were washed with sat. NH4Cl (2 x 400 mL) followed by brine (400 mL), dried over Na2SO4, filtered, and concentrated in vacuo. Purification by silica gel chromatography (10 to 50% EtOAc in hexanes) afforded bis-ketal 93 (50.4 g, 141 mmol, 91% yield) as a colorless oil. 1 H NMR (500 MHz, CDCl3): δ 4.83 (td, J = 4.5, 1.9 Hz, 1H), 3.97 – 3.81 (m, 8H), 3.71 (s, 3H), 3.71 (s, 3H), 2.70 – 2.60 (m, 1H), 2.05 – 1.95 (m, 3H), 1.91 – 1.80 (m, 2H), 1.80 – 1.66 (m, 2H), 1.64 – 1.50 (m, 3H). 13 C NMR (126 MHz, CDCl3): δ 171.2, 171.1, 116.6, 103.9, 64.9, 64.9, 64.2, 64.1, 59.8, 52.2, 52.1, 40.4, 38.0, 35.3, 29.0, 27.9, 25.4. FTIR (NaCl, thin film): 2954, 2915, 2877, 1728, 1434, 1227, 1141, 1024 cm-1. HRMS: (FAB) calc’d for C17H27O8 [M + H]+ 359.1706, found 359.1716. [𝜶]𝟐𝟓 𝐃 = –0.69° (c = 0.60, CHCl3). TLC (1:1 Hexanes:EtOAc), Rf: 0.3 (turquoise in p-anisaldehyde) Chapter 2 – Convergent Approach to C19-Diterpenoid Alkaloids 27 Preparation of enone 94: O O HCl (aq) O MeO2C MeO2C H O acetone, 70 °C MeO2C MeO2C H O 74% yield 93 94 In a 1L, round-bottom flask equipped with a reflux condenser, bis-dioxolane 93 (25.2 g, 70.3 mmol, 1.0 equiv) was dissolved in acetone (280 mL). Aqueous HCl (70 mL, 2 N, 140 mmol, 2.0 equiv) was added, and the reaction was heated to 70 °C with stirring for 16 h. The reaction was cooled to room temperature and quenched with sat. NaHCO3 (300 mL) and partially concentrated in vacuo to remove acetone. The resulting mixture was extracted with CH2Cl2 (3 x 450 mL), and the combined organic extracts were dried over Na2SO4, filtered, and concentrated in vacuo. The crude residue was purified by silica gel chromatography (10 to 40% EtOAc in hexanes) to afford enone 94 (13.03 g, 51.6 mmol, 74% yield) as a white solid. 1 H NMR (500 MHz, CDCl3): δ 6.69 (q, J = 3.8 Hz, 1H), 3.78 (s, 3H), 3.67 (s, 3H), 3.10 (qdd, J = 7.4, 3.5, 2.1 Hz, 1H), 2.53 – 2.37 (m, 4H), 2.32 – 2.22 (m, 2H), 2.11 –1.90 (m, 2H). 13 C NMR (126 MHz, CDCl3): δ 204.6, 171.8, 169.5, 137.1, 131.3, 55.0, 52.7, 52.2, 42.8, 37.7, 29.4, 23.9, 23.2. FTIR (NaCl, thin film): 3022, 2956, 2898, 2848, 1732, 1659, 1451, 1434, 1283, 1256, 1214, 1174, 1143, 1064 cm–1. HRMS: (FAB) calc’d for C13H17O5 [M + H]+ 253.1076, found 253.1076. [𝜶]𝟐𝟓 𝐃 = –34° (c = 1.7, CHCl3). TLC (1:1 Hexanes:EtOAc), Rf: 0.6 (UV, blue in p-anisaldehyde) Chapter 2 – Convergent Approach to C19-Diterpenoid Alkaloids 28 Preparation bromohydrin 96: O O S N Br O MeO2C MeO2C H 94 O MeCN/H2O, 0 to 20 °C 62% yield OH 95 MeO2C MeO2C H Br O 96 A 250 mL, round-bottom flask was charged with enone 94 (3.91 g, 15.5 mmol, 1.0 equiv), MeCN (64 mL), and H2O (12 mL), and the mixture was cooled to 0 °C. Nbromosaccharin (5.28 g, 20.1 mmol, 1.3 equiv) was added slowly in portions over 30 minutes. The reaction was warmed to 20 °C and stirred for 6.5 h, then quenched by addition of sat. Na2S2O3 (150 mL). The mixture was extracted with Et2O (3 x 250 mL), and the combined organic extracts were washed with sat. NaHCO3 (2 x 150 mL), brine (150 mL), dried over MgSO4, filtered, and concentrated in vacuo to afford a crude white solid. This was dissolved in Et2O (600 mL) at 35 °C. The solution was concentrated until ca. 300 mL of solvent remained, and then hexanes (300 mL) was added. The solution was concentrated until ca. 300 mL of solvent remained, with white solids suspended within. The solvent was decanted into a Büchner funnel, washing the flask carefully with hexanes. The residual solids in the flask were collected, affording bromohydrin 96 (3.37 g, 9.65 mmol, 62% yield) as a white solid. Note: An NMR sample of the crude reaction mixture shows ~3:1 mixture of diastereomers. 1 H NMR (500 MHz, Chloroform-d): δ 4.37 (t, J = 2.8 Hz, 1H), 3.77 (s, 3H), 3.74 (s, 3H), 3.10 (ddd, J = 12.3, 5.9, 1.6 Hz, 1H), 2.81 (d, J = 1.7 Hz, 1H), 2.79 – 2.71 (m, 1H), 2.51 (dd, J = 18.0, 7.3 Hz, 1H), 2.41 – 2.32 (m, 2H), 2.30 – 2.17 (m, 2H), 2.07 – 1.98 (m, 2H). 13 C NMR (101 MHz, CDCl3): δ 211.6, 171.3, 171.1, 76.5, 55.5, 53.0, 52.5, 47.9, 42.2, 36.2, 28.2, 27.6, 21.3. Chapter 2 – Convergent Approach to C19-Diterpenoid Alkaloids 29 FTIR (NaCl, thin film): 3436, 2998, 2955, 2847, 1748, 1732, 1435, 1403, 1252, 1213, 1152, 1091, 1073, 1056, 980 cm-1. HRMS: (FAB) calc’d for C13H18BrO6 [M + H]+ 349.0287, found 349.0275. [𝜶]𝟐𝟓 𝐃 = –49° (c = 0.17, CHCl3). TLC (1:1 Hexanes:EtOAc), Rf: 0.6 (orange in p-anisaldehyde) Preparation of epoxyketone 71: OH Et3N Br MeO2C MeO2C H O CH2Cl2 O MeO2C MeO2C H O 99% yield 96 71 In a 250 mL, round-bottom flask, bromohydrin 96 (6.66 g, 19.1 mmol, 1.0 equiv) was dissolved in CH2Cl2 191 mL). Et3N (3.2 mL, 22.9 mmol, 1.2 equiv) was added, and the reaction was stirred at 20 °C for 9 h. The mixture was loaded directly onto a column of silica gel (slurried in hexanes), and purification by chromatography (100% hexanes, then 40% EtOAc in hexanes) afforded epoxyketone 71 (5.10 g, 19.1 mmol, 99% yield) as a white solid. The resulting solid was recrystallized from acetone layered with hexanes to afford epoxyketone 71 in >99% ee. 1 H NMR (500 MHz, CDCl3): δ 3.76 (s, 1H), 3.76 – 3.73 (m, 4H), 2.93 (dd, J = 12.5, 5.8 Hz, 1H), 2.68 (qd, J = 11.7, 7.4 Hz, 1H), 2.58 (ddd, J = 18.7, 7.6, 1.6 Hz, 1H), 2.48 – 2.28 (m, 4H), 2.12 – 2.01 (m, 1H), 1.72 – 1.61 (m, 1H). 13 C NMR (126 MHz, CDCl3): δ 212.0, 171.4, 169.8, 63.0, 56.1, 55.2, 53.1, 52.4, 43.8, 38.4, 29.3, 23.0, 21.7. FTIR (NaCl/thin film): 3006, 2956, 2903, 2848, 1755, 1732, 1453, 1434, 1408, 1372, 1306, 1251, 1214, 1176, 1139, 1058, 890 cm-1. Chapter 2 – Convergent Approach to C19-Diterpenoid Alkaloids HRMS: (FAB) calc’d for C13H17O6 [M + H]+ 269.1025, found 269.1050. [𝜶]𝟐𝟓 𝐃 = –171° (c = 0.60, CHCl3). TLC (1:1 Hexanes:EtOAc), Rf: 0.6 (yellow/orange in p-anisaldehyde) SFC data for racemic epoxyketone 9: SFC data for enantiopure epoxyketone 9: 30 Chapter 2 – Convergent Approach to C19-Diterpenoid Alkaloids 31 Preparation of Diol (2R, 4R)-(–)–84:4 Me Me O O RuCl2[(R)-BINAP] (0.1 mol %) H2 (1100 psi) EtOH, 30 °C 83 76% yield Me Me OH OH (2R,4R)-(–)84 A flame-dried 100 mL Schlenk flask was charged with acetylacetone (83, 32.3 mL, 315 mmol, 1.0 equiv) and 200-proof EtOH (32.3 mL). The solution was sparged with N2 for 1 hour. A parr bomb, an oven-dried 150 mL glass jar with a stirbar, and the Schlenk flask of acac/EtOH solution were brought into a N2-filled glovebox. The solution of acac in EtOH was transferred to the glass jar and RuCl2[(R)-BINAP] (0.250 g, 0.315 mmol, 0.1 mol %) was added. The jar of reaction mixture was lowered into the bomb, which was then sealed and brought out of the glovebox. The bomb was purged with H2 by pressurizing to 100 psi then releasing the pressure (3 cycles). The bomb was pressurized with H2 to 1100 psi, lowered into an oil bath at 30 °C and secured with a chain clamp. After 1 day, the pressure had dropped to ~850 psi, so the vessel was pressurized to 1100 psi. After another 1 day, the pressure had dropped to ~900 psi and was again pressurized to 1100 psi. The reaction was stirred an additional 4 days (6 days total). The pressure was released and the reaction mixture was transferred to a 250 mL roundbottom flask and sparged with N2 for 1 hour. The mixture was concentrated in vacuo. The product was purified by distillation using a short-path distillation head under reduced pressure (120 °C oil bath, 15 torr). (2R,4R)-2,4-pentanediol ((2R,4R)-(–)-84) was isolated as a colorless oil which crystallized on standing (b.p. 90-100 °C (15 torr), 24.9 g, 239 mmol, 76% yield). 1 H NMR (500 MHz, Chloroform-d) δ 4.18 (q, J = 6.0 Hz, 2H), 2.26 (br, 2H), 1.62 (dd, J = 6.0, 5.1 Hz, 2H), 1.26 (d, J = 6.3 Hz, 6H). Chapter 2 – Convergent Approach to C19-Diterpenoid Alkaloids 32 Preparation of aryl ether 100: Me Me HO Me DIAD, PPh3 + OH 84 Me O OH THF, 0 to 21 °C OH 85 65% yield 100 In a flame dried 1 L, round-bottom flask equipped with an addition funnel, (2R, 4R)pentanediol (84, 10.42 g, 100 mmol, 1.0 equiv), phenol (85, 10.35 g, 110 mmol, 1.1 equiv), and PPh3 (31.47 g, 120 mmol, 1.2 equiv) were dissolved in THF (450 mL) and the mixture was cooled to 0 °C. A solution of DIAD (23.6 mL, 120 mmol, 1.2 equiv) in THF (100 mL) was prepared in another flask, and transferred to the addition funnel. The DIAD solution was added dropwise to the vigorously stirring reaction over 40 min at 0 °C. After complete addition of the DIAD solution, the reaction was allowed to warm to 20 °C and stirred for 11 h before being partially concentrated in vacuo to a volume of ~200 mL. The mixture was filtered over celite to remove (O)PPh3, and then further concentrated in vacuo. Purification of the crude residue by silica gel chromatography (10 to 45% EtOAc in hexanes) afforded arene 100 (11.7 g, 64.9 mmol, 65% yield). Spectroscopic data matched that reported by Sugimura et al.5 1 H NMR (500 MHz, CDCl3): δ 7.31 – 7.26 (m, 2H), 6.99 – 6.91 (m, 3H), 4.61 (dqd, J = 8.6, 6.0, 4.4 Hz, 1H), 4.06 (dqt, J = 8.9, 6.1, 2.7 Hz, 1H), 2.56 (s, 1H), 1.95 (dt, J = 14.4, 8.8 Hz, 1H), 1.70 (ddd, J = 14.4, 4.5, 3.1 Hz, 1H), 1.31 (d, J = 6.0 Hz, 3H), 1.23 (d, J = 6.2 Hz, 3H). TLC (2:1 Hexanes:EtOAc), Rf: 0.4 (UV, purple in p-anisaldehyde) Chapter 2 – Convergent Approach to C19-Diterpenoid Alkaloids 33 Preparation of arene olefin 78: Me Me Me O OH Hg(OAc)2 (20 mol%) Me O O OEt 45 °C 100 >95% yield 78 In a 250 mL, round-bottom flask equipped with a reflux condenser, arene 100 (3.90 g, 21.6 mmol, 1.0 equiv) was dissolved in ethyl vinyl ether (108 mL), and Hg(OAc)2 (689 mg, 2.16 mmol, 0.10 equiv) was added. The mixture was heated to reflux and stirred for 18 h, at which point, the reaction was cooled to room temperature, and another portion of Hg(OAc)2 (689 mg, 2.16 mmol, 0.10 equiv) was added, and the reaction was brought back to reflux. After stirring for another 15 h at reflux, the reaction was quenched by addition of sat. NaHCO3 (100 mL). The layers were separated, and the aqueous phase was extracted with Et2O (3 x 200 mL). The combined organic extracts were washed with brine (200 mL), dried over MgSO4, filtered, and concentrated in vacuo. The crude residue was purified by filtering over a short plug of silica (eluting with 10% EtOAc/1% Et3N in hexanes) to provide arene olefin 78 (4.5 g, 21.8 mmol, 100% yield) as a colorless oil. Spectroscopic data matched that reported by Sugimura et al.5 1 H NMR (400 MHz, CDCl3): δ 7.31 – 7.24 (m, 2H), 6.97 – 6.86 (m, 3H), 6.34 (dd, J = 14.2, 6.6 Hz, 1H), 4.52 (h, J = 6.2 Hz, 1H), 4.33 (dd, J = 14.2, 1.6 Hz, 1H), 4.12 (h, J = 6.3 Hz, 1H), 4.03 (dd, J = 6.7, 1.6 Hz, 1H), 2.20 (dt, J = 13.9, 6.9 Hz, 1H), 1.65 (dt, J = 14.0, 6.0 Hz, 1H), 1.33 (d, J = 6.1 Hz, 3H), 1.25 (d, J = 6.2 Hz, 3H). TLC (2:1 Hexanes:EtOAc), Rf: 0.7 (UV, purple in p-anisaldehyde) Chapter 2 – Convergent Approach to C19-Diterpenoid Alkaloids 34 Preparation of 7-substituted meta-photoadduct 79: H hν (254 nm) O O Me pentane 25 °C Me 78 1.5 g batches, 67% avg. yield O Me H O H Me 79 In a 1 L, round-bottom quartz flask, arene olefin 78 (1.5 g, 7.3 mmol, 1.0 equiv) was dissolved in pentane (700 mL). This solution was sparged with Ar for 75 min, then irradiated with stirring using a Honeywell 254 nm lamp for 27 h. The temperature was deliberately maintained at 25 °C using ventilation fans. Upon completion, the reaction was concentrated in vacuo to afford a crude residue. This procedure was repeated for a total of 5 batches. All 5 crude batches were combined and purified by silica gel chromatography (9% EtOAc in hexanes) to afford 7-substituted photoadduct 79 (5.0 g, 24.2 mmol, 67% average yield over 5 batches) as a white solid. Spectroscopic data matched that reported by Sugimura et al.5 1 H NMR (400 MHz, Chloroform-d): δ 5.66 – 5.58 (m, 1H), 5.43 (dd, J = 5.6, 1.8 Hz, 1H), 4.48 (dd, J = 6.9, 2.5 Hz, 1H), 4.31 (p, J = 6.5 Hz, 1H), 4.05 (p, J = 6.6 Hz, 1H), 3.25 (dd, J = 8.3, 2.6 Hz, 1H), 2.50 – 2.42 (m, 3H), 2.34 (dt, J = 17.4, 7.4 Hz, 1H), 2.09 (ddd, J = 14.6, 6.9, 1.1 Hz, 1H), 1.59 (d, J = 17.4 Hz, 1H), 1.23 (s, 3H), 1.21 (s, 3H). TLC (3:1 Hexanes:EtOAc), Rf: 0.5 (UV, purple in p-anisaldehyde) Chapter 2 – Convergent Approach to C19-Diterpenoid Alkaloids 35 Preparation of allylic alcohol 82: H Me m-CPBA CH2Cl2, 0 to 19 °C O Me H O Me H Me OH O then HCl (aq) 82% yield 79 O OH 82 A 2 L, round-bottom flask was charged with photoadduct 79 (11.84 g, 57.4 mmol, 1.0 equiv) and CH2Cl2 (575 mL), and cooled to 0 °C. m-CPBA (77% by wt., 14.79 g, 66.0 mmol, 1.15 equiv) was added, and the reaction was stirred for 30 min at 0 °C, then warmed to room temperature. After stirring for an additional 5 h at room temperature, 2 N HCl (115 mL, 230 mmol, 4.0 equiv) was added, and the biphasic mixture was stirred vigorously. After 40 min, the reaction was quenched by careful addition of sat. NaHCO3 (330 mL), and stirred until bubbling ceased (ca. 30 min). The layers were separated, and the aqueous phase was extracted with 3:1 CHCl3:i-PrOH (3 x 600 mL). The combined organic extracts were dried over Na2SO4, filtered, and concentrated in vacuo. Purification by silica gel chromatography (40 to 45% acetone in hexanes) afforded allylic alcohol 82 (11.34 g, 47.2 mmol, 82% yield) as a colorless oil. 1 H NMR (400 MHz, CDCl3): δ 5.97 (dd, J = 9.0, 7.2 Hz, 1H), 5.77 (ddd, J = 9.0, 3.8, 1.2 Hz, 1H), 4.58 (s, 1H), 4.02 (dd, J = 6.1, 2.2 Hz, 1H), 3.90 (dqd, J = 9.2, 6.2, 3.0 Hz, 1H), 3.70 (dqd, J = 8.8, 6.0, 4.3 Hz, 1H), 2.79 (dd, J = 7.2, 1.5 Hz, 1H), 2.67 – 2.53 (m, 2H), 2.29 (br s, 1H), 2.18 – 2.05 (m, 2H), 1.61 (dt, J = 14.4, 8.9 Hz, 1H), 1.48 (ddd, J = 14.5, 4.4, 3.1 Hz, 1H), 1.16 (d, J = 3.8 Hz, 3H), 1.14 (d, J = 4.1 Hz, 3H). 13 C NMR (101 MHz, CDCl3): δ 213.9, 131.4, 129.6, 81.1, 75.2, 73.9, 67.0, 51.6, 49.5, 45.9, 32.2, 23.6, 20.0. Chapter 2 – Convergent Approach to C19-Diterpenoid Alkaloids 36 FTIR (NaCl, thin film): 3400, 2969, 2933, 1753, 1458, 1447, 1376, 1329, 1120, 1080, 1048, 926 cm-1. HRMS: (FAB) calc’d for C13H21O4 [M + H]+ 241.1440, found 241.1421. [𝜶]𝟐𝟓 𝐃 = –6.2° (c = 1.0, CHCl3). TLC (1:1 Hexanes:acetone), Rf: 0.3 (KMnO4) Preparation of triol 83: Me Me OH Me NaBH4, CeCl3•7H2O O Me OH O MeOH, –78°C O OH 89% yield 82 OH OH 83 In a 1 L, round-bottom flask, ketone 82 (6.7 g, 28 mmol, 1.0 equiv) and CeCl3•7H2O (15.6 g, 41.8 mmol, 1.5 equiv) were dissolved in MeOH (280 mL). The solution was then cooled to –78 °C, and NaBH4 (1.27 g, 33.5 mmol, 1.2 equiv) was added. After stirring for 2 h at –78 °C, the reaction was quenched with 1 M NaOH (200 mL), and concentrated in vacuo to remove MeOH. The aqueous phase was extracted with EtOAc (8 x 250 mL), and the combined organic extracts were dried over Na2SO4, filtered, and concentrated in vacuo. Filtration of the crude residue over a short plug of silica (eluting with 50% acetone in hexanes) afforded triol 83 (6.1 g, 25 mmol, 89% yield) as a white solid. Note: The product is highly water soluble, and thus requires rigorous extraction with a polar solvent such as EtOAc. Chapter 2 – Convergent Approach to C19-Diterpenoid Alkaloids 1 37 H NMR (400 MHz, CDCl3): δ 5.88 (ddt, J = 9.9, 3.7, 1.7 Hz, 1H), 5.74 (dd, J = 9.4, 7.0 Hz, 1H), 4.46 (s, 1H), 3.97 (dqd, J = 8.9, 6.3, 2.7 Hz, 1H), 3.83 (br s, 1H), 3.77 – 3.67 (m, 3H), 3.56 (br s, 2H), 2.63 – 2.58 (m, 1H), 2.46 (appar s, 1H), 1.77 (td, J = 4.7, 4.1, 1.7 Hz, 2H), 1.63 – 1.47 (m, 2H), 1.15 (d, J = 6.1 Hz, 3H), 1.12 (d, J = 6.0 Hz, 3H). 13 C NMR (101 MHz, CDCl3): δ 129.7, 126.5, 78.3, 75.4, 72.0, 71.7, 68.1, 45.5, 45.4, 40.8, 33.8, 23.4, 20.1. FTIR (NaCl, thin film): 3369, 3032, 2967, 2934, 1757, 1642, 1447, 1420, 1376, 1318, 1180, 1084, 1033, 970, 934 cm-1. HRMS: (FAB) calc’d for C13H23O4 [M + H]+ 243.1596, found 243.1616. [𝜶]𝟐𝟓 𝐃 = –6.2° (c = 1.0, CHCl3). TLC (1:1 Hexanes:acetone), Rf: 0.4 (UV, blue/green in p-anisaldehyde) Preparation of siliconide 84: Me Me Me OH tBu Si(OTf) 2 2 O Me OH O 2,6-lutidine CH2Cl2, –78 °C OH 83 OH 93% yield O Si O 84 t-Bu t-Bu A flame-dried 1 L, round-bottom flask was charged with triol 83 (9.0 g, 37.1 mmol, 1.0 equiv), which was then azeotroped with anhydrous PhMe to remove any trace moisture. CH2Cl2 (370 mL) and 2,6-lutidine (10.3 mL, 89.1 mmol, 2.40 equiv) were added, and the mixture was cooled to –78 °C. To this solution was added t-Bu2Si(OTf)2 (15.0 mL, 44.6 mmol, 1.20 equiv), and the reaction was stirred for 1 h. The reaction was quenched with sat. NaHCO3 (150 mL) and H2O (150 mL), and the layers were separated. The aqueous phase was extracted with CH2Cl2 (3 x 180 mL), and the combined organic extracts were washed with 0.1 M HCl Chapter 2 – Convergent Approach to C19-Diterpenoid Alkaloids 38 (300 mL), washed with sat. NaHCO3 (100 mL), dried over Na2SO4, filtered, and concentrated in vacuo. The crude residue was purified by silica gel chromatography (10 to 25% acetone in hexanes) to afford siliconide 84 (13.27 g, 34.7 mmol, 93% yield) as a colorless oil. 1 H NMR (400 MHz, CDCl3): δ 5.83 (ddd, J = 9.5, 4.4, 1.6 Hz, 1H), 5.74 (ddd, J = 9.6, 6.3, 1.2 Hz, 1H), 4.59 (t, J = 4.8 Hz, 1H), 4.04 – 3.91 (m, 2H), 3.80 – 3.70 (m, 2H), 3.39 (s, 1H), 2.78 (dd, J = 6.3, 4.1 Hz, 1H), 2.77 – 2.67 (m, 1H), 1.82 (ddd, J = 15.2, 7.5, 1.3 Hz, 1H), 1.71 (dd, J = 15.0, 7.9 Hz, 1H), 1.59 (dt, J = 14.5, 9.2 Hz, 1H), 1.51 (ddd, J = 14.5, 4.0, 2.6 Hz, 1H), 1.16 (d, J = 6.2 Hz, 3H), 1.12 (d, J = 6.0 Hz, 3H), 1.05 (s, 9H), 0.96 (s, 9H). 13 C NMR (101 MHz, CDCl3): δ 129.8, 129.1, 76.4, 74.7, 73.7, 71.0, 67.6, 46.4, 45.9, 38.6, 33.0, 28.7, 28.2, 23.6, 21.2, 20.7, 20.0. FTIR (NaCl, thin film): 3436, 3032, 2968, 2934, 2900, 2859, 1476, 1388, 1364, 1326, 1196, 1174, 1058, 1036, 1019, 998, 826 cm-1. HRMS: (FAB) calc’d for C21H39O4Si [M + H]+ 383.2618, found 383.2630. [𝜶]𝟐𝟓 𝐃 = +59° (c = 0.90, CHCl3). TLC (1:1 Hexanes:EtOAc), Rf: 0.7 (green in p-anisaldehyde) Preparation of alcohol 86:6 Me Me Me OH Cu(MeCN)4OTf ABNO MeOBpy, NMI O 84 MeCN O Si O t-Bu t-Bu Me O HO O then K2CO3 85 MeOH O Si O t-Bu not isolated O Si O t-Bu t-Bu 93% yield t-Bu 86 To a 500 mL, round-bottom flask was added alcohol 84 (3.92 g, 10.25 mmol, 1 equiv) and MeCN (102 mL). To this solution was added 4,4-dimethoxy-2,2’-bipyridine (111 mg, 0.513 mmol, 5 mol %), N-methylimidazole (82.5 µL, 1.03 mmol, 10 mol %), and ABNO (72 Chapter 2 – Convergent Approach to C19-Diterpenoid Alkaloids 39 mg, 0.513 mmol, 5 mol %). Finally, Cu(MeCN)4OTf (193 mg, 0.513 mmol, 5 mol %) was added last, causing the solution to turn red/brown. The reaction was vigorously stirred for 60 minutes at room temperature open to air, during which time the reaction mixture turns blue, indicating its completion. At this time, MeOH (205 mL) was added, followed by powdered K2CO3 (powdered with mortar and pestle 7.84 g), causing the solution to turn brown, and the mixture was allowed to stir for 20 hours at room temperature. The mixture was filtered through a plug of silica gel and celite (to remove copper and solid K2CO3), flushed with excess ethyl acetate and concentrated in vacuo. The crude residue was purified by silica gel chromatography (20% ethyl acetate in hexanes, note: streaky on the column) to afford alcohol 86 (2.82 g, 9.53 mmol, 93% yield) as a colorless oil. 1 H NMR (400 MHz, CDCl3): δ 5.82 (ddd, J = 9.4, 4.3, 1.6 Hz, 1H), 5.77 (ddd, J = 9.5, 6.1, 1.2 Hz, 1H), 4.78 (dd, J = 5.5, 4.1 Hz, 1H), 4.06 (dd, J = 6.6, 1.6 Hz, 1H), 4.01 (t, J = 3.9 Hz, 1H), 2.81 – 2.72 (m, 1H), 2.63 (ddd, J = 6.0, 4.3, 1.2 Hz, 1H), 1.82 (dd, J = 14.8, 6.8 Hz, 1H), 1.70 (ddt, J = 15.0, 7.9, 1.2 Hz, 1H), 1.07 (s, 9H), 0.98 (s, 9H). 13 C NMR (101 MHz, CDCl3): δ 129.6, 129.0, 73.6, 71.8, 71.0, 50.4, 38.7, 34.1, 28.7, 28.2, 21.2, 20.7. FTIR (NaCl, thin film): 3401, 3033, 2968, 2935, 2896, 2860, 1476, 1442, 1388, 1364, 1321, 1280, 1224, 1175, 1032, 998, 920, 825 cm-1. HRMS: (FAB) calc’d for C16H27O3Si [M + H – H2]+ 295.1747, found 295.1730. [𝜶]𝟐𝟓 𝐃 = +89° (c = 1.3, CHCl3). TLC (20%EtOAc in Hexanes), Rf: 0.4 (blue in p-anisaldehyde) Chapter 2 – Convergent Approach to C19-Diterpenoid Alkaloids 40 Preparation of ketone 87: O HO Cu(MeCN)4OTf, ABNO, MeOBpy, NMI MeCN O Si O t-Bu t-Bu O Si O t-Bu t-Bu 91% yield 86 87 To a 500 mL, round-bottom flask was added alcohol 86 (2.82 g, 9.53 mmol, 1 equiv) and MeCN (95 mL). To this solution was added 4,4-dimethoxy-2,2’-bipyridine (103 mg, 0.476 mmol, 5 mol %), N-methylimidazole (73.6 µL, 0.953 mmol, 10 mol %), and ABNO (66.7 mg, 0.476 mmol, 5 mol %). Finally, Cu(MeCN)4OTf (179 mg, 0.476 mmol, 5 mol %) was added last, causing the solution to turn red/brown. The reaction was vigorously stirred for 60 minutes at room temperature open to air, during which time the reaction mixture turns blue, indicating its completion. The mixture was then filtered through a plug of silica gel, flushed with excess ethyl acetate, and concentrated in vacuo. The crude residue was purified by silica gel chromatography (15% ethyl acetate in hexanes) to afford ketone 87 (2.55 g, 8.67 mmol, 91% yield) as a white crystalline solid. 1 H NMR (400 MHz, CDCl3): δ 6.08 (ddd, J = 9.3, 4.6, 1.6 Hz, 1H), 5.76 (ddd, J = 9.2, 6.9, 1.4 Hz, 1H), 4.59 (ddt, J = 5.6, 4.1, 1.1 Hz, 1H), 4.27 (ddd, J = 4.5, 3.4, 0.9 Hz, 1H), 3.15 (dddd, J = 6.8, 4.2, 1.4, 0.7 Hz, 1H), 3.02 (dddq, J = 6.8, 5.0, 3.3, 1.6 Hz, 1H), 2.37 – 2.31 (m, 1H), 2.27 (dd, J = 19.4, 6.4 Hz, 1H), 1.09 (s, 9H), 0.99 (s, 9H). 13 C NMR (101 MHz, CDCl3): δ 206.7, 131.3, 124.8, 71.7, 69.8, 55.1, 38.5, 37.6, 28.6, 28.2, 21.1, 20.8. FTIR (NaCl, thin film): 3036, 2968, 2935, 2859, 1743, 1629, 1476, 1404, 1387, 1365, 1298, 1221, 1187, 1145, 1115, 997, 790 cm-1. HRMS: (FAB) calc’d for C16H27O3Si [M + H]+ 295.1730, found 295.1731. Chapter 2 – Convergent Approach to C19-Diterpenoid Alkaloids 41 [𝜶]𝟐𝟓 𝐃 = +51° (c = 1.1, CHCl3). TLC (20%EtOAc in Hexanes), Rf: 0.4 (KMnO4) Note: Does not stain in p-anisaldehyde. Preparation of enol triflate 88: TfO O KHMDS, THF, –78 °C O Si O t-Bu t-Bu then Comins’ reagent 96% yield 87 O Si O t-Bu t-Bu 88 A flame-dried 250 mL, round-bottom flask was charged with ketone 87 (2.00 g, 6.80 mmol, 1.0 equiv), which was azeotroped with anhydrous PhMe to remove trace moisture. THF (58 mL) was added, and the mixture was cooled to –78 °C in a dry ice/acetone bath. KHMDS (16.3 mL, 0.5 M in toluene, 8.15 mmol, 1.2 equiv) was added, causing the solution to turn yellow, and the reaction was stirred for 45 minutes while kept at –78 °C. In a separate flask, a solution of Comins’ reagent (2.93 g, 7.47 mmol, 1.1 equiv) in THF (10 mL) was prepared and cannulated into the first reaction flask. The resulting mixture was stirred for an additional hour at –78 °C, and then quenched with sat. NaHCO3 (40 mL), and allowed to warm to room temperature. The biphasic mixture was then transferred to a separatory funnel with Et2O (75 mL), and the layers were separated. The aqueous layer was extracted with more Et2O (3 x 75 mL). The combined organic layers were then washed with aq. 0.5 M NaOH solution (4 x 75 mL), followed by brine (1 x 100 mL). The combined organic extracts were dried over Na2SO4, filtered, and concentrated in vacuo. Purification of the crude residue by silica gel chromatography (0 to 10% EtOAc in hexanes) afforded vinyl triflate 88 (2.77 g, 6.50 mmol, 96% yield) as a yellow oil which solidified on standing. Chapter 2 – Convergent Approach to C19-Diterpenoid Alkaloids 1 42 H NMR (400 MHz, CDCl3): δ 6.21 (ddd, J = 9.6, 6.0, 1.1 Hz, 1H), 5.84 (dddd, J = 9.6, 4.5, 2.0, 0.8 Hz, 1H), 5.49 (dd, J = 4.1, 0.8 Hz, 1H), 4.69 (tt, J = 4.8, 1.0 Hz, 1H), 4.28 (ddd, J = 4.0, 2.9, 0.8 Hz, 1H), 3.13 (ddddd, J = 4.8, 3.9, 2.8, 1.9, 0.9 Hz, 1H), 2.91 (ddq, J = 5.6, 4.7, 0.9 Hz, 1H), 1.08 (s, 9H), 1.01 (s, 9H). 13 C NMR (101 MHz, CDCl3): δ 163.3, 131.7, 130.8, 118.5 (q, JC–F =321.3 Hz, SO2CF3), 113.2, 75.7, 65.0, 45.2, 44.2, 28.8, 28.2, 21.1, 20.7. FTIR (NaCl, thin film): 3042, 2976, 2938, 2906, 2852, 1640, 1478, 1429, 1380, 1365, 1258, 1249, 1214, 1141, 1123, 1074, 1001, 926, 901 cm-1. HRMS: (FAB) calc’d for C17H26F3SSiO5 [M + H]+ 427.1222, found 427.1205. [𝜶]𝟐𝟓 𝐃 = +20° (c = 1.2, CHCl3). TLC (4:1 Hexanes:EtOAc), Rf: 0.8 (KMnO4) Preparation of alkenyl bromide 90: Br TfO Ni(OAc)2 • 4 H2O (5 mol %) Zn (10 mol %) NMI (10 mol %) O Si O t-Bu t-Bu 88 LiBr (1.5 equiv.) 3:1 THF:DMA 87% yield O Si O t-Bu 90 t-Bu An oven-dried 20 mL scintillation vial with a cross-shaped star bar was charged with Ni(OAc)2•4H2O (29.2 mg, 117 µmol, 0.05 equiv), Zn (15.3 mg, 234 µmol, 0.10 equiv), and LiBr (305 mg, 3.52 mmol, 1.50 equiv). The vial was brought into a nitrogen-filled glovebox, and 1-methylimidazole (18.7 uL, 234 µmol), DMA (1.34 mL) and THF (5.0 mL) were added. After stirring for 15 minutes, the enol triflate 88 (1.00 g, 2.34 mmol, 1.0 equiv) was added as a solid, and the vial of 88 was rinsed with a 3:1 mixture of THF:DMA (4 mL). The vial was brought out of the glovebox. The reaction was stirred (1500 rpm) at 21 °C Chapter 2 – Convergent Approach to C19-Diterpenoid Alkaloids 43 for 16 hours. Water (20 mL) was added, and the mixture was filtered through cotton. The mixture was extracted with Et2O (4 x 30 mL) and the combined organic extracts were dried (MgSO4), filtered and concentrated in vacuo to give the crude product as a yellow solid. The product was purified by column chromatography (SiO2, 1-2-3-4% Et2O in hexanes) to afford alkenyl bromide 90 (0.730 g, 2.04 mmol, 87% yield) as a waxy white solid. Note: The reaction is quite sensitive to stirring, presumably due to the heterogeneous reductant. The above procedure represents the largest scale we achieved without a significant reduction in yield (due primarily to incomplete conversion). To improve material throughput, 10 reactions on the scale presented above (10.156 g 10 total, 23.8 mmol) were run in parallel and combined for workup and purification. In this instance, we obtained an 81% yield (6.93 g, 19.3 mmol). 1 H NMR (400 MHz, Chloroform-d) δ 6.24 (ddd, J = 9.6, 6.0, 1.1 Hz, 1H), 5.86 (d, J = 3.9 Hz, 1H), 5.80 (dddd, J = 9.5, 4.4, 2.0, 0.8 Hz, 1H), 4.64 (tt, J = 4.7, 1.0 Hz, 1H), 4.23 (ddd, J = 4.2, 3.0, 0.9 Hz, 1H), 3.06 – 3.03 (m, 1H), 2.83 (ddd, J = 6.1, 4.5, 0.8 Hz, 1H), 1.07 (s, 9H), 1.01 (s, 9H). 13 C NMR (101 MHz, CDCl3) δ 135.9, 132.9, 130.4, 128.2, 76.6, 65.1, 51.6, 47.3, 28.8, 28.2, 21.2, 20.7. FTIR (NaCl, thin film): 3424, 2970, 2935, 2860, 2709, 1733, 1632, 1476, 1386, 1364, 1315, 1301, 1288, 1250, 1204, 1118, 1085, 1057, 1028, 994, 973, 938, 914, 849, 826, 809, 797, 780, 761, 726, 761, 701, 692, 682, 645, 613 cm-1. HRMS: (ESI-TOF) calc’d for C16H26BrO2Si [M + H]+ 357.0880, found 357.0873. [𝜶]𝟐𝟐 𝐃 = +14.7° (c = 0.52, CHCl3). TLC (5% Et2O in Hexanes), Rf: 0.4 (KMnO4) Chapter 2 – Convergent Approach to C19-Diterpenoid Alkaloids 44 Preparation of silyl ether 98: Br MeO2C MeO2C H i. t-BuLi THF, –78 °C O Si O t-Bu t-Bu 89 O OTMS ii. 71, THF, –94 to –78°C iii. TMSCl –78 to 21 °C O Si O t-Bu 77% yield MeO2C MeO2C H 98 t-Bu O O 71 In a 250 mL, round-bottom flask, alkenyl bromide 90 (2.18 g, 6.10 mmol, 1.0 equiv) was dissolved in THF (61 mL) and cooled to –78 °C. To this solution was added t-BuLi (7.2 mL, 1.7 M in pentane, 12.2 mmol, 2.0 equiv) drop wise via syringe. During the last few drops of the addition of t-BuLi, the yellow color of t-BuLi in THF persisted, indicating that all equivalents of the alkenyl bromide had been consumed. The resulting mixture was stirred for an additional 15 minutes at –78 °C. Meanwhile, in a separate 500 mL, round-bottom flask, epoxyketone 71 (2.13 g, 7.93 mmol, 1.3 equiv) was dissolved in THF (79 mL). 9 requires vigorous stirring or sonication at room temperature in order to dissolve in THF. After the epoxyketone had dissolved, it was cooled –94 °C (acetone/liquid N2 bath) and stirred for 10 minutes at this temperature to ensure thorough cooling (some precipitate of 71 forms during this time, but this is not a problem). After the alkenyl lithium had been stirred for 15 minutes at –78 °C, the solution of alkenyl lithium was then cannulated over 10 minutes into the –94 °C solution of 71. The reaction was stirred for an additional 30 minutes, allowing the reaction to warm to –78 °C (the acetone/liquid N2 bath was replaced with a dry ice/acetone bath). TMSCl (1.55 mL, 12.2 mmol, 2.0 equiv) was added, and the mixture was warmed to 21 °C and stirred for 1 hour. The reaction was then quenched with saturated aqueous NaHCO3 (150 mL), and Chapter 2 – Convergent Approach to C19-Diterpenoid Alkaloids 45 was allowed to warm to room temperature. The biphasic mixture was transferred to a separatory funnel, and the aqueous layer was extracted with EtOAc (3 x 200 mL), and the combined organic extracts were dried over Na2SO4, filtered, and concentrated in vacuo. The crude residue was purified by silica gel chromatography (5-10-20-30% EtOAc in hexanes) to afford silyl ether 98 (2.89 g, 4.67 mmol, 77% yield) as a white crystalline solid. 1 H NMR (400 MHz, CDCl3): δ 6.03 (ddd, J = 9.6, 6.1, 1.1 Hz, 1H), 5.68 (ddd, J = 9.5, 4.3, 1.5, 1H), 5.57 (d, J = 3.7 Hz, 1H), 4.48 (t, J = 4.7 Hz, 1H), 4.26 (ddd, J = 4.1, 3.0, 0.7 Hz, 1H), 3.74 (s, 3H), 3.67 (s, 3H), 3.30 (d, J = 3.5 Hz, 1H), 3.08 – 3.03 (m, 1H), 2.90 (dd, J = 5.5, 4.8 Hz, 1H), 2.74 (t, J = 9.3 Hz, 1H), 2.61 (dddd, J = 12.8, 11.0, 8.9, 3.9 Hz, 1H), 2.43 – 2.25 (m, 2H), 2.12 (ddd, J = 12.7, 8.7, 3.9 Hz, 1H), 2.04 (ddd, J = 12.7, 11.0, 7.5 Hz, 1H), 1.96 (ddd, J = 15.1, 8.2, 3.7 Hz, 1H), 1.83 (ddt, J = 12.7, 9.4, 8.2, 1H), 1.45 (ddd, J = 13.3, 10.1, 8.4 Hz, 1H), 1.09 (s, 9H), 1.02 (s, 9H), 0.05 (s, 9H). 13 C NMR (101 MHz, CDCl3): δ 172.0, 170.4, 163.2, 135.1, 129.0, 122.5, 77.7, 77.6, 70.1, 66.1, 55.0, 54.0, 52.8, 52.1, 45.8, 44.9, 41.4, 34.5, 29.7, 28.9, 28.3, 22.2, 21.6, 21.2, 20.7, 2.2. FTIR (NaCl, thin film): 3028, 2952, 2904, 2860, 1732, 1477, 1462, 1434, 1364, 1250, 1175, 1110, 1058, 992, 881, 842 cm-1. HRMS: (PPM) calc’d for C32H51O8Si2 [M + H]+ 619.3117, found 619.3106. [𝜶]𝟐𝟓 𝐃 = +36° (c = 0.35, CHCl3). TLC (10%EtOAc/ 90% Hexanes), Rf: 0.4 (green/black in p-anisaldehyde) Notes: (1) It is extremely important for this reaction to be rigorously dry. Trace water diminishes yield. To achieve this, both substrates were separately azeotroped with PhMe (from the solvent system, 3x) prior to use and dried under high vacuum. Additionally, the THF Chapter 2 – Convergent Approach to C19-Diterpenoid Alkaloids 46 from the solvent system was titrated. As long as the THF titrated at 7 ppm (or lower), good yields were achieved. (2) Typically a smaller scale (300 mg) test reaction was done with the same reagent batches, solvent, and substrate prior to scaling up, to ensure that the scale-up run would proceed successfully. The structure was confirmed via X-ray crystallography of the corresponding epoxy alcohol (S11). O MeO2C MeO2C H OH O Si O tBu tBu S11 Characterization of epoxy-alcohol S11: 1 H NMR (400 MHz, CDCl3): δ 6.10 (ddd, J = 9.6, 6.1, 1.1 Hz, 1H), 5.70 (ddd, J = 9.4, 4.3, 1.6 Hz, 1H), 5.65 (d, J = 3.8 Hz, 1H), 4.48 (t, J = 4.7 Hz, 1H), 4.25 (dd, J = 4.2, 2.8 Hz, 1H), 3.75 (s, 3H), 3.70 (s, 3H), 3.32 (d, J = 3.6 Hz, 1H), 3.05 – 3.01 (m, 1H), 2.83 (ddd, J = 5.4, 4.9 Hz, 1H), 2.71 (dd, J = 10.7, 7.5 Hz, 1H), 2.53 – 2.42 (m, 2H), 2.42 – 2.29 (m, 2H), 2.10 – 1.98 (m, 2H), 1.97 – 1.86 (m, 2H), 1.61 – 1.47 (m, 1H), 1.08 (s, 9H), 1.02 (s, 9H). 13 C NMR (101 MHz, CDCl3): δ 171.7, 170.2, 161.6, 134.5, 129.2, 123.4, 77.7, 75.3, 71.2, 66.1, 56.5, 55.0, 52.9, 52.2, 45.6, 45.2, 44.4, 37.3, 29.3, 28.9, 28.3, 23.3, 21.6, 21.2, 20.7. FTIR (NaCl, thin film): 3496, 3032, 2950, 2860, 1732, 1476, 1458, 1434, 1383, 1364, 1306, 1244, 1176, 1108, 991 cm-1. HRMS: (PPM) calc’d for C29H43O8Si [M + H]+ 547.2722, found 547.2713. [𝜶]𝟐𝟓 𝐃 = –5.1° (c = 0.30, CHCl3). Chapter 2 – Convergent Approach to C19-Diterpenoid Alkaloids 47 TLC (33%EtOAc/ 67% Hexanes), Rf: 0.5 (blue in p-anisaldehyde) Preparation of ketone 99: MeO2C MeO2C H O 11 17 O TMS 10 TMSNTf2 (10 mol %) 2,6-t-Bu2-4-MePy MeO2C 11 OTMS CO2Me O 10 17 CH2Cl2, –78 °C O Si O t-Bu t-Bu 97% yield 98 O Si O t-Bu t-Bu 99 A 1 L, round-bottom flask was charged with epoxide 98 (4.27 g, 6.90 mmol, 1.0 equiv), 2,6-(t-Bu)2-4-MePy (1.56 g, 7.59 mmol, 1.1 equiv) and CH2Cl2 (430 mL), and the solution was cooled to –78 °C in a dry ice/acetone bath. In an N2-filled glovebox, a solution TMSNTf2 (244 mg, 0.69 mmol, 0.10 equiv) in CH2Cl2 (2 mL) was taken up into a 3 mL syringe, which was plugged with a rubber stopper and removed from the glovebox. The TMSNTf2 solution was immediately added dropwise over 2 minutes to the reaction mixture, causing the solution to turn yellow. After stirring for an additional 50 minutes at –78 °C, the reaction was quenched with sat. NaHCO3 (250 mL), and the solution was allowed to warm to room temperature. The biphasic mixture was transferred to a separatory funnel and the layers were separated. The aqueous phase was extracted with CH2Cl2 (3 x 250 mL), and the combined organic extracts were dried over Na2SO4, filtered, and concentrated in vacuo. The resulting crude residue was purified by silica gel chromatography (3% to 4% acetone in hexanes to elute the 2,6-t-Bu2-4MePy followed by 8% to 10% acetone in hexanes to elute the product) to afford ketone 99 (4.23 g, 6.70 mmol, 97% yield) as a white solid. Notes: (1) 2,6-(t-Bu)2-4-MePy was purchased from Combi Blocks and repurified before use. The crude brown oil was dissolved in pentanes, and filtered over a plug of silica gel that Chapter 2 – Convergent Approach to C19-Diterpenoid Alkaloids 48 had been pre-equilibrated with pentanes, eluted with more pentanes and concentrated in vacuo, and dried under high-vacuum. Upon standing, the material turned into a white crystalline solid. This repurification step is crucial for high yields. (2) If the starting material is not rigorously dry, the reaction may not reach full conversion. It is possible that trace amounts of water can quench TMSNTf2 and result in early termination. In this case, the crude product can simply be resubjected to the reaction conditions. In order to ensure the starting material is sufficiently free of water, it should be azeotroped with toluene or benzene (1–3x) prior to the reaction. 1 H NMR (500 MHz, CDCl3): δ 5.88 (ddd, J = 9.5, 6.1, 1.1 Hz, 1H), 5.65 (ddd, J = 9.5, 4.3, 1.9 Hz, 1H), 5.40 (d, J = 3.8 Hz, 1H), 4.66 (t, J = 4.7 Hz, 1H), 4.31 (br s, 1H), 4.23 (dd, J = 4.0, 3.1 Hz, 1H), 3.74 (s, 3H), 3.67 (s, 3H), 3.40 (dd, J = 11.7, 7.3 Hz, 1H), 3.03 – 2.94 (m, 2H), 2.48 (td, J = 14.1, 3.4 Hz, 1H), 2.38 – 2.26 (m, 1H), 2.17 – 2.05 (m, 3H), 1.80 – 1.71 (m, 1H), 1.65 – 1.60 (m, 1H), 1.52 (tdd, J = 14.3, 2.9, 1.9 Hz, 1H), 1.08 (s, 9H), 1.00 (s, 9H), 0.05 (s, 9H). 13 C NMR (126 MHz, CDCl3): δ 215.9, 170.7, 170.6, 158.9, 134.8, 128.2, 124.4, 76.7, 69.9, 66.2, 58.0, 56.2, 52.8, 52.6, 46.2, 46.1, 41.8, 38.8, 28.9, 28.3, 27.4, 23.7, 21.2, 20.7, 19.3, 0.0. FTIR (NaCl, thin film): 3032, 2952, 2896, 2859, 1741, 1477, 1462, 1443, 1384, 1363, 1252, 1170, 1097, 991, 840 cm-1. HRMS: (PMM) calc’d for C32H51O8Si2 [M + H]+ 619.3117, found 619.3105. [𝜶]𝟐𝟓 𝐃 = +92° (c = 0.73, CHCl3). TLC (10%EtOAc/ 90% Hexanes), Rf: 0.4 (blue in p-anisaldehyde) Chapter 2 – Convergent Approach to C19-Diterpenoid Alkaloids 49 2.7 NOTES AND REFERENCES (1) (2) (3) (4) (5) (6) (7) (8) (9) (10) (11) Filippini, M.-H.; Rodriguez, J. Chem. Rev. 1999, 99 (1), 27–76. Presset, M.; Coquerel, Y.; Rodriguez, J. Chem. Rev. 2013, 113 (1), 525–595. Hagiya, K.; Yamasaki, A.; Okuyama, T.; Sugimura, T. Tetrahedron: Asymmetry 2004, 15 (9), 1409–1417. Sugimura, T.; Yamasaki, A.; Okuyama, T. Tetrahedron: Asymmetry 2005, 16 (3), 675–683. Kitamura, Masato.; Ohkuma, Takeshi.; Inoue, Shinichi.; Sayo, Noboru.; Kumobayashi, Hidenori.; Akutagawa, Susumu.; Ohta, Tetsuo.; Takaya, Hidemasa.; Noyori, Ryoji. J. Am. Chem. Soc. 1988, 110 (2), 629–631. Hofstra, J. L.; Poremba, K. E.; Shimozono, A. M.; Reisman, S. E. Angew. Chem. Int. Ed. 2019, 58 (42), 14901–14905. Arai, T.; Yamada, Y. M. A.; Yamamoto, N.; Sasai, H.; Shibasaki, M. Chem. - Eur. J. 1996, 2 (11), 1368–1372. Fastuca, N. J.; Wong, A. R.; Mak, V. W.; Reisman, S. E. Org. Synth. 2020, 97, 327– 338. Ye, W.; Xu, J.; Tan, C.-T.; Tan, C.-H. Tetrahedron Lett. 2005, 46 (40), 6875–6878. Hagiya, K.; Yamasaki, A.; Okuyama, T.; Sugimura, T. Tetrahedron: Asymmetry 2004, 15 (9), 1409–1417. Steves, J. E.; Stahl, S. S. J. Am. Chem. Soc. 2013, 135 (42), 15742–15745. Chapter 3 – Towards C19-Diterpenoid Alkaloids: Approaches to the C4 Chiral Quaternary Center 50 Chapter 3 Towards C19-Diterpenoid Alkaloids: Approaches to the C4 Chiral Quaternary Center 3.1 INTRODUCTION Having accessed the product of the semipinacol rearrangement product 99, which contains all of the carbon atoms of the aconitine core, our efforts turned to introducing the proper functionality for synthesis of the natural product. Functionalization of the C18 and C19 positions has presented a significant challenge in our efforts towards 1. Our enantioselective synthesis of epoxy ketone fragment 71 relies on an enantioselective conjugate addition of dimethyl malonate (74) to 2-cyclopenten-1-one (73), followed by enolate alkylation to form the C4 quaternary center. This approach brings C18 and C19 in as diastereotopic esters which require diastereoselective functionalization to set the C4 chiral quaternary center. Our firstgeneration retrosynthesis included an intramolecular lactamization between a C17 amine and the C19 methyl ester (Figure 3.1). Efforts to introduce nitrogen through reductive amination at C17 were unsuccessful, which led us to modify our strategy such that the C19–N bond would Chapter 3 – Towards C19-Diterpenoid Alkaloids: Approaches to the C4 Chiral Quaternary Center 51 be formed before the C17–N bond.1,2 This required selective incorporation of an amine by reaction with the C19 ester over its diastereotopic C18 ester. This chapter discusses efforts to differentiate the reactivity of these two sites in order to set the C4 chiral quaternary center, as well as an alternative approach to set that chiral center at an early stage in the synthesis, prior to the fragment coupling. Figure 3.1 Functionalization of C18 and C19 for the synthesis of talatisamine (1), liljestrandinine (2), and liljestrandisine (3).1,2 C17 Reductive amination unsuccessful 18 MeO2C 4 OTMS CO2Me 19 O BOMO 101 18 MeO2C 4 OTMS CO2Me 19 NHEt 17 17 O BOMO 102 O 18 4 MeO2C OTMS NEt 19 O 17 BOMO 103 O 2nd Generation approach: Form C19–N bond first MeO2C 18 4 OTMS CO2Me 19 O O Si O t-Bu t-Bu 99 Functionalization of C18 and C19 Esters MeO 4 19 18 O t-Bu Si OMe NHEt O t-Bu 104 3.2 SEQUENTIAL REDUCTION OF C18 AND C19 ESTERS Our first-generation approach to form the the C19-N bond was to reduce the C18 ester selectively (Scheme 3.1). In order to differentiate the reactivity of the two esters, the C1 TMSether of 99 was deprotected with trichloroacetic acid and subsequent treatment with potassium carbonate and methanol in dichloromethane effected lactonization with C19 to give 106. The C17 ketone ins converted to the corresponding enol triflate 107, which is then reduced under palladium-catalyzed conditions to cyclopentene 108. The C18 ester of 108 can be selectively reduced to the alcohol using lithium tri-tert-butoxyaluminum hydride, a reducing agent with Chapter 3 – Towards C19-Diterpenoid Alkaloids: Approaches to the C4 Chiral Quaternary Center 52 unique reactivity with 1,3-diesters,3 leaving the C19 lactone intact. Methylation of the C18 alcohol followed by reduction of the C19 lactone 110 with lithium borohydride furnishes the C1/C19 diol 111. Selective TES protection of the primary C19 alcohol and methylation of the remaining C1 alcohol affords 113. To introduce the amine at C19, the triethylsilyl ether of 113 is deprotected, and the resulting alcohol, 114, is cleanly oxidized to the C19 aldehyde 115 using Stahl’s conditions.4 This aldehyde serves as a handle for reductive amination with Nallylamine followed by cleavage of the N-allyl group gives C19 primary amine 116. Scheme 3.1 Sequential reduction of C18 and C19 esters to incorporate nitrogen at C19. MeO2C 4 1 18 OTMS CO2Me 19 O 4 MeO2C i. TCA CH2Cl2, –20 °C 19 O 18 O 17 O O Si O t-Bu O Si O t-Bu 95% yield 1 O O Si O t-Bu OH 1 MeO OH Me3OBF4 Proton Sponge MeO 4 Å MS CH2Cl2, 0 to 20 °C OMe MeO 19 OTES TFA 96% yield 1 OMe O Si O t-Bu t-Bu 113 90% yield 4 Å MS CH2Cl2, 0 to 20 °C MeO 19 OH Cu(MeCN)4OTf (5 mol %) MeObpy (5 mol %), CH2Cl2, 0 °C 90% yield O Si O t-Bu t-Bu 110 O Si O t-Bu t-Bu 114 O 18 Me3OBF4 Proton Sponge O THF, 20 °C O Si O t-Bu t-Bu 111 98% yield 1 HO O 18 LiBH4 CH2Cl2, 0 °C O Si O t-Bu t-Bu 112 O Si O t-Bu t-Bu 108 MeO TESCl imidazole H O 107 19 OH O MeO2C 93% yield t-Bu 106 19 OTES OTf Pd(OAc) (7.5 mol %) 2 PPh3 (15 mol %) HCO2H, NEt3 DMF, 60 °C 95% yield t-Bu 99 MeO O MeO2C THF, -78 °C ii. K2CO3 MeOH, CH2Cl2 t-Bu KHMDS; Comins’ rgt. NMI (5 mol %), ABNO (5 mol % MeCN, air 94% yield 95% yield O Li(t-BuO)3AlH THF, rt THF, 0 to 20 °C O Si O t-Bu t-Bu 109 97% yield MeO OMe O 18 1. allylamine, Ti(Oi-Pr)4; then NaBH4 EtOH 2. Pd(PPh3)4 (10 mol %) 1,3-dimethylbarbituric acid O Si O CH2Cl2, 21 °C t-Bu t-Bu 90% yield 115 (2 steps) 4 OMe 19 NH2 O Si O t-Bu t-Bu 116 This route from semipinacol product 99 to primary amine 116 amounts to twelve steps, with nine of these twelve representing modification of functionality at C18 and C19. Four of the twelve steps represent redox manipulations at C18 or C19. We sought a more direct route to access amine 116 through a lactone-selective aminolysis. Chapter 3 – Towards C19-Diterpenoid Alkaloids: Approaches to the C4 Chiral Quaternary Center 53 3.3 SELECTIVE AMINOLYSIS OF C19 LACTONE We wondered if we could leverage the ring strain of the [2.2.2]-bridged bicyclic lactone 108 for selective C19 ester aminolysis. Gratifyingly, we found that treatment of 108 with solvent quantities of amine and an ammonium carboxylate catalyst at elevated temperatures gave moderate yields of the desired N-allyl and N-ethyl amide products (117 and 118) (Scheme 3.2).5,6 These reactions were stopped before complete conversion, as undesired aminolysis of the C18 methyl ester of the product occurred at longer reaction times. The C18 ester of 117 and 118 was reduced with lithium triethylborohydride to afford C1/C18 diols 119/120 in good yield. Accessing these diols enables methylation of the C1 and C18 alcohols in a single step with trimethyloxonium tetrafluoroborate. A small amount of mono-methylated products were isolated as well. Extending the reaction time led to lower yields and formation of the C19 methyl ester, presumably through amide O-methylation and imidate hydrolysis on workup. Reduction of the C19 amide was achieved using Brookhart’s iridium-catalyzed hydrosilylation conditions.7 These conditions proved uniquely effective for substrate 121 in reducing the amide without concomitant reduction of the N-allyl group to an N-propyl group. N-ethyl amine 123 was isolated in 87% yield, while the corresponding N-allyl amine was deprotected using the same palladium-catalyzed conditions as before to give primary amine 116. This lactone aminolysis route limits the redox manipulations necessary to access 116 and123. No protecting group manipulations are necessary on this route, and the diol intermediate 119/120 allows for methylation of the C1 and C19 alcohols in a single step. This route saves four steps over the first-generation route. Chapter 3 – Towards C19-Diterpenoid Alkaloids: Approaches to the C4 Chiral Quaternary Center 54 Scheme 3.2 Lactone-selective aminolysis route to 116 and 123. OH 4 O MeO2C RNH2•HCl sodium 2-ethylhexanoate O 18 THF, -20 to 10 °C O Si O R = allyl 54% yield t-Bu (64% BRSM) O Si O t-Bu t-Bu t-Bu R = Et 117 R = allyl 118 R = Et 51% yield Lactone Aminolysis Route O 124 O N O Me 91% yield R = Et 86% yield MeO OMe NH2 4 4 Å MS CH2Cl2, 0 to 20 °C O Si O t-Bu O Si O t-Bu R = allyl 64% yield t-Bu 119 R = allyl 120 R = Et R = Et 25% yield (49% after 1 recycle) 121 R = allyl 122 R = Et MeO Pd(PPh3)4 (10 mol %) 124 (6 equiv.) 4 18 OMe NHR 19 [Ir(COE)2Cl]2 (0.5 mol %) Et2SiH2 (4 equiv.) CH2Cl2, 21 °C; O Si O R = allyl t-Bu t-Bu 116 O t-Bu 19 18 • Reduction reactions only • No protecting groups • Single O-methylation step • 4 steps shorter than firstgeneration route MeN R = allyl NHR Me3OBF4 Proton Sponge O LiBHEt3 OMe MeO NHR 18 O THF/RNH2 60–80 °C 108 OH 1 HO 19 NHR MeO2C 90% yield (2 steps) PhH, 21 °C; then LiBHEt3 O Si O t-Bu t-Bu 123 R = Et 87% yield 3.4 TOWARDS A C18/C19 FUNCTIONALIZED EPOXY KETONE FRAGMENT Figure 3.2 Opportunity to increase convergency of synthesis by properly functionalizing C18 and C19 prior to fragment coupling. [6 steps from cyclopentenone] 1. MeO C 4 A 11 O Br 2 MeO2C 10 C O F H 17 18 71 D 1,2-addition/ semipinacol sequence O Si O t-Bu 4 MeO2C t-Bu A OTMS CO2Me 19 O 11 MeO 12 steps 17 F 10 C [10 steps from phenol] (9 steps modifying C18 + C19) D O Si O t-Bu t-Bu 116 99 1. R 2N 19 4 MeO 18 H A O 11 F 125 O 17 1,2-addition/ semipinacol sequence MeO 4 18 OMe 19 NH 17 O Si O t-Bu t-Bu 90 4 18 A 11 OTMS NR2 O 19 17 F 10 C D O Si O t-Bu t-Bu 124 2 Chapter 3 – Towards C19-Diterpenoid Alkaloids: Approaches to the C4 Chiral Quaternary Center 55 For the key fragment coupling in our approach to the C19-diterpenoid alkaloids, the epoxy ketone fragment 71 and alkenyl bromide fragment 90 are prepared in six and ten steps LLS, respectively. This disparity in step count required for the preparation of each of the fragments highlights an opportunity to make the synthesis more convergent by preparation of a more appropriately functionalized epoxy ketone fragment, such as 125 (Figure 3.2). In our elaboration of the fragment coupling product 99 to primary amine 116, six of the eight steps required are spent modifying C18 and C19. Since these carbons are a part of the epoxy ketone fragment 71, if these sites were properly functionalized prior to the fragment coupling to the synthesis would be more convergent, and therefore more efficient. We set out to evaluate approaches to differentiate the reactivity at C18 and C19 and introduce proper functionality at these positions for the epoxy ketone fragment. Figure 3.3 Development of more complex A/F-ring fragment. MeO2C 4 MeO2C A F H Ester Differentiation O 11 O 17 71 3.4.1 R 2N 19 4 18 O A MeO F H O 125 Selective Functionalization of a C18/C19 Diol Key to our enantioselective synthesis of epoxy ketone 71 is an asymmetric Michael addition of dimethyl malonate (74) to 2-cyclopenten-1-one (73) using a chiral bimetallic catalyst reported by Shibasaki and coworkers.8,9 This brings C18 and C19 in at the ester oxidation state while the natural products are at the alcohol oxidation state at those positions. As a result, this strategy inherently requires reduction at those positions. For synthesis of of our desired epoxy ketone fragment, we sought to reduce at both positions and selectively functionalize one diastereotopic alcohol over the other. Chapter 3 – Towards C19-Diterpenoid Alkaloids: Approaches to the C4 Chiral Quaternary Center 56 Scheme 3.3 Synthesis of C18/C19 diol 127 and unexpected oxy-Michael product 128. O O O LAH MeO2C MeO2C O H 93 THF, reflux O single diastereomer! HCl HO O HO H O 70% yield O COMe2 (2.0 equiv.) THF/H2O, 70°C HO O H (87% combined yield) 126 HO HO + O O H 127 128 68% yield 19% yield HO MeO O H conditions MeO O H 130 129 O Me3OBF4 Proton Sponge 4Å MS, CH2Cl2 72% yield To start, previously-synthesized diester 93 was reduced to 1,3-diol 126 with lithium aluminum hydride. Acid-catalyzed dioxolane deprotection and aldol condensation gave enone product, 127, along with the unexpected oxy-Michael product 128 as a single diastereomer. We were excited by this result, as it amounts to a selective protection of the C18 alcohol. Methylation of the C19 alcohol of 128 affords 129. Unfortunately, attempts to effect retro-oxyMichael to regenerate the enone were plagued by low yields (Table 3.1). We believe this is due to the instability of the resulting enone 130 in the presence of base. Chapter 3 – Towards C19-Diterpenoid Alkaloids: Approaches to the C4 Chiral Quaternary Center 57 Table 3.1 Retro-oxy-Michael conditions for synthesis of 130. MeO O Conditions O H HO MeO 129 O H 130 Entry Scale Solvent Base Temp. Yield comments 1 2 mg THF LiHMDS 50 °C 0% no rxn 2 2 mg THF NaHMDS 50 °C 0% — 3 2 mg THF KHMDS 0 °C 0% no rxn 4 2 mg THF TMPMgCl•LiCl -78 to 0 °C 0% no rxn 5 2 mg MeOH 0.2M LiOH 50 °C 0% — 6 2 mg MeOH 0.2M KOH 50 °C 0% — 7 2 mg MeOH 0.1 M NaOH 20 °C 10% — 8 9 mg MeOH 0.2 M NaOH 50 °C 24% — We turned back to a diastereoselective protection of C18/C19 diol 127. Using an optimized ratio of triethylamine to imidazole with TBSCl, C18 TBS-ether 131 was isolated in 48% yield (Table 3.2). To install the epoxide, Luche reduction of 131 followed by alcohol directed epoxidation affords 134 (Scheme 3.4). At this point, we realized further elaboration with this route would result in a longer step count and less efficient synthesis of 116 than the routes described in sections 3.2 and 3.3 where C18 and C19 are functionalized after the fragment coupling. As such, we turned to an alternative approach to set the C4 quaternary center. Chapter 3 – Towards C19-Diterpenoid Alkaloids: Approaches to the C4 Chiral Quaternary Center 58 Table 3.2 Diastereoselective TBS protection of 127. HO HO TBSO 3.5 equiv. TBSCl HO O TBSO base THF, rt H HO O O H H 131 major 127 132 minor entry scale conditions yield 131 1 50 mg 3.5 equiv. imid. 35%b 2 50 mg 5.3 equiv. imid. 28%b 3 50 mg 1.0 equiv imid, 3.5 equiv NEt3 48%b 4 50 mg 0.5 equiv imid, 4.0 equiv NEt3 66%, 2.6:1 d.r.a 5 50 mg 4.5 equiv. NEt3 82%, 1.6:1 d.r.a 6 150 mg 1.0 equiv imid, 2.5 equiv. NEt3 42%b a NMR yield b Isolated major diast. yield Scheme 3.4 Elaboration of 131 to epoxide 134. HO NaBH4 CeCl3 TBSO O H 131 3.4.2 MeOH, -78 °C 42% yield [3:1 crude d.r.] HO V(O)(acac)2 TBHP TBSO OH H 133 PhH, rt 99% yield R 2N HO O TBSO O OH H 134 MeO O H 125 Setting C4 Quaternary Center by Michael Addition of a Prochiral Nucleophile Previously discussed approaches relied on differentiation of diastereotopic C18 and C19 carbon centers about the C4 quaternary center. As an alternative, we considered the possibility of setting the C4 stereocenter directly through asymmetric catalysis. We were inspired by a report from Maruoka and coworkers detailing an enantioselective conjugate addition of prochiral α-cyanoacetate tertiary enolates to ynoate electrophiles to form quaternary centers (Figure 3.4).10 They also reported a single example of a doubly stereoselective conjugate addition of these nucleophiles to 2-cyclohexen-1-one (140) to form Chapter 3 – Towards C19-Diterpenoid Alkaloids: Approaches to the C4 Chiral Quaternary Center 59 a β/γ-stereodiad in 141 with excellent yield, enantioselectivity and diastereoselectivity. Enticed by this result, we looked to apply it to our system. Figure 3.4 Maruoka’s enantioselective conjugate addition catalyzed by chiral ammonium salt. NC 1 mol % (S)-PTC (138) CO2t-Bu CO2tBu Cs2CO3, PhMe -40 °C R 135 136 NC CO2t-Bu R * CO2t-Bu 137 11 examples 90-99% yield 92-99% ee (S)-PTC CF3 Ar N O CO2tBu NC Ar 138 O 1 mol % (S)-PTC (138) * * 140 Br F3C CF3 141 99% yield 91% ee, 5.7:1 d.r. Cs2CO3, PhMe 0 °C Ph 139 NC tBuO C 2 CF3 O Ar Ph We started with Maruoka’s optimal conditions using α-cyanoacetate 142 as the Michael donor and 2-cyclopenten-1-one (73) as the acceptor (Table 3.3, entry 1). To our delight, the major diastereomer was isolated in 50% ee and 1.5:1 d.r. Increasing the catalyst loading to 2 mol% gave significant improvements in enantioselectivity and diastereoselectivity, though any further increase in catalyst loading did not have any effect. Decreasing the loading of base improved the yield of the reaction with slight increase in ee. Decreasing the temperature had little effect on the reaction. Optimal conditions in entry 6 gave the product in quantitative yield, 3.3:1 d.r. and 74% ee. Control reactions show that base is necessary for the reaction (entry 9). Leaving out chiral catalyst, the reaction still proceeds in 80% yield (entry 10), suggesting that uncatalyzed reaction could be a problem. The relative stereochemistry of the product was determined by X-ray crystallography of the minor diastereomer. The major diastereomer has the ester at C19 and nitrile at C18. We could not envision a straightforward route from 143 which would incorporate the amine at C19, so we decided to move away from this strategy. Chapter 3 – Towards C19-Diterpenoid Alkaloids: Approaches to the C4 Chiral Quaternary Center 60 Table 3.3 Doubly stereoselective Michael addition of α-cyanoacetate 142 to 2cyclopenten-1-one (73) NC CO2t-Bu O O O O (S)-PTC (138, x mol%) O 142 O O 73 Cs2CO3 (x equiv.) PhMe 19 19 t-BuO2C NC 18 NC 4 O H t-BuO2C 18 143 major Entry scale (mmol) mol% (S)-PTC equiv. Cs2CO3 4 O H 144 minor T (°C) Yield (%) d.r. ee of 143 1.5:1 50% 1 0.20 1 0.5 0 98a 2 0.05 2 0.5 0 63a 3.0:1 72% 3 0.05 5 0.5 0 80a 2.9:1 70% 4 0.05 2 0.5 -20 93b 3.0:1 71% 5 0.05 1 0.25 0 65b 2.9:1 77% 6 0.20 2 0.25 0 99a 3.3:1 74% 7 0.05 2 0.10 0 99 3.0:1 75% 8 0.20 1 0.05 0 56 3.6:1 67% 9 0.05 1 0 0 0 — — 10 0.05 0 0.25 0 80 4.0:1 n/a [x-ray] (S)-PTC Ar N O Ar 138 Br CF3 CF3 Ar F3C CF3 aNMR yield; bisolated yield incomplete conversion 3.5 CONCLUDING REMARKS Our synthesis of epoxy ketone fragment 71 requires the use of dimethyl malonate to introduce C18 and C19. Differentiating the reactivity at these two centers to introduce the proper functionality proved to be a significant challenge. Two routes to introduce the amine at C19 are described, one by sequential reduction of the two esters and one using a selective aminolysis of a strained C19 lactone. Strategies to incorporate the proper functionality at C18 and C19 prior to the fragment coupling are also described. Chapter 3 – Towards C19-Diterpenoid Alkaloids: Approaches to the C4 Chiral Quaternary Center 61 3.6 EXPERIMENTAL SECTION Preparation of lactone 106: OTMS CO2Me O MeO2C O MeO2C i. TCA, DCM –20 °C O O ii. K2CO3, MeOH, CH2Cl2 O Si O t-Bu t-Bu 99 95% yield O Si O t-Bu t-Bu 106 A 250 mL, round-bottom flask was charged with silyl ether 99 (1.5 g, 2.43 mmol, 1.0 equiv), and CH2Cl2 (50 mL), and cooled to –20 °C in a cryocool. Trichloroacetic acid (15.86 g, 97.04 mmol, 40 equiv) was added, and the reaction vessel was sealed and allowed to continue stirring at –20 °C for 36 hours. The reaction was then carefully quenched with sat. NaHCO3 (100 mL) at –20 °C, and let warm to room temperature. The biphasic mixture was then transferred to a 500 mL Erlenmeyer flask, and vigorously stirred for an additional 10 minutes. The aqueous phase was ensured to be basic with pH paper. The mixture was then transferred to a separatory funnel and the layers were separated. Aqueous phase was extracted with CH2Cl2 (3 x 50 mL), and added to a round-bottom flask, and MeOH (20 mL) was added followed by K2CO3 (1.5 g). The mixture was allowed to stir at room temperature for 2 hours. The reaction was monitored by TLC and indicated full conversion. If the reaction is going slowly, more K2CO3 and MeOH can be added without deleterious effects. At this time, H2O (20 mL) was added, and the layers were separated. The aqueous phase was extracted with CH2Cl2 (3 x 30 mL), and the organic extracts were dried over Na2SO4, filtered, and concentrated in vacuo. The resulting crude residue was purified by silica gel chromatography (11% acetone in hexanes to 12.5% acetone in hexanes) to afford lactone 106 (1.185 g, 2.303 mmol, 95% yield) as a white foam. Chapter 3 – Towards C19-Diterpenoid Alkaloids: Approaches to the C4 Chiral Quaternary Center 1 62 H NMR (400 MHz, Chloroform-d): δ 6.10 (ddd, J = 9.5, 6.2, 1.1 Hz, 1H), 5.79 (d, J = 3.8 Hz, 1H), 5.75 (ddd, J = 9.5, 4.3, 1.9 Hz, 1H), 4.93 (dd, J = 3.4, 1.8 Hz, 1H), 4.35 (t, J = 4.7 Hz, 1H), 4.21 (ddd, J = 4.1, 3.1, 0.8 Hz, 1H), 3.83 (s, 3H), 3.13 – 3.07 (m, 2H), 3.03 (ttd, J = 4.2, 2.5, 1.1 Hz, 1H), 2.52 (dddd, J = 14.4, 10.6, 9.2, 4.1 Hz, 1H), 2.41 – 2.29 (m, 2H), 2.18 (ddd, J = 17.7, 8.8, 4.1 Hz, 1H), 2.04 (ddt, J = 13.2, 10.3, 1.9 Hz, 1H), 1.95 (ddd, J = 14.3, 6.7, 3.4 Hz, 1H), 1.91 – 1.82 (m, 1H), 1.75 (dddd, J = 14.3, 10.4, 8.9, 6.7 Hz, 1H), 1.05 (s, 9H), 0.99 (s, 9H). 13 C NMR (101 MHz, CDCl3): δ 212.0, 169.8, 169.6, 156.4, 134.1, 130.1, 127.0, 78.6, 77.5, 65.7, 58.0, 53.5, 52.9, 45.9, 44.8, 43.0, 35.3, 28.8, 28.2, 26.1, 23.5, 22.3, 21.2, 20.7. FTIR (NaCl, thin film): 3030, 2974, 2937, 2895, 2860, 1766, 1743, 1476, 1464, 1364, 1271, 1106, 1063, 995, 826 cm-1. HRMS: (ESI-TOF) calc’d for C28H42O7SiN [M + NH4]+ 532.2725, found 532.2720. [𝜶]𝟐𝟓 𝐃 = +9.9° (c = 0.66, CHCl3). TLC (30%EtOAc/70%Hexanes), Rf: 0.4 (UV, teal in p-anisaldehyde) Preparation of enol triflate 107 from 106: O MeO2C O O MeO2C O i. KHMDS, THF, –78 °C OTf O ii. Comins’ reagent O Si O t-Bu t-Bu 106 95% yield O Si O t-Bu t-Bu 107 A 250 mL, round-bottom flask was charged with ketone 106 (2.99 g, 5.81 mmol, 1.0 equiv), and azeotroped PhMe from the solvent system (3 x 20 mL), and put under high vacuum overnight. The following day, the flask was equipped with a stir-bar and rubber septum, purged Chapter 3 – Towards C19-Diterpenoid Alkaloids: Approaches to the C4 Chiral Quaternary Center 63 with N2, THF (58 mL) was added, and the solution was cooled to –78 °C in a dry ice/acetone bath. KHMDS solution (0.5 M in PhMe, 13.9 mL, 6.97 mmol, 1.2 equiv) was added dropwise via syringe causing the reaction mixture to turn yellow. Meanwhile, in a separate flame-dried 50 mL conical flask, Comins’ reagent (2.74 g, 6.97 mmol, 1.2 equiv) was dissolved in THF (15 mL) under N2. After the substrate and KHMDS solution had been stirring for 30 minutes at –78 °C, the Comins’ reagent solution was added dropwise via cannula, and the solution was allowed to stir for an additional 20 minutes at –78 °C. The reaction was quenched with sat. NaHCO3 (50 mL), the bath was removed and allowed to warm to room temperature. Diluted with Et2O (75 mL), transferred to a separatory funnel, and the layers were separated. The aqueous phase was extracted with Et2O (3 x 75 mL). The organic extracts were dried over MgSO4, filtered, and concentrated in vacuo. The resulting crude residue was purified by silica gel chromatography (11% acetone in hexanes to 12.5% acetone in hexanes) to afford enol triflate 107 (3.55 g, 5.52 mmol, 95% yield) as a white foam. 1 H NMR (400 MHz, Chloroform-d): δ 6.06 (ddd, J = 9.5, 6.1, 1.2 Hz, 1H), 5.73 (ddd, J = 9.5, 4.4, 1.6 Hz, 1H), 5.70 (t, J = 2.6 Hz, 1H), 5.59 (dd, J = 3.8, 0.7 Hz, 1H), 4.78 (dd, J = 3.7, 1.7 Hz, 1H), 4.49 (t, J = 4.7 Hz, 1H), 4.23 (ddd, J = 4.2, 3.0, 0.8 Hz, 1H), 3.82 (s, 3H), 3.10 (m, 1H), 2.95 (ddd, J = 17.8, 9.3, 2.3 Hz, 1H), 2.89 – 2.83 (m, 2H), 2.50 (dt, J = 17.8, 2.7 Hz, 1H), 2.34 (ddd, J = 13.4, 11.4, 3.5 Hz, 1H), 2.20 – 2.04 (m, 2H), 1.79 (ddd, J = 13.4, 10.9, 6.6 Hz, 1H), 1.08 (s, 9H), 1.00 (s, 9H). 13 C NMR (101 MHz, CDCl3): δ 169.5, 169.0, 156.9, 145.2, 133.4, 129.9, 126.1, 118.3 (q, J=321 Hz, SO2CF3), 114.3, 77.2, 75.3, 65.2, 56.8, 53.0, 52.7, 46.2, 46.2, 43.9, 32.5, 28.8, 28.2, 26.6, 21.9, 21.2, 20.7. Chapter 3 – Towards C19-Diterpenoid Alkaloids: Approaches to the C4 Chiral Quaternary Center 64 FTIR (NaCl, thin film): 3036, 2938, 2896, 2860, 1769, 1739, 1668, 1476, 1426, 1251, 1217, 1142, 1112, 998, 827 cm-1. HRMS: (ESI-TOF) calc’d for C29H41O9SF3SiN [M + NH4]+ 664.2218, found 664.2216. [𝜶]𝟐𝟓 𝐃 = +123° (c = 0.67, CHCl3). TLC (20%EtOAc/80%Hexanes), Rf: 0.4 (UV, blue in p-anisaldehyde) Preparation of cyclopentene 108: O MeO2C O OTf Pd(OAc)2, PPh3 NEt3, HCO2H O MeO2C H O DMF, 60 °C O Si O t-Bu t-Bu 107 93% yield O Si O t-Bu t-Bu 108 A 200 mL, round-bottom flask was charged with enol triflate 107 (2.28 g, 3.53 mmol, 1.0 equiv), PPh3 (139 mg, 0.53 mmol, 15 mol %), and Pd(OAc)2 (60 mg, 0.265 mmol, 7.5 mol %). The flask was equipped with a rubber septum, purged with N2, and dissolved in DMF (35 mL). Then, the reaction mixture was sparged with an Ar balloon for 5 minutes. NEt3 (3.94 mL, 28.27 mmol, 8 equiv) was added followed by HCO2H (0.66 mL, 17.67 mmol, 5 equiv) – a cloudy gas was observed upon addition. The reaction was lowered into a 60 °C oil bath, and allowed to continue to stir at that temperature for 20 minutes. The reaction turns black during this time, indicating its completion. The reaction was cooled to room temperature, diluted with sat. NH4Cl (30 mL), transferred to a separatory funnel and diluted with Et2O (30 mL). The layers were separated and the aqueous extracted with Et2O (3 x 30 mL). The organic extracts were dried over Na2SO4, filtered, and concentrated in vacuo. The resulting crude residue was purified by silica gel chromatography (10% acetone in hexanes to 12.5% acetone in hexanes, Chapter 3 – Towards C19-Diterpenoid Alkaloids: Approaches to the C4 Chiral Quaternary Center 65 the product is very crystalline, it was loaded in PhMe where some of the product elutes but is collected) to afford olefin 108 (1.64 g, 3.28 mmol, 93% yield) as a crystalline solid. Melting Point: 225–230 °C 1 H NMR (400 MHz, Chloroform-d): δ 6.06 (ddd, J = 9.5, 6.1, 1.2 Hz, 1H), 5.76 – 5.68 (m, 2H), 5.51 (d, J = 3.8 Hz, 1H), 5.48 – 5.44 (m, 1H), 4.61 (dd, J = 3.9, 1.6 Hz, 1H), 4.46 (td, J = 4.2, 0.9 Hz, 1H), 4.22 (ddd, J = 4.2, 3.1, 0.7 Hz, 1H), 3.80 (s, 3H), 3.04 (ttd, J = 4.2, 2.4, 1.0 Hz, 1H), 2.95 – 2.85 (m, 2H), 2.79 (dd, J = 4.9, 5.7, 1H), 2.54 – 2.42 (m, 1H), 2.27 (ddd, J = 13.1, 11.7, 3.2 Hz, 1H), 2.17 (dddd, J = 14.2, 11.1, 3.3, 1.6 Hz, 1H), 1.99 (dddd, J = 13.9, 11.6, 6.2, 3.8 Hz, 1H), 1.85 (ddd, J = 13.1, 11.0, 6.2 Hz, 1H), 1.07 (s, 9H), 1.00 (s, 9H). 13 C NMR (101 MHz, CDCl3): δ 170.4, 170.3, 161.3, 134.7, 132.0, 131.3, 129.1, 121.8, 78.7, 77.3, 65.9, 59.8, 53.4, 52.7, 46.4, 45.7, 44.4, 38.2, 28.8, 28.2, 26.8, 22.4, 21.2, 20.7. A 13C signal is observed to be overlapping with residual CDCl3 at 77.3, resolved peaks are seen in the HSQC spectrum, provided below. FTIR (NaCl, thin film): 3053, 2937, 2898, 2256, 1760, 1748, 1732, 1476, 1463, 1444, 1364, 1283, 1109, 1063, 993, 911 cm-1. HRMS: (ESI-TOF) calc’d for C28H39O6Si [M + H]+ 499.2510, found 499.2512. [𝜶]𝟐𝟓 𝐃 = +166° (c = 0.40, CHCl3). TLC (20%EtOAc/80%Hexanes), Rf: 0.4 (UV, teal in p-anisaldehyde) Chapter 3 – Towards C19-Diterpenoid Alkaloids: Approaches to the C4 Chiral Quaternary Center 66 Preparation of alcohol 109: HO O MeO2C O O O Li(OtBu)3AlH THF O Si O t-Bu t-Bu 97% yield 108 O Si O t-Bu t-Bu 109 A 250-mL, round-bottomed flask was charged with methyl ester 108 (1.56 g, 3.13 mmol, 1.0 equiv), equipped with a rubber septum, purged with N2, and dissolved in THF (31 mL). Then, the reaction was cooled to 0 °C in an ice bath Li(Ot-Bu)3AlH (2.39 g, 9.39 mmol, 3 equiv) was added, and the ice bath was removed. The reaction was monitored by TLC, and after 100 minutes, trace starting material remained, so an additional portion of Li(Ot-Bu)3AlH (0.95 g, 3.66 mmol, 1.17 equiv) was added, and allowed to stir for an additional 70 minutes. The reaction was carefully quenched with water (10 mL) followed by sat. Rochelle’s salt (30 mL). The biphasic mixture was allowed to stir for 30 minutes, then transferred to a separatory funnel with Et2O (30 mL), and the layers were separated. The aqueous layer was extracted Et2O (3 x 30 mL). The combined organics were washed with brine (1 x 30 mL), dried over Na2SO4, filtered, and concentrated in vacuo. The resulting crude residue was purified by silica gel chromatography (20% acetone in hexanes to 25 % acetone in hexanes) to afford alcohol 109 (1.42 g, 3.02 mmol, 97% yield) as a white solid. 1 H NMR (400 MHz, Chloroform-d): δ 6.06 (ddd, J = 9.5, 6.1, 1.2 Hz, 1H), 5.74 – 5.68 (m, 2H), 5.51 – 5.48 (m, 2H), 4.62 (dd, J = 3.8, 1.6 Hz, 1H), 4.45 (tt, J = 4.4, 1.0 Hz, 1H), 4.23 (ddd, J = 4.2, 2.9, 0.6 Hz, 1H), 4.03 (dd, J = 11.8, 3.7 Hz, 1H), 3.43 (dd, J = 11.8, 9.1 Hz, 1H), 3.04 (ddtt, J = 4.9, 2.8, 2.0, 1.0 Hz, 1H), 2.77 (dd, J = 5.5, 4.7 Hz, 1H), 2.58 (ddt, J = Chapter 3 – Towards C19-Diterpenoid Alkaloids: Approaches to the C4 Chiral Quaternary Center 67 17.9, 9.3, 2.3 Hz, 1H), 2.48 – 2.34 (m, 3H), 2.20 – 2.03 (m, 2H), 1.90 (dddd, J = 13.7, 11.4, 6.2, 3.7 Hz, 1H), 1.57 (ddd, J = 13.2, 11.1, 6.3 Hz, 1H), 1.07 (s, 9H), 1.00 (s, 9H). 13 C NMR (101 MHz, CDCl3): δ 176.6, 161.4, 134.7, 131.9, 131.2, 129.0, 121.6, 78.6, 65.9, 64.2, 60.4, 53.7, 46.4, 46.2, 45.7, 44.5, 35.0, 28.8, 28.2, 24.6, 22.3, 21.2, 20.7. FTIR (NaCl, thin film): 3458, 2935, 2859, 2245, 1738, 1748, 1732, 1476, 1382, 1364, 1364, 1107, 1038, 995, 911 cm-1. HRMS: (ESI-TOF) calc’d for C27H39O5Si [M + H]+ 471.2561, found 471.2555. [𝜶]𝟐𝟓 𝐃 = +168° (c = 0.51, CHCl3). TLC (50%EtOAc/50%Hexanes), Rf: 0.5 (UV, blue in p-anisaldehyde) Preparation of methyl ether 110: HO MeO O O O Si O t-Bu CH2Cl2 4 Å MS 0 °C to rt t-Bu 109 O Me3OBF4 Proton Sponge O O Si O t-Bu t-Bu 95% yield 110 A 250-mL, round-bottomed flask was charged with alcohol 109 (1.41 g, 3.00 mmol, 1.0 equiv), equipped with a rubber septum, purged with N2, and cooled to 0 °C in an ice bath. Meanwhile, Me3OBF4 (1.33 g, 9.00 mmol, 3 equiv), Proton Sponge (1.93 g, 9.00 mmol, 3 equiv), and activated 4 Å MS (2.8 g) were added to a 40 mL vial in the glovebox, brought out of the glovebox, and added to the flask containing the substrate. The mixture was then suspended in CH2Cl2 (30 mL), and allowed to stir for 24 hours, allowing the reaction to warm up to room temperature over this period. The reaction mixture was filtered through a plug of silica gel and celite that had been pre-packed with a 1:1 Hex/EtOAc mixture and rinsed further Chapter 3 – Towards C19-Diterpenoid Alkaloids: Approaches to the C4 Chiral Quaternary Center 68 with 1:1 Hex/EtOAc and concentrated in vacuo. The resulting crude residue was purified by silica gel chromatography (15% ethyl acetate in hexanes to 18% ethyl acetate in hexanes) to afford methyl ether 110 (1.39 g, 2.85 mmol, 95% yield) as a white crystalline solid. 1 H NMR (400 MHz, Chloroform-d): δ 5.99 (ddd, J = 9.5, 6.1, 1.1 Hz, 1H), 5.67 – 5.59 (m, 2H), 5.43 (d, J = 3.7 Hz, 1H), 5.40 (dt, J = 5.7, 2.1 Hz, 1H), 4.54 – 4.52 (m, 1H), 4.39 (t, J = 4.7 Hz, 1H), 4.16 (ddd, J = 4.2, 2.9, 0.6 Hz, 1H), 3.58 (d, J = 10.0 Hz, 1H), 3.38 (d, J = 10.0 Hz, 1H), 3.30 (s, 3H), 2.96 (dtq, J = 5.0, 2.9, 0.8 Hz, 1H), 2.71 (dd, J = 5.6, 4.8 Hz, 1H), 2.62 – 2.49 (m, 2H), 2.28 (dq, J = 17.3, 2.2 Hz, 1H), 2.10 – 2.00 (m, 1H), 1.94 – 1.77 (m, 2H), 1.61 – 1.54 (m, 1H), 1.01 (s, 9H), 0.94 (s, 9H). 13 C NMR (101 MHz, CDCl3): δ 175.1, 161.6, 134.9, 132.2, 131.3, 128.8, 121.4, 77.9, 77.3, 72.8, 66.0, 59.8, 59.5, 46.4, 45.8, 44.8, 43.2, 35.0, 28.8, 28.2, 25.5, 21.9, 21.2, 20.7. A 13C signal is observed to be overlapping with residual CDCl3 at 77.3, resolved peaks are seen in the HSQC spectrum, provided below. FTIR (NaCl, thin film): 2894, 2933, 2858, 1748, 1475, 1393, 1364, 1109, 994, 825 cm-1. HRMS: (ESI-TOF) calc’d for C28H41O5Si [M + H]+ 485.2718, found 485.2716. [𝜶]𝟐𝟓 𝐃 = +156° (c = 0.75, CHCl3). TLC (20%EtOAc/80%Hexanes), Rf: 0.5 (UV, blue in p-anisaldehyde) Chapter 3 – Towards C19-Diterpenoid Alkaloids: Approaches to the C4 Chiral Quaternary Center 69 Preparation of diol 111: OH OH MeO MeO O O LiBH4 THF O Si O t-Bu t-Bu 110 O 96% yield Si t-Bu O t-Bu 111 A 250-mL, round-bottomed flask was charged with lactone 110 (1.39 g, 2.88 mmol, 1.0 equiv), was equipped with a rubber septum, purged with N2, and dissolved in THF (29 mL). LiBH4 solution (2 M in THF, 7.2 mL, 14.4 mmol, 5 equiv) was added via syringe, and the reaction was allowed to continue stirring for 20 hours. The reaction was monitored by TLC, which indicated that it had not gone to completion. An additional portion of LiBH4 solution (2 M in THF, 7.2 mL, 14.4 mmol, 5 equiv) was added via syringe. After an additional 3 hours another portion of LiBH4 solution (2 M in THF, 7.2 mL, 14.4 mmol, 5 equiv) was added via syringe (15 equiv total). The reaction was allowed to continue to stir for another 24 hours at room temperature. The reaction was carefully quenched (bubbles evolve) with sat. NaHCO3 (30 mL), and diluted with Et2O (30 mL), and transferred to a separatory funnel. The layers were separated, and the aqueous layer was extracted with 20%IPA/80%CHCl3 (3 x 30 mL). The combined organics were dried over Na2SO4, filtered, and concentrated in vacuo. The resulting crude residue was purified by silica gel chromatography (15% acetone in hexanes to 20% acetone in hexanes) to afford diol 111 (1.34 g, 2.76 mmol, 96% yield) as a white solid. 1 H NMR (400 MHz, Chloroform-d): δ 6.19 (ddd, J = 5.8, 2.9, 1.7 Hz, 1H), 6.08 (ddd, J = 9.5, 6.1, 1.1 Hz, 1H), 5.67 (ddd, J = 9.3, 4.4, 1.9 Hz, 1H), 5.49 (d, J = 3.7 Hz, 1H), 5.44 (ddd, J = 5.8, 2.7, 1.1 Hz, 1H), 4.44 (td, J = 4.3, 0.9 Hz, 1H), 4.22 (ddd, J = 4.2, 3.0, 0.7 Hz, 1H), 4.03 (d, J = 4.1 Hz, 1H), 3.58 – 3.52 (m, 2H), 3.49 – 3.42 (m, 1H), 3.31 (s, 3H), 3.24 (d, J = Chapter 3 – Towards C19-Diterpenoid Alkaloids: Approaches to the C4 Chiral Quaternary Center 70 8.8 Hz, 1H), 3.08 – 3.01 (m, 2H), 2.91 (t, J = 5.3 Hz, 1H), 2.71 – 2.64 (m, 1H), 2.51 (dddd, J = 16.0, 8.4, 2.9, 1.1 Hz, 1H), 2.44 – 2.35 (m, 1H), 1.79 – 1.55 (m, 4H), 1.13 – 1.10 (m, 1H), 1.08 (s, 9H), 1.01 (s, 9H). 13 C NMR (101 MHz, CDCl3): δ 165.4, 135.9, 135.6, 133.6, 128.3, 120.9, 79.7, 77.1, 72.6, 68.2, 66.3, 59.1, 56.1, 46.5, 46.2, 41.5, 39.6, 35.0, 28.9, 28.3, 24.6, 21.2, 20.7, 20.7. FTIR (NaCl, thin film): 2950, 2910, 2876, 1748, 1476, 1362, 1105, 996, 826 cm-1. HRMS: (ESI-TOF) calc’d for C28H45O5Si [M + H]+ 489.3031, found 489.3020. [𝜶]𝟐𝟓 𝐃 = +152° (c = 0.42, CHCl3). TLC (50%EtOAc/50%Hexanes), Rf: 0.5 (UV, brown/pink in p-anisaldehyde) Preparation of silyl ether 112: OH OH MeO OH OSiEt3 MeO TESCl imidazole CH2Cl2 0 °C O Si t-Bu 111 O O t-Bu 98% yield Si t-Bu O t-Bu 112 A 200-mL, round-bottomed flask was charged with diol 111 (1.35 g, 2.76 mmol, 1.0 equiv), imidazole (0.42 g, 6.20 mmol, 2.25 equiv), dissolved in CH2Cl2 (55 mL), and cooled to 0 °C in an ice bath. TESCl (0.69 mL, 4.14 mmol, 1.5 equiv) added via syringe, and was allowed to continue to stir for 25 minutes at 0 °C. The reaction was monitored by TLC, which showed completion of the reaction. The reaction was quenched with sat. NaHCO3 (30 mL), and transferred to a separatory funnel. The layers were separated, and the aqueous was extracted with CH2Cl2 (3 x 30 mL). The combined organic extracts were dried over Na2SO4, filtered, and concentrated in vacuo. The resulting crude residue was purified by silica gel Chapter 3 – Towards C19-Diterpenoid Alkaloids: Approaches to the C4 Chiral Quaternary Center 71 chromatography (10% ethyl acetate in hexanes to 15% ethyl acetate in hexanes) to afford silyl ether 112 (1.63 g, 2.70 mmol, 98% yield) as a white solid. 1 H NMR (400 MHz, Chloroform-d): δ 6.18 (dt, J = 5.9, 2.3 Hz, 1H), 6.07 (ddd, J = 9.5, 6.2, 1.1 Hz, 1H), 5.66 (ddd, J = 9.4, 4.3, 1.9 Hz, 1H), 5.44 (d, J = 3.8 Hz, 1H), 5.40 (dt, J = 5.9, 1.8 Hz, 1H), 4.49 (t, J = 4.7 Hz, 1H), 4.25 – 4.18 (m, 1H), 4.03 (t, J = 3.4 Hz, 1H), 3.49 (d, J = 9.3 Hz, 1H), 3.29–3.26 (m, 2H), 3.25 (s, 3H), 3.19 (d, J = 8.7 Hz, 1H), 3.08 – 3.04 (m, 1H), 2.87 (t, J = 5.3 Hz, 1H), 2.42 – 2.33 (m, 2H), 2.22 (t, J = 9.5 Hz, 1H), 1.80 (s, 1H), 1.71 (dt, J = 7.6, 3.9 Hz, 2H), 1.46 (dt, J = 13.6, 8.2 Hz, 1H), 1.31 (dt, J = 13.7, 4.0 Hz, 1H), 1.09 (s, 9H), 1.01 (s, 9H), 0.95 (t, J = 7.9 Hz, 9H), 0.62 – 0.54 (m, 6H). 13 C NMR (101 MHz, CDCl3): δ 165.5, 136.1, 135.7, 133.9, 128.2, 120.9, 77.1, 72.3, 68.1, 66.3, 66.1, 58.8, 55.9, 46.3, 46.2, 43.1, 40.5, 34.4, 28.9, 28.3, 24.1, 21.3, 20.7, 18.6, 6.9, 4.3. FTIR (NaCl, thin film): 2950, 2910, 2876, 1748, 1476, 1362, 1105, 996, 826 cm-1. HRMS: (ESI-TOF) calc’d for C34H59O5Si2 [M + H]+ 603.3896, found 603.3881. [𝜶]𝟐𝟓 𝐃 = +103° (c = 0.38, CHCl3). TLC (15%EtOAc/85%Hexanes), Rf: 0.4 (UV, orange/pink in p-anisaldehyde) Preparation of methyl ether 113: OH OSiEt3 MeO OMe OSiEt3 MeO Me3OBF4 Proton Sponge O Si t-Bu 112 O CH2Cl2 4 Å MS 0 °C to rt O t-Bu 90% yield t-Bu Si O t-Bu 113 A 250-mL, round-bottomed flask was charged with alcohol 112 (1.63 g, 2.71 mmol, 1.0 equiv), equipped with a rubber septum, purged with N2, and cooled to 0 °C in an ice bath. Chapter 3 – Towards C19-Diterpenoid Alkaloids: Approaches to the C4 Chiral Quaternary Center 72 Meanwhile, Me3OBF4 (2.00 g, 13.55 mmol, 5 equiv), Proton Sponge (2.91 g, 13.55 mmol, 5 equiv), and activated 4 Å MS (3.3 g) were added to a 40 mL vial in the glovebox, brought out of the glovebox, and added to the flask containing the substrate. The mixture was then suspended in CH2Cl2 (27.1 mL), and allowed to stir for 24 hours, allowing the reaction to warm up to room temperature over this period. The reaction mixture was filtered through a plug of silica gel and celite that had been pre-packed with a 15% ethyl acetate/ 85% hexanes mixture and rinsed further with 15% ethyl acetate/ 85% hexanes and concentrated in vacuo. The resulting crude residue was purified by silica gel chromatography (2% to 3% to 4% to 5% to 6% ethyl acetate in hexanes to) to afford methyl ether 113 (1.5 g, 2.43 mmol, 90% yield) as a white solid. 1 H NMR (400 MHz, Chloroform-d): δ 6.08 (ddd, J = 9.5, 6.2, 1.1 Hz, 1H), 5.88 (dt, J = 5.8, 2.2 Hz, 1H), 5.65 (ddd, J = 9.3, 4.4, 1.9 Hz, 1H), 5.42 – 5.35 (m, 2H), 4.47 (td, J = 4.2, 0.9 Hz, 1H), 4.22 (ddd, J = 4.4, 3.0, 0.7 Hz, 1H), 3.55 (dd, J = 6.9, 2.7 Hz, 1H), 3.45 (d, J = 9.2 Hz, 1H), 3.30 (d, J = 9.3 Hz, 1H), 3.24 (s, 3H), 3.24 (s, 3H), 3.21 (d, J = 8.7 Hz, 1H), 3.16 (d, J = 8.7 Hz, 1H), 3.04 (ddddt, J = 5.8, 3.8, 2.9, 2.0, 1.0 Hz, 1H), 2.87 (dd, J = 6.4, 4.1 Hz, 1H), 2.35 – 2.27 (m, 2H), 2.23 (dd, J = 10.2, 7.8 Hz, 1H), 1.67 (dddd, J = 13.9, 8.4, 5.6, 2.7 Hz, 1H), 1.57 (dtd, J = 13.6, 6.8, 5.1 Hz, 1H), 1.40 – 1.24 (m, 2H), 1.09 (s, 9H), 1.01 (s, 9H), 0.95 (t, J = 7.9 Hz, 9H), 0.63 – 0.52 (m, 6H). 13 C NMR (101 MHz, CDCl3): δ 166.6, 136.0, 134.0, 131.6, 128.0, 119.8, 79.7, 77.1, 73.4, 66.5, 66.0, 58.7, 57.2, 56.5, 46.2 (2 overlapping 13C signals), 46.2, 43.7, 40.6, 34.8, 28.9, 28.3, 21.4, 21.3, 20.9, 20.7, 6.8, 4.3. Two 13C signals are observed to be overlapping at 46.2, resolved peaks are seen in the HSQC spectrum, provided below. Chapter 3 – Towards C19-Diterpenoid Alkaloids: Approaches to the C4 Chiral Quaternary Center 73 FTIR (NaCl, thin film): 3049, 2936, 2911, 2976, 2808 1476, 1103, 994 cm-1. HRMS: (ESI-TOF) calc’d for C35H61O5Si2 [M + H]+ 617.4052, found 617.4045. [𝜶]𝟐𝟓 𝐃 = +87° (c = 0.57, CHCl3). TLC (5%EtOAc/95%Hexanes), Rf: 0.4 (UV, green in p-anisaldehyde) Preparation of alcohol 114: MeO OMe OTES MeO OMe OH TFA CH2Cl2, 0 °C O Si O t-Bu t-Bu 113 90% yield O Si O t-Bu t-Bu 114 A 500-mL, round-bottomed flask was charged with silyl ether 113 (1.60 g, 2.59 mmol, 1.0 equiv), and CH2Cl2 (52 mL) and cooled to 0 °C in an ice bath. TFA (0.40 mL, 5.18 mmol, 2 equiv) was added via syringe. The reaction was monitored by TLC, and after 45 minutes at 0 °C, additional TFA (0.10 mL, 1.30 mmol, 0.5 equiv) was added. After a further 30 minutes, additional TFA (0.10 mL, 1.30 mmol, 0.5 equiv) was added (3 equiv total). After 15 more minutes, the reaction was done by TLC. The reaction was quenched with sat. NaHCO3 (50 mL), and the pH of the aqueous layer was checked to make sure it was basic. The biphasic mixture was transferred to a separatory funnel, the layers were separated, and the aqueous layer was extracted with CH2Cl2 (3 x 30 mL). The combined organic extracts were dried over Na2SO4, filtered, and concentrated in vacuo. The resulting crude residue was purified by silica gel chromatography (25% ethyl acetate in hexanes to 30% ethyl acetate in hexanes) to afford alcohol 114 (1.17 g, 2.33 mmol, 90% yield) as a white solid. 1 H NMR (400 MHz, Chloroform-d): δ 6.08 (ddd, J = 9.5, 6.2, 1.1 Hz, 1H), 5.90 (ddd, J = 5.8, 2.7, 1.9 Hz, 1H), 5.66 (ddd, J = 9.3, 4.4, 1.9 Hz, 1H), 5.45 – 5.42 (m, 2H), 4.44 (td, J = Chapter 3 – Towards C19-Diterpenoid Alkaloids: Approaches to the C4 Chiral Quaternary Center 74 4.2, 0.9 Hz, 1H), 4.26 – 4.19 (m, 1H), 3.57 – 3.52 (m, 2H), 3.46 (dd, J = 8.9, 1.3 Hz, 2H), 3.27 (s, 3H), 3.26 (s, 3H), 3.21 (d, J = 8.7 Hz, 1H), 3.05 – 2.99 (m, 2H), 2.91 (dd, J = 5.9, 4.6 Hz, 1H), 2.64 (t, J = 9.2 Hz, 1H), 2.46 (dddd, J = 15.7, 8.7, 2.8, 1.4 Hz, 1H), 2.30 (ddt, J = 15.7, 9.9, 2.1 Hz, 1H), 1.67 – 1.61 (m, 1H), 1.61 – 1.42 (m, 2H), 1.17 – 1.10 (m, 1H), 1.08 (s, 9H), 1.01 (s, 9H). 13 C NMR (101 MHz, CDCl3): δ 166.5, 135.8, 133.7, 131.5, 128.1, 119.6, 80.5, 79.7, 77.0, 72.4, 66.5, 58.9, 57.2, 56.8, 46.4, 46.2, 42.3, 39.7, 35.5, 28.9, 28.3, 23.2, 21.6, 21.2, 20.7. FTIR (NaCl, thin film): 3472, 3048, 2934, 2894, 2859, 1475, 1380, 1101, 991 cm-1. HRMS: (ESI-TOF) calc’d for C29H47O5Si [M + H]+ 503.3187, found 503.3184. [𝜶]𝟐𝟓 𝐃 = +136° (c = 0.58, CHCl3). TLC (30%EtOAc/70%Hexanes), Rf: 0.4 (UV, pink/orange in p-anisaldehyde) Preparation of aldehyde 115: MeO OMe OH MeO OMe O Cu(MeCN)4OTf (5 mol %) MeObpy (5 mol %), O Si O t-Bu t-Bu 114 NMI (5 mol %), ABNO (5 mol % MeCN, air 94% yield O Si O t-Bu t-Bu 115 A 100-mL, round-bottomed flask was charged with alcohol 114 (218 mg, 0.434 mmol, 1.0 equiv) and MeCN (8.6 mL). 4,4’-dimethoxy-2,2’-bipyridine (MeObpy, 4.7 mg, 21.7 µmol, 5 mol %), N-methylimidazole (6.9 µL, 86.8 µmol, 20 mol %), and ABNO (3.0 mg, 21.7 µmol, 5 mol %). Lastly, Cu(MeCN)4OTf (8.2 mg, 21.7 µmol, 5 mol %) was added, and the clear red/brown reaction mixture was stirred until slightly yellow green, and the TLC indicated the reaction had gone to completion (ca. 90 min), at which point the solution was filtered through a short plug of silica, flushed with ethyl acetate, and concentrated in vacuo. Purification of the Chapter 3 – Towards C19-Diterpenoid Alkaloids: Approaches to the C4 Chiral Quaternary Center 75 crude residue by silica gel chromatography (10 to 12% EtOAc in hexanes) afforded aldehyde 115 (204 mg, 0.408 mmol, 94% yield) as a white foam. Note: The product aldehyde 115 is prone to decomposition, prolonged storage should be avoided if possible. If necessary, it can be stored as a solid in the freezer and should be fine for at least a few weeks. Residual solvent accelerates the decomposition process. It is also possible to run the subsequent step without purification, and using the crude from the silica plug directly. Once the decomposition pathway was discovered, the aldehyde was reproducibly carried through as the crude in the subsequent reductive amination step, to keep the aldehyde for a minimum amount of time. 1 H NMR (400 MHz, Chloroform-d): δ 9.53 (s, 1H), 6.09 (ddd, J = 9.5, 6.1, 1.1 Hz, 1H), 5.85 (dt, J = 5.8, 2.3 Hz, 1H), 5.68 (dddd, J = 9.5, 4.3, 1.9, 0.6 Hz, 1H), 5.62 (dt, J = 5.8, 1.9 Hz, 1H), 5.40 (dd, J = 3.9, 0.7 Hz, 1H), 4.45 (tt, J = 4.3, 0.9 Hz, 1H), 4.21 (ddd, J = 4.3, 3.0, 0.7 Hz, 1H), 3.49 (dd, J = 8.9, 3.3 Hz, 1H), 3.39 (d, J = 9.2 Hz, 1H), 3.31 (d, J = 9.2 Hz, 1H), 3.27 (s, 3H), 3.26 (s, 3H), 3.02 (ddddd, J = 4.9, 3.8, 2.9, 2.0, 0.8 Hz, 1H), 2.89 – 2.86 (m, 1H), 2.44 (t, J = 7.0 Hz, 1H), 2.29 (ddt, J = 16.0, 7.5, 2.2 Hz, 1H), 2.22 (ddt, J = 16.0, 6.6, 2.2 Hz, 1H), 2.12 (ddd, J = 14.2, 6.7, 4.5 Hz, 1H), 1.83 (dddd, J = 13.4, 6.7, 4.6, 3.3 Hz, 1H), 1.55 (dtd, J = 13.6, 9.3, 4.6 Hz, 1H), 1.40 (ddd, J = 14.3, 9.7, 4.6 Hz, 1H), 1.08 (s, 9H), 1.02 (s, 9H). 13 C NMR (101 MHz, CDCl3): δ 204.0, 164.5, 135.2, 134.0, 132.0, 128.5, 121.8, 79.5, 77.3, 76.1, 66.3, 59.3, 58.1, 56.9, 50.8, 46.2, 46.0, 44.1, 34.5, 28.9, 28.3, 21.5, 21.2, 21.1, 20.7. A 13C signal is observed to be overlapping with residual CDCl3 at 77.3, resolved peaks are seen in the HSQC spectrum, provided below. FTIR (NaCl, thin film): 2933, 2894, 2859, 2859, 1724, 1476, 1103, 993 cm-1. HRMS: (ESI-TOF) calc’d for C29H45O5Si [M + H]+ 501.3031, found 501.3038. Chapter 3 – Towards C19-Diterpenoid Alkaloids: Approaches to the C4 Chiral Quaternary Center 76 [𝜶]𝟐𝟓 𝐃 = +87° (c = 0.60, CHCl3). TLC (20%EtOAc/80%Hexanes), Rf: 0.5 (UV, blue in p-anisaldehyde) Preparation of N-allylamine 145: OMe O MeO Ti(Oi-Pr)4 OMe N MeO NH2 H EtOH then NaBH4 O O O Si t-Bu t-Bu 114 (quantitative) O Si t-Bu t-Bu 145 A 100-mL, round-bottomed flask was charged with aldehyde 114 (311 mg, 0.621 mmol, 1 equiv), and azeotroped 3x with toluene (from the solvent system) to remove water, and put under high vacuum for 2 hours. The flask was equipped with a rubber septum, purged with N2, and dissolved in EtOH (stored over 3 Å MS, 21 mL). Allylamine (freshly distilled over CaCl2, 0.19 mL, 248 mmol, 4 equiv) was added followed by Ti(Oi-Pr)4 (0.39 mL, 1.24 mmol, 2 equv) and let stir for 90 minutes at room temperature. At this time, NaBH4 (117 mg, 3.10 mmol, 5 equiv) was added in one portion, and let stir for 15 minutes further. The reaction was carefully quenched with aqueous 28–30% NH3 solution (10 mL), and let stir for 10 minutes at room temperature. The reaction was then filtered through a basic alumina plug that had been pre-washed with EtOH, then flushed with ethyl acetate, and concentrated in vacuo. Then, the crude mixture was filtered through a silica plug, flushed with 100% CH2Cl2 to remove nonpolar impurities, and then with 5% 7 NH3/95% CH2Cl2 to elute the product N-allyl amine 145. A sample of this material was purified by silica gel chromatography (to 2% to 3% to 4% 7 N NH3/MeOH solution/ CH2Cl2) to afford analytically pure material for characterization: Chapter 3 – Towards C19-Diterpenoid Alkaloids: Approaches to the C4 Chiral Quaternary Center 1 77 H NMR (400 MHz, Chloroform-d): δ 6.06 (ddd, J = 9.5, 6.2, 1.1 Hz, 1H), 5.93 – 5.81 (m, 2H), 5.64 (ddd, J = 9.5, 4.4, 1.9 Hz, 1H), 5.43 – 5.38 (m, 2H), 5.16 (dq, J = 17.2, 1.7 Hz, 1H), 5.08 (dq, J = 10.3, 1.3 Hz, 1H), 4.47 – 4.42 (m, 1H), 4.22 (ddd, J = 4.2, 3.0, 0.7 Hz, 1H), 3.55 (dd, J = 7.5, 2.8 Hz, 1H), 3.29 – 3.15 (m, 10H), 3.02 (tdd, J = 5.8, 2.8, 1.4 Hz, 1H), 2.90 – 2.86 (m, 1H), 2.49 (s, 2H), 2.41 (dd, J = 10.3, 8.0 Hz, 1H), 2.33 – 2.26 (m, 2H), 1.71 – 1.61 (m, 1H), 1.61 – 1.49 (m, 2H), 1.41 – 1.31 (m, 1H), 1.07 (s, 9H), 1.00 (s, 9H). 13 C NMR (101 MHz, CDCl3): δ 166.5, 136.8, 135.8, 134.0, 131.3, 128.0, 119.6, 116.1, 79.6, 77.0 (overlapping with CDCl3), 76.5, 66.5, 58.8, 57.2, 56.8, 56.3, 53.0, 46.3, 46.2, 44.5, 39.3, 35.4, 28.9, 28.3, 24.3, 21.7, 21.2, 20.7. FTIR (NaCl, thin film): 3049, 2934, 2898, 2859, 2859, 1475, 1102, 1106, 992, 825, 732 cm-1. HRMS: (ESI-TOF) calc’d for C32H52NO4Si [M + H]+ 542.3660, found 542.3653. [𝜶]𝟐𝟓 𝐃 = +104° (c = 2.20, CHCl3). TLC (5% 7N NH3 in MeOH/95%CH2Cl2), Rf: 0.5 (UV, blue in p-anisaldehyde) Preparation of primary amine 116 via deprotection of N-allyl 145: O NMe OMe N MeO H O N Me O OMe NH2 MeO Pd(PPh)3 (10 mol %) CH2Cl2 O O Si t-Bu t-Bu 145 90% yield (2 steps) O O Si t-Bu t-Bu 116 A 100-mL, round-bottomed flask was charged with the crude allylamine 145 and 3,5dimethylbarbituric acid (0.582 g, 3.72 mmol, 6 equiv), and dissolved in CH2Cl2 (12.4 mL) under N2. Meanwhile, a solution of Pd(PPh3)4 (72 mg, 0.0621 mmol, 10 mol % in 3 mL Chapter 3 – Towards C19-Diterpenoid Alkaloids: Approaches to the C4 Chiral Quaternary Center 78 CH2Cl2) was made inside the glovebox, brought out, and added via syringe to the solution of substrate. The reaction was allowed to stir for 1 hour at room temperature, and was then quenched with sat. NaHCO3 (20 mL). The mixture was then transferred to a separatory funnel, and extracted with 20%IPA/80% CHCl3 (3 x 30 mL). The combined organic extracts were dried over Na2SO4, filtered, and concentrated in vacuo. The resulting crude residue was purified by silica gel chromatography (1% to 2% to 3% to 4% 7 N NH3/MeOH solution/ CH2Cl2) to afford primary amine 116 (280 mg, 0.559 mmol, 90% yield) as a clear oil. 1 H NMR (400 MHz, Chloroform-d): δ 6.06 (ddd, J = 9.5, 6.1, 1.1 Hz, 1H), 5.87 (dt, J = 5.9, 2.3 Hz, 1H), 5.65 (ddd, J = 9.4, 4.4, 1.9 Hz, 1H), 5.44 – 5.39 (m, 2H), 4.46 (t, J = 4.7 Hz, 1H), 4.23 (ddd, J = 4.3, 3.0, 0.7 Hz, 1H), 3.56 (dd, J = 7.7, 2.8 Hz, 1H), 3.26 – 3.24 (m, 6H), 3.22 – 3.19 (m, 1H), 3.18 (d, J = 9.2 Hz, 1H), 3.05 – 3.00 (m, 1H), 2.88 (dd, J = 5.9, 4.6 Hz, 1H), 2.63 (d, J = 13.1 Hz, 1H), 2.58 (d, J = 13.1 Hz, 1H), 2.39 (dd, J = 10.0, 8.4 Hz, 1H), 2.31 – 2.26 (m, 2H), 1.68 (dddd, J = 13.3, 8.0, 5.4, 2.7 Hz, 1H), 1.61 – 1.37 (m, 4H), 1.34 – 1.21 (m, 1H), 1.08 (s, 9H), 1.00 (s, 9H). 13 C NMR (101 MHz, CDCl3): δ 166.4, 135.8, 134.1, 131.1, 128.1, 119.7, 79.7, 77.0, 76.1, 66.5, 58.8, 57.2, 56.9, 49.5, 46.3, 46.2, 44.0, 39.7, 35.2, 28.9, 28.3, 23.9, 21.8, 21.2, 20.7. A 13C signal is observed to be overlapping with residual CDCl3 at 77.00, the peak can be seen in the HSQC spectrum, provided below. FTIR (NaCl, thin film): 3386, 3050, 2933, 2859, 2362, 1476, 1463, 1106, 992 cm-1. HRMS: (ESI-TOF) calc’d for C29H48NO4Si [M + H]+ 502.3347, found 502.3344. [𝜶]𝟐𝟓 𝐃 = +105° (c = 0.67, CHCl3). TLC (10%MeOH/90%CH2Cl2), Rf: 0.15 (UV, blue in p-anisaldehyde) Chapter 3 – Towards C19-Diterpenoid Alkaloids: Approaches to the C4 Chiral Quaternary Center 79 Preparation of N-ethyl amide 118: O MeO2C O O Si O t-Bu t-Bu 108 Na-2-ethylhexanoate NEtH2·HCl OH NHEt MeO2C O THF/NEtH2 60 °C to 70 °C [sealed pressure vessel] O 51% yield Si t-Bu O t-Bu 118 To a 75 mL pressure vessel containing lactone 108 (437 mg, 0.877 mmol, 1 equiv), sodium 2-ethylhexanoate (657 mg, 3.95 mmol, 4.5 equiv), ethylamine hydrochloride (161 mg, 1.97 mmol, 2.25 equiv) were added, followed by THF (9 mL) and neat NEtH2 (4 mL). The vessel was sealed (with a Teflon cap that had a perfluoro O-ring), and heated to 60 °C in an oil bath. The reaction was allowed to stir at 60 °C for 2 days, and the reaction progress was monitored. At this time, the reaction had not gone to completion, so the flask was re-sealed and the bath temperature was raised to 70 °C and stirred for another 2 days at this temperature. The reaction was then cooled to room temperature, and transferred to a separatory funnel with EtOAc (30 mL) and aqueous 0.5 M NaOH (20 mL). The layers were separated, and the aqueous phase was extracted with EtOAc (3 x 20 mL) and CH2Cl2 (2 x 20 mL). The combined organic extracts were dried over Na2SO4, filtered, and concentrated in vacuo. The crude residue was purified by silica gel chromatography (20% to 30% to 40% ethyl acetate in hexanes) to afford secondary amide 118 as a white foam (243 mg, 0.447 mmol, 51% yield). Note: Neat NH2Et was obtained from distillation of 70%NH2Et/ 30% H2O solution, using a cold finger (with dry ice/acetone) to condense the very volatile NH2Et, and stored over KOH pellets in the freezer. 1 H NMR (400 MHz, Chloroform-d): δ 6.22 (t, J = 5.6 Hz, 1H), 6.16 – 6.05 (m, 2H), 5.65 (ddd, J = 9.4, 4.4, 1.9 Hz, 1H), 5.50 (dt, J = 5.9, 1.9 Hz, 1H), 5.33 (s, 1H), 4.54 (t, J = 4.8 Hz, Chapter 3 – Towards C19-Diterpenoid Alkaloids: Approaches to the C4 Chiral Quaternary Center 80 1H), 4.17 (ddd, J = 4.0, 2.9, 0.7 Hz, 1H), 3.91 (t, J = 4.4 Hz, 1H), 3.68 (s, 3H), 3.28 (dqd, J = 14.5, 7.2, 5.7 Hz, 1H), 3.22 – 3.10 (m, 2H), 3.01 – 2.96 (m, 1H), 2.90 (t, J = 5.3 Hz, 1H), 2.37 – 2.26 (m, 3H), 2.09 – 2.02 (m, 1H), 1.80 – 1.69 (m, 3H), 1.11 – 1.06 (m, 12H), 1.00 (s, 9H). 13 C NMR (101 MHz, CDCl3): δ 174.9, 168.6, 163.6, 135.6, 135.1, 132.8, 128.5, 122.0, 76.6, 66.9, 66.2, 56.9, 55.5, 52.9, 46.4, 46.3, 46.2, 35.2, 34.7, 28.9, 28.3, 25.6, 21.2, 20.7, 18.0, 14.6. FTIR (NaCl, thin film): 3409, 3033, 3969, 6935, 2896, 2859, 2242, 1713, 1668, 1520, 1229, 1101, 826, 732 cm-1. HRMS: (ESI-TOF) calc’d for C30H46O6NSi [M + H]+ 544.3089, found 544.3102. [𝜶]𝟐𝟓 𝐃 = +137° (c = 1.00, CHCl3). TLC (50%EtOAc/ 50% Hexanes), Rf: 0.25 (UV, yellow in p-anisaldehyde) Preparation of amide 117: OH O MeO2C O (allyl)NH2•HCl sodium 2-ethylhexanoate NH(allyl) MeO2C O THF/allylamine (1:1.5) 80 °C, 3 d O Si O t-Bu t-Bu 108 54% yield (64% BRSM) O Si O t-Bu t-Bu 117 To a 150 mL pressure vessel containing lactone 108 (1.36 g, 2.73 mmol, 1.0 equiv), sodium 2-ethylhexanoate (2.04 g, 12.3 mmol, 4.5 equiv), allylamine hydrochloride (0.574 g, 6.14 mmol, 2.25 equiv) were added, followed by THF (10.7 mL) and neat N-allylamine (16 mL). The vessel was sealed (with a Teflon cap that had a perfluoro O-ring), and heated to 80 °C in an oil bath. The reaction was allowed to stir at 80 °C for 3 days. The reaction was then cooled to room temperature, and transferred to a separatory funnel with EtOAc (100 mL) and aqueous 0.5 M NaOH (30 mL). The layers were separated, and the aqueous phase was extracted with EtOAc (4 x 50 mL). The combined organic extracts were dried (Na2SO4), filtered, and Chapter 3 – Towards C19-Diterpenoid Alkaloids: Approaches to the C4 Chiral Quaternary Center 81 concentrated in vacuo. The crude residue was purified by silica gel chromatography (20-3040% EtOAc in hexanes) to afford N-allyl amide 117 (0.803 g, 1.47 mmol, 54% yield, 64% BRSM) and starting material 108 (0.203 g, 0.41 mmol, 15%). Note: N-allylamine was purified by distillation over CaCl2 prior to use. 1 H NMR (500 MHz, Chloroform-d): δ 6.33 (t, J = 5.7 Hz, 1H), 6.13 (dt, J = 5.9, 2.3 Hz, 1H), 6.10 (ddd, J = 9.5, 6.2, 1.1 Hz, 1H), 5.82 – 5.73 (m, 1H), 5.68 – 5.63 (m, 1H), 5.51 (dt, J = 5.9, 1.9 Hz, 1H), 5.34 (d, J = 3.9 Hz, 1H), 5.14 (t, J = 1.5 Hz, 1H), 5.11 (dq, J = 6.7, 1.4 Hz, 1H), 4.56 – 4.51 (m, 1H), 4.18 (ddd, J = 4.1, 3.0, 0.7 Hz, 1H), 3.92 (dd, J = 5.3, 3.4 Hz, 1H), 3.88 – 3.77 (m, 2H), 3.70 (s, 3H), 3.17 (td, J = 9.1, 1.5 Hz, 1H), 3.00 (dddp, J = 5.0, 3.1, 2.1, 1.0 Hz, 1H), 2.92 – 2.89 (m, 1H), 2.39 – 2.30 (m, 3H), 2.09 (dddd, J = 14.4, 5.6, 4.0, 1.5 Hz, 1H), 1.86 – 1.70 (m, 3H), 1.08 (s, 9H), 1.01 (s, 9H). 13 C NMR (126 MHz, CDCl3): δ 174.9, 168.6, 163.5, 135.6, 135.2, 133.7, 132.8, 128.5, 122.0, 116.7, 76.6, 66.9, 66.2, 57.0, 55.6, 52.9, 46.4, 46.3, 46.2, 42.1, 35.3, 28.9, 28.3, 25.5, 21.2, 20.7, 18.1. FTIR (NaCl, thin film): 3413, 2934, 3858, 1714, 1674, 1519, 1476, 1252, 1230, 1101, 992, 827, 731 cm-1. HRMS: (ESI-TOF) calc’d for C31H46NO6Si [M + H]+ 556.3089, found 556.3085 [𝜶]𝟐𝟓 𝐃 = +129° (c = 1.34, CHCl3). TLC (70% EtOAc/30% Hexanes), Rf: 0.5 (brown in p-anisaldehyde) Chapter 3 – Towards C19-Diterpenoid Alkaloids: Approaches to the C4 Chiral Quaternary Center 82 Preparation of diol 120: OH O OH HO NHEt MeO2C NHEt LiBHEt3 O THF, -20 to 10 °C O Si O t-Bu t-Bu 86% yield 118 O Si O t-Bu t-Bu 120 A flame-dried 100 mL round-bottom flask was charge with ester 118 (360 mg, 0.67 mmol, 1.0 equiv). The starting material was dried by azeotroping with toluene (3 x 25 mL). The flask was put under N2 before THF (13 mL) was added and the solution cooled to -20 °C. In a separate flame-dried 50 mL pear-shaped flask, a solution of LiBHEt3 (1M in THF, 3.3 mL, 3.3 mmol, 5.0 equiv) was diluted with THF (5 mL). The solution of LiBHEt3 was cooled to 20 °C and transferred to the flask of 118 by cannula over 10 minutes. After stirring at -20 °C for an additional 5 minutes, the reaction was warmed to 0 °C and stirred for 45 min. The reaction quenched with 0.5 M NaOH (aq., 15 mL) at 0 °C. The mixture was warmed to room temperature and stirred for 10 minutes. The mixture was transferred to a separatory funnel and extracted with EtOAc (6 x 50 mL). The combined organic extracts were dried (Na2SO4), filtered, and concentrated in vacuo to give the crude product as a colorless oil. The product was purified by column chromatography (SiO2, 75-100% EtOAc in hexanes) to give diol 120 (293 mg, 0.58 mmol, 86% yield) as a white foam. 1 H NMR (400 MHz, Chloroform-d) δ 6.18 (dt, J = 5.8, 2.3 Hz, 1H), 6.11 – 5.98 (m, 2H), 5.65 (ddd, J = 9.4, 4.4, 2.0 Hz, 1H), 5.54 (d, J = 3.7 Hz, 1H), 5.44 (dt, J = 5.8, 1.8 Hz, 1H), 4.58 – 4.49 (m, 1H), 4.21 (ddt, J = 3.8, 3.0, 1.4 Hz, 1H), 4.04 – 3.98 (m, 1H), 3.75 (dd, J = 10.9, 7.8 Hz, 1H), 3.52 (dd, J = 10.9, 3.8 Hz, 1H), 3.41 – 3.16 (m, 3H), 3.07 – 2.99 (m, 1H), 2.95 (t, J = 5.3 Hz, 1H), 2.86 (t, J = 9.6 Hz, 1H), 2.33 – 2.12 (m, 3H), 1.98 – 1.90 (m, 1H), 1.90 – 1.78 (m, 2H), 1.46 – 1.34 (m, 1H), 1.13 (t, J = 7.3 Hz, 3H), 1.07 (s, 9H), 1.00 (s, 9H). Chapter 3 – Towards C19-Diterpenoid Alkaloids: Approaches to the C4 Chiral Quaternary Center 13 83 C NMR (101 MHz, CDCl3) δ 176.44, 164.58, 136.42, 135.74, 133.95, 128.66, 121.89, 77.04, 67.34, 66.40, 65.63, 56.13, 47.38, 46.64, 46.40, 41.63, 36.17, 34.45, 29.06, 28.45, 24.23, 21.35, 20.85, 18.20, 14.89. FTIR (NaCl, thin film): 3386, 2934, 2861, 2356, 1635, 1529, 1476, 1332, 1226, 1102, 1060, 1012, 993, 824, 810, 757, 732, 643 cm-1. HRMS: (ESI-TOF) calc’d for C29H46NO5Si [M + H]+ 516.3140, found 516.3148 [𝜶]𝟐𝟐 𝐃 = +111° (c = 1.0, CHCl3). TLC (100%EtOAc): Rf: 0.4 (brown/purple in p-anisaldehyde). Preparation of diol 119: OH MeO2C O OH HO NH(allyl) NH(allyl) LiBHEt3 O THF, -20 to 10 °C O Si O t-Bu t-Bu 117 91% yield O Si O t-Bu t-Bu 119 A flame-dried 250 mL round-bottom flask was charge with ester 117 (1.830 g, 3.29 mmol, 1.0 equiv). The starting material was dried by azeotroping with toluene (3 x 25 mL). The flask was put under N2 before THF (45 mL) was added and the solution cooled to -20 °C. In a separate flame-dried 100 mL pear-shaped flask, a solution of LiBHEt3 (1M in THF, 16.5 mL, 16.5 mmol, 5.0 equiv) was diluted with THF (20 mL). The solution of LiBHEt3 was cooled to -20 °C and transferred to the flask of 30 by cannula over 30 minutes. After stirring at -20 °C for an additional 15 minutes, the reaction was warmed to 10 °C and stirred for 1h. The reaction was cooled to 0 °C before quenching with 0.5 M NaOH (aq., 20 mL). The mixture was warmed to room temperature and stirred for 1 hour. The mixture was extracted with EtOAc (2 x 100 mL; 6 x 40 mL). The combined organic extracts were dried (Na2SO4), filtered, and Chapter 3 – Towards C19-Diterpenoid Alkaloids: Approaches to the C4 Chiral Quaternary Center 84 concentrated in vacuo to give the crude product as a colorless oil. The product was purified by column chromatography (SiO2, 25-50-75-100% EtOAc in hexanes) to give diol 119 (1.574 g, 2.99 mmol, 91% yield) as a white foam. 1 H NMR (400 MHz, Chloroform-d): δ 6.19 (dt, J = 5.9, 2.3 Hz, 1H), 6.13 (t, J = 5.7 Hz, 1H), 6.06 (ddd, J = 9.5, 6.1, 1.1 Hz, 1H), 5.82 (ddt, J = 17.2, 10.2, 5.7 Hz, 1H), 5.66 (ddd, J = 9.3, 4.4, 1.9 Hz, 1H), 5.54 (d, J = 3.7 Hz, 1H), 5.44 (dt, J = 5.8, 1.9 Hz, 1H), 5.22 –5.13 (m, 2H), 4.53 (td, J = 4.3, 0.9 Hz, 1H), 4.21 (ddd, J = 4.2, 3.0, 0.7 Hz, 1H), 4.02 (t, J = 3.1 Hz, 1H), 3.94 (dtt, J = 15.6, 5.7, 1.6 Hz, 1H), 3.84 (dtt, J = 15.7, 5.7, 1.5 Hz, 1H), 3.76 (d, J = 10.8 Hz, 1H), 3.55 (d, J = 10.9 Hz, 1H), 3.14 – 3.01 (m, 2H), 2.95 (dd, J = 5.9, 4.6 Hz, 1H), 2.86 (t, J = 9.7 Hz, 1H), 2.32 – 2.18 (m, 3H), 1.88 – 1.81 (m, 2H), 1.48 – 1.38 (m, 2H), 1.08 (s, 9H), 1.00 (s, 9H). 13 C NMR (101 MHz, CDCl3): δ 176.2, 164.4, 136.3, 135.6, 133.9, 133.8, 128.5, 121.8, 116.8, 76.9, 67.2, 66.2, 65.5, 56.0, 47.6, 46.5, 46.2, 41.8, 41.6, 36.1, 28.9, 28.3, 24.1, 21.2, 20.7, 18.0. FTIR (NaCl, thin film): 3352, 2931, 2858, 1634, 1539, 1475, 1103, 994, 827, 733 cm-1. HRMS: (ESI-TOF) calc’d for C30H46NO5Si [M + H]+ 528.3140, found 528.3130 [𝜶]𝟐𝟓 𝐃 = +104° (c = 1.08, CHCl3). TLC (100%EtOAc): Rf: 0.5 (pink/brown in p-anisaldehyde). Chapter 3 – Towards C19-Diterpenoid Alkaloids: Approaches to the C4 Chiral Quaternary Center 85 Preparation of Amide 122: OH HO OMe MeO NHEt NHEt Me3OBF4 Proton Sponge O OH MeO NHEt O OMe HO NHEt O O 4ÅMS, CH2Cl2 O Si O t-Bu t-Bu 25% yield 121 (49% after 1 recycle) 120 O Si O t-Bu t-Bu 122 25% yield O Si O t-Bu t-Bu O Si O t-Bu t-Bu 146 147 38% yield (~1:1) An oven-dried 20 mL scintillation vial with a cross stir bar was charged with diol 120 (122 mg, 0.24 mmol, 1.0 equiv). CH2Cl2 (4.8 mL) was added, followed by Proton Sponge® (406 mg, 1.9 mmol, 8.0 equiv), trimethyloxonium tetrafluoroborate (210 mg, 14.2 mmol, 6.0 equiv), and 4Å molecular sieve powder (120 mg). The vial was stirred (1000 rpm) at 20 °C for 14 hours. HOAc (0.1 mL) was added, and the mixture was passed over a plug of silica gel, eluting with 50-100% EtOAc in hexanes. The filtrate was concentrated to give the crude product as a yellow oil. The product was purified by column chromatography (SiO2, 20-3040-50-60-70-80% EtOAc in hexane) to give di-methylated product 122 (32.2 mg, 0.059 mmol, 25% yield) and a ~1:1 mixture of isomeric mono-methylated products 146 and 147 (47.6 mg, 0.090 mmol, 38% yield). The mixture of 146 and 147 (47.6 mg) was resubjected to the same set of conditions to yield an additional 31.0 mg (0.057 mmol, 65% yield) of 122. This represents a combined 49% yield of 122 after 1 recycle of 146 and 147. Characterization data for 122: 1 H NMR (400 MHz, Chloroform-d) δ 6.66 (t, J = 5.5 Hz, 1H), 6.06 (ddd, J = 9.4, 6.1, 0.6 Hz, 1H), 5.84 (dt, J = 4.7, 2.3 Hz, 1H), 5.66 (ddd, J = 9.2, 4.4, 1.8 Hz, 1H), 5.42 (d, J = 3.8 Chapter 3 – Towards C19-Diterpenoid Alkaloids: Approaches to the C4 Chiral Quaternary Center 86 Hz, 1H), 5.40 – 5.32 (m, 1H), 4.49 (t, J = 4.7 Hz, 1H), 4.29 – 4.19 (m, 1H), 3.59 (dd, J = 8.2, 2.8 Hz, 1H), 3.40 (d, J = 9.4 Hz, 1H), 3.27 (s, 3H), 3.26 – 3.21 (m, 5H), 3.18 (d, J = 9.3 Hz, 1H), 3.06 – 3.00 (m, 1H), 2.87 (t, J = 5.3 Hz, 1H), 2.65 (t, J = 9.4 Hz, 1H), 2.57 (dt, J = 13.7, 6.5 Hz, 1H), 2.33 – 2.24 (m, 1H), 2.24 – 2.14 (m, 1H), 1.81 – 1.66 (m, 2H), 1.66 – 1.55 (m, 1H), 1.09 (d, J = 7.7 Hz, 12H), 1.01 (s, 9H). 13 C NMR (101 MHz, CDCl3) δ 174.99, 166.16, 135.80, 133.51, 131.78, 128.36, 119.52, 79.04, 76.98, 76.14, 66.61, 58.53, 57.32, 57.03, 46.93, 46.52, 46.35, 43.58, 36.59, 34.40, 29.03, 28.44, 21.66, 21.39, 21.14, 20.81, 15.01. FTIR (NaCl, thin film): 3355, 2932, 2902, 2859, 1648, 1556, 1534, 1476, 1380, 1265, 1228, 1102, 991, 935, 912, 843, 829, 755, 642 cm-1. HRMS: (ESI-TOF) calc’d for C31H50NO5Si [M + H]+ 544.3453, found 544.3451 [𝜶]𝟐𝟐 𝐃 = +94° (c = 1.07, CHCl3). TLC (2:1 EtOAc:Hexanes), Rf: 0.6 (blue in p-anisaldehyde) Characterization data for C1-O methyl ether 147: 1 H NMR (400 MHz, Chloroform-d) δ 6.16 (t, J = 5.6 Hz, 1H), 6.07 (ddd, J = 9.5, 6.2, 1.1 Hz, 1H), 5.87 (dt, J = 6.0, 2.3 Hz, 1H), 5.65 (ddd, J = 9.5, 4.5, 1.9 Hz, 1H), 5.46 (d, J = 3.7 Hz, 1H), 5.39 (ddd, J = 5.8, 2.3, 1.4 Hz, 1H), 4.51 (t, J = 4.7 Hz, 1H), 4.26 – 4.18 (m, 1H), 3.71 (d, J = 10.9 Hz, 1H), 3.59 – 3.42 (m, 2H), 3.40 – 3.26 (m, 1H), 3.25 (s, 4H), 3.10 – 2.98 (m, 2H), 2.97 – 2.89 (m, 1H), 2.71 (t, J = 9.3 Hz, 1H), 2.30 – 2.05 (m, 3H), 1.75 (q, J = 4.7 Hz, 2H), 1.41 – 1.32 (m, 1H), 1.13 (t, J = 7.3 Hz, 3H), 1.07 (s, 9H), 1.00 (s, 9H). Chapter 3 – Towards C19-Diterpenoid Alkaloids: Approaches to the C4 Chiral Quaternary Center 13 87 C NMR (101 MHz, CDCl3) δ 176.34, 165.99, 135.90, 133.95, 131.96, 128.40, 120.89, 78.80, 66.47, 66.23, 57.96, 56.46, 47.74, 46.56, 46.35, 42.82, 36.22, 34.46, 29.06, 28.45, 21.91, 21.35, 20.84, 19.66, 14.89. FTIR (NaCl, thin film): 3354, 2933, 1633, 1462, 1358, 1101, 991, 825, 755, 637 cm-1. HRMS: (ESI-TOF) calc’d for C30H48NO5Si [M + H]+ 530.3296, found 530.3311. [𝜶]𝟐𝟐 𝐃 = +103° (c = 0.755, CHCl3). TLC (2:1 EtOAc:Hexanes), Rf: 0.3 (blue in p-anisaldehyde) Characterization data for C18-O methyl ether 146: 1 H NMR (600 MHz, Chloroform-d) δ 6.48 (t, J = 5.5 Hz, 1H), 6.11 (dt, J = 5.3, 2.2 Hz, 1H), 6.05 (dd, J = 9.4, 6.1 Hz, 1H), 5.66 (ddd, J = 9.5, 4.4, 1.9 Hz, 1H), 5.49 (d, J = 3.8 Hz, 1H), 5.40 (dd, J = 6.0, 2.5 Hz, 1H), 4.46 (t, J = 4.6 Hz, 1H), 4.21 (t, J = 3.6 Hz, 1H), 4.03 (dd, J = 5.2, 2.4 Hz, 1H), 3.48 (d, J = 9.2 Hz, 1H), 3.32 – 3.21 (m, 6H), 3.06 – 3.01 (m, 1H), 2.89 (t, J = 5.3 Hz, 1H), 2.62 (t, J = 9.6 Hz, 1H), 2.40 (ddd, J = 13.6, 11.4, 4.7 Hz, 1H), 2.36 – 2.29 (m, 1H), 2.24 (ddd, J = 16.6, 8.9, 3.0 Hz, 1H), 1.80 – 1.69 (m, 2H), 1.33 (dt, J = 13.8, 4.5 Hz, 1H), 1.10 (t, J = 7.3 Hz, 3H), 1.08 (s, 9H), 1.00 (s, 9H). 13 C NMR (101 MHz, CDCl3) δ 175.01, 165.12, 135.84, 135.66, 133.52, 128.56, 121.07, 77.03, 75.39, 67.69, 66.41, 58.88, 56.48, 47.00, 46.58, 46.38, 42.75, 36.18, 34.34, 29.01, 28.43, 24.78, 21.38, 20.81, 18.64, 15.00. FTIR (NaCl, thin film): 3363, 2934, 2861, 1653, 1530, 1475, 1378, 1265, 1103, 1058, 993, 827, 812, 779, 760, 640 cm-1. HRMS: (ESI-TOF) calc’d for C30H48NO5Si [M + H]+ 530.3296, found 530.3306 [𝜶]𝟐𝟐 𝐃 = +134° (c =1.15, CHCl3). Chapter 3 – Towards C19-Diterpenoid Alkaloids: Approaches to the C4 Chiral Quaternary Center 88 TLC (2:1 EtOAc:Hexanes), Rf: 0.2 (purple in p-anisaldehyde) Preparation of Amide 148: OH HO NH(allyl) O OMe MeO 17. Me3OBF4 Proton Sponge NH(allyl) OH MeO NH(allyl) OMe HO NH(allyl) O O O O Si O tBu tBu O Si O tBu tBu O Si O tBu tBu 148 64% yield 149 150 4ÅMS, CH2Cl2 O Si O tBu tBu 121 69% yield 148 (1 recycle) 14% yield (~1:1) 5 oven-dried 20 mL scintillation vials with cross stir bars were charged with 0.250 g (1.25 g total, 2.37 mmol, 1.0 equiv) of diol 121, each. CH2Cl2 (9.5 mL) was added to each vial, followed by Proton Sponge® (0.610 g each, 0.35 g total, 14.2 mmol, 6.0 equiv), trimethyloxonium tetrafluoroborate (0.420 g each, 2.10 g total, 14.2 mmol, 6.0 equiv), and 4Å molecular sieve powder (250 mg to each vial). The vials were stirred (1000 rpm) at 20 °C for 18 hours. The contents of the vials were combined, HOAc (0.5 mL) was added, and the mixture was passed over a plug of silica gel, eluting with 50-100% EtOAc in hexanes. The eluent was concentrated to give the crude product as a yellow oil. The product was purified by column chromatography (SiO2, 20-30-40-50-60-70-80% EtOAc in hexane) to give di-methylated product 148 (0.839 g, 1.52 mmol, 64% yield) and a ~1:1 mixture of isomeric mono-methylated products 149 and 150 (0.175 g, 0.33 mmol, 14% yield). The mixture of 149 and 150 (0.175 g) was resubjected to the same set of conditions to yield an additional 77.0 mg (0.15 mmol, 43%) of 148, along with 70.0 mg (40%) of a mixture of 149 and 150. This represents a 69% yield of 148 after 1 recycle of 149 and 150. Chapter 3 – Towards C19-Diterpenoid Alkaloids: Approaches to the C4 Chiral Quaternary Center 89 Characterization data for 148: 1 H NMR (400 MHz, Chloroform-d): δ 6.80 (t, J = 5.7 Hz, 1H), 6.06 (ddd, J = 9.5, 6.2, 1.1 Hz, 1H), 5.88 – 5.77 (m, 2H), 5.67 (ddd, J = 9.4, 4.4, 1.9 Hz, 1H), 5.44 (d, J = 3.8 Hz, 1H), 5.39 (ddd, J = 5.9, 2.6, 1.4 Hz, 1H), 5.19 – 5.08 (m, 2H), 4.49 (t, J = 4.7, 1H), 4.24 (ddd, J = 4.1, 3.0, 0.7 Hz, 1H), 3.94 – 3.78 (m, 2H), 3.60 (dd, J = 8.2, 2.8 Hz, 1H), 3.42 (d, J = 9.4 Hz, 1H), 3.28 (s, 3H), 3.26 (s, 3H), 3.21 (d, J = 9.4 Hz, 1H), 3.06 – 3.01 (m, 1H), 2.87 (dd, J = 5.9, 4.6 Hz, 1H), 2.69 (t, J = 9.4 Hz, 1H), 2.59 (ddd, J = 13.9, 7.5, 5.9 Hz, 1H), 2.30 (ddt, J = 16.1, 9.8, 2.2 Hz, 1H), 2.20 (dddd, J = 16.1, 9.0, 2.7, 1.4 Hz, 1H), 1.79 – 1.69 (m, 1H), 1.66 – 1.62 (m, 1H), 1.22 – 1.13 (m, 1H), 1.08 (s, 9H), 1.01 (s, 9H). 13 C NMR (101 MHz, CDCl3): δ 174.9, 166.0, 135.6, 134.6, 133.4, 131.6, 128.2, 119.4, 115.8, 78.9, 76.8, 76.0, 66.5, 58.4, 57.2, 56.9, 47.0, 46.4, 46.2, 43.3, 41.8, 36.5, 28.9, 28.3, 21.5, 21.3, 21.1, 20.7. FTIR (NaCl, thin film): 3362, 2932, 2859, 1652, 1531, 1476, 1380, 1258, 1102, 991, 911, 825, 732 cm-1. HRMS: (ESI-TOF) calc’d for C32H50NO5Si [M + H]+ 556.3453, found 556.3454 [𝜶]𝟐𝟓 𝐃 = +103° (c = 0.50, CHCl3). TLC (30% EtOAc/70% Hexanes), Rf: 0.4 (blue in p-anisaldehyde) Characterization data for C1-O methyl ether 150: 1 H NMR (400 MHz, Chloroform-d) δ 6.30 (t, J = 5.8 Hz, 1H), 6.07 (ddd, J = 9.5, 6.1, 1.1 Hz, 1H), 5.92 – 5.73 (m, 2H), 5.65 (ddd, J = 9.5, 4.5, 2.1 Hz, 1H), 5.46 (d, J = 3.7 Hz, 1H), 5.39 (ddd, J = 5.9, 2.4, 1.4 Hz, 1H), 5.19 (dq, J = 17.2, 1.6 Hz, 1H), 5.14 (dq, J = 10.2, 1.3 Hz, Chapter 3 – Towards C19-Diterpenoid Alkaloids: Approaches to the C4 Chiral Quaternary Center 90 1H), 4.50 (t, J = 4.7 Hz, 1H), 4.27 – 4.19 (m, 1H), 3.95 – 3.78 (m, 2H), 3.73 (d, J = 11.0 Hz, 1H), 3.65 – 3.47 (m, 2H), 3.24 (s, 3H), 3.07 – 2.98 (m, 1H), 2.92 (t, J = 5.3 Hz, 1H), 2.70 (t, J = 9.3 Hz, 1H), 2.33 – 2.13 (m, 3H), 1.79 – 1.69 (m, 2H), 1.40 (dt, J = 13.0, 4.8 Hz, 1H), 1.08 (s, 9H), 1.00 (s, 9H). 13 C NMR (101 MHz, CDCl3) δ 176.1, 165.8, 135.7, 134.1, 133.8, 131.8, 128.3, 120.8, 116.6, 78.6, 66.3, 66.1, 57.8, 56.3, 47.9, 46.4, 46.2, 42.8, 41.8, 36.2, 28.9, 28.3, 21.7, 21.2, 20.7, 19.5. FTIR (NaCl, thin film): 3359, 2934, 2860, 1641, 1526, 1475, 1461, 1382, 1228, 1168, 1102, 1060, 1012, 994, 931, 852, 826, 810, 756, 724, 682, 665, 642 cm-1. HRMS: (ESI-TOF) calc’d for C31H48NO5Si [M + H]+ 542.3302, found 542.3326 [𝜶]𝟐𝟐 𝐃 = +96.9° (c = 1.19, CHCl3). Characterization data for C18-O methyl ether 149: 1 H NMR (400 MHz, Chloroform-d) δ 6.62 (t, J = 5.6 Hz, 1H), 6.11 (ddd, J = 5.9, 2.8, 1.7 Hz, 1H), 6.06 (ddd, J = 9.5, 6.2, 1.1 Hz, 1H), 5.82 (ddt, J = 17.2, 10.2, 5.6 Hz, 1H), 5.66 (ddd, J = 9.7, 4.5, 1.9 Hz, 1H), 5.50 (d, J = 3.8 Hz, 1H), 5.41 (ddd, J = 5.9, 2.7, 1.2 Hz, 1H), 5.16 (dq, J = 17.2, 1.7 Hz, 1H), 5.11 (dq, J = 10.3, 1.4 Hz, 1H), 4.46 (td, J = 4.2, 0.9 Hz, 1H), 4.28 – 4.15 (m, 1H), 4.04 (dd, J = 5.0, 2.5 Hz, 1H), 3.98 – 3.76 (m, 2H), 3.50 (d, J = 9.2 Hz, 1H), 3.29 (s, 4H), 3.15 – 2.98 (m, 1H), 2.98 – 2.81 (m, 1H), 2.65 (t, J = 9.6 Hz, 1H), 2.53 – 2.14 (m, 3H), 1.95 – 1.62 (m, 2H), 1.47 – 1.27 (m, 1H), 1.16 – 1.04 (m, 9H), 1.00 (s, 9H). 13 C NMR (101 MHz, CDCl3) δ 175.0, 165.0, 135.7, 135.5, 134.5, 133.4, 128.5, 121.0, 116.0, 76.9, 75.3, 67.6, 66.3, 58.8, 56.4, 47.1, 46.5, 46.3, 42.5, 41.8, 36.2, 28.9, 28.9, 28.3, 24.6, 21.3, 20.7, 18.6. Chapter 3 – Towards C19-Diterpenoid Alkaloids: Approaches to the C4 Chiral Quaternary Center 91 FTIR (NaCl, thin film): 3362, 2934, 2897, 2859, 1651, 1520, 1476, 1461, 1382, 1258, 1227, 1168, 1102, 1058, 1012, 993, 934, 888, 853, 826, 811, 755, 736, 682, 642 cm-1. HRMS: (ESI-TOF) calc’d for C31H48NO5Si [M + H]+ 542.3302, found 542.3305 [𝜶]𝟐𝟐 𝐃 = +123° (c = 1.19, CHCl3). Preparation of amine 123:7 OMe MeO NHEt O OMe MeO NHEt 1. [Ir(COE)2Cl]2 (0.5 mol %) Et2SiH2 (4 equiv.) PhH, 21 °C; then LiBHEt3, 6 °C O Si O t-Bu t-Bu 122 87% yield O Si O t-Bu t-Bu 123 Following a modified procedure from Brookhart and coworkers, a 2-dram vial was charged with amide 122 (40.2 mg, 74 µmol, 1.0 equiv.) and brought into an N2-filled glovebox. Benzene (1.0 mL) was added. Also in the glovebox, a stock solution of catalyst was prepared:[Ir(COE)2Cl]2 (1.0 mg, 1.1 µmol, 0.015 equiv.) was suspended in benzene (0.9 mL) and Et2SiH2 (6.0 mg) was added. After stirring for 5 minutes, the portion of the catalyst solution (0.3 mL, 0.5 mol % [Ir(COE)2)Cl]2) was added to the reaction vial. An additional 24 mg of and Et2SiH2 (300 µmol, 4.0 equiv.) in benzene (0.1 mL) was added, and the reaction was stirred brought out of the glovebox and stirred for 12 hours at 21 °C. The reaction was cooled to 6 °C before a solution of LiBHEt3 (1 M in THF, 0.08 mL, 80 µmol, 1.10 equiv.) was added dropwise. The solution was stirred at 6 °C for 15 minutes before the reaction was quenched at 6 °C with 0.5 M NaOH (aq., 1 mL). The mixture was warmed to room temperature and stirred for 30 minutes. The mixture was extracted with EtOAc (6 x 5 mL) and the combined organic phases were dried (Na2SO4), filtered and concentrated in vacuo to give the crude product as a Chapter 3 – Towards C19-Diterpenoid Alkaloids: Approaches to the C4 Chiral Quaternary Center 92 yellow foam. The product was purified by silica gel chromatography (SiO2, 1-2-3% (7N NH3 in MeOH) in CH2Cl2) to give amine 123 (34.2 mg, 64 µmol, 87% yield) as a white foam. 1 H NMR (400 MHz, Chloroform-d) δ 6.06 (ddd, J = 9.5, 6.2, 1.1 Hz, 1H), 5.88 (dt, J = 5.9, 2.2 Hz, 1H), 5.65 (ddd, J = 9.3, 4.4, 1.9 Hz, 1H), 5.41 (dd, J = 5.5, 2.7 Hz, 2H), 4.46 (t, J = 4.7 Hz, 1H), 4.32 – 4.06 (m, 1H), 3.56 (dd, J = 7.7, 2.8 Hz, 1H), 3.25 (s, 3H), 3.24 (s, 3H), 3.19 (d, J = 1.5 Hz, 2H), 3.05 – 2.99 (m, 1H), 2.93 – 2.85 (m, 1H), 2.65 – 2.53 (m, 2H), 2.46 (s, 2H), 2.40 (dd, J = 10.5, 7.7 Hz, 1H), 2.36 – 2.26 (m, 2H), 1.74 – 1.61 (m, 1H), 1.61 – 1.44 (m, 2H), 1.44 – 1.31 (m, 1H), 1.07 (d, J = 7.7 Hz, 12H), 1.01 (s, 9H). 13 C NMR (101 MHz, CDCl3) δ 166.80, 136.06, 134.18, 131.52, 128.20, 119.72, 79.92, 76.52, 66.69, 58.93, 57.31, 56.98, 56.89, 46.43, 46.35, 45.06, 44.81, 39.48, 35.51, 29.06, 28.46, 24.58, 21.94, 21.40, 20.83, 15.57. FTIR (NaCl, thin film): 2936, 2860, 2338, 1606, 1476, 1359, 1107, 993, 860, 825, 816, 782, 744, 730, 668, 644 cm-1. HRMS: (ESI-TOF) calc’d for C31H52NO4Si [M + H]+ 530.3660, found 530.3652 [𝜶]𝟐𝟐 𝐃 = +112° (c = 0.95, CHCl3). Preparation of primary amine 116:7,11 OMe MeO NH(allyl) O MeO OMe N 1. [Ir(COE)2Cl]2 (0.5 mol %) Et2SiH2 (4 equiv.) H OMe MeO NH2 2. Pd(PPh3)4 (10 mol %) 1,3-dimethylbarbituric acid PhH, 21 °C; then LiBHEt3, 6 °C CH2Cl2, 23 °C O Si O t-Bu t-Bu O Si O t-Bu t-Bu 121 145 95% yield (2 steps) O Si O t-Bu t-Bu 116 Following a modified procedure from Brookhart and coworkers, a 50 mL round-bottom flask was charged with N-allylamide 121 (0.839 g, 1.51 mmol, under N2 and before benzene Chapter 3 – Towards C19-Diterpenoid Alkaloids: Approaches to the C4 Chiral Quaternary Center 93 (22 mL) was added. In a nitrogen-filled glovebox, [Ir(COE)2Cl]2 (6.8 mg, 7.6 µmol, 0.005 equiv) was suspended in benzene (5 ml) and Et2SiH2 (0.533 g, 6.04 mmol, 4.0 equiv) was added dropwise. After stirring for 5 minutes, the catalyst solution was drawn up into a syringe, taken out of the glovebox and added to the reaction dropwise at 21 °C. The reaction was stirred for 16 hours at 21 °C. The reaction was cooled to 6 °C before a solution of LiBHEt3 (1M in THF, 1.66 mL, 1.66 mmol, 1.10 equiv) was added dropwise over 10 minutes. The solution was stirred at 6 °C for 30 minutes before the reaction was quenched at 6 °C with 0.5 M NaOH (aq., 15 mL). The mixture was warmed to room temperature and stirred for 30 minutes. The mixture was extracted with EtOAc (6 x 30 mL) and the combined organic phases were dried (Na2SO4), filtered and concentrated in vacuo to give the crude N-allylamine 145 as a yellow foam. 145 was carried on to the following step without purification. A 50 mL round-bottom flask charged with crude S17 and 1,3-dimethylbarbituric acid (1.41 g, 9.06 mmol, 6.0 equiv), put under N2, and CH2Cl2 (30 mL) was added. In a nitrogenfilled glovebox, Pd(PPh3)4 (0.174 g, 0.15 mmol, 0.10 equiv) was taken up in CH2Cl2 (3 mL), and this solution was added to the reaction mixture. The reaction was stirred at 23 °C for 1h before quenching with saturated aq. NaHCO3 (25 mL). The mixture was extracted with 3:1 CHCl3:iPrOH (5 x 30 mL). The combined organic phases were dried (Na2SO4), filtered and concentrated in vacuo to give the crude product as a red foam. The product was purified by column chromatography (SiO2, 1-2-3-4-5-6-7% 7N NH3 in MeOH/CH2Cl2) to give primary amine 116 (0.720 g, 1.43 mmol, 95% yield, 2 steps) as a white foam. 1 H NMR (400 MHz, Chloroform-d): δ 6.06 (ddd, J = 9.5, 6.1, 1.1 Hz, 1H), 5.87 (dt, J = 5.9, 2.3 Hz, 1H), 5.65 (ddd, J = 9.4, 4.4, 1.9 Hz, 1H), 5.44 – 5.39 (m, 2H), 4.46 (t, J = 4.7 Hz, 1H), 4.23 (ddd, J = 4.3, 3.0, 0.7 Hz, 1H), 3.56 (dd, J = 7.7, 2.8 Hz, 1H), 3.26 – 3.24 (m, 6H), 3.22 Chapter 3 – Towards C19-Diterpenoid Alkaloids: Approaches to the C4 Chiral Quaternary Center 94 – 3.19 (m, 1H), 3.18 (d, J = 9.2 Hz, 1H), 3.05 – 3.00 (m, 1H), 2.88 (dd, J = 5.9, 4.6 Hz, 1H), 2.63 (d, J = 13.1 Hz, 1H), 2.58 (d, J = 13.1 Hz, 1H), 2.39 (dd, J = 10.0, 8.4 Hz, 1H), 2.31 – 2.26 (m, 2H), 1.68 (dddd, J = 13.3, 8.0, 5.4, 2.7 Hz, 1H), 1.61 – 1.37 (m, 4H), 1.34 – 1.21 (m, 1H), 1.08 (s, 9H), 1.00 (s, 9H). 13 C NMR (101 MHz, CDCl3): δ 166.4, 135.8, 134.1, 131.1, 128.1, 119.7, 79.7, 77.0, 76.1, 66.5, 58.8, 57.2, 56.9, 49.5, 46.3, 46.2, 44.0, 39.7, 35.2, 28.9, 28.3, 23.9, 21.8, 21.2, 20.7. A 13C signal is observed to be overlapping with residual CDCl3 at 77.00, the peak can be seen in the HSQC spectrum, provided below. FTIR (NaCl, thin film): 3386, 3050, 2933, 2859, 2362, 1476, 1463, 1106, 992 cm-1. HRMS: (ESI-TOF) calc’d for C29H48NO4Si [M + H]+ 502.3347, found 502.3344. [𝜶]𝟐𝟓 𝐃 = +105° (c = 0.67, CHCl3). TLC (10%MeOH/90%CH2Cl2), Rf: 0.15 (UV, blue in p-anisaldehyde) Preparation of diol 126: O O O LAH MeO2C MeO2C O H 93 THF, reflux O HO O HO H O 70% yield O 126 A flame-dried 2 L round-bottom flask was charged with diester 93 (24.0 g, 66.9 mmol, 1.0 equiv.). THF (670 mL) added, and the solution was cooled to 0 °C. LAH (12.7 g, 335 mmol, 5.0 equiv.) was added slowly in portions over 30 minutes. The suspension was warmed to room temperature and then heated to reflux for 6 hours. The reaction was cooled to 0 °C and quenched using the Fieser procedure: the mixture was diluted with ether (500 mL) before water (13 mL) was added slowly. 15 % aqueous NaOH (13 mL) was added, followed by water (50 Chapter 3 – Towards C19-Diterpenoid Alkaloids: Approaches to the C4 Chiral Quaternary Center 95 mL). The mixture was warmed to room temperatrure and stirred vigorously for 15 minutes. MgSO4 was added and the mixture was filtered to remove the solids. The filtrate was concentrated in vacuo. The product was purified by silica gel chromatography (50-70% acetone in hexanes) to give 126 (14.5 g, 47.6 mmol, 70% yield) as a colorless oil. 1 H NMR (400 MHz, Chloroform-d) δ 4.86 (t, J = 4.5 Hz, 1H), 4.03 – 3.95 (m, 2H), 3.95 – 3.81 (m, 6H), 3.74 – 3.64 (m, 2H), 3.65 – 3.50 (m, 2H), 2.60 (q, J = 7.7, 5.5 Hz, 2H), 2.14 (dddd, J = 19.3, 10.6, 6.3, 4.0 Hz, 1H), 1.96 – 1.79 (m, 2H), 1.76 – 1.66 (m, 5H), 1.60 – 1.45 (m, 3H). 13 C NMR (101 MHz, CDCl3) δ 117.35, 104.95, 67.20, 65.11, 64.49, 64.34, 42.10, 39.59, 36.75, 35.82, 27.33, 24.46, 23.90. FTIR (NaCl, thin film): 3450, 2949, 2882, 2340, 1407, 1332, 1115, 1030, 947 cm-1. HRMS: (ESI-TOF) calc’d for C15H26O6 [M + H]+ 325.1622, found 325.1510. [𝜶]𝟐𝟐 𝐃 = –1.3° (c = 1.0, CHCl3). Preparation of 127 and 128: O O HCl HO O HO H O 126 COMe2 (2.0 equiv.) THF/H2O, 70°C (87% combined yield) HO HO HO O H + O O H 127 128 68% yield 19% yield A 250 mL round-bottom flask was charged with diol 126 (3.00 g, 9.9 mmol, 1.0 equiv.). THF (40 mL), 2 M HCl (aq., 10 mL) and acetone (1.5 mL) were added. The flask was fitted with a reflux condenser and heated to reflux for 2 days. The reaction was cooled to room temperature before quenching with saturated aqueous NaHCO3 (50 mL). The mixture was extracted with EtOAc (4 x 60 mL). The combined organic extracts were dried (Na2SO4), Chapter 3 – Towards C19-Diterpenoid Alkaloids: Approaches to the C4 Chiral Quaternary Center 96 filtered and concentrated in vacuo. The product was purified by silica gel chromatography (2030-40-50-60-70% acetone in hexanes) to give enone 127 (1.30 g, 7.8 mmol, 79% yield) and oxy-Michael product 128 (0.381 g, 1.9 mmol, 19% yield). Characterization data for enone 127: 1 H NMR (500 MHz, Chloroform-d) δ 6.82 (q, J = 3.5 Hz, 1H), 3.96 (d, J = 10.4 Hz, 1H), 3.82 (d, J = 10.8 Hz, 1H), 3.72 – 3.67 (m, 1H), 3.54 (d, J = 10.8 Hz, 1H), 2.63 – 2.56 (m, 1H), 2.41 – 2.32 (m, 2H), 2.29 – 2.17 (m, 6H), 1.58 – 1.49 (m, 1H), 1.31 (dd, J = 12.4, 6.3 Hz, 1H). 13 C NMR (101 MHz, CDCl3) δ 205.86, 138.24, 132.79, 71.63, 63.51, 43.61, 39.43, 38.33, 27.04, 23.11, 22.61. FTIR (NaCl, thin film): 3415, 2940, 2883, 1707, 1648, 1423, 1216, 1178, 1098, 1032, 924, 865, 789, 754, 666 cm-1. [𝜶]𝟐𝟏 𝐃 = –93° (c = 1.0, CHCl3). TLC (8%MeOH/92%CH2Cl2), Rf: 0.3 (UV, blue in p-anisaldehyde) Characterization data for oxy-Michael product 128: 1 H NMR (500 MHz, Chloroform-d) δ 4.16 (td, J = 2.7, 1.4 Hz, 1H), 3.77 (dd, J = 9.4, 3.4 Hz, 1H), 3.63 (dd, J = 9.3, 1.6 Hz, 1H), 3.48 (d, J = 10.9 Hz, 1H), 3.43 (d, J = 10.9 Hz, 1H), 2.61 (dddd, J = 11.1, 9.1, 7.3, 1.6 Hz, 1H), 2.38 – 2.27 (m, 2H), 2.24 (d, J = 11.6 Hz, 1H), 2.15 – 2.02 (m, 2H), 1.97 (dtd, J = 13.6, 9.6, 7.6 Hz, 1H), 1.78 (ddd, J = 12.9, 10.9, 2.1 Hz, 1H), 1.72 – 1.67 (m, 1H), 1.55 – 1.48 (m, 1H). 13 C NMR (101 MHz, CDCl3) δ 211.14, 67.96, 66.34, 65.66, 50.37, 38.85, 37.69, 36.18, 27.66, 25.66, 21.32. Chapter 3 – Towards C19-Diterpenoid Alkaloids: Approaches to the C4 Chiral Quaternary Center 97 FTIR (NaCl, thin film): 3445, 2940, 2868, 2352, 1735, 1456, 1407, 1268, 1168, 1140, 1095, 1021, 922, 847, 822 cm-1. [𝜶]𝟐𝟏 𝐃 = +59° (c = 1.0, CHCl3). TLC (30% acetone/70% hexanes), Rf: 0.15 (blue/green in p-anisaldehyde) Preparation of C18 Methyl Ether 129: HO O H 128 Me3OBF4 MeO O O Proton Sponge 4Å MS, CH2Cl2 O H 129 72% yield An oven-dried 20 mL scintillation vial was charged with 128 (30.5 mg, 0.16 mmol, 1.0 equiv.) and brought into a N2 filled glovebox. CH2Cl2 (6.2 mL) was added, followed by Me3OBF4 (69.0 mg, 0.47 mmol, 3.0 equiv.), Proton Sponge (100 mg, 0.47 mmol, 3.0 equiv.), and 4 Å molecular sieve powder (61 mg). The vial was capped, removed from the glovebox and stirred for 3 hours at 20 °C. The reaction was quenched with saturated aqueous NaHCO3 (5 mL) and extracted with EtOAc (4 x 5 mL). The combined organic extracts were dried (Na2SO4), filtered and concentrated in vacuo. The crude product was purified by silica gel chromatography (10-30-40-60% EtOAc in hexanes) to afford the product 129 as a colorless oil (23.6 mg, 0.11 mmol, 72% yield). 1 H NMR (500 MHz, Chloroform-d) δ 4.17 (td, J = 2.7, 1.4 Hz, 1H), 3.80 (dd, J = 9.4, 3.3 Hz, 1H), 3.65 (dd, J = 9.5, 1.5 Hz, 1H), 3.34 (s, 3H), 3.21 – 3.13 (m, 2H), 2.65 – 2.53 (m, 1H), 2.42 – 2.29 (m, 2H), 2.24 (d, J = 11.6 Hz, 1H), 2.16 – 2.05 (m, 2H), 2.05 – 1.96 (m, 1H), 1.82 (ddd, J = 12.9, 10.9, 2.1 Hz, 1H), 1.74 – 1.65 (m, 1H), 1.56 – 1.42 (m, 1H). Chapter 3 – Towards C19-Diterpenoid Alkaloids: Approaches to the C4 Chiral Quaternary Center 13 98 C NMR (101 MHz, CDCl3) δ 220.43, 76.24, 67.99, 66.57, 59.67, 50.48, 38.89, 38.36, 35.57, 28.32, 25.76, 21.53. FTIR (NaCl, thin film): 2940, 2869, 1737, 16541, 1457, 1389, 1268, 1195, 1167, 1106, 1044, 902, 848 cm-1. [𝜶]𝟐𝟏 𝐃 = +83° (c = 1.0, CHCl3). TLC (30% acetone/70% hexanes), Rf: 0.5 (green in p-anisaldehyde) Preparation of C18 silyl ether 131: HO HO 3.5 equiv. TBSCl HO O H 127 1.0 equiv. imid. 3.5 equiv. Et3N THF, 20 °C TBSO O H 131 major 48% yield An oven-dried 20 mL scintillation vial was charged with 127 (50 mg, 0.25 mmol, 1.0 equiv.) and THF (3.5 mL). Imidazole (17.0 mg, 0.25 mmol, 1.0 equiv.), triethylamine (0.12 mL, 0.89 mmol, 3.5 equiv.), and TBSCl (134 mg, 0.89 mmol, 3.5 equiv.) were added. The reaction was stirred at 20 °C for 4 hours. The reaction was quenched with saturated aqueous NaHCO3 (5 mL) and extracted with EtOAc (4 x 5 mL). The combined organic extracts were dried (Na2SO4), filtered and concentrated in vacuo. The crude product was purified by silica gel chromatography (10-15-20% EtOAc in hexanes) to afford the product 131 as a colorless oil (37.3 mg, 0.12 mmol, 48% yield). 1 H NMR (600 MHz, Chloroform-d) δ 6.80 (q, J = 3.5 Hz, 1H), 3.89 (d, J = 9.5 Hz, 1H), 3.74 (dd, J = 11.1, 3.2 Hz, 1H), 3.56 (dd, J = 9.6, 1.4 Hz, 1H), 3.42 (ddd, J = 10.8, 8.7, 1.4 Hz, 1H), 2.65 (dd, J = 8.5, 3.2 Hz, 1H), 2.48 (tt, J = 7.1, 3.7 Hz, 1H), 2.31 (qd, J = 14.1, 13.4, 6.9 Hz, Chapter 3 – Towards C19-Diterpenoid Alkaloids: Approaches to the C4 Chiral Quaternary Center 99 3H), 2.23 – 2.12 (m, 2H), 2.06 (dt, J = 12.1, 7.6 Hz, 1H), 1.47 (qd, J = 12.4, 7.8 Hz, 1H), 1.20 (ddd, J = 13.5, 11.1, 6.3 Hz, 1H), 0.93 (d, J = 1.0 Hz, 9H), 0.11 (d, J = 2.3 Hz, 5H). 13 C NMR (101 MHz, CDCl3) δ 205.77, 138.04, 133.18, 72.30, 63.43, 43.33, 39.30, 38.30, 27.09, 26.00, 23.27, 22.10, 18.32, -5.45, -5.53. FTIR (NaCl, thin film): 3449, 2951, 2892, 2855, 2340, 1716, 1651, 1470, 1424, 1255, 1216, 1176, 1084, 1049, 836, 777, 680, 667 cm-1. [𝜶]𝟐𝟏 𝐃 = +59° (c = 1.0, CHCl3). TLC (40%COMe2/60%hexanes), Rf: 0.6 (UV, blue in p-anisaldehyde) Preparation of allylic alcohol 133: HO NaBH4 CeCl3 TBSO O H 131 MeOH, -78 °C 39% yield [3:1 crude d.r.] HO TBSO OH H 133 A 50 mL round-bottom flask was charged with 131 (0.217 mg, 0.70 mmol, 1.0 equiv.), MeOH (14 mL) and CeCl3 • 7H2O (0.391 g, 1.05 mmol, 1.5 equiv.). The mixture was cooled to –78 °C, and NaBH4 (32 mg, 0.84 mmol, 1.2 equiv.) was added in one portion. The reaction was stirred for 45 minutes. The reaction was quenched with saturated aqueous 0.1 M NaOH (10 mL), warmed to room temperature and stirred for 20 minutes. The mixture was extracted with EtOAc (4 x 5 mL). The combined organic extracts were dried (Na2SO4), filtered and concentrated in vacuo. The crude product was purified by silica gel chromatography (30-4050% EtOAc in hexanes) to afford the product 133 as a colorless oil (84.5 mg, 0.27 mmol, 39% yield). 1 H NMR (400 MHz, Chloroform-d) δ 5.84 (t, J = 3.4 Hz, 1H), 4.48 – 4.40 (m, 1H), 3.85 (d, J = 9.5 Hz, 1H), 3.68 (d, J = 10.9 Hz, 1H), 3.45 (ddd, J = 18.9, 10.2, 1.7 Hz, 2H), 2.88 (s, 1H), Chapter 3 – Towards C19-Diterpenoid Alkaloids: Approaches to the C4 Chiral Quaternary Center 100 2.45 – 2.26 (m, 1H), 2.23 – 2.10 (m, 3H), 2.09 – 1.98 (m, 1H), 1.87 – 1.75 (m, 1H), 1.52 – 1.37 (m, 1H), 1.32 – 1.10 (m, 3H), 0.91 (s, 9H), 0.09 (s, 3H), 0.09 (s, 3H). 13 C NMR (101 MHz, CDCl3) δ 144.24, 122.53, 74.08, 73.39, 64.08, 43.11, 39.14, 34.23, 27.22, 26.01, 24.01, 22.74, 18.31, -5.48, -5.55. FTIR (NaCl, thin film): 3347, 2952, 2928, 2856, 1461, 1252, 1085, 1006, 813, 776 cm-1. HRMS: (ESI-TOF) calc’d for C17H33O3Si [M + H]+ 313.2193, found 313.2192. [𝜶]𝟐𝟏 𝐃 = +37° (c = 0.75, CHCl3). TLC (50% EtOAc/50%hexanes), Rf: 0.5 (purple in p-anisaldehyde) Preparation of epoxide 134: HO V(O)(acac)2 TBHP TBSO OH H 133 PhH, rt 99% yield HO O TBSO OH H 134 Alcohol 133 (4.2 mg, 13 mmol, 1.0 equiv.) was added to an oven-dried 1-dram vial. Benzene (1 mL) was added, followed by V(O)(acac)2 (0.7 mg, 2.6 mmol, 20 mol %) and TBHP (5 M in decane, 5 mL, 26 mmol, 2.0 equiv.). The reaction was stirred at 20 °C for 1.5 hours. The reaction was quenched with saturated aqueous Na2SO3 (1 mL) and the mixture was extracted with EtOAc (4 x 2 mL). The combined organic extracts were dried (Na2SO4), filtered and concentrated in vacuo. The crude product was purified by silica gel chromatography (50% EtOAc in hexanes) to afford the product 134 as a colorless oil (4.5 mg, 13 µmol, 99% yield). 1 H NMR (400 MHz, Chloroform-d) δ 4.26 (dd, J = 6.0, 3.1 Hz, 1H), 4.17 (d, J = 4.2 Hz, 1H), 3.71 – 3.60 (m, 2H), 3.61 – 3.50 (m, 2H), 3.31 (br, 1H), 2.58 (t, J = 8.7 Hz, 1H), 2.33 – 2.18 (m, 1H), 1.99 – 1.75 (m, 2H), 1.70 – 1.60 (m, 3H), 1.56 – 1.41 (m, 2H), 0.87 (s, 9H), 0.06 – 0.02 (m, 6H). Chapter 3 – Towards C19-Diterpenoid Alkaloids: Approaches to the C4 Chiral Quaternary Center 13 101 C NMR (101 MHz, CDCl3) δ 94.54, 79.43, 72.14, 70.62, 65.94, 50.62, 46.46, 35.88, 29.89, 27.79, 25.99, 22.57, 18.38, -5.41, -5.44. FTIR (NaCl, thin film): 3354, 2950, 2929, 2858, 1462, 1360, 1255, 1223, 1098, 1070, 1002, 958, 838, 776, 668 cm-1. [𝜶]𝟐𝟏 𝐃 = +37° (c = 0.75, CHCl3). TLC (30% acetone in hexanes), Rf: 0.61 (green in p-anisaldehyde) Preparation of S1: OH OH (S)–100 NaH, MOMCl OMOM OMOM THF, 0 to 21 °C 93% yield S1 A flame-dried 1L round-bottom flask was charged with (S)–BINOL ((S)–100) (14.3 g, 50 mmol, 1.0 equiv.) and THF (500 mL). The solution was cooled to 0 °C and NaH (60 wt.% dispersion in mineral oil, 5.0 g, 125 mmol, 2.5 equiv.) was added slowly in portions. After complete addition of NaH, the mixture was stirred at 0 °C for 15 minutes. MOMCl (9.5 mL, 125 mmol, 2.5 equiv.) was added dropwise and the mixture was warmed to 21 °C. The mixture was stirred for 4.5 hours before quenching with water (200 mL) and saturated aqueous NaHCO3 (100 mL). The mixture was extracted with EtOAc (3 x 400 mL). The combined organic extracts were dried (Na2SO4), filtered, and concentrated in vacuo. The crude product was recrystallized from hot MeOH (~400 mL) to give S1 (17.4 g, 46.5 mmol, 93% yield) as white, needle-shaped crystals. 1H NMR of S1 matches previously reported spectroscopic data.12 Chapter 3 – Towards C19-Diterpenoid Alkaloids: Approaches to the C4 Chiral Quaternary Center 102 Preparation of S2: OMOM OMOM S1 1. n-BuLi; B(OMe)3 THF -78 to 21 °C 2. H2O2 PhH, 80 °C OMe 3. K2CO3, MeI COMe2, 56 °C 4. HCl MeOH, 50 °C OH 94% yield (4 steps) OH OMe S2 Directed lithiation of S1 was employed to introduce oxidation at C3/3’.13 A flame-dried 250 mL round-bottom flask was charged with S1 (3.74 g, 10 mmol, 1.0 equiv.) and put under N2. THF (30 mL) was added and the solution was cooled to –78 °C. A solution of n-BuLi (2.5M in hexanes, 9.6 mL, 24 mmol, 2.4 equiv.) was added dropwise. The mixture was warmed to 0 °C and stirred for 1 hour. The solution was then cooled to –78 °C before B(OMe)3 (3.3 mL, 30 mmol, 3.0 equiv.) was added dropwise. The reaction was warmed to 21 °C and stirred for 24 hours. The reaction was concentrated in vacuo to give a white solid. Benzene (35 mL) was added, and the solution was cooled to 0 °C. H2O2 (50 wt.% in water, 2.9 mL) was added dropwise and the reaction was then heated to reflux. After stirring for 2 hours, the reaction was cooled to 0 °C and quenched with saturated aqueous Na2S2O3 (100 mL). The reaction was extracted with Et2O (4 x 100 mL). The combined organic extracts were dried (Na2SO4), filtered, and concentrated in vacuo. The crude product was carried on to the next step. The crude product of the boronic ester oxidation was dissolved in acetone (135 mL) in a 250 mL round-bottom flask. K2CO3 (4.15 g, 30 mmol, 3.0 equiv.) was added, followed by MeI (3.1 mL, 50 mmol, 5.0 equiv.). The reaction flask was fitted with a reflux condenser and the reaction was heated to reflux for 14 hours. The reaction was cooled to room temperature and quenched with water (100 mL). The reaction was extracted with Et2O (4 x 100 mL). The combined organic extracts were dried (Na2SO4), filtered, and concentrated in vacuo. The crude product was carried on to the next step. Chapter 3 – Towards C19-Diterpenoid Alkaloids: Approaches to the C4 Chiral Quaternary Center 103 The crude methyl ether product was dissolved in MeOH (20 mL) in a 100 mL roundbottom flask. HCl (4M, aq.) (10 mL, 40 mmol, 4 equiv.) was added and the reaction flask was fitted with a reflux condenser. The mixture was heated to 50 °C for 48 hours. The reaction was then cooled to room temperature and quenched with saturated aqueous NaHCO3 (100 mL). The mixture was extracted with EtOAc (4 x 100 mL). The combined organic extracts were dried (Na2SO4), filtered, and concentrated in vacuo. The crude product was purified by silica gel chromatography (50% EtOAc in hexanes) to afford the product S2 (2.821 g, 8.1 mmol, 81% yield) as a white solid. The 1H NMR of S2 matches previously reported spectroscopic data.14 1 H NMR (600 MHz, Chloroform-d) δ 7.78 (d, J = 8.2 Hz, 2H), 7.32 (ddd, J = 8.1, 6.3, 1.8 Hz, 2H), 7.30 (s, 2H), 7.18 – 7.12 (m, 4H), 4.10 (s, 6H). Preparation of aryl triflate S3: OMe OH OH OMe S2 OMe Tf2O, Et3N OTf OTf CH2Cl2, –78 to 21 °C 96% yield OMe S3 A flame-dried 250 mL round-bottom flask was charged with S2 (1.88 g, 5.42 mmol, 1.0 equiv.) and put under N2 atmosphere. CH2Cl2 (108 mL) and Et3N (2.3 mL, 16.3 mmol, 3.0 equiv.) were added, and the mixture was cooled to –78 °C. Tf2O (2.0 mL, 11.9 mmol, 2.2 equiv.) was added dropwise, and the reaction was warmed to 21 °C. After stirring for 4 hours, the reaction mixture was poured into a stirring solution of HCl (1M, aq.) (100 mL). Et2O (100 mL) was added, and the mixture was transferred to a separatory funnel. The phases were separated, and the organic phase was extracted with Et2O (2 x 100 mL). The combined organic Chapter 3 – Towards C19-Diterpenoid Alkaloids: Approaches to the C4 Chiral Quaternary Center 104 phases were washed with saturated aqueous NaHCO3 (2 x 100 mL), then with brine (100 mL). The organic extracts were dried (MgSO4), filtered, and concentrated in vacuo to yield the crude product as an orange foam. Purification by silica gel chromatography (10-20% EtOAc in hexanes) afforded S3 (3.17 g, 5.20 mmol, 96% yield) as a white solid. The 1H NMR of S3 matches previously reported spectroscopic data.14 1 H NMR (500 MHz, Chloroform-d) δ 7.87 (d, J = 8.4 Hz, 2H), 7.52 (ddd, J = 8.2, 6.8, 1.2 Hz, 2H), 7.49 (s, 2H), 7.24 (ddd, J = 8.2, 6.8, 1.2 Hz, 2H), 7.13 (d, J = 8.5 Hz, 2H), 4.11 (s, 6H). TLC (25%EtOAc/ 75% Hexanes), Rf: 0.3 (UV) Preparation of S4:14,15 OMe OTf OTf OMe S3 OMe MeMgI NiCl2(dppp) (5 mol %) Me Me THF/Et2O, 0 to 40 °C 79% yield OMe S4 A solution of MeMgI was prepared as follows: A flame-dried 250 mL 3-neck roundbottom flask was charged with oven-dried Mg (4.56 g, 188 mmol, 2.5 equiv.), fitted with a reflux condenser, and put under argon atmosphere. Et2O (5 mL) was added, followed by neat DIBAL (0.02 mL), and the mixture was stirred vigorously for 15 minutes. The flask was heated to 30 °C in an oil bath before a solution of MeI (4.7 mL, 75 mmol, 1.0 equiv.) in Et2O (75 mL) was added slowly until the reaction began to reflux. At this point, the oil bath was removed, and the solution of MeI was added at a rate just fast enough to maintain reflux. After addition of MeI was complete, the mixture was heated to reflux in an oil bath for 1 hour. The mixture was cooled to room temperature. Titration with salicylaldehyde phenylhydrazone indicated the Chapter 3 – Towards C19-Diterpenoid Alkaloids: Approaches to the C4 Chiral Quaternary Center 105 concentration of the MeMgI solution was 0.8 M. The solution was transferred to a flame-dried Schlenk flask under argon for storage. For the Kumada coupling, a flame-dried 50 mL round-bottom flask was charged with aryl triflate S3 (1.10 g, 1.8 mmol, 1.0 equiv.) and NiCl2•dppp (49.0 mg, 0.09 mmol, 5 mol %). The flask was fitted with a reflux condenser and put under argon atmosphere. Et2O (1.8 mL) was added, followed by a solution of MeMgI (0.8 M in Et2O, 11.3 mL, 9.0 mmol, 5.0 equiv.). The reaction was heated to reflux for 11 hours. The reaction was quenched with 1M HCl (aq.) (15 mL). The phases were separated, and the aqueous phase was extracted with Et2O (3 x 25 mL). The combined organic extracts were washed with saturated aqueous NaHCO3 (2 x 50 mL) then with brine (50 mL), dried (MgSO4), filtered, and concentrated in vacuo. Purification by silica gel chromatography (5-10-20% EtOAc in hexanes) afforded S4 (0.492 g, 1.44 mmol, 80% yield) as a white powder. The 1H NMR of S4 matches previously reported spectroscopic data.14 1 H NMR (500 MHz, Chloroform-d) δ 7.79 (d, J = 8.3 Hz, 2H), 7.35 (ddd, J = 8.1, 6.8, 1.3 Hz, 2H), 7.23 (s, 2H), 7.06 (ddd, J = 8.1, 6.7, 1.2 Hz, 2H), 6.96 (d, J = 8.5 Hz, 2H), 4.04 (s, 6H), 1.93 (s, 5H). TLC (25%EtOAc/ 75% Hexanes), Rf: 0.5 (UV) Preparation of bis-aryltriflate S5: OMe OTf 1. Me Me OMe S4 2. BBr3 CH2Cl2, 0 °C Me Me Tf2O, NEt3 CH2Cl2, –78 to 20 °C 85% yield (2 steps) OTf S5 Chapter 3 – Towards C19-Diterpenoid Alkaloids: Approaches to the C4 Chiral Quaternary Center 106 A flame-dried 10 mL round-bottom flask was charged with bis-methylether S4 (0.200 g, 0.58 mmol, 1.0 equiv.) and put under a N2-atmosphere. CH2Cl2 (2.4 mL) was added, and the solution was cooled to 0 °C. BBr3 (0.13 mL, 1.4 mmol, 2.4 equiv.) was added dropwise, and the reaction was stirred at 0 °C for 4.5 hours. Water (5 mL) was added, and the mixture was filtered through a pad of celite. The phases were separated, and the aqueous phase was extracted with CH2Cl2 (3 x 10 mL). The combined organic phases were dried (Na2SO4), filtered, and concentrated in vacuo. The crude product was carried on to the next reaction The crude product from the demethylation reaction was dissolved in CH2Cl2 (in a flame-dried 50 mL round-bottom flask. Triethylamine (0.24 mL, 1.7 mmol, 3.0 equiv.) was added, and the solution was cooled to –78 °C. Triflic anhydride (0.21 mL, 1.3 mmol, 2.2 equiv.) was added dropwise and the mixture was stirred at –78 °C for 10 minutes before warming to room temperature. After stirred for 3 hours, the reaction was quenched with 1M HCl (aq.) (10 mL). The phases were separated, and the aqueous phase was extracted with Et2O (4 x 20 mL). The combined organic phases were washed with saturated aqueous NaHCO3 (2 x 50 mL) then with brine (50 mL), dried (MgSO4), filtered, and concentrated in vacuo. Purification by silica gel chromatography (5:1 hexanes:EtOAc) afforded S5 (0.287 g, 0.50 mmol, 85% yield over 2 steps) as a white foam. The 1H NMR of S5 matches previously reported spectroscopic data.14 1 H NMR (500 MHz, Chloroform-d) δ 7.99 – 7.90 (m, 4H), 7.53 (ddd, J = 8.1, 6.8, 1.2 Hz, 2H), 7.33 (ddd, J = 8.3, 6.8, 1.3 Hz, 2H), 6.98 (dd, J = 8.5, 1.0 Hz, 2H), 2.05 (s, 6H). TLC (25%EtOAc/ 75% Hexanes), Rf: 0.6 (UV) Chapter 3 – Towards C19-Diterpenoid Alkaloids: Approaches to the C4 Chiral Quaternary Center 107 Preparation of 1,8-diaminonaphthalene-protected boronic acid S8: NH2 Br B(OH)2 NH2 S6 S7 N H PhMe, 110 °C Dean-Stark Br An oven-dried 250 mL B Br S8 Br S8 quant. yield Br (dan)B NH Br round-bottom flask was charged with 3,5- dibromophenylboronic acid (3.55 g, 12.7 mmol, 1.0 equiv.), 1,8-diaminonaphthalene (2.21 g, 14.0 mmol, 1.1 equiv.) and toluene (160 mL). The flask was fitted with a Dean-Stark apparatus, and the mixture was heated to a vigorous reflux for 5 hours. The reaction was concentrated in vacuo and the product was purified by silica gel chromatography (50% CH2Cl2 in hexanes) to afford S8 (5.10 g, 12.7 mmol., 100% yield) as a light orange solid. The 1H NMR of S8 matches previously reported spectroscopic data.16 Note: S8 was stored in a N2-filled glovebox, as it begins to change color to purple under air. 1 H NMR (500 MHz, Chloroform-d) δ 7.76 (t, J = 1.8 Hz, 1H), 7.68 (d, J = 1.8 Hz, 2H), 7.15 (dd, J = 8.3, 7.2 Hz, 2H), 7.08 (dd, J = 8.3, 1.0 Hz, 2H), 6.43 (dd, J = 7.2, 1.1 Hz, 2H), 5.94 (s, 2H). Preparation of boronic ester S10: (HO)2B S9 CF3 CF3 (pin)B CF3 pinacol MgSO4 Et2O, 20 °C S10 CF3 quant. yield A 250 mL round-bottom flask was charged with 3,5-bis(trifluoromethyl)phenylboronic acid (3.82 g, 14.8 mmol, 1.0 equiv.), pinacol (1.92 g, 16.3 mmol, 1.1 equiv.), MgSO4 (10 g), Chapter 3 – Towards C19-Diterpenoid Alkaloids: Approaches to the C4 Chiral Quaternary Center 108 and Et2O (130 mL). The mixture was stirred vigorously at 20 °C for 20 hours. The mixture was filtered and concentrated in vacuo. The crude product was purified by silica gel chromatography (20% EtOAc in hexanes) to give boronic ester S10 (4.98 g, 14.8 mmol, 100% yield) as a white solid. The 1H NMR of S8 matches previously reported spectroscopic data.17 1 H NMR (500 MHz, Chloroform-d) δ 8.24 (s, 2H), 7.94 (s, 1H), 1.38 (s, 12H). Preparation of protected boronic acid S11: CF3 (pin)B CF3 Br (dan)B + S10 CF 3 3 equiv. S8 Br Pd(OAc)2 (5 mol %) SPhos (10 mol %) K3PO4, H2O (10 equiv.) PhMe, 90 °C (dan)B CF3 S11 97% yield F3C CF3 A flame-dried 50 mL round-bottom flask was charged with aryl bis-bromide S8 (0.804 g, 2.0 mmol, 1.0 equiv.) and boronic ester S10 (2.040 g, 6.0 mmol, 3.0 equiv.). The flask was brought into a N2-filled glovebox. Pd(OAc)2 (22.5 mg, 0.10 mmol, 5 mol %), SPhos (82.1 mg, 0.20 mmol, 10 mol %), and K3PO4 (1.274 g, 6.0 mmol, 3.0 equiv.). The flask was sealed with a rubber septum and removed from the glovebox. Toluene (20 mL) was added along with water (0.36 mL, 20 mmol, 10 equiv.) and the mixture was heated to 90 °C. After stirring for 30 hours, the reaction was cooled to room temperature before water (80 mL) was added. The phases were separated, and the aqueous phase was extracted with Et2O (3 x 100 mL). The combined organic phases were dried (MgSO4), filtered, and concentrated in vacuo. Purification by silica gel chromatography (10:1 hexanes:EtOAc) afforded S11 (1.298 g, 1.94 mmol, 97% yield) as an orange powder. Boronic ester S10 (152 mg, 0.45 mmol, 0.22 equiv.) was also recovered. Chapter 3 – Towards C19-Diterpenoid Alkaloids: Approaches to the C4 Chiral Quaternary Center 109 Note: This procedure is modified from a literature procedure which used THF as solvent.17 In our hands, only mono-cross-coupled product with protodebromination was obtained under the previously reported conditions. 1 H NMR (500 MHz, Chloroform-d) δ 8.09 (s, 4H), 7.95 (s, 2H), 7.90 (d, J = 1.8 Hz, 2H), 7.83 (t, J = 1.8 Hz, 1H), 7.18 (dd, J = 8.3, 7.2 Hz, 2H), 7.11 (dd, J = 8.4, 1.0 Hz, 2H), 6.51 (dd, J = 7.3, 1.0 Hz, 2H), 6.13 (s, 2H). Preparation of boronic acid S12: CF3 (dan)B CF3 CF3 S11 (HO)2B CF3 HCl THF/H2O, 20 °C S12 80% yield F3C CF3 F3C CF3 A 50 mL round-bottom flask was charged with S11 (0.780 g, 1.17 mmol, 1.0 equiv.). THF (25 mL) and 5M HCl (aq.) were added, and the mixture was stirred at 20 °C for 22 hours. 3M HCl (aq.) (35 mL) was added, and the mixture was extracted with Et2O (4 x 40 mL). The combined organic extracts were dried (MgSO4), filtered, and concentrated in vacuo to yield S12 (0.518 g, 0.95 mmol, 80% yield) as a white solid. 1 H NMR (500 MHz, Chloroform-d) δ 8.07 (s, 4H), 8.04 (d, J = 1.8 Hz, 2H), 7.93 (s, 2H), 7.86 (t, J = 1.9 Hz, 1H). Chapter 3 – Towards C19-Diterpenoid Alkaloids: Approaches to the C4 Chiral Quaternary Center 110 Suzuki Coupling to synthesize S13: Ar OTf ArB(OH)2 (S12, 2.4 equiv.) Pd(OAc)2 (5 mol %) PPh3 (11 mol %) Me Me Me Me K3PO4 THF, 65 °C OTf Ar 99% yield S5 S13 CF3 CF3 Ar F3C CF3 An oven–dried 1-dram vial with a stir bar was charged with S5 (75 mg, 0.13 mmol, 1.0 equiv.), S12 (170 mg, 0.31 mmol, 2.4 equiv.), K3PO4 (90 mg, 0.39 mmol, 3.0 equiv.), and PPh3 (3.8 mg, 14 µmol, 11 mol %). The vial was brought into a N2–filled glovebox. Pd(OAc)2 (1.5 mg, 6.6 µmol, 5 mol %) and THF (1.3 mL) were added. The vial was capped and removed from the glovebox, and the reaction was heated to 65 °C for 20 hours. Saturated aqueous NH4Cl (2 mL) was added, and the mixture was filtered over a short pad of celite. The aqueous phase was extracted with Et2O (5 x 2 mL), and the combined extracts were dried (MgSO4), filtered, and concentrated in vacuo. The product was purified via silica gel chromatography (16:4:1 hexanes:PhMe:EtOAc) to afford S13 (164 mg, 0.13 mmol, 99% yield) as an colorless, amorphous solid. 1 H NMR (500 MHz, Chloroform-d) δ 8.11 (s, 8H), 7.97 (s, 4H), 7.93 (s, 4H), 7.80 – 7.76 (m, 6H), 7.53 – 7.45 (m, 2H), 7.33 (ddd, J = 8.2, 6.8, 1.3 Hz, 2H), 7.18 (d, J = 8.6 Hz, 2H), 2.07 (s, 6H). Chapter 3 – Towards C19-Diterpenoid Alkaloids: Approaches to the C4 Chiral Quaternary Center 111 Synthesis of bis-benzylic bromide S14: CF3 Ar Me Me Ar S13 Ar Br NBS CF3 Br AIBN PhH, reflux 44% yield Ar Ar S14 F3C CF3 An oven–dried 1-dram vial with a stir bar was charged with S13 (163 mg, 0.13 mmol, 1.0 equiv.), N-bromosuccinimide (50 mg, 0.28 mmol, 2.2 equiv.), AIBN (2.1 mg, 13 µmol, 0.10 equiv.). The vial was brought into a N2–filled glovebox, and benene was added. The vial was capped and removed from the glovebox. The reaction was heated to 80 °C for 3 hours. After cooling to room temperature, water (1.5 mL) was added, and the aqueous phase was extracted with EtOAc (5 x 2 mL), and the combined extracts were dried (Na2SO4), filtered, and concentrated in vacuo. The product was purified via silica gel chromatography (16:4:1 hexanes:PhMe:EtOAc) to afford S14 (80 mg, 56 µmol, 44% yield) as a white solid. 1 H NMR (500 MHz, Chloroform-d) δ 8.16 (s, 8H), 8.06 (s, 2H), 8.01 – 7.97 (m, 6H), 7.94 (s, 4H), 7.87 (t, J = 1.7 Hz, 2H), 7.60 (ddd, J = 8.1, 6.9, 1.2 Hz, 2H), 7.38 (ddd, J = 8.4, 6.9, 1.3 Hz, 2H), 7.18 (d, J = 7.3 Hz, 2H), 4.37 (d, J = 10.3 Hz, 2H), 4.32 (d, J = 10.1 Hz, 2H). Chapter 3 – Towards C19-Diterpenoid Alkaloids: Approaches to the C4 Chiral Quaternary Center 112 Preparation of quaternary ammonium bromide catalyst (S)–138: CF3 Ar Ar Br HN O N Br Ar S14 K2CO3 MeCN, 85 °C 49% yield Ar O CF3 Ar Br (S)–138 F3C CF3 An oven-dried 1-dram vial with a stir bar was charged with S14 (80 mg, 56 µmol, 1.0 equiv.), K2CO3 (39 mg, 0.28 mmol, 5.0 equiv.), MeCN (1.1 mL), and freshly-distilled morpholine (15 mg, 0.17 mmol, 3.0 equiv.). The mixture was heated to 85 °C for 14 hours. After the reaction was allowed to cool to room temperature, the mixture was filtered over celite, and the filtrate was concentrated in vacuo. The product was purified by silica gel chromatography, taking precautions to avoid contaminating the product with impurities from the silica gel: 3 column volumes of MeOH was passed through a column pre-equilibrated with MeOH. The column was then flushed with the mobile phase before loading the crude product. Purification (2-4-5-10-20% MeOH in CH2Cl2) afforded (S)–138 (39.4 mg, 28 µmol, 49% yield) as a white solid. 1 H NMR (500 MHz, Chloroform-d) δ 8.21 (s, 2H), 8.16 – 8.08 (m, 10H), 7.98 (s, 2H), 7.94 (s, 2H), 7.90 (t, J = 1.7 Hz, 2H), 7.89 (s, 2H), 7.75 – 7.67 (m, 4H), 7.55 (d, J = 8.4 Hz, 2H), 7.48 (ddd, J = 8.4, 6.8, 1.3 Hz, 2H), 5.28 (d, J = 12.5 Hz, 1H), 5.24 (s, 1H), 4.04 (d, J = 13.4 Hz, 2H), 3.95 – 3.84 (m, 2H), 3.56 – 3.44 (m, 2H), 3.18 – 3.06 (m, 2H), 3.07 – 2.93 (m, 2H). Chapter 3 – Towards C19-Diterpenoid Alkaloids: Approaches to the C4 Chiral Quaternary Center 113 Stereoselective conjugate addition: NC CO2t-Bu O O O O (S)-PTC (138, 2 mol%) O 142 O O 73 Cs2CO3 (0.25 equiv.) PhMe, 0 °C 19 19 t-BuO2C NC 99% yield 3.3:1 d.r. 18 NC 4 O H 143 4 t-BuO2C 18 O H major minor 74% ee 23% ee 144 An oven-dried 1-dram vial was charged with α-cyanoacetate 141 (12.1 mg, 0.05 mmol, 1.0 equiv.), 2-cyclopenten-1-one (8.2 mg, 0.10 mmol, 2.0 equiv.), and quaternary ammonium catalyst (S)–138 (1.4 mg, 0.001 mmol, 2 mol %). Toluene (1 mL) was added, and the vial was put under Argon with a balloon and cooled to 0 °C in a cryocool. After 15 minutes of cooling, Cs2CO3 (1.6 mg, 0.005 mmol, 0.10 equiv.) was added, and the mixture was stirred at 0 °C for 11 hours. The reaction was quenching with saturated aqueous NH4Cl (1 mL) and extracted with EtOAc (5 x 2 mL). The combined organic extracts were dried (Na2SO4), filtered and concentrated in vacuo. Quantitative NMR with dimethyl fumarate as an internal standard showed a 3.3:1 ratio of 143:144 in 99% yield. Clean samples of each diastereomer were obtained by silica gel chromatography (5-10-15-20% EtOAc in hexanes). Enantiopurity of 143 and 144 were determined to be 74% ee and 23% ee, respectively, using chiral SFC. Characterization data for major diastereomer 143: 1 H NMR (500 MHz, Chloroform-d) δ 4.92 (t, J = 4.2 Hz, 1H), 4.01 – 3.96 (m, 2H), 3.91 – 3.84 (m, 2H), 2.65 (tdd, J = 11.7, 7.8, 6.2 Hz, 1H), 2.50 – 2.43 (m, 1H), 2.37 – 2.27 (m, 3H), 2.27 – 2.15 (m, 1H), 2.06 – 1.93 (m, 3H), 1.92 – 1.83 (m, 1H), 1.74 – 1.64 (m, 1H), 1.52 (s, 9H). 13 C NMR (101 MHz, CDCl3) δ 167.03, 117.69, 103.11, 85.04, 65.21, 65.19, 54.21, 43.03, 41.27, 38.22, 29.89, 29.55, 27.95, 25.19. Chapter 3 – Towards C19-Diterpenoid Alkaloids: Approaches to the C4 Chiral Quaternary Center 114 HRMS: (ESI-TOF) calc’d for C13H17NO5 [M + H – CH2C(CH3)2]+ 268.1179, found 268.1179. [𝜶]𝟐𝟐 𝐃 = +39° (c = 0.33, CHCl3). TLC (50%EtOAc/50%Heaxanes), Rf: 0.35 (blue in p-anisaldehyde) Characterization data for minor diastereomer 144: 1 H NMR (500 MHz, Chloroform-d) δ 4.90 (t, J = 4.2 Hz, 1H), 4.02 – 3.93 (m, 2H), 3.92 – 3.80 (m, 2H), 2.73 – 2.62 (m, 1H), 2.55 – 2.40 (m, 2H), 2.33 – 2.16 (m, 2H), 2.16 – 2.05 (m, 1H), 2.05 – 1.83 (m, 4H), 1.74 – 1.64 (m, 1H), 1.54 (s, 9H). 13 C NMR (101 MHz, CDCl3) δ 166.95, 117.63, 103.02, 85.01, 65.19, 65.17, 54.57, 43.32, 40.64, 38.27, 30.83, 29.89, 28.01, 27.94, 25.84. TLC (50%EtOAc/50%Heaxanes), Rf: 0.35 (blue in p-anisaldehyde) Chapter 3 – Towards C19-Diterpenoid Alkaloids: Approaches to the C4 Chiral Quaternary Center Chiral SFC Traces: Major diastereomer 143, 74% ee. Conditions: 10% iPrOH, OD-H column, 2.5 mL/min, 5 minute run. Major diastereomer 143, racemic trace. 115 Chapter 3 – Towards C19-Diterpenoid Alkaloids: Approaches to the C4 Chiral Quaternary Center Minor diastereomer 144, 23% ee: Conditions: 10% iPrOH, OD-H column, 2.5 mL/min, 8 minute run. Minor diastereomer 144, racemic trace: 116 Chapter 3 – Towards C19-Diterpenoid Alkaloids: Approaches to the C4 Chiral Quaternary Center 117 3.7 NOTES AND REFERENCES (1) (2) (3) (4) (5) (6) (7) (8) (9) (10) (11) (12) (13) (14) (15) (16) (17) Mak, V. W. California Institute of Technology, Pasadena, California, USA, 2017. Wong, A. R. California Institute of Technology, Pasadena, California, USA, 2019. Ayers, T. A. Tetrahedron Lett. 1999, 40 (30), 5467–5470. Steves, J. E.; Stahl, S. S. J. Am. Chem. Soc. 2013, 135 (42), 15742–15745. Liu, W.; Xu, D. D.; Repicˇ, O.; Blacklock, T. J. Tetrahedron Lett. 2001, 3. Foley, M. A.; Jamison, T. F. Org. Process Res. Dev. 2010, 14 (5), 1177–1181. Cheng, C.; Brookhart, M. J. Am. Chem. Soc. 2012, 134 (28), 11304–11307. Arai, T.; Yamada, Y. M. A.; Yamamoto, N.; Sasai, H.; Shibasaki, M. Chem. - Eur. J. 1996, 2 (11), 1368–1372. Fastuca, N. J.; Wong, A. R.; Mak, V. W.; Reisman, S. E. Org. Synth. 2020, 97, 327– 338. Wang, X.; Kitamura, M.; Maruoka, K. J. Am. Chem. Soc. 2007, 129 (5), 1038–1039. Garro-Helion, F.; Merzouk, A.; Guibe, F. J Org Chem 1993, 58, 6109–6113. Simonin, C.; Awale, M.; Brand, M.; van Deursen, R.; Schwartz, J.; Fine, M.; Kovacs, G.; Häfliger, P.; Gyimesi, G.; Sithampari, A.; Charles, R.; Hediger, M. A.; Reymond, J. Angew. Chem. Int. Ed. 2015, 54 (49), 14748–14752. Cox, P. J.; Wang, W.; Snieckus, V. Tetrahedron Lett. 33 (17), 2253–2256. Ooi, T.; Kameda, M.; Maruoka, K. J. Am. Chem. Soc. 2003, 125 (17), 5139–5151. Yanagisawa, A.; Satou, T.; Izumiseki, A.; Tanaka, Y.; Miyagi, M.; Arai, T.; Yoshida, K. Chem. - Eur. J. 2009, 15 (43), 11450–11453. Chianese, A. R.; Mo, A.; Datta, D. Organometallics 2009, 28 (2), 465–472. Wilckens, K.; Lentz, D.; Czekelius, C. Organometallics 2011, 30 (6), 1287–1290. Chapter 4 – Completion of Aconitine Core and Synthesis of (–)-Talatisamine, (–)-Liljestrandisine, and (–)-Liljestrandinine 118 Chapter 4 Completion of Aconitine Core and Synthesis of (–)-Talatisamine, (–)-Liljestrandisine, and (–)-Liljestrandinine 4.1 INTRODUCTION This chapter describes studies on the completion of the B and E rings of the aconitine core, as well as the completion of the total synthesis of (–)-talatisamine, (–)-liljestrandisine, and (–)-liljestrandinine. 4.2 RADICAL CYCLIZATION CASCADE With all carbon atoms of the natural product framework in place, all that remains to access the core is C17–N and C7–C8 bond formation. To do so, we envisioned an olefin carboamination with anti-stereoselectivity to form two rings in a single step (Figure 4.1). In order to effect this transformation, we investigated a radical cascade approach, wherein an Ncentered radical would undergo 6-exo-trig cyclization to form the E-ring and generate a C7radical which would engage in Giese addition to a D-ring enone. To test this approach, we targeted N-chloro amines as N-centered radical precursors. Chapter 4 – Completion of Aconitine Core and Synthesis of (–)-Talatisamine, (–)-Liljestrandisine, and (–)-Liljestrandinine 119 Figure 4.1 Proposed radical cyclization cascade to form aconitine core. MeO2C OR NHEt 17 MeO2C NEt 6-exo MeO2C OR OR OR OR NEt 6-exo NEt MeO2C MeO2C NEt 1 B 7 7 E 2 8 OR 170 4.2.1 O RO 172 O RO 171 O RO 173 O RO 174 O Reaction of Neutral Aminyl Radical Starting from N-ethyl amide 118, O-methylation with trimethyloxonium tetrafluoroborate proceeds in moderate yield to afford 175 along with recovered starting material. The amide was reduced selectively using iridium-catalyzed hydrosilylation to afford amine 176. Next, the D-ring was functionalized to introduce the radical acceptor enone. The siliconide is deprotected with HF•pyridine to afford a diol, which can undergo selective oxidation at the allylic site under Stahl’s conditions. The remaining C16 alcohol is protected as the triethylsilyl ether to give 177. Treatment of 177 with NCS affords the N-chloroamine 178. In the key attempted radical cascade reaction, when 178 was treated with AIBN and tributyltin hydride, an unexpected C6-functionalized product 179 was obtained. Chapter 4 – Completion of Aconitine Core and Synthesis of (–)-Talatisamine, (–)-Liljestrandisine, and (–)-Liljestrandinine 120 Scheme 4.1 Synthesis of N-centered radical precursor 178 and unexpected reactivity. OH NHEt MeO2C Me3OBF4 Proton Sponge O O t-Bu O t-Bu Si t-Bu Unexpected C6functionalized product O MeO2C OMe NHEt 6 TES O OMe NEt Cl MeO2C 59% yield OMe NHEt MeO2C NCS 7 6 CH2Cl2 0 °C 8 8 O 63% yield (3 steps) 176 PHMe, 110 °C 7 3. TESOTf, 2,6-(t-Bu)2-4-Me-Py CH2Cl2, –78 °C O Si O tBu tBu 99% yield t-Bu n-Bu3SnH AIBN 179 1. HF·pyr, MeCN 2. Cu(MeCN)4OTf, ABNO NMI, MeObpy, MeCN NHEt PhH, 45 °C, 4 d; then NaBH(OAc)3 175 50% yield (90% BRSM) 118 OMe MeO2C [Ir(COE)2Cl]2 (1 mol %) Et2SiH2 (8 equiv.) O 4 Å MS CH2Cl2 0 to 21 °C O Si OMe NHEt MeO2C OTES O OTES 50% yield O 177 178 Our proposed mechanism for the formation of 179 is a Hoffmann-Löffler-Freytag type reaction, the mechanism of which is shown in Scheme 4.2. N-centered radical intermediate 180 undergoes a 1,5-hydrogen atom transfer at C6 to afford allyl-radical intermediate 181. The radical then undergoes Giese addition to the D-ring enone at C6 to afford the observed product 179 after quenching of the resulting α-radical. It is worth noting that there are very few examples of 6-exo cyclizations of neutral aminyl radicals with olefins, especially those with allylic H-atoms. This observed reactivity led us to consider less nucleophilic N-centered radical intermediates with the hope that this would minimize HAT pathways. Scheme 4.2 Hoffmann-Löffler-Freytag type reaction of aminyl radical 180 to afford 179. MeO2C OMe NEt Cl AIBN n-Bu3SnH PhMe 110 °C OTES 178 O 59% yield MeO2C OMe NEt MeO2C 1,5-HAT 6 OMe NEt H 6 H 6-exo-trig; nBu3SnH OTES 180 O OTES 181 O MeO2C O OMe NEt H TES 179 O Chapter 4 – Completion of Aconitine Core and Synthesis of (–)-Talatisamine, (–)-Liljestrandisine, and (–)-Liljestrandinine 121 Reactivity of Aminyl Radical Cations 4.2.2 We turned to methods of generating aminyl radical cations. Lewis-acidic singleelectron reducing catalysts have been used to effect 1,2-aminochlorination of alkenes.1–5 Especially relevant are examples of 6-exo-trig cyclizations on substrates with allylic hydrogen atoms with the potential for 1,5-HAT to occur (Figure 4.2). To quickly evaluate conditions for radical cyclization to form the E-ring piperidine, we turned to a simplified model system. Figure 4.2 Aminyl radical cations generated from Lewis-acidic single-electron reductants engaging in intramolecular aminochlorination of alkenes. Stella et al., 1970 Cl MX N Cl Pr 182 HOAc/H2O N Pr 183 MX Yield CuCl 64% CuCl-CuCl2 62% FeSO4-FeCl3 70% TiCl3 81% Stella et al., 1977 Cl N Me Ph 184 Cl TiCl3 H 2O 6-exo-trig Ph 185 TiCl3 N Me NMe Ph Cl 186 92% yield 4:1 cis:trans We chose to test conditions for N-centered radical conditions using a simplified model system synthesized via a route analogous to the lactone-aminolysis route presented in Chapter 3 (Scheme 4.3). The [3.2.1]-bicyclooctene C/D-ring bicycle substituent off of the C11 quaternary center was replaced with a simple cyclopentene. 1,2-addition of an cyclopentenylmagnesium bromide (182) to epoxy ketone 71 affords 1,2-addition product 189. Subjection to TMSOTf effects semipinacol rearrangement to afford 190. C1 silyl-ether deprotection and in situ lactonization was achieved by treatment with trifluoroacetic acid to give 191. The ketone of 191 was converted to the enol triflate 192 and then reduced under Chapter 4 – Completion of Aconitine Core and Synthesis of (–)-Talatisamine, (–)-Liljestrandisine, and (–)-Liljestrandinine 122 palladium-catalyzed conditions to afford 193. Lactone-opening aminolysis with ethylamine affords C19 amide 194. The C1 alcohol was converted to the methyl ether 195 upon treatment with trimethyloxonium tetrafluoroborate. The amide of 195 was reduced to the amine via rhodium-catalyzed hydrosilylation.6 Lastly, the amine 196 was N-chlorinated to give the radical cyclization substrate 197. Scheme 4.3 Synthesis of model system for N-centered radical cyclization cascade. MgBr 188 O MeO2C MeO2C H O 71 THF, -94 °C NHEt O 194 EtNH2•HCl sodium 2-ethylhexanoate OH O MeO2C O THF/EtNH2 80 °C 193 53% yield H 87% yield 4 Å MS CH2Cl2, 20 °C 90% yield 90% yield Pd(OAc)2 (5 mol %) PPh3 (10 mol %) MeO2C O O HCO2H, NEt3 DMF, 60 °C 192 96% yield NHEt MeO2C O 195 TFA CH2Cl2 20 °C 190 OMe Me3OBF4 Proton Sponge OTMS CO2Me O MeO2C CH2Cl2, -78 to 0 °C 189 61% yield OH MeO2C O MeO2C MeO2C H TMSOTf 2,6-di-t-Bu-4-Me-pyr OTf KHMDS PhNTf2 CH2Cl2 –78 to 0 °C PhSiH3 THF, 21 °C 52% yield O O 191 78% yield OMe Rh(acac)(COD) (10 mol%) MeO2C O MeO2C NHEt OMe NCS MeO2C NEt Cl CH2Cl2, 0 °C 196 197 71% yield A small series of single-electron reducing metal reagents were tested on our model substrate 197 to see if 6-exo-trig cyclization to afford the E-ring piperidine could occur (Table 4.1). While Stella’s stoichiometric TiCl3 conditions gave only protodechlorination, treatment with catalytic copper (I) chloride yielded 199 which represents cyclization and formal loss of HCl. With this result in hand, we sought to apply these conditions to a substrate which would lead to the aconitine core. Chapter 4 – Completion of Aconitine Core and Synthesis of (–)-Talatisamine, (–)-Liljestrandisine, and (–)-Liljestrandinine 123 Table 4.1 Conditions for N-centered radical cyclization cascade evaluated on model substrate 197. OMe OMe NEt MeO2C Conditions Cl OMe NEt MeO C MeO2C OMe NEt MeO C 2 2 NHEt Cl 197 198 199 200 Entry Reductant Temperature Solvent 1 TiCl3 (1.1 equiv) 0 °C 1:1 HOAc:H2O Result 200 2 CuCl (0.2 equiv.) 70 °C THF 60% yield 198 N-chlorination of 123 gave a radical cyclization substrate with an unactivated D-ring olefin (201, Scheme 4.4). Subjection of 201 to catalytic copper (I) chloride again resulted in E-ring piperidine formation, although the final C7–C8 bond of the aconitine core was not formed. The product isolated is tentatively assigned as olefin 1,2-aminochlorination product 203, though we have been unable to confirm the relative stereochemistry at C7. We sought to oxidize the C16 alcohol to the ketone in order to increase the reactivity of the C8-C15 alkene. To do so, the siliconide of 123 was deprotected, the allylic C16 alcohol was oxidized selectively, and the C14 alcohol was protected as the triethylsilyl ether (Scheme 4.4). Unfortunately, this 3 step sequence resulted in epimerization at C14 to give an inseparable 1.7:1 mixture of diastereomers (204). N-chlorination of 204 gave 205. Subjecting this mixture to a variety of single-electron reductant catalysts again did not result in formation of the aconitine core, as evidenced by the diagnostic 1H NMR signal from the β-position of the Dring enone (C8) (Table 4.2). The lack of reactivity of these substrates led us to question the mechanism of this reaction. Chapter 4 – Completion of Aconitine Core and Synthesis of (–)-Talatisamine, (–)-Liljestrandisine, and (–)-Liljestrandinine 124 Scheme 4.4 Synthesis of radical cascade substrates 201 and 205. O MeO OMe N Cl NEt 202 O H OMe MeO NEt Cl N CuCl (0.2 equiv) CH2Cl2 0 °C O Si O t-Bu t-Bu 66% yield 203 NEt Et H O Si O t-Bu t-Bu 123 OMe MeO Cl THF, 70 °C 73% yield O Si O t-Bu OMe MeO O Si O t-Bu t-Bu 201 t-Bu 203 123 O OMe MeO NEt H O Si O t-Bu 1. HF·pyr, MeCN 2. Cu(MeCN)4OTf, ABNO NMI, MeObpy, MeCN 3. TESOTf, 2,6-(t-Bu)2-4-Me-Py CH2Cl2, –78 °C t-Bu OMe MeO H O OMe MeO N Cl NEt 202 OMe MeO NEt NEt Conditions Cl CH2Cl2 0 °C TESO 46% yield (3 steps) O X TESO 43% yield 204 O TESO 205 O 206, X = Cl or H 123 Table 4.2 Evaluating radical cyclization cascade conditions on a substrate with a D-ring enone. MeO OMe N Cl OMe MeO N Et Conditions O TESO NMe 210 2 N Et OMe MeO N Et H Et Cl O TESO 205 NMe2 OMe MeO O TESO 207 O TESO 208 209 Entry Catalyst Temperature Solvent product 1 CuCl (0.5 equiv.) 70 °C THF 208 + 209 208 + 209 2 CuCl (0.2 equiv.) 50 °C THF 3 CuCl (0.2 equiv.), hv (halogen lamp) ~35 °C THF decomp 4 CuI (0.5 equiv.) 70 °C PhMe 208 + 209 5 [Cu(MeCN)4]PF6, (±)–210 (0.1 equiv. each) 70 °C THF 208 + 209 6 FeCl2 (0.2 equiv.) 50 °C THF 208 + 209 7 FeCl2 (0.2 equiv.) 50 °C THF/H2SO4 decomp 8 FeCl2 (0.2 equiv.), hv (halogen lamp) ~35 °C THF 208 + 209 9 FeCl2 (0.2 equiv.), hv (halogen lamp) ~35 °C THF/H2SO4 decomp 10 TiCl3 (0.2 equiv.) 35 °C THF 209 Previous synthetic work from Gin and coworkers as well as work from our lab (vide infra) suggests that a C7 radical should readily cyclize with a D-ring enone to close the central B-ring (Figure 4.3).7 The lack of C7–C8 bond formation under copper (I) catalyzed conditions Chapter 4 – Completion of Aconitine Core and Synthesis of (–)-Talatisamine, (–)-Liljestrandisine, and (–)-Liljestrandinine 125 suggests that this system does not go through a C7 radical intermediate. One potential mechanism which accounts for these observations is shown in Figure 4.4. After the copper (I) catalyst reduces the N-chloroamine substrate, aminyl radical cation 213 may undergo a 6-exotrig cyclization with the undesired regioselectivity, forming a C7–N bond to give C17 radical 214. The regioselectivity of this cyclization may be the result of steric hindrance at the neopentyl C17 position. If the C17 radical 214 is unable to cyclize with the D-ring enone, it may abstract a chlorine atom from another equivalent of substrate or from copper (II) chloride to turn over the catalyst. This would give the 1,2-aminochlorination product 215, which may undergo intramolecular SN2 displacement of the C17 chloride by the amine to give aziridinium 216. SN2 aziridinium opening by chloride at C7 would give the observed 1,2aminochlorination product 207. Alternatively, deprotonation of aziridinium 216 at C6 would effect E2-elimination to give the observed C6–C7 olefin product 208. This proposal is purely hypothetical at this point, and computational and synthetic investigations are ongoing to shed light on this mechanism. Figure 4.3 Previous example of radical C7–C8 bond formation to close central B-ring. Gin, 2013 O O NBn MeO2C O Br O NBn MeO2C O Bu3SnH, AIBN O 7 PhH, 80 °C OMe O 86% yield OMe 58 O OMe NEt CuCl Cl THF OTES 205 O O OMe 211 MeO NR MeO2C 59 OMe NEt MeO OTES 212 O Chapter 4 – Completion of Aconitine Core and Synthesis of (–)-Talatisamine, (–)-Liljestrandisine, and (–)-Liljestrandinine 126 Figure 4.4 Possible mechanism to explain observed reactivity of N-chloroamines 197, 191 and 205. MeO OMe N Et MeO OMe 6-exo-trig N Et MeO CuCl Cl R MeO Cl CuCl R N Et C17-radical Cl 213 OMe N Et CuCl R 205 OMe O OTES 214 OMe N Et MeO R OMe N Et MeO SN2 R 7 207 7 Cl OMe N Et MeO Chlorine-atom abstraction 217 Cl Intramolecular MeO SN2 OMe N Et MeO E2 R OMe Cl N Et R 215 R B 208 H 218 4.3 SYNTHESIS OF C19 DITERPENOID ALKALOIDS (–)-TALATISAMINE (1), (–)-LILJESTRANDISINE (238), AND (–)-LILJESTRANDININE (3) The following sections describe the successful completion of the total syntheses of C19 diterpenoid alkaloids (–)-talatisamine (1), (–)-liljestrandisine (238), and (–)-liljestrandinine (3). A stepwise approach was taken for the formation of the final two rings of the aconitine core. Reduction/oxidation manipulations of the D-ring at a late stage allow for divergent synthesis of the three natural products named above. The synthesis of talatisamine (1) and liljestrandisine from the same C16 alcohol diastereomer led to a revision of the structure of liljestrandisine from 2 to 238. 4.3.1 Completion of the Aconitine Core Unable to effect the desired radical cyclization cascade discussed above, we turned to a stepwise approach to closing the E-ring piperidine and central B-ring cyclohexane. A report Chapter 4 – Completion of Aconitine Core and Synthesis of (–)-Talatisamine, (–)-Liljestrandisine, and (–)-Liljestrandinine 127 from Xu, Wang and coworkers describing a synthesis of the A/E/F-tricycle of the aconitinetype diterpenoid alkaloids detailed a strategy for E-ring piperidine formation which mapped on well to intermediates we had been able to access (Figure 4.5).8 Intramolecular aziridination between the C19 primary amine and C7–C17 alkene of 219 efficiently closes the E-ring. In addition to forming the C17–N bond of the aconitine core, this strategy allows for functionalization of C7. Activation of aziridine 220 with ethyl iodide forms intermediate aziridinium 221, which is subsequently opened via SN2 displacement by acetate at the lesshindered C7. We imagined changing the nucleophile in this aziridinium opening from acetate to a halide would provide a functional group handle for C7–C8 bond formation. Figure 4.5 Functionalization at C7 and C17 enabled by intramolecular aziridination. Xu and Wang, 2013 MeO TBSO NH2 R PhI(OAc)2 K2CO3, SiO2 DCE, 50 °C R = CO2Me 70% yield A MeO E TBSO R N F DMF R = CO2Me 219 MeO EtI; then NaOAc TBSO MeO N Et E TBSO R R 7 7 77% yield 221 AcO 220 R = CO2Me 222 R = CO2Me N Et 17 OAc olefin amino-acetoxylation Alternative Aziridinium Opening OMe MeO NH2 OMe N MeO PhI(OAc)2 K2CO3, SiO2 E OMe NAc MeO Ac E Br 223 O OR 224 E 17 Br DCE, 50 °C O OMe NAc MeO 7 7 OR AIBN Bu3SnH O OR 225 B 7 O OR 226 olefin carbo-amination We were pleased to find that subjecting intermediate 116 to Xu and Wang’s aziridination conditions, (diacetoxyiodo)benzene (PIDA) with potassium carbonate and silica gel, resulted in clean conversion to our desired aziridine 227 (Scheme 4.5). Gratifyingly, treating 27 with acetyl bromide gave aziridine-opened product 228 in good yield. In addition to forging the Ering piperidine, this two-step sequence efficiently introduces a radical-precursor at C7 and an Chapter 4 – Completion of Aconitine Core and Synthesis of (–)-Talatisamine, (–)-Liljestrandisine, and (–)-Liljestrandinine 128 N-acetyl group, which will later be reduced to the N-ethyl group found in many C19-diterpenoid alkaloids. Having achieved E-ring formation, we turned to closing the B-ring and completing the aconitine core. Scheme 4.5 Completion of the aconitine core. OMe MeO E AcBr K2CO3, SiO2 DCE, 50 °C CH2Cl2, 0 °C OMe MeO H H2, Pd/C 232 EtOH, 21 °C 84% yield OMOM O 10 O Si O t-Bu t-Bu 228 MeO OMOM O Br MeCN 100% yield OH OH 229 OMe NAc AIBN nBu SnH 3 99% yield NAc HF•pyr PhH, 80 °C B 231 OMe MeO NAc Br OMe NAc OMe 7 83% yield O Si O t-Bu t-Bu 227 MeO NAc 16 MeO N PhI(OAc)2 74% yield O Si O t-Bu t-Bu 116 OMe MeO NH2 Br 2. MOMCl, TBAI DMF, 60 °C O 75% yield 2 steps 230 OMOM 1. Cu(MeCN)4OTf NMI, MeObpy ABNO, MeCN To form the final C7–C8 bond of the aconitine core, we envisioned a Giese addition of a C7 radical generated from 230 to a D-ring enone. To introduce the D-ring enone, the siliconide was deprotected with HF•pyridine to yield C14/C16 diol 229. The allylic C16 alcohol was oxidized, selectively, and the remaining C14 alcohol was protected at its MOM ether to give Giese reaction substrate 230. We were elated to find that subjecting 230 to AIBN and tributyltin hydride resulted in formation of 231 in quantitative yield, validating our fragment coupling approach to this family of natural products. The C10–C12 olefin was hydrogenated with Pd on carbon. Having demonstrated retrosynthetic disconnection through the central Bring as a viable strategy to access the aconitine core, reduction/oxidation manipulations of the D-ring were the only remaining challenges to accessing natural products 1, 238, and 3. Chapter 4 – Completion of Aconitine Core and Synthesis of (–)-Talatisamine, (–)-Liljestrandisine, and (–)-Liljestrandinine 129 Reduction/Oxidation of D-ring for Divergent Syntheses of (–)- 4.3.2 Talatisamine (1), (–)-Liljestrandisine (238) and (–)-Liljestrandinine (3) Scheme 4.6 Total synthesis of (–)-talatisamine (1). LiHMDS OMe MeO Cl NAc t-Bu H N S OMe NAc MeO Ph 16 OMOM O 97% yield 232 OMe NEt RedAl® 233 THF, –78 to 21 °C; then H2O, pyr 8 MeO 8 MOMO OH 234 O Et2O 0 to 21 °C 58% yield 16 MOMO HO OH 245 1. BF3•OEt2 Me3OBF4 DTBMP MeO OMe NEt 2. H2SO4 (aq) 40 °C 77% yield (2 steps) OH HO OMe 1 (–)-talatisamine For the synthesis of 1, 2, and 238, Giese reaction product 232 is dehydrogenated using Mukaiyama’s reagent to form a strained enone which then undergoes spontaneous oxyMichael reaction when treated with water and pyridine to give 234 with the proper oxidation at C8 (Scheme 4.6).9. From there, conditions for diastereoselective reduction of the C16 ketone were evaluated (Table 4.3). Lithium aluminum hydride and sodium borohydride each favored formation of the axial alcohol 235, while samarium (II) iodide favored the equatorial alcohol 236 2:1 in excellent yield. Talatisamine (1) and the reported structure of liljestrandisine (2) are epimeric at C16, so preferential access to either diastereomer is valuable. The optimal reagent for accessing talatisamine was found to be sodium bis(2-methoxyethoxy)aluminum hydride (RedAl ®), as it also reduced the acetamide to the N-ethylamine to give a 58% yield of 245. Using an equivalent of BF3•OEt2 to protect the basic amine, the C8/C16 diol was permethylated, then treated with aqueous sulfuric acid at 40 °C to effect selective hydrolysis at C8 and afford (–)-talatisamine (1) in 77% yield over two steps. This represents a 31 step synthesis in the longest linear sequence (LLS) starting from phenol, the shortest synthesis reported to date. Chapter 4 – Completion of Aconitine Core and Synthesis of (–)-Talatisamine, (–)-Liljestrandisine, and (–)-Liljestrandinine 130 Table 4.3 Evaluating conditions for diastereoselective C16 ketone reduction. OMe MeO NAc OMe MeO NAc [H] OMe MeO NAc H O MOM O OH O O H MOM 234 O H O OH MOM O 235 H H 236 Entry Reductant Solvent Temp S20:S21 yield 1 SmI2, Et3N, H2O THF -78 to 21 °C 1:2 87% 2 NaBH4 MeOH 0 °C 2:1 62% 3 LAH THF 0 °C 1.2:1 n.d. 4 LAH THF -78 to 0 °C 1.8:1 n.d. 5 LAH Et2O -78 to 0 °C 2.4:1 n.d. 6 LAH DME 0 °C 1.3:1 n.d. To access the reported structure of liljestrandisine (2), the equatorial C16 alcohol 236 was treated with lithium aluminum hydride to reduce the acetamide to the N-ethylamine 237 (Scheme 4.7). Deprotection of the C14 MOM ether of 237 gave a product with 1H NMR spectrum which varied significantly from that of the material isolated from nature.10 The reported structure 2 represents the only reported C19-diterpenoid alkaloid natural product with the equatorial alcohol stereochemistry at C16. As such, we hypothesized that the reported structure of liljestrandisine is epimeric at C16 of the actual natural product structure. Indeed, when C16 axial alcohol was treated with sulfuric acid to deprotect the C14 MOM ether, the material isolated had characterization data which matched that reported by the isolation chemists. As such, we propose the structure of (–)-liljestrandisine be revised from 2 to 238. Chapter 4 – Completion of Aconitine Core and Synthesis of (–)-Talatisamine, (–)-Liljestrandisine, and (–)-Liljestrandinine 131 Scheme 4.7 Total synthesis and structural revision of (–)-liljestrandisine (238). OMe NAc MeO OMe NEt MeO MOMO HO 236 MeO H2SO4 (aq) LiAlH4 Et2O OH 0 to 40 °C 27% yield OMe NEt MeO H2SO4 (aq) (–)-liljestrandisine 40 °C OH MOMO HO 237 NMR spectroscopic data doesn’t match OMe NEt 40 °C 16 OH HO OH (–)-liljestrandisine (238) revised 96% yield MOMO HO OH 235 The sequence to access (–)-liljestrandinine (3) varies slightly from the one above (Scheme 4.8). 232 is treated with Mukaiyama’s reagent again, now quenching with methanol and pyridine to afford the C8 methyl ether. The C16 ketone is converted to the corresponding enoltriflate, then reduced under palladium-catalyzed conditions to afford 240. Amide reduction with lithium aluminum hydride followed by heating to 110 °C in aqueous sulfuric acid effects C14 MOM ether deprotection and SN1 substitution with water at C8 to afford (–)liljestrandinine (3) in 60% yield over 2 steps. Scheme 4.8 Total synthesis of (–)-liljestrandinine (3). OMe MeO LiHMDS Cl NAc t-Bu OMOM 232 OMe NAc MeO S N Ph 233 16 THF, –78 to 21 °C then MeOH, pyr O 66% yield 1. LiHMDS Comins’ Reagent THF, –78 to 21 °C 55% yield 2. Pd(OAc)2 (40 mol %) PPh3 (80 mol %) OMe O NEt3, HCO2H, DMF, 60 °C OMe NAc MeO 8 MOMO 239 79% yield 8 MOMO OMe 240 MeO 1. LiAlH4 Et2O, 40 °C OMe NEt 2. H2SO4 (aq) 110 °C 60% yield (2 steps) HO OH (–)-liljestrandinine (3) 4.4 CONCLUDING REMARKS In summary, we report syntheses of the C19 diterpenoid alkaloids (–)-talatisamine (1), (– )-liljestrandisine (238), and (–)-liljestrandinine (3) using a fragment coupling strategy. A twostep sequence of 1,2-addition of an alkenyl lithium to an epoxy ketone electrophile followed by a Lewis Acid-catalyzed semipinacol rearrangement enables this fragment coupling and forms the key C11 all-carbon quaternary center. Creative syntheses of C19 diterpenoid alkaloids Chapter 4 – Completion of Aconitine Core and Synthesis of (–)-Talatisamine, (–)-Liljestrandisine, and (–)-Liljestrandinine 132 have been reported for nearly 50 years at the time of this publication. We believe the success of this novel approach demonstrates that there are still opportunities for strategic innovation to access these highly-bridged natural product scaffolds. This project has exposed opportunities for innovations in methodology as well. Significant synthetic effort was spent on functional group interconversion at the diastereotopic C18/C19 methyl esters after the key fragment coupling. The synthesis could be more convergent if the epoxy ketone coupling partner which bears these centers had the proper functionality installed prior to the coupling event. Work is ongoing to develop a doubly-stereoselective Michaelreaction of prochiral tertiary enolates to α,β-unsaturated ketones to form β/γ-stereodiads. We will seek to apply our fragment coupling strategy to the synthesis of other subfamilies of diterpenoid alkaloids, the C20 denudatine and C20 napelline-type natural products in particular, by varying the bridged bicyclic coupling partner. We also hope to learn from our experience studying radical cyclization cascades to potentially try revised radical cascade approaches in pursuit of these C20 cores. Chapter 4 – Completion of Aconitine Core and Synthesis of (–)-Talatisamine, (–)-Liljestrandisine, and (–)-Liljestrandinine 133 4.5 EXPERIMENTAL SECTION Preparation of methyl ether 178: OH NHEt MeO2C Me3OBF4 Proton Sponge O O Si t-Bu 118 OMe NHEt MeO2C O t-Bu 4 Å MS CH2Cl2 0 to 21 °C 50% yield (90% BRSM) O O Si t-Bu O t-Bu 175 A 250 mL, round-bottom flask was charged with alcohol 26 (787 mg, 1.447 mmol, 1.0 equiv.), equipped with a rubber septum, purged with N2, and cooled to 0 °C in an ice bath. Meanwhile, Me3OBF4 (0.856 g, 5.79 mmol, 4 equiv.), Proton Sponge (1.24 g, 5.79 mmol, 4 equiv.), and activated 4 Å MS (2.36 mg) were added to a 40 mL vial in the glovebox, brought out of the glovebox, and added to the flask containing the substrate. The mixture was then suspended in CH2Cl2 (14.5 mL), and allowed to stir for 48 hours, allowing the reaction to warm up to room temperature over this period. The reaction mixture was filtered through a plug of silica gel and celite that had been pre-packed with EtOAc and rinsed further with EtOAc and concentrated in vacuo. The resulting crude residue was purified by silica gel chromatography (15% in hexanes to elute the product to 20% acetone in hexanes to elute the starting material) to afford methyl ether 175 (399 mg, 71.6 mmol, 50% yield) as a white solid and recovered alcohol 26 (350 mg, 0.637 mmol, 44% yield). 13 C NMR (101 MHz, CDCl3): δ 174.9, 168.7, 164.6, 135.9, 132.8, 131.7, 128.2, 121.1, 77.2, 76.8, 66.4, 57.5, 57.0, 55.7, 52.8, 46.6, 46.1, 46.0, 35.4, 34.8, 28.9, 28.3, 22.0, 21.2, 20.7, 19.2, 14.6. 1 H NMR (400 MHz, Chloroform-d): δ 6.18 (ddd, J = 9.5, 6.1, 1.1 Hz, 1H), 6.11 (t, J = 5.6 Hz, 1H), 5.87 (dt, J = 5.9, 2.3 Hz, 1H), 5.66 (ddd, J = 9.3, 4.4, 1.9 Hz, 1H), 5.45 (dt, J = 5.9, Chapter 4 – Completion of Aconitine Core and Synthesis of (–)-Talatisamine, (–)-Liljestrandisine, and (–)-Liljestrandinine 134 2.0 Hz, 1H), 5.25 (d, J = 3.5 Hz, 1H), 4.51 (td, J = 4.2, 0.9 Hz, 1H), 4.19 (ddd, J = 4.2, 3.1, 0.8 Hz, 1H), 3.70 (s, 3H), 3.38 (dd, J = 7.8, 4.0 Hz, 1H), 3.34 – 3.24 (m, 1H), 3.21 (s, 3H), 3.19 – 3.14 (m, 1H), 3.14 – 3.05 (m, 1H), 3.00 – 2.96 (m, 1H), 2.89 – 2.81 (m, 1H), 2.37 (ddt, J = 16.4, 8.7, 2.2 Hz, 1H), 2.28 – 2.10 (m, 3H), 1.79 (dddd, J = 13.2, 9.0, 7.0, 4.0 Hz, 1H), 1.62 – 1.54 (m, 1H), 1.11 – 1.06 (m, 12H), 1.01 (s, 9H). FTIR (NaCl, thin film): 3375, 2970, 2936, 2896, 2860, 1731 1715, 1668, 1652, 1519, 1476, 1463, 1255, 1780, 1104, 991, 826, 732 cm-1. HRMS: (ESI-TOF) calc’d for C31H48O6NSi [M + H]+ 558.3245, found 558.3240. [𝜶]𝟐𝟓 𝐃 = +122° (c = 0.4, CHCl3). TLC (50%EtOAc/ 50% Hexanes), Rf: 0.6 (UV, blue in p-anisaldehyde) Preparation of N-ethylamine 176:11 OMe NHEt MeO2C [Ir(COE)2Cl]2 (1 mol %) Et2SiH2 (8 equiv.) O O Si t-Bu 175 OMe O t-Bu NHEt MeO2C PhH, 45 °C, 4 d; then NaBH(OAc)3 99% yield O Si O tBu tBu 176 A 2-dram vial charged with amide 175 (30 mg. 54 µmol, 1.0 equiv.) was brought into a N2 filled glovebox. In a separate vial, a stock solution of [Ir(COE)2Cl]2 (1.5 mg, 1.7 µmol, 0.03 equiv.) and Et2SiH2 (7 mg, 79 µmol, 1.5 equiv.) in PhH (1.5 mL) was prepared. Amide 175 was dissolved in PhH (2.0 mL) before [Ir] and Et2SiH2 solution (0.5 mL, 0.01 equiv. [Ir(COE)2Cl]2, 0.5 equiv. Et2SiH2) was added, followed by a solution of E2SiH2 (16.6 mg, 0.19 mmol, 3.5 equiv.) in PhH (0.5 mL). The reaction was stirred at 45 °C for 1 day before more Et2SiH2 (9.2 mg, 0.10 mmol, 2.0 equiv.) was added. The reaction was stirred for an additional 1 day before more Et2SiH2 (9.2 mg, 0.10 mmol, 2.0 equiv.) was added. The reaction was stirred Chapter 4 – Completion of Aconitine Core and Synthesis of (–)-Talatisamine, (–)-Liljestrandisine, and (–)-Liljestrandinine 135 an additional 2 days (4 days total) before TLC indicated conversion to the imine was complete. The reaction was cooled to 19 °C and NaBH(OAc)3 was prepared from NaBH4 (20.3 mg, 0.54 mmol, 10 equiv.) and HOAc (0.09 mL, 1.50 mmol, 30 equiv.). The NaBH(OAc)3 was added dropwise by pipette to the reaction, and the mixture stirred for 10 minutes. The reaction was quenched with 0.5 M NaOH (3 mL) and stirred for 30 minutes. The reaction was extracted with 3:1 CHCl3:iPrOH (4 x 5 mL). The combined organic extracts were dried (Na2SO4), filtered and concentrated in vacuo. The crude mixture was purified by column chromatography to give amine 176 (29.0 mg, 54 µmol, 99% yield) as a colorless oil. 1 H NMR (400 MHz, Chloroform-d): δ 6.05 (ddd, J = 9.5, 6.1, 1.1 Hz, 1H), 5.89 (dt, J = 5.9, 2.3 Hz, 1H), 5.64 (ddd, J = 9.3, 4.4, 1.9 Hz, 1H), 5.46 (ddd, J = 5.8, 2.4, 1.4 Hz, 1H), 5.27 (d, J = 3.7 Hz, 1H), 4.45 – 4.37 (m, 1H), 4.23 – 4.19 (m, 1H), 3.58 (s, 3H), 3.54 (dd, J = 9.2, 3.3 Hz, 1H), 3.23 (s, 3H), 2.99 – 2.95 (m, 1H), 2.82 – 2.73 (m, 3H), 2.63 – 2.49 (m, 3H), 2.43 (dddd, J = 16.0, 9.1, 2.7, 1.5 Hz, 1H), 2.36 – 2.25 (m, 2H), 1.88 – 1.79 (m, 1H), 1.59 – 1.40 (m, 3H), 1.07 (s, 9H), 1.04 – 0.98 (m, 12H). 13 C NMR (101 MHz, CDCl3): δ 176.9, 164.6, 135.6, 133.4, 131.1, 128.2, 119.8, 77.7, 76.7, 66.6, 58.7, 57.7, 57.1, 51.5, 47.9, 46.2, 46.1, 46.0, 44.5, 35.4, 28.9, 28.3, 23.7, 21.8, 21.2, 20.6, 15.1. FTIR (NaCl, thin film): 3449, 2965, 2935, 2898, 2859, 1732, 1476, 1103, 992, 827, 731 cm-1. HRMS: (ESI-TOF) calc’d for C31H50O5NSi [M + H]+ 544.3453, found 544.3463. [𝜶]𝟐𝟓 𝐃 = +84.5° (c = 1.26, CHCl3). TLC (5% 7 N NH3 in MeOH/ 95% CH2Cl2), Rf: 0.5 (UV, blue in p-anisaldehyde) Chapter 4 – Completion of Aconitine Core and Synthesis of (–)-Talatisamine, (–)-Liljestrandisine, and (–)-Liljestrandinine 136 Preparation of enone 177: OMe NHEt MeO2C O Si t-Bu 176 O t-Bu 1. HF·pyr, MeCN 2. Cu(MeCN)4OTf, ABNO NMI, MeObpy, MeCN OMe NHEt MeO2C 3. TESOTf, 2,6-(t-Bu)2-4-Me-Py CH2Cl2, –78 °C OTES O 63% yield (3 steps) 177 A 2-dram vial was charged with silylene 176 (20.2 mg, 0.037 mmol, 1 equiv.) and MeCN (1 mL). A solution of HF•Py (pyridine ~30%, HF ~70%, 20.2 mg) in MeCN (0.2 mL) was added, and the reaction was allowed to stir for 40 minutes at 20 °C. The reaction was quenched by filtering through a silica gel plug and flushing with lots of 10% 7N NH3 in MeOH/ 90% CH2Cl2 solution, and concentrated in vacuo. The crude material was used without further purification. A 2-dram vial was charged with the crude diol, and dissolved in MeCN (1 mL). A homemade stock-solution [see note below] 0.05M 4,4’-dimethoxy-2,2’-bipyridine (MeObpy), 0.05 M in ABNO, and 0.2 M N-methylimidazole was added to the solution of diol (74 µL, 0.0037 mmol, 0.1 equiv.) followed by Cu(MeCN)4OTf (1.4 mg, 0.0037 mmol, 0.1 equiv.) was added, and the clear red/brown reaction mixture was stirred at 20 °C until slightly yellow green, and the TLC indicated the reaction had gone to completion (ca. 90 min), at which point the solution was filtered through a short plug of silica that had been pre-packed with 10% 7N NH3 in MeOH/ 90% CH2Cl2 and flushed with more 10% 7N NH3 in MeOH/ 90% CH2Cl2 to elute the product, and concentrated in vacuo. The crude enone was subjected to the next step without further purification. A 2-dram vial was charged with crude enone, and azeotroped with PhMe (from the solvent system, 2 x 1 mL), and dried under high vacuum. 2,6-(t-Bu)2-4-MePy (76 mg, 0.371 mmol, 10 equiv.) was added, followed by CH2Cl2 (2 mL). The solution was cooled to –78 °C, Chapter 4 – Completion of Aconitine Core and Synthesis of (–)-Talatisamine, (–)-Liljestrandisine, and (–)-Liljestrandinine 137 and TESOTf (40 µL, 0.186 mmol, 5 equiv.) was added dropwise via microsyringe. After 10 minutes of stirring at –78 °C, the reaction was quenched via the addition of aqueous sat. NaHCO3 solution 2 mL), and was allowed to warm to room temperature. The layers were separated, and the aqueous layer was extracted with CH2Cl2 (5 x 3 mL). The combined organic extracts were dried over Na2SO4, filtered, and concentrated in vacuo. The resulting crude residue was filtered through a plug of silica gel and filtered with hexanes (to remove the 2,6(t-Bu)2-4-MePy) and then 3% 7N NH3 solution in MeOH/ 97% CH2Cl2 to elute the product, and then concentrated in vacuo. The crude residue was purified by preparative thin layer chromatography (3% 7N NH3 solution in MeOH/ 97% CH2Cl2) to afford enone 177 (12 mg, 0.023 mmol, 63% yield) as a clear oil. Notes for the Stahl oxidation step: (1) A 0.05 M solution of 4,4’-dimethoxy-2,2’-bipyridine, 0.05M ABNO and 0.2M NMI solution in MeCN could be pre-made and kept in an 8 °C freezer and was stable and good to use for an extended time (>4 months). This homemade stock solution was used in this reaction, and makes it easy to add more catalyst. (2) This selective oxidation to the enone is very clean and performs well, but oxidation to the diketone has been observed if too much catalyst is added. It is important to monitor this reaction by TLC. It is best to run this reaction carefully, and start with less catalyst, because more can be added if needed to reach full conversion to the desired product. 1 H NMR (400 MHz, Chloroform-d): δ 6.94 (ddd, J = 9.8, 6.4, 1.6 Hz, 1H), 5.98 – 5.94 (m, 1H), 5.76 (ddd, J = 9.8, 2.0, 0.7 Hz, 1H), 5.50 (d, J = 3.7 Hz, 1H), 5.38 – 5.35 (m, 1H), 4.65 Chapter 4 – Completion of Aconitine Core and Synthesis of (–)-Talatisamine, (–)-Liljestrandisine, and (–)-Liljestrandinine 138 (td, J = 4.5, 1.6 Hz, 1H), 3.61 (s, 3H), 3.46 (dd, J = 7.0, 3.0 Hz, 1H), 3.19 (s, 3H), 3.13 – 3.05 (m, 2H), 2.92 (t, J = 9.2 Hz, 1H), 2.77 (d, J = 11.3 Hz, 1H), 2.64 – 2.52 (m, 3H), 2.43 (dddd, J = 15.7, 8.7, 2.6, 1.5 Hz, 1H), 2.33 (ddt, J = 15.6, 9.7, 2.2 Hz, 1H), 2.24 – 2.16 (m, 1H), 1.83 – 1.73 (m, 1H), 1.65 – 1.54 (m, 3H), 1.03 (t, J = 7.1 Hz, 3H), 0.93 (t, J = 7.9 Hz, 9H), 0.64 – 0.53 (m, 6H). 13 C NMR (101 MHz, CDCl3): δ 197.0, 176.8, 161.4, 149.2, 132.8, 132.2, 126.6, 120.6, 84.6, 78.2, 62.1, 58.5, 57.4, 57.2, 51.7, 48.2, 47.9, 45.6, 44.5, 34.9, 22.9, 22.5, 15.1, 6.7, 4.7. HRMS: (ESI-TOF) calc’d for C29H46O5NSi [M + H]+ 516.3140, found 516.3122. [𝜶]𝟐𝟓 𝐃 = +163° (c = 0.085, CHCl3). TLC (5% 7 N NH3 in MeOH/ 95% CH2Cl2), Rf: 0.5 (UV, blue in p-anisaldehyde) Preparation of N-chloroamine 178: OMe NHEt MeO2C OMe NEt Cl MeO2C NCS CH2Cl2 0 °C OTES 177 O 50% yield OTES O 178 A 10 mL round-bottom flask was charged with amine 177 (12 mg, 0.0233 mmol, 1 equiv.), and CH2Cl2 (2.3 mL). The solution was cooled to 0 °C, and N-chlorosuccinimide (3.7 mg, 0.028 mmol, 1 equiv.) was added. The reaction was allowed to stir for 1 hour at 0 °C, and then the reaction was filtered through a plug of silica gel and flushed with EtOAc. The crude residue was purified by preparative thin layer chromatography (20% EtOAc/ 80% Hexanes) to afford N-chloroamine 178 (6 mg, 0.011 mmol, 47% yield) as a clear oil. Note: This reaction is fairly clean and the poor yield in this case resulted from incomplete conversion (procedure is not optimized). Some starting material was recovered. Chapter 4 – Completion of Aconitine Core and Synthesis of (–)-Talatisamine, (–)-Liljestrandisine, and (–)-Liljestrandinine 1 139 H NMR (400 MHz, Chloroform-d): δ 6.93 (ddd, J = 9.8, 6.4, 1.6 Hz, 1H), 5.98 – 5.90 (m, 1H), 5.76 (dd, J = 9.5, 1.8 Hz, 1H), 5.51 (d, J = 3.8 Hz, 1H), 5.37 (ddd, J = 5.9, 2.6, 1.5 Hz, 1H), 4.68 (td, J = 4.6, 1.6 Hz, 1H), 3.60 (s, 3H), 3.49 (dd, J = 7.9, 3.0 Hz, 1H), 3.28 (d, J = 14.0 Hz, 1H), 3.19 (s, 3H), 3.13 – 3.01 (m, 3H), 2.97 – 2.88 (m, 3H), 2.46 – 2.26 (m, 3H), 1.84 – 1.75 (m, 1H), 1.70 (dt, J = 13.9, 6.6 Hz, 1H), 1.62 – 1.56 (m, 1H), 1.16 (t, J = 6.9 Hz, 3H), 0.93 (t, J = 7.9 Hz, 9H), 0.58 (q, J = 7.7 Hz, 6H). 13 C NMR (101 MHz, CDCl3): δ 196.9, 175.9, 161.2, 149.1, 133.0, 131.8, 126.6, 120.6, 84.6, 77.9, 72.3, 62.0, 60.4, 57.3, 57.3, 51.7, 48.4, 47.9, 45.9, 35.3, 23.1, 22.3, 12.8, 6.7, 4.7. FTIR (NaCl, thin film): 2953, 2933, 2877, 1738, 1731, 1722, 1682, 1455, 1176, 1121, 1100, 1013, 739 cm-1. HRMS: (ESI-TOF) calc’d for C29H45O5NSiCl [M + H]+ 550.2750, found 550.2774. [𝜶]𝟐𝟓 𝐃 = +122° (c = 0.2, CHCl3). TLC (20%EtOAc/ 80%Hexanes), Rf: 0.5 (UV, blue in p-anisaldehyde) Preparation of cyclization product 179: OMe NEt Cl MeO2C n-Bu3Sn–H AIBN PHMe, 110 °C OTES O 59% yield MeO2C OMe NHEt O TES O 179 178 A 1-dram vial was charged with N-chloroamine 178 (5 mg, 9.1 µmol, 1 equiv.) and PhMe (1 mL). The reaction was heated to 110 °C in an oil bath, and n-Bu3SnH (1 M in cyclohexane, 0.05 mL, 0.05 mmol, 5 equiv.) and AIBN (0.7 mg, 4.3 µmol, 0.5 equiv.) was added dropwise as a PhMe solution (0.5 mL). The reaction was heated to 110 °C for 30 min. The reaction was then cooled to room temperature, loaded onto a plug of silica gel which was Chapter 4 – Completion of Aconitine Core and Synthesis of (–)-Talatisamine, (–)-Liljestrandisine, and (–)-Liljestrandinine 140 flushed with CH2Cl2 to remove excess Bu3SnH, followed by 10% 7N NH3 in MeOH/90% CH2Cl2 to elute the product. The crude residue was purified by column chromatography (0.51-2-3-4% 7N NH3 in MeOH/CH2Cl2) to afford cyclized 179 (2.8 mg, 5.4 µmol, 63% yield) as a clear oil. 1 H NMR (600 MHz, Chloroform-d) δ 5.78 – 5.73 (m, 1H), 5.70 (dt, J = 5.9, 1.8 Hz, 1H), 4.92 – 4.87 (m, 1H), 4.78 (dd, J = 6.3, 5.0 Hz, 1H), 3.76 (d, J = 6.6 Hz, 1H), 3.71 (s, 3H), 3.54 (s, 1H), 3.39 (dd, J = 11.7, 5.0 Hz, 1H), 3.28 (s, 1H), 3.20 (s, 3H), 3.16 – 3.13 (m, 1H), 2.90 (t, J = 6.0 Hz, 1H), 2.72 – 2.69 (m, 1H), 2.63 – 2.59 (m, 2H), 2.50 (dd, J = 18.6, 9.2 Hz, 1H), 2.41 (dd, J = 18.6, 1.9 Hz, 1H), 2.27 – 2.21 (m, 1H), 2.15 – 2.10 (m, 1H), 2.06 – 2.00 (m, 1H), 1.43 – 1.39 (m, 1H), 1.25 (s, 1H), 1.06 (t, J = 7.1 Hz, 3H), 0.93 (t, J = 8.0 Hz, 9H), 0.59 (q, J = 8.0 Hz, 6H). 13 C NMR (101 MHz, CDCl3) δ 205.8, 177.4, 156.7, 135.2, 131.1, 114.7, 74.7, 71.7, 63.0, 61.1, 56.1, 54.9, 53.7, 52.1, 48.7, 48.0, 44.5, 41.9, 33.9, 25.2, 21.3, 15.0, 6.8, 4.7. FTIR (NaCl, thin film): 3340, 2932, 1722, 1714, 1514, 1462, 1234, 1102, 1013, 743 cm-1. [𝜶]𝟐𝟏 𝐃 = +78° (c = 0.22, CHCl3). HRMS: (ESI-TOF) calc’d for C29H46O5NSi [M + H]+ 516.3140, found 516.3144. TLC (5% 7 N NH3 in MeOH/ 95% CH2Cl2), Rf: 0.5 (UV, yellow in p-anisaldehyde) Notes: 1) PhMe was degassed via Freeze-Pump-Thaw (3 cycles) prior to use. Chapter 4 – Completion of Aconitine Core and Synthesis of (–)-Talatisamine, (–)-Liljestrandisine, and (–)-Liljestrandinine 141 Preparation of epoxy alcohol 189: MeO2C MeO2C H 71 MgBr 188 O O THF, -94 °C 61% yield O MeO2C MeO2C H OH 189 In a 200-mL, round-bottomed flask, epoxyketone 71 (1.59 g, 5.93 mmol, 1.0 equiv.) was dissolved in THF (59 mL) and cooled to –78 °C. To this solution was added cyclopentyl magnesium bromide (prepared from cyclopentenyl bromide, Mg(0) and I2 activation, 0.3M in THF, 25 mL, 7.50 mmol, 1.27 equiv.) via cannula. The reaction was allowed to stir for an additional 10 minutes at –78 °C, then quenched with sat. NH4Cl solution (30 mL) and H2O (30 mL), and was allowed to warm to room temperature. The biphasic mixture was then transferred to a separatory funnel with Et2O (50 mL), and the layers were separated. The aqueous layer was extracted with more Et2O (3 x 75 mL). The combined organics were washed with brine (1 x 100 mL), dried over Na2SO4, filtered, and concentrated in vacuo. The crude residue was purified by silica gel chromatography (12% to 13% to 14% acetone in hexanes to elute the product and then 20% acetone in hexanes to recover the unreacted epoxy ketone starting material) to afford epoxy alcohol 189 (1.20 g, 3.61 mmol, 61% yield) as a white solid. 1 H NMR (400 MHz, Chloroform-d): δ 5.67 (t, J = 1.8 Hz, 1H), 3.75 (s, 3H), 3.70 (s, 3H), 3.34 (d, J = 3.8 Hz, 1H), 2.74 (dd, J = 10.5, 7.8 Hz, 1H), 2.52 – 2.40 (m, 2H), 2.39 – 2.28 (m, 6H), 2.19 – 2.08 (m, 1H), 2.05 – 1.96 (m, 1H), 1.95 – 1.85 (m, 4H), 1.62 – 1.52 (m, 1H). 13 C NMR (101 MHz, CDCl3): δ 171.9, 170.3, 145.4, 126.0, 75.4, 71.4, 56.5, 55.1, 52.9, 52.1, 44.0, 37.8, 32.3, 31.7, 29.3, 23.4, 23.2, 21.4. FTIR (NaCl, thin film): 3503, 2952, 1732, 1433, 1244, 1218, 1174 cm-1. HRMS: (ESI-TOF) calc’d for C18H25O6 [M + H]+ 337.1646, found 337.1635. TLC (33%EtOAc/ 67% Hexanes), Rf: 0.40 (blue/purple in p-anisaldehyde) Chapter 4 – Completion of Aconitine Core and Synthesis of (–)-Talatisamine, (–)-Liljestrandisine, and (–)-Liljestrandinine 142 Semipinacol rearrangement of 189 to prepare 190: O MeO2C MeO2C H OH TMSOTf 2,6-di-t-Bu-4-Me-pyr MeO2C OTMS CO2Me O CH2Cl2, -78 to 0 °C 189 190 87% yield A 100-mL, round-bottomed flask was charged with epoxy alcohol 189 (1.37 g, 4.07 mmol, 1.0 equiv.), 2,6-t-Bu2-4-MePy (2.51 g, 12.2 mmol, 3 equiv.) and CH2Cl2 (41 mL), and the solution was cooled to –78 °C in a dry/ice acetone bath. TMSOTf added dropwise via syringe (1.47 mL, 8.15 mmol, 2 equiv.), causing the solution to turn a faint pink color. The reaction was allowed to stir at – 78 °C for 5 minutes, and then the dry//ice acetone bath was replaced with an ice bath, allowing the reaction to warm to 0 °C. The reaction was allowed to stir for an additional 10 minutes at 0 °C, then the reaction was quenched with sat. NaHCO3 (50 mL), and the mixture was allowed to warm to room temperature. The biphasic mixture was transferred to a separatory funnel and the layers were separated. The aqueous phase was extracted with CH2Cl2 (3 x 75 mL), and the combined organic extracts were dried over Na2SO4, filtered, and concentrated in vacuo. The resulting crude residue was purified by silica gel chromatography (10% to 12% to 15% ethyl acetate in hexanes) to afford ketone 190 (1.45 g, 4.07 mmol, 87% yield) as a white solid. 1 H NMR (500 MHz, Chloroform-d): δ 5.38 (t, J = 2.2 Hz, 1H), 4.33 (t, J = 2.5 Hz, 1H), 3.74 (s, 3H), 3.64 (s, 3H), 3.37 (dd, J = 12.5, 6.9 Hz, 1H), 2.57 – 2.49 (m, 1H), 2.45 (td, J = 13.9, 3.9 Hz, 1H), 2.41 – 2.23 (m, 3H), 2.22 – 2.08 (m, 3H), 1.97 – 1.89 (m, 1H), 1.87 – 1.81 (m, 2H), 1.81 – 1.71 (m, 2H), 1.68 (dq, J = 14.3, 3.6 Hz, 1H), 0.06 (s, 9H). 13 C NMR (126 MHz, CDCl3): δ 219.2, 170.8, 170.5, 142.9, 127.3, 70.3, 57.8, 56.1, 52.7, 52.5, 41.9, 39.1, 32.9, 32.7, 27.8, 23.5, 22.8, 19.6, -0.0. FTIR (NaCl, thin film): 2952, 2846, 1740, 1434, 1252, 1171, 1058, 841 cm-1. Chapter 4 – Completion of Aconitine Core and Synthesis of (–)-Talatisamine, (–)-Liljestrandisine, and (–)-Liljestrandinine 143 HRMS: (ESI-TOF) calc’d for C21H33O6Si [M + H]+ 409.2041, found 409.2056. TLC (15%EtOAc/ 85% Hexanes), Rf: 0.4 (grey/blue/black in p-anisaldehyde) Preparation of lactone 191: MeO2C OTMS CO2Me O TFA CH2Cl2 20 °C 190 O MeO2C O O 191 90% yield In a 250 mL round-bottom flask, silyl ether 190 (1.445 g, 3.54 mmol, 1.0 equiv.) was dissolved in CH2Cl2 (70 mL) and TFA (0.82 mL, 10.6 mmol, 3 equiv.) was added dropwise. The reaction was stirred for 7 hours when TLC indicated complete conversion of starting material. The reaction was quenched with saturated aqueous NaHCO3 (50 mL) and the phases were separated. The aqueous phase was extracted with CH2Cl2 (3 x 50 mL) and the combined extracts were dried (Na2SO4), filtered, and concentrated in vacuo. The reaction was purified by silica gel chromatography (30% EtOAc/70% hexanes) to afford lactone 191 (0.956 g, 3.14 mmol, 90% yield) as a white powder. 1 H NMR (600 MHz, Chloroform-d) δ 5.78 (t, J = 2.2 Hz, 1H), 5.10 – 4.99 (m, 1H), 3.84 (s, 3H), 3.23 (t, J = 9.2, 7.5 Hz, 1H), 2.63 – 2.52 (m, 1H), 2.49 – 2.40 (m, 2H), 2.21 – 2.12 (m, 2H), 2.01 – 1.76 (m, 5H), 1.75 – 1.64 (m, 1H). TLC (30%EtOAc/ 70% Hexanes), Rf: 0.4 (blue in p-anisaldehyde) Chapter 4 – Completion of Aconitine Core and Synthesis of (–)-Talatisamine, (–)-Liljestrandisine, and (–)-Liljestrandinine 144 Preparation of enol triflate 192: O MeO2C O O KHMDS; PhNTf2 CH2Cl2, –78 to 0 °C 191 O MeO2C OTf O 192 78% yield An oven-dried 100 mL round-bottom flask was charged with 191 (0.946 g, 3.11 mmol, 1.0 equiv.) and put under N2. THF (31 mL) was added, and the solution was cooled to –78 °C. A solution of KHMDS (0.5 M in PhMe, 7.5 mL, 3.8 mmol, 1.2 equiv.) was added dropwise by syringe and the mixture was stirred at –78 °C for 40 minutes. In an oven-dried 50 mL pearshaped flask, PhNTf2 (1.33 g, 3.73 mmol, 1.2 equiv.) was put under N2 and dissolved in THF (10 mL). The solution of PhNTf2 was transferred to the reaction flask by cannula, and the reaction was stirred at –78 °C for 10 minutes. The reaction was quenched with saturated aqueous NaHCO3 (50 mL), and the mixture was allowed to warm to room temperature. The mixture was transferred to a separatory funnel and was extracted with Et2O (3 x 50 mL). The combined organic extracts were washed with 0.5 M NaOH (aq.) (3 x 50 mL), dried (Na2SO4), filtered, and concentrated in vacuo. The reaction was purified by silica gel chromatography (20% EtOAc/80% hexanes) to afford enol triflate 192 (1.059 g, 2.43 mmol, 78% yield) as a white solid. 1 H NMR (500 MHz, Chloroform-d) δ 5.68 (t, J = 2.6 Hz, 1H), 5.64 (t, J = 2.0 Hz, 1H), 4.91 (dd, J = 3.8, 1.7 Hz, 1H), 3.83 (s, 3H), 3.05 (dd, J = 9.4, 2.9 Hz, 1H), 2.92 (ddd, J = 18.2, 9.4, 2.4 Hz, 1H), 2.51 – 2.40 (m, 3H), 2.39 – 2.31 (m, 2H), 2.18 – 2.07 (m, 2H), 1.96 – 1.79 (m, 3H). 13 C NMR (101 MHz, CDCl3) δ 169.98, 169.79, 146.02, 139.86, 129.91, 116.90, 116.34, 76.26, 56.45, 53.11, 53.03, 43.60, 33.14, 33.05, 32.35, 26.65, 22.83, 21.89. HRMS: (ESI-TOF) calc’d for C18H19O7F3SNa [M + Na]+ 459.0696, found 459.0708. Chapter 4 – Completion of Aconitine Core and Synthesis of (–)-Talatisamine, (–)-Liljestrandisine, and (–)-Liljestrandinine 145 TLC (20%EtOAc/ 80% Hexanes), Rf: 0.2 (brown in p-anisaldehyde) Preparation of cyclopentene 193: O MeO2C O 192 OTf Pd(OAc)2 (5 mol %) MeO2C PPh3 (10 mol %) HCO2H, NEt3 DMF, 60 °C O H O 193 96% yield A 100 mL, round-bottom flask was charged with enol triflate 192 (1.046 g, 2.40mmol, 1.0 equiv.), PPh3 (63 mg, 0.24 mmol, 10 mol %), and Pd(OAc)2 (27 mg, 0.12 mmol, 5 mol %). The flask was equipped with a rubber septum, purged with N2, and DMF (24 mL) was added. Then, the reaction mixture was sparged with N2 for 5 minutes. NEt3 (2.68 mL, 19.2 mmol, 8 equiv.) was added, followed by HCO2H (0.45 mL, 12.0 mmol, 5 equiv.) – a cloudy gas was observed upon addition. The reaction was heated to 60 °C allowed stirred at that temperature for 15 minutes. The reaction turns black during this time, indicating its completion. The reaction was cooled to room temperature, diluted with sat. NH4Cl (20 mL), transferred to a separatory funnel and diluted with Et2O (30 mL). The layers were separated and the aqueous extracted with Et2O (5 x 30 mL). The organic extracts were dried over Na2SO4, filtered, and concentrated in vacuo. The resulting crude residue was purified by silica gel chromatography (15-16% EtOAc in hexanes) to afford olefin 193 (0.657 g, 2.3 mmol, 96% yield) as a white crystalline solid. 1 H NMR (600 MHz, Chloroform-d) δ 5.69 (dt, J = 5.7, 2.3 Hz, 1H), 5.59 (t, J = 2.1 Hz, 1H), 5.51 (dt, J = 5.8, 2.3 Hz, 1H), 4.70 (dd, J = 3.2, 2.2 Hz, 1H), 3.81 (s, 3H), 3.08 (dd, J = 9.5, 3.5 Hz, 1H), 2.91 (ddt, J = 18.7, 9.4, 2.3 Hz, 1H), 2.45 (ddt, J = 18.7, 3.5, 2.3 Hz, 1H), 2.40 – 2.32 (m, 3H), 2.30 – 2.22 (m, 2H), 2.01 – 1.97 (m, 2H), 1.95 – 1.83 (m, 3H). Chapter 4 – Completion of Aconitine Core and Synthesis of (–)-Talatisamine, (–)-Liljestrandisine, and (–)-Liljestrandinine 13 146 C NMR (101 MHz, CDCl3) δ 170.86, 170.78, 145.07, 132.39, 130.90, 125.13, 79.50, 59.31, 54.02, 52.79, 44.87, 38.25, 32.92, 32.45, 26.62, 23.65, 22.50. HRMS: (ESI-TOF) calc’d for C17H21O4 [M + H]+ 289.1434, found 289.1441. TLC (20%EtOAc/ 80% Hexanes), Rf: 0.2 (brown in p-anisaldehyde) Preparation of N-ethyl amide 194: O MeO2C O 193 H EtNH2•HCl sodium 2-ethylhexanoate THF/EtNH2 80 °C OH NHEt MeO2C O 194 53% yield To a 75 mL pressure vessel containing lactone 193 (0.647 g, 2.24 mmol, 1.0 equiv.), sodium 2-ethylhexanoate (1.680 g, 10.1 mmol, 4.5 equiv.), ethylamine hydrochloride (0.412 g, 5.1 mmol, 2.25 equiv.) were added, followed by THF (5 mL) and neat EtNH2 (10 mL). The vessel was sealed (with a Teflon cap that had a perfluoro O-ring), and heated to 80 °C in an oil bath. The reaction was allowed to stir at 80 °C for 2 days, and the reaction progress was monitored. The reaction was then cooled to room temperature and transferred to a separatory funnel with EtOAc (30 mL) and aqueous 0.5 M NaOH (20 mL). The layers were separated, and the aqueous phase was extracted with Et2O (3 x 20 mL) and EtOAc (2 x 20 mL). The combined organic extracts were dried over Na2SO4, filtered, and concentrated in vacuo. The crude residue was purified by silica gel chromatography (30% to 50% to 100% EtOAc in hexanes) to afford secondary amide 194 as a white foam (400 mg, 1.20 mmol, 53% yield). Note: Neat NH2Et was obtained from distillation of 70%NH2Et/ 30% H2O solution, using a cold finger (with dry ice/acetone) to condense the very volatile NH2Et, and stored over KOH pellets in the freezer. Chapter 4 – Completion of Aconitine Core and Synthesis of (–)-Talatisamine, (–)-Liljestrandisine, and (–)-Liljestrandinine 1 147 H NMR (500 MHz, Chloroform-d) δ 6.23 (s, 1H), 6.10 (dt, J = 5.9, 2.3 Hz, 1H), 5.50 (dt, J = 5.9, 2.0 Hz, 1H), 5.41 (t, J = 2.1 Hz, 1H), 3.96 (t, J = 4.5 Hz, 1H), 3.35 – 3.15 (m, 3H), 2.47 (m, 1H), 2.39 – 2.16 (m, 6H), 2.13 – 2.02 (m, 1H), 1.95 – 1.77 (m, 5JH), 1.11 (t, J = 7.2 Hz, 3H). 13 C NMR (101 MHz, CDCl3) δ 174.64, 169.11, 146.86, 134.74, 134.38, 125.44, 67.89, 57.00, 55.75, 52.87, 46.45, 35.46, 34.85, 33.24, 33.14, 25.75, 23.38, 18.60, 14.81. HRMS: (ESI-TOF) calc’d for C19H27NO4Na [M + Na]+ 356.1832, found 356.1850. TLC (50%EtOAc/ 50% Hexanes), Rf: 0.2 (teal in p-anisaldehyde) Preparation of methyl ether 195: OH NHEt MeO2C O 194 OMe Me3OBF4 Proton Sponge 4 Å MS CH2Cl2, 20 °C NHEt MeO2C O 195 90% yield In a N2-filled glovebox, an oven-dried 15 mL round-bottom flask with a stir bar was charged with 194 (150 mg, 0.45 mmol, 1.0 equiv.). CH2Cl2 (4.5 mL) was added, followed by Proton Sponge® (289 mg, 1.35 mmol, 3.0 equiv.), trimethyloxonium tetrafluoroborate (200 mg, 1.35 mmol, 3.0 equiv.), and 4Å molecular sieve powder (750 mg). The reaction was stirred at 20 °C for 18 hours. The mixture was passed over a plug of silica gel, eluting with 50-100% EtOAc in hexanes. The filtrate was concentrated to give the crude product as a yellow oil. The product was purified by column chromatography (SiO2, 20-30-40-50% EtOAc in hexanes) to give methyl ether 195 (140 mg, 0.41 mmol, 90% yield). 1 H NMR (500 MHz, Chloroform-d) δ 6.10 (s, 1H), 5.85 (dt, J = 5.8, 2.4 Hz, 1H), 5.49 (dt, J = 5.9, 2.1 Hz, 1H), 5.34 (t, J = 2.0 Hz, 1H), 3.70 (s, 3H), 3.44 (dd, J = 8.4, 4.2 Hz, 1H), 3.34 Chapter 4 – Completion of Aconitine Core and Synthesis of (–)-Talatisamine, (–)-Liljestrandisine, and (–)-Liljestrandinine 148 – 3.27 (m, 1H), 3.25 (s, 3H), 3.23 – 3.05 (m, 2H), 2.52 – 2.08 (m, 8H), 1.97 – 1.89 (m, 1H), 1.85 (p, J = 7.5 Hz, 2H), 1.63 – 1.57 (m, 1H), 1.10 (t, J = 7.2 Hz, 3H). 13 C NMR (101 MHz, CDCl3) δ 174.77, 169.33, 148.58, 134.36, 130.81, 124.10, 78.47, 57.46, 57.06, 55.98, 52.82, 47.32, 35.72, 34.87, 32.84, 32.53, 23.52, 22.12, 19.95, 14.75. FTIR (NaCl, thin film): 3353, 2933, 1727, 1654, 1522, 1439, 1377, 1203, 1101, 754 cm-1. HRMS: (ESI-TOF) calc’d for C20H29NO4 [M + H]+ 370.1989, found 370.1994. TLC (2:1 Hexanes:EtOAc), Rf: 0.4 (blue in p-anisaldehyde) Preparation of amine 196: OMe NHEt MeO2C O 195 OMe Rh(acac)(COD) (10 mol%) MeO2C PhSiH3 THF, 21 °C NHEt 196 52% yield A flame-dried 100 mL round-bottom flask was charged with amide 195 (100 mg, 0.29 mmol, 1.0 equiv.) and brought into a N2-filled glovebox. THF was added, followed by Rh(acac)cod (8.9 mg, 0.029 mmol, 10 mol%) and phenylsilane (0.71 mL, 5.8 mmol, 20 equiv.). The flask was sealed with a rubber septum, brought out of the glovebox and put under Ar atmosphere with a balloon. The reaction was stirred at room temperature for 16 hours. The reaction was diluted with THF (5 mL) and quenched with careful addition of saturated aqueous NH4F (5 mL), at which point reaction bubbles vigorously. The mixture was stirred for 90 minutes before diluting with water (5 mL) and EtOAc (15 mL). The phases were separated and the aqueous phase was extracted with 4:1 CHCl3 : iPrOH mixture (5 x 15 mL). The organic extracts were dried (Na2SO4), filtered and concentrated in vacuo. The crude residue was purified by silica gel chromatography (1-2% (7N NH3 in MeOH) in CH2Cl2) to afford the amine 196 as a colorless oil (49.1 mg, 0.15 mmol, 52 % yield). Chapter 4 – Completion of Aconitine Core and Synthesis of (–)-Talatisamine, (–)-Liljestrandisine, and (–)-Liljestrandinine 1 149 H NMR (400 MHz, Benzene-d6) δ 5.83 (dt, J = 5.9, 2.3 Hz, 1H), 5.65 (dt, J = 5.8, 2.0 Hz, 1H), 5.45 (p, J = 1.9 Hz, 1H), 3.69 (dd, J = 7.7, 3.1 Hz, 1H), 3.36 (s, 3H), 3.11 (s, 3H), 3.10 – 3.04 (m, 1H), 2.77 (d, J = 11.1 Hz, 1H), 2.60 – 2.50 (m, 1H), 2.48 (d, J = 11.2 Hz, 1H), 2.45 – 2.37 (m, 3H), 2.36 – 2.24 (m, 5H), 2.05 (dddd, J = 13.8, 7.8, 6.2, 3.1 Hz, 1H), 1.81 (p, J = 7.4 Hz, 2H), 1.66 (ddd, J = 13.7, 7.7, 6.5 Hz, 1H), 1.61 – 1.48 (m, 1H), 0.91 (t, J = 7.1 Hz, 4H). 13 C NMR (101 MHz, C6D6) δ 176.56, 148.73, 136.23, 130.36, 123.68, 79.90, 59.16, 57.52, 57.16, 51.16, 48.39, 46.74, 44.91, 35.40, 33.43, 33.35, 23.93, 23.72, 23.06, 15.52. HRMS: (ESI-TOF) calc’d for C20H32NO3 [M + H]+ 334.2377, found 334.2377. TLC (8% MeOH/ 92% CH2Cl2), Rf: 0.4 (blue in p-anisaldehyde) Preparation of N-chloroamine 197: OMe MeO2C NHEt OMe NCS MeO2C NEt Cl CH2Cl2, 0 °C 196 197 71% yield A solution of secondary amine 196 (20 mg, 60 µmol, 1.0 equiv.) in CH2Cl2 (2.0 mL) was cooled to 0 °C before NCS (8.0 mg, 60 µmol, 1.0 equiv.) was added in one portion. The reaction was stirred for 60 minutes before filtering over a short pad of silica gel, eluting with 1:1 hexanes:EtOAc. The filtrate was concentrated in vacuo. The crude product was purified by column chromatography (10-20-30-40% EtOAc in hexanes) to afford N-chloroamine 197 (15.6 mg, 42 µmol, 71% yield) as a white powder. 1 H NMR (500 MHz, Chloroform-d) δ 5.85 (dt, J = 5.8, 2.3 Hz, 1H), 5.44 (ddd, J = 5.8, 2.5, 1.4 Hz, 1H), 5.41 – 5.38 (m, 1H), 3.70 (dd, J = 9.4, 3.3 Hz, 1H), 3.57 (s, 3H), 3.33 (d, J = 14.0 Hz, 1H), 3.28 (s, 3H), 3.00 (d, J = 14.0 Hz, 1H), 2.94 (qd, J = 6.8, 2.9 Hz, 2H), 2.86 (td, J = Chapter 4 – Completion of Aconitine Core and Synthesis of (–)-Talatisamine, (–)-Liljestrandisine, and (–)-Liljestrandinine 150 9.2, 1.3 Hz, 1H), 2.62 – 2.49 (m, 1H), 2.46 – 2.22 (m, 5H), 2.23 – 2.07 (m, 1H), 1.94 (dddd, J = 13.1, 7.0, 5.6, 3.3 Hz, 1H), 1.90 – 1.68 (m, 3H), 1.56 – 1.42 (m, 1H), 1.15 (t, J = 6.9 Hz, 3H). Reaction of N-chloroamine 197 with Copper (I) Chloride: OMe MeO2C NEt Cl OMe CuCl (20 mol %) MeO2C NEt THF, 70 °C 197 199 60% yield CuCl (0.5 mg, 5.4 µmol, 0.2 equiv.) was added to a 1-dram vial followed by a solution of N-chloroamine 197 (10.0 mg, 27 µmol, 1.0 equiv.) in THF (2.0 mL), and the mixture was heated to 70 °C for 12 hours. Saturated aqueous NaHCO3 (2 mL) was added, and the mixture was extracted with 4:1 CHCl3 : iPrOH mixture (3 x 5 mL). The organic extracts were dried (Na2SO4), filtered and concentrated in vacuo. Purification by preparative TLC (1% (7 N NH3 in MeOH) in CH2Cl2) gave a product tentatively assigned as 199 (5.2 mg, 16 µmol, 60% yield). 1 H NMR (400 MHz, Benzene-d6) δ 6.29 (dd, J = 5.9, 1.5 Hz, 1H), 5.77 (dd, J = 5.8, 1.5 Hz, 1H), 5.51 (p, J = 2.1 Hz, 1H), 4.31 (dt, J = 7.2, 1.6 Hz, 1H), 3.67 (d, J = 7.1 Hz, 1H), 3.43 – 3.38 (m, 1H), 3.37 (d, J = 3.2 Hz, 4H), 3.29 (d, J = 8.8 Hz, 1H), 3.12 (d, J = 1.0 Hz, 4H), 2.69 – 2.57 (m, 1H), 2.54 – 2.39 (m, 5H), 2.31 (dtq, J = 11.0, 6.5, 2.1 Hz, 4H), 2.09 – 1.76 (m, 4H), 1.70 (dq, J = 12.5, 4.2 Hz, 2H), 1.64 – 1.43 (m, 2H), 1.43 – 1.30 (m, 2H), 1.03 (t, J = 7.2 Hz, 3H). Chapter 4 – Completion of Aconitine Core and Synthesis of (–)-Talatisamine, (–)-Liljestrandisine, and (–)-Liljestrandinine 151 Preparation of N-chloroamine 201: O MeO OMe N Cl NEt 202 O H OMe MeO NEt Cl CH2Cl2 0 °C O Si O t-Bu t-Bu 73% yield 123 O Si O t-Bu t-Bu 201 A solution of secondary amine 123 (24 mg, 65 µmol, 1.0 equiv.) in CH2Cl2 (2.2 mL) was cooled to 0 °C before NCS (17 mg, 130 µmol, 2.0 equiv.) was added in one portion. The reaction was stirred for 90 minutes before filtering over a short pad of silica gel, eluting with 1:1 hexanes:EtOAc. The filtrate was concentrated in vacuo. The crude product was purified by column chromatography (10-20-30-40% EtOAc in hexanes) to afford N-chloroamine 201 (27 mg, 47 µmol, 73% yield) as a white powder. 1 H NMR (400 MHz, Chloroform-d) δ 6.07 (ddd, J = 9.5, 6.1, 1.1 Hz, 1H), 5.87 (dt, J = 5.8, 2.2 Hz, 1H), 5.65 (ddd, J = 9.3, 4.4, 1.9 Hz, 1H), 5.46 – 5.38 (m, 2H), 4.47 (t, J = 4.8 Hz, 1H), 4.26 – 4.17 (m, 1H), 3.57 (dd, J = 7.5, 2.6 Hz, 1H), 3.29 – 3.24 (m, 5H), 3.22 (s, 3H), 3.08 – 2.93 (m, 5H), 2.93 – 2.84 (m, 1H), 2.47 (t, J = 9.2 Hz, 1H), 2.42 – 2.20 (m, 2H), 1.71 – 1.52 (m, 3H), 1.49 – 1.40 (m, 1H), 1.19 (t, J = 6.9 Hz, 3H), 1.09 (s, 9H), 1.01 (s, 9H). 13 C NMR (101 MHz, CDCl3) δ 166.56, 136.04, 134.27, 131.37, 128.25, 119.94, 79.64, 77.36, 74.37, 69.55, 66.68, 60.90, 58.33, 57.36, 57.26, 46.45, 46.36, 44.59, 40.82, 35.61, 29.07, 28.48, 24.01, 21.86, 21.41, 20.84, 13.25. FTIR (NaCl, thin film): 2933, 1476, 1102, 992, 812, 780, 762, 641cm-1. HRMS: (ESI-TOF) calc’d for C31H51NO4ClSi [M + H]+ 564.3270, found 564.3279 [𝜶]𝟐𝟐 𝐃 = +126° (c = 0.40, CHCl3). Chapter 4 – Completion of Aconitine Core and Synthesis of (–)-Talatisamine, (–)-Liljestrandisine, and (–)-Liljestrandinine 152 TLC (3:1 Hexanes:EtOAc), Rf: 0.8 (UV, white/green in p-anisaldehyde) Reaction of N-chloroamine 201 with Copper (I) Chloride: OMe MeO NEt Cl OMe MeO N CuCl (0.2 equiv) Cl THF, 70 °C O Si O t-Bu t-Bu 201 66% yield 203 Et O Si O t-Bu t-Bu 203 CuCl (0.18 mg, 1.8 µmol, 0.2 equiv.) was dispensed as a stock solution in MeCN to a ½ dram vial. The MeCN was removed in vacuo. The vial of CuCl was brought into a N2 filled glovebox and a solution of N-chloroamine 201 (5.0 mg, 8.9 µmol, 1.0 equiv.) in THF (0.5 mL) was added, and the mixture was heated to 70 °C for 4 hours. Saturated aqueous NaHCO3 (2 mL) was added, and the mixture was extracted with 4:1 CHCl3 : iPrOH mixture (3 x 5 mL). The organic extracts were dried (Na2SO4), filtered and concentrated in vacuo. Purification by preparative TLC (5% (7 N NH3 in MeOH) in CH2Cl2) gave a product tentatively assigned as 203 (3.3 mg, 5.9 µmol, 66% yield). Assignments were made using 1H, COSY, HSQC, HMBC and TOCSY NMR experiments. 1 H NMR (400 MHz, Chloroform-d) δ 6.07 (ddd, J = 9.5, 6.0, 1.1 Hz, 1H), 5.74 (ddd, J = 9.5, 4.4, 1.9 Hz, 1H), 5.64 (d, J = 3.9 Hz, 1H), 4.57 (d, J = 4.9 Hz, 1H), 4.47 (t, J = 4.7 Hz, 1H), 4.25 (t, J = 3.7 Hz, 1H), 3.94 – 3.83 (m, 1H), 3.81 – 3.72 (m, 3H), 3.41 – 3.36 (m, 1H), 3.35 (s, 3H), 3.32 (s, 3H), 3.27 – 3.19 (m, 3H), 3.16 (d, J = 13.1 Hz, 1H), 3.08 (s, 1H), 2.83 (t, J = 5.2 Hz, 1H), 2.58 (d, J = 14.9 Hz, 1H), 2.37 – 2.26 (m, 2H), 2.11 – 2.05 (m, 1H), 1.41 – 1.38 (m, 3H), 1.08 (s, 9H), 1.00 (s, 9H). Chapter 4 – Completion of Aconitine Core and Synthesis of (–)-Talatisamine, (–)-Liljestrandisine, and (–)-Liljestrandinine 153 Preparation of diol 241: OMe NHEt MeO OMe NHEt MeO HF • pyr MeCN, 20 °C O Si t-Bu O 71% yield t-Bu OH OH 241 123 A 20 mL Teflon vial was charged with siliconide 123 (0.116 g, 0.22 mmol, 1.0 equiv.) and MeCN (8 mL). HF•pyridine (0.116 g, ~70% HF) was diluted with MeCN (1.6 mL) and this solution was added dropwise to the vial of 123 and the reaction was stirred at 20 °C for 1 hour. The reactions was quenched with saturated aqueous NaHCO3 (10 mL) and the mixture was extracted with 4:1 CHCl3 : iPrOH mixture (5 x 10 mL). The organic extracts were dried (Na2SO4), filtered and concentrated in vacuo. The crude residue was purified by silica gel chromatography (2-3-4-5-6% (7N NH3 in MeOH) in CH2Cl2) to afford the diol 241 as a colorless oil (60.0 mg, 0.16 mmol, 71 % yield). 1 H NMR (400 MHz, Chloroform-d) δ 5.96 (dd, J = 9.5, 6.4 Hz, 1H), 5.88 (dt, J = 5.7, 2.2 Hz, 1H), 5.76 (ddd, J = 9.6, 3.6, 2.0 Hz, 1H), 5.47 (d, J = 3.6 Hz, 1H), 5.37 (dt, J = 5.8, 1.8 Hz, 1H), 4.38 (t, J = 4.7 Hz, 1H), 4.06 – 3.88 (m, 1H), 3.54 (dd, J = 7.3, 2.8 Hz, 1H), 3.25 (s, 3H), 3.24 (s, 3H), 3.19 (s, 2H), 2.81 (dd, J = 6.3, 4.3 Hz, 1H), 2.76 – 2.67 (m, 1H), 2.67 – 2.53 (m, 2H), 2.47 (d, J = 1.6 Hz, 2H), 2.39 (dd, J = 10.7, 7.7 Hz, 1H), 2.31 (dt, J = 8.3, 2.3 Hz, 2H), 1.70 – 1.45 (m, 3H), 1.38 – 1.28 (m, 1H), 1.07 (t, J = 7.0 Hz, 3H). 13 C NMR (101 MHz, CDCl3): δ 165.03, 134.31, 132.42, 131.58, 129.34, 120.14, 79.80, 77.36, 76.51, 74.85, 66.16, 59.05, 57.37, 56.97, 56.72, 47.78, 44.99, 44.74, 39.38, 35.45, 24.47, 21.91, 15.35. FTIR (NaCl, thin film): 3374, 2923, 2352, 1453, 1362, 1218, 1100, 1010, 960, 935, 817, 758, 733 cm-1. Chapter 4 – Completion of Aconitine Core and Synthesis of (–)-Talatisamine, (–)-Liljestrandisine, and (–)-Liljestrandinine 154 HRMS: (ESI-TOF) calc’d for C23H35NO4 [M + H]+ 390.2639, found 390.2621 [𝜶]𝟐𝟓 𝐃 = +98.2° (c = 0.82, CHCl3). Preparation of enone 242: OMe NHEt MeO OH 241 OH MeO 1. Cu(MeCN)4OTf, ABNO NMI, MeObpy, MeCN 2. TESOTf, imidazole CH2Cl2, –78 °C 73% yield (2 steps) OMe NHEt TESO O 1.5:1 mixture of diastereomers 204 0.64 mL of a stock solution of ABNO (1 mM), MeObpy (1 mM) and NMI (6 mM) (0.05 equiv. ABNO; 0.05 equiv. MeObpy; 0.40 equiv. NMI) was added to a ½ dram vial. Cu(MeCN)4OTf (0.2 mg, 0.64 µmol, 0.05 equiv.) was added, and the catalyst solution was stirred for 5 minutes. Diol 241 (5.0 mg, 13 µmol, 1.0 equiv.) in MeCN (0.5 mL) was added, and the mixture was stirred for 90 minutes at room temperature. The reaction mixture was filtered over a short pad of silica gel, eluting with 10% (7N NH3 in MeOH) in CH2Cl2. The filtrate was concentrated and taken on crude to the next step. The crude isolate from the Stahl oxidation was added to an oven-dried 10 mL roundbottom flask along with imidazole (3.3 mg, 48 µmol, 3.8 equiv.). The flask was put under N2 atmosphere before THF (1.0 mL) was added and the mixture was cooled to –78 °C. TESCl (4.9 mg, 32 µmol, 2.5 equiv.) in THF (0.1 mL) was added dropwise, and the mixture was stirred at –78 °C for 10 minutes. The reaction was quenched with saturated aqueous NaHCO3 (2 mL) and the mixture was extracted with 4:1 CHCl3 : iPrOH mixture (3 x 5 mL). The organic extracts were dried (Na2SO4), filtered and concentrated in vacuo. The crude residue was purified by silica gel chromatography (2-3% (7N NH3 in MeOH) in CH2Cl2) to afford 204 (4.7 Chapter 4 – Completion of Aconitine Core and Synthesis of (–)-Talatisamine, (–)-Liljestrandisine, and (–)-Liljestrandinine 155 mg, 9.5 µmol, 73% yield) as a ~1.5:1 mixture of diastereomers. The 1H NMR spectrum of the major diastereomer is reported below: 1 H NMR (600 MHz, Chloroform-d) δ 6.94 (ddd, J = 9.8, 6.4, 1.7 Hz, 1H), 5.94 (dt, J = 5.9, 2.3 Hz, 1H), 5.78 (dd, J = 9.7, 1.9 Hz, 1H), 5.66 (d, J = 3.8 Hz, 1H), 5.34 (dt, J = 5.9, 1.9 Hz, 1H), 4.66 (td, J = 4.5, 1.7 Hz, 1H), 3.52 (dd, J = 7.3, 2.7 Hz, 1H), 3.24 (d, J = 2.3 Hz, 3H), 3.22 (s, 3H), 3.21 – 3.16 (m, 2H), 3.13 (t, J = 5.5 Hz, 1H), 2.67 – 2.57 (m, 2H), 2.57 – 2.43 (m, 3H), 2.36 (dt, J = 10.1, 2.2 Hz, 2H), 1.71 – 1.62 (m, 1H), 1.57 (dtt, J = 12.7, 8.4, 3.8 Hz, 4H), 1.34 (q, J = 7.1, 6.1 Hz, 1H), 1.08 (t, J = 7.1 Hz, 3H), 0.92 (t, J = 7.9 Hz, 9H), 0.57 (q, J = 7.9 Hz, 6H). Preparation of N-chloroamine 205: MeO OMe NHEt MeO NCS OMe NEt Cl CH2Cl2, 0 °C TESO O 1.5:1 mixture of diastereomers 204 42% yield TESO O 1.5:1 mixture of diastereomers 205 A solution of secondary amine 204 (4.7 mg, 9.5 µmol, 1.0 equiv.) in CH2Cl2 was cooled to 0 °C before NCS (3.1 mg, 24 µmol, 2.5 equiv.) was added in one portion. The reaction was stirred for 30 minutes before filtering over a short pad of silica gel, eluting with EtOAc. The filtrate was concentrated in vacuo. The crude product was purified by column chromatography (10-20-30-40% EtOAc in hexanes) to afford N-chloroamine 205 (2.1 mg, 3.9 µmol, 42% yield) as a mixture of diastereomers. The 1H NMR spectrum of the major diastereomer is reported below: Chapter 4 – Completion of Aconitine Core and Synthesis of (–)-Talatisamine, (–)-Liljestrandisine, and (–)-Liljestrandinine 1 156 H NMR (600 MHz, Chloroform-d) δ 6.94 (ddd, J = 9.8, 6.5, 1.7 Hz, 1H), 5.93 (dt, J = 5.8, 2.3 Hz, 1H), 5.78 (dd, J = 9.6, 1.4 Hz, 1H), 5.66 (d, J = 3.7 Hz, 1H), 5.39 – 5.33 (m, 1H), 4.66 (td, J = 4.6, 1.7 Hz, 1H), 3.53 (dd, J = 7.6, 2.4 Hz, 1H), 3.29 (s, 2H), 3.25 – 3.21 (m, 6H), 3.18 (s, 1H), 3.14 – 3.09 (m, 1H), 3.10 – 3.05 (m, 1H), 3.02 – 2.93 (m, 3H), 2.54 (t, J = 9.3 Hz, 1H), 2.39 – 2.23 (m, 2H), 1.80 – 1.64 (m, 2H), 1.64 – 1.56 (m, 1H), 1.50 – 1.42 (m, 1H), 1.20 (t, J = 6.9 Hz, 3H), 0.92 (t, J = 8.0 Hz, 9H), 0.57 (q, J = 8.0 Hz, 6H). Tentative identification of cyclized product 208: MeO OMe NEt Cl OMe NEt MeO CuCl (20 mol %) tentative assignment THF, 50 °C TESO O 1.5:1 mixture of diastereomers 205 OTES 208 O CuCl (0.04 mg, 0.4 µmol, 0.2 equiv.) was dispensed as a stock solution in MeCN to a ½ dram vial. The MeCN was removed in vacuo. A solution of N-chloroamine 205 (1.0 mg, 1.9 µmol, 1.0 equiv.) in THF (0.2 mL) was added, and the mixture was heated to 50 °C for 12 hours. Saturated aqueous NaHCO3 (1 mL) was added, and the mixture was extracted with 4:1 CHCl3 : iPrOH mixture (3 x 5 mL). The organic extracts were dried (Na2SO4), filtered and concentrated in vacuo. Purification by preparative TLC (5% (7 N NH3 in MeOH) in CH2Cl2) gave a trace amount of a product tentatively assigned as 208 based on HRMS and 1H NMR. 1 H NMR (400 MHz, Chloroform-d) δ 7.06 (s, 1H), 5.96 (s, 1H), 5.92 (s, 1H), 5.77 (d, J = 9.8 Hz, 1H), 5.47 (s, 1H), 4.64 (s, 1H), 4.14 (s, 1H), 3.65 (s, 2H), 3.37 (s, 4H), 3.33 (s, 4H), 3.17 (s, 2H), 3.11 (s, 1H), 3.04 (s, 1H), 2.65 (s, 1H), 2.47 (d, J = 9.9 Hz, 2H), 2.38 (d, J = 6.6 Hz, Chapter 4 – Completion of Aconitine Core and Synthesis of (–)-Talatisamine, (–)-Liljestrandisine, and (–)-Liljestrandinine 157 1H), 2.19 – 2.08 (m, 2H), 1.12 – 1.05 (m, 4H), 0.92 (t, J = 7.8 Hz, 9H), 0.57 (q, J = 7.9 Hz, 6H). HRMS: (ESI-TOF) calc’d for C29H46NO4Si [M + H]+ 500.3196, found 500.3172 Preparation of aziridine 227:8 MeO OMe MeO NH2 OMe E N PhI(OAc)2 K2CO3, SiO2 DCE, 50 °C O Si O t-Bu t-Bu 116 74% yield O Si O t-Bu t-Bu 227 A 100 mL, round-bottom flask was charged with primary amine 116 (0.348 g, 0.694 mmol, 1 equiv.), PhI(OAc)2 (0.357 g, 1.11 mmol, 1.6 equiv.), K2CO3 (0.256 g, 1.85 mmol, 2.7 equiv.), SiO2 gel (17.3 g), and suspended in DCE (70 mL) under N2. The mixture was stirred for 30 minutes at room temperature, and then heated to 50 °C in an oil bath and was allowed to stir at 50 °C for 1 hour. At this time, the reaction was allowed to cool to room temperature, and was filtered through a plug of SiO2 and flushed with 8% 7 N NH3/ 92% CH2Cl2 solution to elute the product aziridine, and concentrated in vacuo. The resulting crude residue was purified by silica gel chromatography (4% to 6% MeOH in CH2Cl2) to afford aziridine 227 (0.257 g, 0.514 mmol, 74% yield) as a clear oil. 1 H NMR (400 MHz, Chloroform-d): δ 6.11 (ddd, J = 9.5, 6.1, 1.1 Hz, 1H), 5.67 (ddd, J = 9.3, 4.4, 1.9 Hz, 1H), 5.45 (dd, J = 3.9, 0.7 Hz, 1H), 4.51 (t, J = 4.7 Hz, 1H), 4.24 – 4.17 (m, 1H), 3.38 – 3.32 (m, 4H), 3.27 (s, 3H), 3.09 (d, J = 8.9 Hz, 1H), 3.05 – 3.01 (m, 1H), 2.99 – 2.91 (m, 2H), 2.88 (dd, J = 6.0, 4.6 Hz, 1H), 2.78 (d, J = 14.3 Hz, 1H), 2.60 – 2.58 (m, 1H), 2.48 – 2.46 (m, 1H), 2.17 – 2.08 (m, 1H), 1.82 (d, J = 13.4 Hz, 1H), 1.69 – 1.56 (m, 2H), 1.50 – 1.43 (m, 2H), 1.30 (td, J = 14.2, 3.7 Hz, 1H), 1.07 (s, 9H), 1.01 (s, 9H). Chapter 4 – Completion of Aconitine Core and Synthesis of (–)-Talatisamine, (–)-Liljestrandisine, and (–)-Liljestrandinine 13 158 C NMR (101 MHz, CDCl3): δ 13C NMR (101 MHz, CDCl3) δ 162.7, 135.0, 128.8, 123.9, 78.9 (2 overlapping 13C signals), 77.3, 66.3, 59.3, 56.8, 50.2, 50.0, 46.0, 45.1, 38.1, 35.9, 35.6, 33.7, 32.9, 28.9, 28.3, 24.0, 22.6, 21.2, 20.7. Two 13C signals are observed to be overlapping at 78.9. Additionally, a 13C signal is observed to be overlapping with residual CDCl3 at 77.3. Resolved peaks are seen in the HSQC spectrum, provided below. FTIR (NaCl, thin film): 2938, 2859, 2824, 2362, 1476, 1463, 1110, 993 cm-1. HRMS: (ESI-TOF) calc’d for C29H46NO4Si [M + H]+ 500.3191, found 500.3178 [𝜶]𝟐𝟓 𝐃 = +24.6° (c = 1.30, CHCl3). TLC (10%MeOH/90%CH2Cl2), Rf: 0.5 (UV, teal in p-anisaldehyde) Preparation of bromide 228: MeO OMe E OMe MeO N NAc AcBr 7 Br CH2Cl2, 0 °C O Si O t-Bu t-Bu 227 83% yield O Si O t-Bu 228 t-Bu A 100 mL, round-bottom flask was charged with aziridine 227 (198 mg, 0.397 mmol, and CH2Cl2 (7.8 mL) under N2, and cooled to 0 °C in an ice bath. Acetyl bromide (86.1 µL, 1.1645 mmol, 3 equiv.) added dropwise via microsyringe, and was allowed to stir for an additional 15 minutes while maintained at 0 °C. Reaction quenched with sat. NaHCO3 (10 mL), transferred to a separatory funnel, and the layers were separated. The aqueous layer was extracted with CH2Cl2 (3 x 20 mL). The combined organic extracts were dried over Na2SO4, filtered, and concentrated in vacuo. The resulting crude residue was purified by silica gel chromatography (30% to 40% to 50% ethyl acetate in hexanes) to afford bromide 228 (206 mg, 0.330 mmol, 83% yield) as a white solid. Chapter 4 – Completion of Aconitine Core and Synthesis of (–)-Talatisamine, (–)-Liljestrandisine, and (–)-Liljestrandinine 1 159 H NMR (400 MHz, Chloroform-d): δ 6.17 (ddd, J = 9.6, 6.1, 1.1 Hz, 1H), 5.73 – 5.68 (m, 2H), 4.76 – 4.71 (m, 1H), 4.65 (s, 1H), 4.24 – 4.19 (m, 1H), 4.09 – 4.03 (m, 2H), 3.29 (s, 3H), 3.23 (s, 3H), 3.14 (d, J = 9.2 Hz, 1H), 3.09 – 3.05 (m, 1H), 3.04 – 2.98 (m, 2H), 2.51 (dd, J = 14.6, 8.8 Hz, 1H), 2.38 (d, J = 14.4 Hz, 1H), 2.34 – 2.23 (m, 2H), 2.18 (s, 3H), 2.07 – 1.98 (m, 1H), 1.74 – 1.67 (m, 1H), 1.50 – 1.32 (m, 2H), 1.16 – 1.08 (m, 1H), 1.08 (s, 9H), 1.03 (s, 9H). 13 C NMR (101 MHz, CDCl3): δ 169.0, 135.9, 128.2, 124.9, 80.6, 78.1, 77.6, 66.6, 66.2, 59.4, 55.7, 53.1, 47.3, 46.3, 46.0, 45.7, 42.8, 35.5, 35.4, 32.2, 29.7, 29.0, 28.3, 24.6, 21.8, 21.3, 20.8. FTIR (NaCl, thin film): 2935, 2867, 2826, 1658, 1631, 1471, 1462, 1427, 1232, 1109 cm-1. HRMS: (ESI-TOF) calc’d for C31H49BrNO5Si [M + H]+ 622.2558, found 622.2540 [𝜶]𝟐𝟓 𝐃 = +50.9° (c = 1.32, CHCl3). TLC (50%EtOAc/50%hexanes), Rf: 0.4 (UV, green in p-anisaldehyde) Preparation of diol 229: MeO OMe NAc HF•pyr 7 Br O Si O t-Bu t-Bu 228 OMe MeO NAc Br MeCN 100% yield OH OH 229 A 100 mL, round-bottom flask was charged with silylene 228 (206 mg, 0.331 mmol, 1 equiv.) and MeCN (6 mL). A solution of HF•Py (pyridine ~30%, HF ~70%, 206 mg) in MeCN (1 mL) was added, and the reaction was allowed to stir for 30 minutes at room temperature. The reaction was quenched with sat. NaHCO3 (20 mL), transferred to a separatory funnel and extracted with 20%IPA/80%CHCl3 (5 x 20 mL). The combined organic extracts were dried over Na2SO4, filtered, and concentrated in vacuo. The resulting crude residue was dissolved in CH2Cl2, filtered through a plug of SiO2, flushed with CH2Cl2 to remove the nonpolar impurities Chapter 4 – Completion of Aconitine Core and Synthesis of (–)-Talatisamine, (–)-Liljestrandisine, and (–)-Liljestrandinine 160 and then 5% MeOH/95% CH2Cl2 to elute the product to afford diol 229 (160 mg, 0.331 mmol, 100% yield) as a clear oil. 1 H NMR (400 MHz, Chloroform-d): δ 6.10 (ddt, J = 9.6, 6.3, 1.1 Hz, 1H), 5.84 – 5.78 (m, 2H), 4.70 – 4.61 (m, 2H), 4.09 – 4.02 (m, 2H), 3.96 (s, 1H), 3.29 (s, 3H), 3.21 (s, 3H), 3.18 (dd, J = 7.8, 2.4 Hz, 1H), 3.13 (d, J = 9.2 Hz, 1H), 3.03 (d, J = 9.1 Hz, 1H), 2.96 – 2.88 (m, 1H), 2.84 – 2.78 (m, 1H), 2.77 – 2.73 (m, 1H), 2.55 – 2.47 (m, 1H), 2.38 (dd, J = 14.4, 2.1 Hz, 1H), 2.31 – 2.24 (m, 2H), 2.19 (s, 3H), 2.06 – 1.98 (m, 1H), 1.74 – 1.66 (m, 2H), 1.46 (dddd, J = 17.2, 12.6, 4.4, 2.2 Hz, 1H), 1.41 – 1.27 (m, 1H). 13 C NMR (101 MHz, CDCl3): δ 169.1, 132.4, 129.0, 125.1, 80.8, 78.0, 75.1, 66.6, 65.6, 59.4, 55.8, 52.9, 47.4, 47.4, 45.5, 44.5, 42.8, 35.5, 35.4, 32.2, 24.5, 21.8. A 13C signal is observed to be missing in the 13C NMR, but can be seen in the HMBC spectrum at 159.5, which is provided below. FTIR (NaCl, thin film): 3367, 2930, 2877, 2826, 2239, 1624, 1426, 1234, 1094, 917 cm–1. HRMS: (ESI-TOF) calc’d for C23H33BrNO5 [M + H]+ 482.1537, found 482.1539 [𝜶]𝟐𝟓 𝐃 = +39.1° (c = 0.55, CHCl3). TLC (100%EtOAc), Rf: 0.25 (UV, green/blue in p-anisaldehyde) Preparation of enone 242:12 OMe NAc MeO Br OH 229 OH Cu(MeCN)4OTf, ABNO, NMI, MeObpy OMe NAc MeO Br MeCN (quantitative, used crude in next step) OH O 242 A 100 mL, round-bottom flask was charged with diol 229 (159 mg, 0.3308 mmol, 1.0 equiv.) and MeCN (3.3 mL). A homemade stock-solution [see note below] 0.05M 4,4’- Chapter 4 – Completion of Aconitine Core and Synthesis of (–)-Talatisamine, (–)-Liljestrandisine, and (–)-Liljestrandinine 161 dimethoxy-2,2’-bipyridine (MeObpy), 0.05 M in ABNO, and 0.2 M N-methylimidazole was added to the solution of diol (0.61 mL, 0.031 mmol, 0.1 equiv.) followed by Cu(MeCN)4OTf (11.6 mg, 0.031 mmol, 0.1 equiv.) was added, and the clear red/brown reaction mixture was stirred until slightly yellow green, and the TLC indicated the reaction had gone to completion (ca. 90 min), at which point the solution was filtered through a short plug of silica that had been pre-packed with 5%MeOH/95% CH2Cl2, and flushed with more 5%MeOH/95% CH2Cl2 to elute the product, and concentrated in vacuo. The crude enone was subjected to the next step without further purification. For characterization purposes, 242 could be purified via silica gel chromatography (1% to 2% to 3% to 4% to 5% MeOH in CH2Cl2). Notes: (1) A 0.05 M solution of 4,4’-dimethoxy-2,2’-bipyridine, 0.05M ABNO and 0.2M NMI solution in MeCN could be pre-made and kept in an 8 °C freezer and was stable and good to use for an extended time (>4 months). This homemade stock solution was used in this reaction, and makes it easy to add more catalyst. (2) This selective oxidation to the enone is very clean and performs well, but oxidation to the diketone has been observed if too much catalyst is added. It is important to monitor this reaction by TLC. It is best to run this reaction carefully, and start with less catalyst, because more can be added if needed to reach full conversion to the desired product. 1 H NMR (400 MHz, Chloroform-d): δ 7.19 (ddd, J = 9.8, 6.4, 1.6 Hz, 1H), 5.95 – 5.90 (m, 2H), 5.18 – 5.11 (m, 1H), 4.71 (s, 1H), 4.13 – 4.04 (m, 2H), 3.46 (dd, J = 6.4, 4.4 Hz, 1H), 3.33 (dddd, J = 4.8, 3.9, 2.0, 1.0 Hz, 1H), 3.29 (s, 3H), 3.29 – 3.20 (m, 1H), 3.21 (s, 3H), 3.14 (d, J = 9.2 Hz, 1H), 3.04 (d, J = 9.2 Hz, 1H), 2.64 – 2.56 (m, 2H), 2.41 (dd, J = 14.6, 2.3 Hz, Chapter 4 – Completion of Aconitine Core and Synthesis of (–)-Talatisamine, (–)-Liljestrandisine, and (–)-Liljestrandinine 162 1H), 2.36 (d, J = 6.3 Hz, 1H), 2.28 (dt, J = 15.3, 6.0 Hz, 1H), 2.21 (s, 3H), 2.05 – 1.98 (m, 1H), 1.75 – 1.69 (m, 1H), 1.51 – 1.32 (m, 2H). 13 C NMR (101 MHz, CDCl3): δ 195.1, 169.0, 160.1 (br), 150.9, 126.3, 124.1, 85.0, 82.0, 77.9, 66.8, 60.9, 59.4, 55.1, 52.9, 47.8, 47.5, 45.4, 42.7, 35.5, 35.3, 32.0, 24.5, 21.9. FTIR (NaCl, thin film): 3367, 2930, 2877, 2826, 2239, 1624, 1426, 1234, 1094, 917 cm-1. HRMS: (ESI-TOF) calc’d for C23H31BrNO5 [M + H]+ 480.1380, found 480.1367 [𝜶]𝟐𝟓 𝐃 = +169° (c = 0.50, CHCl3). TLC (100%EtOAc), Rf: 0.25 (UV, pink in p-anisaldehyde) Preparation of methoxymethyl ether 230: OMe NAc MeO OH 242 OMe NAc MeO MOMCl i-PrNEt, TBAI Br DMF, 60 °C O 76% yield (2 steps) Br OMOM O 230 A 100 mL, round-bottom flask was charged with crude enone 242 (159 mg, 0.3308 mmol, 1.0 equiv.), TBAI (61 mg, 0.165 mmol, 0.50 mol %), and DMF (6.6 mL). i-Pr2NEt (0.87 mL, 4.96 mmol, 15.0 equiv.) added followed by dropwise addition of MOMCl (0.25 mL, 3.31 mmol, 10 equiv.), causing a smokiness to evolve from the reaction. The flask was then lowered into a 60 °C oil bath, and let stir for 16 hours at this temperature. The reaction was then cooled to room temperature, diluted with sat. NH4Cl (20 mL), transferred to a separatory funnel and extracted with EtOAc (4 x 20 mL). The combined organic extracts were dried over Na2SO4, filtered, and concentrated in vacuo. The resulting crude residue was purified by silica gel chromatography (10% to 20% to 30% to 40% acetone in hexanes) to afford methoxy methyl ether 230 (131 mg, 0.251 mmol, 76% yield over two steps) as a white foam. Chapter 4 – Completion of Aconitine Core and Synthesis of (–)-Talatisamine, (–)-Liljestrandisine, and (–)-Liljestrandinine 1 163 H NMR (400 MHz, Chloroform-d): δ 7.11 (ddd, J = 9.8, 6.4, 1.6 Hz, 1H), 5.95 (d, J = 4.0 Hz, 1H), 5.81 (ddd, J = 9.8, 1.9, 0.6 Hz, 1H), 4.92 (q, J = 4.5 Hz, 1H), 4.70 (s, 1H), 4.65 (s, 2H), 4.12 – 4.04 (m, 2H), 3.50 – 3.46 (m, 1H), 3.41 – 3.37 (m, 1H), 3.32 (s, 3H), 3.27 (s, 3H), 3.27 – 3.19 (m, 1H), 3.19 (s, 3H), 3.13 (d, J = 9.2 Hz, 1H), 3.03 (d, J = 9.2 Hz, 1H), 2.62 – 2.54 (m, 1H), 2.42 – 2.33 (m, 2H), 2.31 – 2.23 (m, 1H), 2.21 (s, 3H), 2.04 – 1.96 (m, 1H), 1.74 – 1.66 (m, 1H), 1.47 – 1.33 (m, 2H). 13 C NMR (101 MHz, CDCl3): δ 195.6, 168.9, 160.0 (br), 150.6, 125.4, 123.9, 96.1, 90.6, 82.0, 77.9, 66.7, 59.4, 59.3, 55.9, 55.1, 52.9, 47.6, 46.2, 45.5, 42.7, 35.4, 35.3, 31.9, 24.5, 21.9. FTIR (NaCl, thin film): 3454, 3056, 2920, 2825, 2244, 1703, 1682, 1654, 1621, 1454, 1109, 1035, 918 cm-1. HRMS: (ESI-TOF) calc’d for C25H35BrNO6 [M + H]+ 524.1642, found 524.1689 [𝜶]𝟐𝟓 𝐃 = +179° (c = 0.50, CHCl3). TLC (100%EtOAc), Rf: 0.5 (UV, pink in p-anisaldehyde) Preparation of hexacycle 231: OMe NAc MeO Br OMOM 230 OMe NAc MeO n-Bu3SnH AIBN benzene 80 °C O 99% yield OMOM O 231 A 100 mL, round-bottom flask was charged with enone 230 (234 mg, 0.45 mmol, 1.0 equiv.) and dissolved in benzene (89 mL). The solution was degassed (freeze-pump-thaw with N2, 3 cycles) and the headspace was purged with argon. Meanwhile, a solution of AIBN (36.6 mg, 0.22 mmol, 0.5 equiv.) and n-Bu3SnH (0.72 mL, 2.68 mmol, 6.0 equiv.) in benzene (7 mL) was degassed in the same manor. The round-bottom flask containing the substrate was Chapter 4 – Completion of Aconitine Core and Synthesis of (–)-Talatisamine, (–)-Liljestrandisine, and (–)-Liljestrandinine 164 then heated to 80 °C in an oil bath, and the AIBN/n-Bu3SnH solution was added over 7 hours to the substrate solution via syringe pump. After the addition of the AIBN/n-Bu3SnH solution was complete, the reaction was cooled to room temperature and concentrated in vacuo. The resulting crude residue was purified by chromatography with a 90% SiO2/ 10% ground K2CO3 solid support13 eluting (20% to 30% to 40% to 50% acetone in hexanes) to afford hexacycle 231 (205 mg, 0.45 mmol, 99% yield) as a clear oil. 1 H NMR (400 MHz, Chloroform-d): δ 5.83 (d, J = 2.6 Hz, 1H), 4.68 – 4.64 (m, 2H), 4.48 (t, J = 5.6 Hz, 1H), 4.07 (s, 1H), 3.74 (d, J = 14.6 Hz, 1H), 3.58 (dd, J = 10.4, 7.2 Hz, 1H), 3.34 (s, 3H), 3.30 (s, 3H), 3.29 (s, 3H), 3.24 – 3.19 (m, 2H), 3.03 (d, J = 9.0 Hz, 1H), 2.91 (t, J = 5.7 Hz, 1H), 2.73 – 2.62 (m, 2H), 2.33 – 2.22 (m, 3H), 2.16 – 2.06 (m, 5H), 2.01 – 1.94 (m, 1H), 1.78 – 1.67 (m, 2H), 1.44 (dddd, J = 14.9, 13.2, 4.0, 1.7 Hz, 1H), 1.35 – 1.30 (m, 1H). 13 C NMR (101 MHz, CDCl3): δ 204.6, 169.8, 153.4, 121.4, 96.3, 80.0, 79.0, 78.2, 65.0, 59.7, 59.4, 55.9, 55.1, 51.5, 46.2, 45.4, 42.5, 42.2, 39.2, 37.7, 33.8, 31.9, 30.5, 24.7, 22.5. 2-dimensional NMR data is included below (HSQC, HMBC, COSY, NOESY). FTIR (NaCl, thin film): 2924, 2825, 1708, 1644, 1418, 1405, 1150, 1112, 1091, 1045, 1001, 919 cm-1. HRMS: (ESI-TOF) calc’d for C25H39N2O6 [M + NH4]+ 463.2803, found 463.2797 [𝜶]𝟐𝟓 𝐃 = +84.5° (c = 1.0, CHCl3). TLC (100%EtOAc), Rf: 0.3 (UV, brown in p-anisaldehyde) Chapter 4 – Completion of Aconitine Core and Synthesis of (–)-Talatisamine, (–)-Liljestrandisine, and (–)-Liljestrandinine 165 Preparation of carbocycle 232: OMe NAc MeO OMe NAc MeO Pd/C H2 H EtOH OMOM O 84% yield 231 OMOM O 232 A 50 mL, round-bottom flask was charged with alkene 231 (100 mg, 0.224 mmol), Pd/C (10% wt % palladium, contains 67% H2O, 300 mg, 0.282 mmol, 1.25 equiv.), and EtOH (5 mL). The flask was then equipped with a rubber septum, and the suspension was sparged with an H2 balloon for 10 minutes, then let stir under H2 for an additional 20 minutes. The reaction was monitored by TLC and LCMS, indicating that the reaction had gone to completion. The reaction mixture was then sparged with an argon balloon for 20 minutes, then filtered through a plug of SiO2 and celite that had been pre-packed with 5%MeOH/95% CH2Cl2, flushed with more 5%MeOH/95% CH2Cl2, and then concentrated in vacuo. The resulting crude residue was purified by silica gel chromatography (20% to 30% to 40% to 50% acetone in hexanes) to afford 232 (84 mg, 0.187 mmol, 84% yield) as a white foam. 1 H NMR (400 MHz, Chloroform-d): δ 4.60 (s, 2H), 4.11 (t, J = 5.0 Hz, 1H), 3.99 (s, 1H), 3.86 (d, J = 14.4 Hz, 1H), 3.31 (s, 3H), 3.28 (s, 3H), 3.20 (m, 5H), 3.09 (dd, J = 10.4, 7.1 Hz, 1H), 2.98 (d, J = 9.1 Hz, 1H), 2.76 (dd, J = 7.8, 5.6 Hz, 1H), 2.70 (dd, J = 19.6, 11.4 Hz, 1H), 2.58 (d, J = 15.0 Hz, 1H), 2.56 – 2.47 (m, 1H), 2.44 – 2.37 (m, 1H), 2.37 – 2.29 (m, 1H), 2.06 (d, J = 8.2 Hz, 7H), 1.92 – 1.85 (m, 2H), 1.71 – 1.64 (m, 2H), 1.43 (tdd, J = 13.4, 4.4, 1.9 Hz, 1H), 1.33 – 1.25 (m, 1H). 13 C NMR (101 MHz, CDCl3): δ 212.9, 169.7, 96.0, 84.0, 79.7, 78.2, 59.4, 56.8, 55.8, 55.6, 52.3, 48.3, 46.8, 46.1, 44.5, 43.9, 39.1, 37.4, 36.6, 33.8, 32.0, 28.6, 27.3, 25.2, 22.5. TLC (100%EtOAc), Rf: 0.3 (UV, yellow in p-anisaldehyde) Chapter 4 – Completion of Aconitine Core and Synthesis of (–)-Talatisamine, (–)-Liljestrandisine, and (–)-Liljestrandinine 166 FTIR (NaCl, thin film): 2913, 2241, 1714, 1698, 1650, 1644, 1634, 1614, 1470, 1454, 1108, 1041, 919 cm-1. HRMS: (ESI-TOF) calc’d for C25H38NO6 [M + H]+ 448.2694, found 448.2687 [𝜶]𝟐𝟓 𝐃 = –57.4° (c = 2.47, CHCl3). Preparation of β-hydroxy ketone 234: OMe MeO NAc H LiHMDS Cl t-Bu N S OMe NAc MeO Ph 233 OMOM 16 THF, –78 to 21 °C; then H2O, pyr O 97% yield MOMO 232 OH O 234 A 50 mL round-bottom flask was charged with ketone 232 (50.0 mg, 0.11 mmol, 1.0 equiv.) and put under N2. THF (4.5 mL) was added and the solution cooled to -78 °C. A solution of LiHMDS (1M in THF, 0.25 mL, 0.25 mmol, 2.2 equiv.) was added dropwise over 10 minutes. The mixture was stirred at -78 °C for 15 minutes before a solution of N-tert-butylphenylsulfinyl chloride (193 mg, 0.89 mmol, 8 equiv.) in THF (2.1 mL) was added dropwise over 5 minutes. The reaction was stirred at -78 °C for 15 minutes, then warmed to 21 °C and stirred 1 hour. Pyridine (0.05 mL) and water (1.7 mL) were added, and the mixture stirred at 21 °C for 16 hours. The reaction was quenched with saturated aqueous NaHCO3 (10 mL), and extracted with 3:1 CHCl3:iPrOH (5 x 10 mL). The combined organic extracts were dried (Na2SO4), filtered and concentrated to give the crude product as a yellow oil. The reaction was purified by column chromatography (1-2-3-4-5-6-7% MeOH in CH2Cl2) to give β-hydroxy ketone 234 (50.0 mg, 0.11 mmol, 97% yield) as a white solid. 1 H NMR (600 MHz, Chloroform-d) δ 4.69 – 4.64 (m, 2H), 4.27 (t, J = 5.1 Hz, 1H), 3.98 (s, 1H), 3.90 (d, J = 14.3 Hz, 1H), 3.35 (s, 3H), 3.31 (s, 3H), 3.25 – 3.10 (m, 6H), 3.03 (s, 1H), Chapter 4 – Completion of Aconitine Core and Synthesis of (–)-Talatisamine, (–)-Liljestrandisine, and (–)-Liljestrandinine 167 2.88 (dd, J = 7.8, 5.2 Hz, 1H), 2.79 (d, J = 2.3 Hz, 1H), 2.63 (d, J = 13.9 Hz, 1H), 2.51 – 2.44 (m, 2H), 2.21 – 2.04 (m, 8H), 2.02 (d, J = 8.0 Hz, 1H), 1.92 (d, J = 7.5 Hz, 1H), 1.80 (dd, J = 15.1, 7.8 Hz, 1H), 1.71 (ddd, J = 13.6, 5.1, 2.8 Hz, 1H), 1.46 (tdd, J = 13.7, 4.7, 1.9 Hz, 1H), 1.34 – 1.26 (m, 1H). 13 C NMR (101 MHz, CDCl3): δ 210.4, 169.7, 96.3, 82.7, 80.5, 78.0, 73.4, 59.5, 58.5, 56.1, 55.5, 52.2, 51.7, 50.6, 48.3, 45.6, 45.3, 44.4, 44.3, 37.4, 31.9, 27.0, 25.3, 25.2, 22.5. FTIR (NaCl, thin film): 3422, 2926, 2891, 2826, 2242, 1713, 1633, 1462, 1434, 1153, 1106, 1043, 920 cm-1. HRMS: (ESI-TOF) calc’d for C25H38NO7 [M + H]+ 464.2643, found 464.2646 [𝜶]𝟐𝟓 𝐃 = –52.6° (c = 0.70, CHCl3). TLC (10%MeOH/90%CH2Cl2), Rf: 0.5 (UV, yellow in p-anisaldehyde) SmI2/NEt3/H2O Reduction of Ketone 234 to diols 235 and 236:14 OMe MeO OMe MeO NAc NAc OMe MeO NAc SmI2 (2.5 equiv.) O MOM O H 234 O H2O (6.25 equiv.) Et3N (5 equiv.) THF, -78 to 0 °C H OH O MOM O H O OH 235 29% yield MOM O H H 236 58% yield Preparation of SmI2: 1,2-diiodoethane was purified prior to use as follows: 1,2-diiodoethane (1.6 g) was dissolved in Et2O (50 mL) and washed with sat. aq. Na2S2O3 (3 x 10 mL) and deionized water (2 x 10 mL), dried over Na2SO4, filtered, and concentrated in vacuo to give 1.41 g of a white solid. A 100 mL Schlenk flask containing a stir bar was charged with freshly filed Sm metal (650 mg). The system was flame-dried under high vacuum then cooled to ambient temperature before adding freshly purified 1,2-diiodoethane (700 mg). The Chapter 4 – Completion of Aconitine Core and Synthesis of (–)-Talatisamine, (–)-Liljestrandisine, and (–)-Liljestrandinine 168 atmosphere was exchanged three times for argon. Subsequently, the flask was charged with anhydrous THF (25 mL) that had been submitted to five freeze-pump-thaw cycles. Note: The THF used for the synthesis of SmI2 must contain <50 ppm H2O; THF containing greater quantities of water resulted in excessive induction times for the synthesis of SmI2. The suspension was stirred for 2 min and the flask was cautiously and briefly (5 s) placed under partial high vacuum, then purged with argon. This process was repeated two additional times to remove ethylene gas formed from insertion of Sm metal into 1,2-diiodoethane. The resulting heterogeneous suspension was rapidly (930 rpm) stirred; after 5 min, the reaction turned dark green, and within 10 min, a dark blue color was observed. After stirring under argon for 3 h at 19 °C, the system was cautiously and briefly placed under high vacuum, then purged with argon. This process was repeated two additional times, then stirring was halted. The mixture was allowed to settle for 15 min prior to use. A 5 mL round-bottom flask charged with ketone 234 (12.0 mg, 26 µmol, 1.0 equiv.) and put under argon. THF (1.2 mL) degassed by 3 freeze-pump-thaw cycles was added, and the mixture was cooled to -78 °C. In a separate flask, a stock solution of NEt3 (0.20 mL, 1.4 mmol, 50 equiv.) and water (30 µL, 1.7 mmol, 65 equiv.) in THF (4.8 mL). 0.50 mL of the Et3N/H2O stock solution (5.0 equiv Et3N, 6.5 equiv H2O) was added to a third flask containing freshly prepared SmI2 solution (~0.1 M in THF, 0.65 mL, 65 µmol, 2.5 equiv.). The SmI2/Et3N/H2O solution was added dropwise to the substrate, and the mixture stirred at -78 °C for 5 minutes before warming to 0 °C and stirring an additional 90 minutes. The reaction was quenched with saturated aqueous NaHCO3 (3 mL). The mixture was extracted with 3:1 CHCl3:iPrOH (5 x 4 mL). The combined organic extracts were dried (Na2SO4), filtered and concentrated in vacuo. The crude residue was purified via column chromatography (SiO2 2-3- Chapter 4 – Completion of Aconitine Core and Synthesis of (–)-Talatisamine, (–)-Liljestrandisine, and (–)-Liljestrandinine 169 4-5-6% MeOH in CH2Cl2) to afford C16-equitorial diol 236 (7.0 mg, 15 µmol, 58% yield) and C16-axial diol 235 (3.5 mg, 7.5 µmol, 29% yield) as colorless oils. NaBH4 Reduction of Ketone 234 to diols 235 and 236: OMe MeO OMe MeO NAc NAc OMe MeO NAc NaBH4 MeOH, 0 °C H O MOM O H OH O O MOM 234 O H O OH MOM 235 41% yield O H H 236 21% yield MeOH (5 mL) was added to a 25 mL round-bottom flask charged with ketone 234 (22 mg, 47 µmol, 1.0 equiv.) and the solution cooled to 0 °C. NaBH4 (36 mg, 0.47 mmol, 10 equiv.) was added slowly in portions (granual by granual). After complete addition of the reductant, the reaction was stirred for an additional 30 minutes before quenching with 0.5 M NaOH (5 mL). The mixture was stirred for 30 minutes, then extracted with 3:1 CHCl3:iPrOH (5 x 10 mL). The combined organic extracts were dried (Na2SO4), filtered and concentrated in vacuo. The crude residue was purified via column chromatography (SiO2 2-3-4-5-6% MeOH in CH2Cl2) to afford C16-axial diol 235 (9.0 mg, 19 µmol, 41% yield) and C16-equitorial diol 236 (4.6 mg, 9.9 µmol, 21% yield) as colorless oils. Characterization of axial C16-alcohol 235: 1 H NMR (600 MHz, Chloroform-d) δ 4.75 (s, 2H), 4.12 (t, J = 5.0 Hz, 1H), 4.01 (s, 1H), 3.87 (d, J = 14.3 Hz, 1H), 3.72 (t, J = 9.7 Hz, 1H), 3.43 (s, 3H), 3.31 (s, 3H), 3.22 (s, 3H), 3.20 (d, J = 1.8 Hz, 1H), 3.20 – 3.05 (m, 2H), 3.00 (t, J = 4.6 Hz, 1H), 2.67 – 2.54 (m, 2H), 2.38 – 2.29 (m, 2H), 2.15 (s, 3H), 2.10 – 2.02 (m, 4H), 1.91 (d, J = 7.7 Hz, 1H), 1.88 – 1.79 (m, 3H), 1.74 – 1.64 (m, 2H), 1.43 (td, J = 14.2, 4.4 Hz, 1H), 1.38 – 1.24 (m, 2H). Chapter 4 – Completion of Aconitine Core and Synthesis of (–)-Talatisamine, (–)-Liljestrandisine, and (–)-Liljestrandinine 13 170 C NMR (101 MHz, CDCl3) δ 169.9, 970, 83.1, 81.8, 78.3, 72.9, 72.4, 59.6, 59.1, 56.20, 55.8, 53.2, 48.4, 45.6, 44.7, 44.0, 41.2, 37.6, 32.2, 27.6, 25.4, 22.7. FTIR (NaCl, thin film): 3480, 3451, 2926, 2824, 2530, 2508, 2490, 2044, 1686, 1618, 1435, 1212, 1152, 1166, 1106, 1058, 1038, 913, 868, 825, 777, 755, 732, 685, 668 cm-1. HRMS: (ESI-TOF) calc’d for C25H39NO7Na [M + Na]+ 488.2619, found 488.2603 diff.: 3.41 ppm. [𝜶]𝟐𝟏 𝐃 = –57.5° (c = 0.21, CHCl3). TLC (6% MeOH/94% CH2Cl2), Rf: 0.5 (Blue in p-anisaldehyde) Characterization of C16-equitorial alcohol 236: 1 H NMR (400 MHz, Chloroform-d) δ 4.67 (d, J = 3.1 Hz, 2H), 4.45 (d, J = 4.7 Hz, 2H), 4.05 (dd, J = 6.0, 4.3 Hz, 1H), 3.79 (d, J = 14.4 Hz, 1H), 3.40 (s, 3H), 3.30 (s, 3H), 3.26 (s, 3H), 3.21 (d, J = 8.9 Hz, 1H), 3.15 (dd, J = 10.6, 7.0 Hz, 1H), 3.00 (d, J = 9.0 Hz, 1H), 2.68 (d, J = 14.3 Hz, 1H), 2.36 – 2.24 (m, 3H), 2.24 – 2.15 (m, 4H), 2.11 – 2.05 (m, 2H), 1.92 (d, J = 7.6 Hz, 1H), 1.89 – 1.76 (m, 3H), 1.72 – 1.63 (m, 3H), 1.44 – 1.38 (m, 1H), 1.32 – 1.29 (m, 1H). 13 C NMR (101 MHz, CDCl3) δ 170.3, 96.1, 83.7, 80.5, 78.3, 75.3, 66.1, 59.7, 59.6, 56.1, 56.0, 52.4, 49.0, 45.4, 45.4, 44.9, 44.5, 41.9, 40.7, 37.8, 32.41, 29.9, 25.7, 25.0, 23.0, 22.6. FTIR (NaCl, thin film): 3897, 3413, 2928, 1721, 1622, 1429, 1379, 1258, 1212, 1166, 1152, 1106, 1061, 1039, 996, 942, 918, 884, 798, 766, 748, 664 cm-1. HRMS: (ESI-TOF) calc’d for C25H39NO7Na [M + Na]+ 488.2619, found 488.2612 diff.: 1.85 ppm [𝜶]𝟐𝟏 𝐃 = –19.4° (c = 0.17, CHCl3). TLC (6% MeOH/94% CH2Cl2), Rf: 0.5 (Blue in p-anisaldehyde) Chapter 4 – Completion of Aconitine Core and Synthesis of (–)-Talatisamine, (–)-Liljestrandisine, and (–)-Liljestrandinine 171 Preparation of amine 245 from amide 235: OMe NAc MeO OMe NEt MeO LiAlH4 Et2O, 0 to 40 °C 75% yield MOMO HO OH MOMO HO OH 235 245 A 1-dram vial charged with amide 235 (4.0 mg, 8.6 µmol, 1.0 equiv.) and Et2O (1 mL) was cooled to 0 °C. A solution of LiAlH4 (1 M in THF, 0.04 mL, 40 µmol, 5 equiv.) was added dropwise. The reaction was heated to 40 °C and stirred for 4 hours. The reaction was cooled to 0 °C before quenching with 0.5 M NaOH (1 mL). The mixture was extracted with 3:1 CHCl3:iPrOH (4 x 2 mL). The combined organic phases were dried (Na2SO4), filtered and concentrated in vacuo. The crude mixture was purified by column chromatography (SiO2, 12-3-4-5% 7N NH3 in MeOH/CH2Cl2) to afford 245 as a colorless oil (3.0 mg, 6.6 µmol, 77% yield). Characterization below. Preparation of amine 237 from amide 236: OMe NAc MeO OMe NEt MeO LiAlH4 OH Et2O, 0 to 40 °C OH 27% yield MOMO HO MOMO HO 236 237 A 1-dram vial charged with amide 236 (5.0 mg, 11 µmol, 1.0 equiv.) and Et2O (1 mL) was cooled to 0 °C. A solution of LiAlH4 (1 M in THF, 0.06 mL, 60 µmol, 5.5 equiv.) was added dropwise. The reaction was heated to 40 °C and stirred for 4 hours. The reaction was cooled to 0 °C before quenching with 0.5 M NaOH (1 mL). The mixture was extracted with Chapter 4 – Completion of Aconitine Core and Synthesis of (–)-Talatisamine, (–)-Liljestrandisine, and (–)-Liljestrandinine 172 3:1 CHCl3:iPrOH (4 x 2 mL). The combined organic phases were dried (Na2SO4), filtered and concentrated in vacuo. The crude mixture was purified by column chromatography (SiO2, 12-3-4-5% 7N NH3 in MeOH/CH2Cl2) to afford amine 237 as a colorless oil (1.3 mg, 2.8 µmol, 27% yield). 1 H NMR (600 MHz, Benzene-d6) δ 4.41 – 4.29 (m, 3H), 3.87 (t, J = 5.3 Hz, 1H), 3.74 (s, 1H), 3.22 (s, 1H), 3.13 (s, 3H), 3.12 – 3.10 (m, 6H), 3.09 – 3.03 (m, 1H), 2.99 – 2.92 (m, 2H), 2.81 (d, J = 8.6 Hz, 1H), 2.56 – 2.43 (m, 4H), 2.35 – 2.26 (m, 3H), 2.20 – 2.12 (m, 2H), 2.10 – 2.01 (m, 3H), 1.99 – 1.95 (m, 1H), 1.92 – 1.87 (m, 1H), 1.62 (d, J = 7.4 Hz, 1H), 1.53 – 1.49 (m, 2H), 1.41 – 1.32 (m, 2H), 1.05 (t, J = 7.1 Hz, 3H). 13 C NMR (101 MHz, C6D6) δ 95.8, 86.2, 81.0, 79.6, 75.7, 66.3, 63.4, 59.1, 55.8, 55.4, 53.8, 49.7, 46.4, 46.1, 45.8, 45.6, 42.7, 41.2, 39.1, 33.4, 30.2, 26.2, 25.8, 23.8, 13.9. FTIR (NaCl, thin film): 3420, 2924, 2820, 1719, 1453, 1388, 1290, 1260, 1203, 1170, 1150, 1110, 1040, 966, 919, 804, 753, 724, 657 cm-1. HRMS: (ESI-TOF) calc’d for C25H42NO6 [M +H]+ 452.3003, found: 452.3007 diff: 0.92 ppm [𝜶]𝟐𝟏 𝐃 = –9.3° (c = 0.18, CHCl3). TLC (6% MeOH/94% CH2Cl2), Rf: 0.2 (KMnO4) Chapter 4 – Completion of Aconitine Core and Synthesis of (–)-Talatisamine, (–)-Liljestrandisine, and (–)-Liljestrandinine 173 Preparation of diol 245: MeO OMe NAc OMe NEt MeO RedAl® Et2O 0 to 21 °C MOMO OH 234 O 58% yield 16 MOMO HO OH 245 β-hydroxy ketone 234 (13 mg, 28 µmol, 1 equiv.) was dissolved in Et2O (1.5 mL) under N2 and cooled to 0 °C. A solution of RedAl in toluene (>46%, ~3M, 0.16 mL, 0.48 mmol, 17 equiv.) was added dropwise over 10 minutes. The reaction was stirred at 0 °C for 30 min. before warming to room temp and stirring an additional 2h. The solution was cooled to 0 °C and quenched with 0.5 M NaOH (aq., 2 mL) and stirred for 15 minutes. The mixture was extracted with 3:1 CHCl3:iPrOH (5 x 3 mL). The combined organic phase was dried (Na2SO4), filtered and concentrated. The crude was purified by column chromatography (1-6% 7N NH3 in MeOH/CH2Cl2) to give 245 (7.3 mg, 16 µmol, 58%) as a thin film. 1 H NMR (600 MHz, C6D6): δ 4.33 (s, 2H), 3.91 (t, J = 5.0 Hz, 1H), 3.79 (t, J = 9.2 Hz, 1H), 3.21 (s, 1H), 3.18 (d, J = 10.2 Hz, 1H), 3.11 (s, 3H), 3.07 (s, 3H), 3.07 (s, 3H), 2.97 (d, J = 8.8 Hz, 1H), 2.87 (dd, J = 10.6, 6.6 Hz, 1H), 2.80 (m, 2H), 2.68 (dd, J = 16.9, 9.2 Hz, 1H), 2.51 (d, J = 11.4 Hz, 1H), 2.36-2.48 (m, 2H), 2.18-2.30 (m, 5H), 2.06-2.10 (m, 2H), 1.94-1.98 (m, 2H), 1.88 (ddd, J = 13.4, 5.0, 2.6 Hz, 1H), 1.48-1.63 (m, 4H), 1.34 (dt, J = 11.4, 6.7 Hz, 1H), 1.03 (t, J = 7.2 Hz, 3H). 13 C NMR (101 MHz, C6D6): δ 96.2, 85.8, 81.4, 79.7, 73.2, 72.7, 62.5, 59.2, 55.7, 55.5, 53.8, 49.5, 49.0, 46.7, 46.2, 46.0, 45.6, 44.9, 41.5, 38.9, 33.3, 28.6, 26.6, 25.5, 13.8. FTIR (NaCl, thin film): cm-1. 3434, 2928, 2822, 1461, 1448, 1375, 1201, 1148, 1110, 1052, 922, 754, 640. HRMS: calc’d for C25H42NO6 [M + H]+ : 452.3003 found: 452.3023 diff: 0.94 ppm Chapter 4 – Completion of Aconitine Core and Synthesis of (–)-Talatisamine, (–)-Liljestrandisine, and (–)-Liljestrandinine 174 [𝜶]𝟐𝟏 𝐃 = -20.1 (c = 0.52, CHCl3) TLC (6% 7N NH3 in MeOH/90%CH2Cl2), Rf: 0.5 (KMnO4) Preparation of (–)-liljestrandisine (238): OMe NEt MeO MeO OMe NEt H2SO4 (0.5 M) 40 °C MOM O HO OH 245 96% yield OH HO OH liljestrandisine (238) revised structure A 1-dram vial was charged with diol 245 (2.5 mg, 5.6 µmol, 1.0 equiv.) and 0.5 M H2SO4 (aq., 1 mL). The solution was heated to 40 °C for 4 hours before quenching with 0.5 M NaOH (1.5 mL) and saturated aqueous NaHCO3 (2 mL). The mixture was extracted with 3:1CHCl3:iPrOH (5 x 3 mL). The combined organic phases were dried (Na2SO4), filtered and concentrated. The product was purified on column chromatography (SiO2, 1-2-3-4-5-6% 7N NH3 in MeOH/CH2Cl2) to give (–)-liljestrandisine (238) (2.2 mg, 5.4 µmol. 96%) as a thin film. 1 H NMR (600 MHz, Chloroform-d) δ 4.25 (t, J = 5.3 Hz, 1H), 4.16 (br, 1H), 3.87 (d, J = 8.7 Hz, 1H), 3.30 (s, 3H), 3.26 (s, 3H), 3.20 (br, 1H), 3.18 (s, 1H), 3.11 (ABq, J = 9.0 Hz, 1H), 3.07 (dd, J = 10.8, 6.5 Hz, 1H), 2.99 (ABq, J = 9.0 Hz, 1H), 2.89 (br, 1H), 2.64 (dd, J = 17.4, 8.9 Hz, 1H), 2.56 – 2.47 (m, 2H), 2.45 – 2.33 (m, 1H), 2.32 – 2.19 (m, 3H), 2.08 (d, J = 7.8 Hz, 1H), 2.05 – 1.86 (m, 4H), 1.83 (dd, J = 10.3, 5.5 Hz, 2H), 1.80 – 1.74 (m, 1H), 1.74 – 1.69 (m, 1H), 1.66 (d, J = 7.4 Hz, 1H), 1.49 (dd, J = 14.9, 7.9 Hz, 1H), 1.45 – 1.35 (m, 1H), 1.06 (t, J = 7.1 Hz, 3H). Chapter 4 – Completion of Aconitine Core and Synthesis of (–)-Talatisamine, (–)-Liljestrandisine, and (–)-Liljestrandinine 13 C NMR (151 MHz, CDCl3) δ 86.2, 79.4, 76.1, 73.6, 72.6, 63.1, 59.5, 56.3, 53.1, 49.55, 48.6, 46.57, 46.47, 46.08, 45.7, 42.5, 40.9, 38.7, 32.8, 27.9, 25.8, 24.6, 13.7. FTIR (NaCl, thin film): 3330, 2923, 2883, 2814, 1455, 1094 cm-1. HRMS: (ESI-TOF) calc’d for C23H38NO5 [M + H]+ 408.2744, found 408.2736 [𝜶]𝟐𝟓 𝐃 = –9.5° (c = 0.15, CHCl3). TLC (10%7N NH3 in MeOH/90%CH2Cl2), Rf: 0.5 (ninhydrin) 1 175 H and 13C line list comparison tables are shown below with natural liljestrandisine.10 The following carbon numbering system is used in the line list comparisons: MeO 2 1’ 3 18’ OMe NEt MeO 18 4 5 19 11 10 17 12 7 8 15 6 9 14 OH OOH H OMe NEt 1 13 16 OH OOH H (–)-liljestrandisine (238) revised Chapter 4 – Completion of Aconitine Core and Synthesis of (–)-Talatisamine, (–)-Liljestrandisine, and (–)-Liljestrandinine 176 authentic liljestrandisine Wang, 2004 (ref. 10) (400 MHz, CHCl3) [α]D=–12 ° (c =0.5, CHCl3) synthetic liljestrandisine this report (600 MHz, CHCl3) [α]D=–9.5 ° (c =0.15, CHCl3) 1H (δ) ppm 1H (δ) ppm Carbon #, multiplicity 1d 3.07, dd(10.4, 6.4) 3.07 (dd (10.8, 6.5), 1H) 2t 1.98, m; 2.19, m 1.96 (m, 4H); 2.25 (m, 3H) 3t 1.38, dd(11.6); 1.74, m 1.39 (m, 1H); 1.77 (m, 1H) 4s – – 5d 1.64, d(7.2) 1.66 (d (7.4), 1H) 6t 1.47, dd(14.4, 7.6; 1.89, d(7.6) 1.49 (dd (14.9, 7.9), 1H); 1.96 (m, 4H) 7d 2.07, d(8.0) 2.08 (d (7.8), 1H) 8s – – 9d 2.25, m 2.25 (m, 3H) 10 d 1.70, m 1.71 (m, 1H) 11 s – – 12 t 1.78, m; 1.78, m 1.83 (m, 2H); 1.83 (m, 2H) 13 d 2.23, m 2.25 (m, 3H) 14 d 4.22, t(5.0) 4.25 (t (5.3), 1H) 15 t 1.95, m; 2.58, m 1.96 (m, 4H); 2.64 (dd (17.4, 8.9), 1H) 16 d 3.82, d(8.0) 3.87 (t (8.7), 1H) 17 d 3.18, s 3.18 (s, 1H) 18 t 2.99, ABq(9.2); 3.11, ABq(8.4) 2.99, ABq (9.0), 1H; 3.11, ABq(9.0), 1H 19 t 2.02, d(11.6); 2.51, m 1.96 (m, 4H); 2.52 (m, 2H) N CH2 2.39, m; 2.56, m 2.38 (m, 1H); 2.52 (m, 2H) 1.05, t(7.2) 1.06 (t (7.1), 3H) 1’ 3.26 s 3.26 (s, 3H) 18’ 3.29 s 3.30 (s, 3H) 8-OH — 2.89 (br, 1H) 14-OH — 4.25 (br, 1H) 16-OH — 3.20 (br, 1H) NCH2CH3 Table 4.4 Comparison of 1H NMR data for Authentic (ref. 10) vs. Synthetic (–)liljestrandisine. Chapter 4 – Completion of Aconitine Core and Synthesis of (–)-Talatisamine, (–)-Liljestrandisine, and (–)-Liljestrandinine 177 authentic liljestrandisine Wang, 2004 (ref. 10) (100 MHz, CHCl3) [α]D=–12 ° (c =0.5, CHCl3) synthetic liljestrandisine this report (151 MHz, CHCl3) [α]D=–9.5 ° (c =0.15, CHCl3) chemical shift difference Carbon #, multiplicity 13C (δ) ppm 13C (δ) ppm 13C (Δδ) ppm 1d 86.2 86.2 0 2t 25.7 25.8 +0.1 3t 32.7 32.8 +0.1 4s 38.7 38.7 0 5d 46.0 46.1 0 6t 24.6 24.6 0 7d 46.4 46.47 +0.1 8s 73.6 73.6 0 9d 46.5 46.57 +0.1 10 d 45.8 45.7 -0.1 11 s 48.7 48.6 -0.1 12 t 27.9 27.9 0 13 d 40.8 40.9 +0.1 14 d 75.8 76.0 +0.2 15 t 42.3 42.5 +0.2 16 d 72.5 72.6 +0.1 17 d 63.0 63.2 +0.2 18 t 79.4 79.4 0 19 t 53.2 53.1 -0.1 N CH2 49.5 49.5 0 NCH2CH3 13.6 13.7 +0.1 1’ 56.2 56.3 +0.1 18’ 59.5 59.5 0 Table 4.5 Comparison of 13C NMR data for Authentic (ref. 10) vs. Synthetic (–)liljestrandisine. Chapter 4 – Completion of Aconitine Core and Synthesis of (–)-Talatisamine, (–)-Liljestrandisine, and (–)-Liljestrandinine 178 Preparation of (–)-talatisamine (1): OMe NEt MeO 1. BF3•OEt2 0 °C; then Me3OBF4, 2,6-tBu2-4-MePyr MeO OMe NEt 2. H2SO4 (0.5 M) 40 °C MOM O HO OH 77% yield 245 OH HO OMe talatisamine (1) A stock solution of BF3•OEt2 (12 µL/mL) in CH2Cl2 was prepared by 10x dilution of a solution of BF3•OEt2 (0.12 mL) in CH2Cl2 (0.88 mL). A 1-dram vial was charged with diol 245 (4.2 mg, 9.3 µmol, 1.0 equiv.) and CH2Cl2 (2.1 mL). The mixture was cooled to 0 °C before 0.10 mL of a stock solution of BF3•OEt2 (12 µL/mL, 0.10 mL ,1.2 µL, 9.3 µmol, 1.0 equiv.) was added dropwise. The mixture was warmed to 20 °C before 2,6-di-tert-butyl-4-methylpyridine (7.6 mg, 37 µmol, 4.0 equiv.) was added, followed by Me3OBF4 (5.5 mg, 37 µmol, 4.0 equiv.). The mixture was stirred (750 rpm) for 2 hours before quenching with saturated aqueous NaHCO3 (1 mL). The mixture was extracted with 3:1 CHCl3:iPrOH (8 x 2 mL) and the combined extracts were dried (Na2SO4), filtered and concentrated in vacuo. The crude product was carried onto the next step without purification. A 1-dram vial was charged with the crude methylation product and 0.5 M H2SO4 (1 mL). The reaction was heated to 40 °C for 5 hours. The reaction was transferred to a 20 mL vial and quenched with saturated aqueous NaHCO3 (10 mL). The mixture was extracted with 3:1 CHCl3:iPrOH (8 x 2 mL) and the combined extracts were dried (Na2SO4), filtered and concentrated in vacuo. The product was purified by column chromatography (SiO2, 0.25-0.51-2% 7N NH3 in MeOH/CH2Cl2) to give (–)-talatisamine (1) (3.0 mg, 7.2 mmol, 77% yield, 2 steps) as a colorless oil. Chapter 4 – Completion of Aconitine Core and Synthesis of (–)-Talatisamine, (–)-Liljestrandisine, and (–)-Liljestrandinine 1 179 H NMR (600 MHz, Benzene-d6) δ 4.82 (d, J = 4.1 Hz, 1H), 4.15 (dd, J = 9.7, 5.1 Hz, 1H), 3.99 (s, 1H), 3.26 (s, 1H), 3.12 (s, 3H), 3.08 (s, 3H), 3.06 – 3.02 (m, 1H), 2.94 (d, J = 8.8 Hz, 1H), 2.89 – 2.86 (m, 1H), 2.87 – 2.84 (m, 4H), 2.84 (d, J = 8.8 Hz, 1H), 2.56 (d, J = 11.3 Hz, 1H), 2.50 – 2.41 (m, 2H), 2.38 (dd, J = 17.2, 8.7 Hz, 1H), 2.35 – 2.29 (m, 2H), 2.27 (d, J = 7.8 Hz, 1H), 2.23 (d, J = 17.4 Hz, 1H), 2.21 – 2.12 (m, 2H), 2.04 (dd, J = 11.3, 2.4 Hz, 1H), 1.99 – 1.90 (m, 2H), 1.85 (dd, J = 15.5, 7.6 Hz, 1H), 1.65 – 1.57 (m, 1H), 1.55 (d, J = 7.5 Hz, 1H), 1.53 – 1.43 (m, 2H), 1.37 (ddd, J = 11.2, 7.6, 6.0 Hz, 1H), 1.06 (t, J = 7.2 Hz, 3H). 13 C NMR (101 MHz, C6D6) δ 86.1, 82.7, 79.8, 76.0, 72.8, 62.8, 59.2, 56.1, 55.7, 53.9, 49.7, 49.0, 47.6, 46.3, 46.2, 39.1, 38.5, 33.2, 28.1, 26.3, 25.6, 13.9. FTIR (NaCl, thin film): 3436, 2926, 2814, 1670, 1495, 1456, 1412, 1392, 1204, 1160, 1091, 770, 751, 682, 666, 650, 624, 614 cm-1 HRMS: (ESI-TOF) calc’d for C24H40NO5 [M + H]+ : 422.2905 found: 422.2894 diff: 2.49 ppm [𝜶]𝟐𝟏 𝐃 = -3.2 (c = 0.23, CHCl3) TLC (8% 7N NH3 MeOH/92%CH2Cl2), Rf: 0.3 (KMnO4) 1 H and 13C line list comparison tables are shown below with natural talatisamine.15,16 The following carbon numbering system is used in the line list comparisons: MeO MeO 18 4 5 6 9 14 OH OOMe H 2 1’ 3 18’ OMe NEt OMe NEt 1 19 11 10 17 12 7 8 15 13 16 OH OOMe H (–)-talatisamine (1) Chapter 4 – Completion of Aconitine Core and Synthesis of (–)-Talatisamine, (–)-Liljestrandisine, and (–)-Liljestrandinine 180 authentic talatisamine Inoue, 2020 (ref. 15 ) (500 MHz, CHCl3) [α]D=0 ° (c =0.6, CHCl3) (ref. 16) synthetic talatisamine this report (600 MHz, C6D6) [α]D=–3.2 ° (c =0.23, CHCl3) 1H (δ) ppm 1H (δ) ppm Carbon #, multiplicity 1d 2.83-2.88 (m, 2H) 2.83-2.88 (m, 2H) 2t 1.92-1.98 (m, 2H) 1.90-1.99 (m, 2H); 2.41-2.50 (m, 2H) 3t 1.43-1.62 (m, 4H); 1.92-1.98 (m, 2H) 1.43-1.53 (m, 2H); 1.90-1.99 (m, 2H) 4s – - 5d 1.43-1.62 (m, 4H) 1.55 (d(7.5), 1H) 6t 1.43-1.62 (m, 4H); 2.15-2.22 (m, 3H) 1.43-1.53 (m, 2H); 2.12-2.21 (m, 2H) 7d 2.25-2.38 (m, 3H) 2.27 (d(7.8), 1H) 8s – - 9d 2.25-2.38 (m, 3H) 2.27-2.35 (m, 2H) 10 d 1.37 (ddd (9.8, 7.5, 7.3), 1H) 1.37 (ddd(11.2, 7.6, 6.0), 1H) 11 s – - 12 t 1.43-1.62 (m, 4H), 1.85 (dd (15.5, 7.5), 1H) 1.57-1.65 (m, 1H); 1.85 (dd(15.5, 7.6), 1H) 13 d 2.15-2.22 (m, 3H) 2.12-2.21 (m, 2H) 14 d 4.15 (dd (5.5, 7.5), 1H) 4.15 (dd(9.7, 5.1), 1H) 15 t 2.15-2.22 (m, 3H), 2.40-2.49 (m, 3H) 2.23 (d(17.4), 1H); 2.38 (dd (8.7, 17.2), 1H) 16 d 3.04 (d (9.2), 1H) 3.02-3.06 (m, 1H) 17 d 3.26 (s, 1H) 3.26 (s, 1H) 18 t 2.83-2.88 (m, 2H); 2.94 (d(8.6), 1H) 2.83-2.88 (m, 2H); 2.94 (d(8.8), 1H) 19 t 2.03 (dd (11.5, 2.3), 1H); 2.56 (d (11.5), 1H) 2.04 (dd(11.3, 2.4), 1H); 2.56 (d(11.3), 1H) N CH2 2.25-2.38 (m, 3H); 2.40-2.49 (m, 3H) 2.27-2.35 (m, 2H); 2.41-2.50 (m, 2H) 1.06 (dd(7.5, 7.5), 3H) 1.06 (t(7.2), 3H) 1’ 3.08 (s, 3H) 3.08 (s, 3H) 16’ 2.85 (s, 3H) 2.85 (s, 3H) 18’ 3.11 (s, 3H) 3.12 (s, 3H) 8-OH 4.01 (s, 1H) 3.99 (s, 1H) 14-OH 4.84 (d(4.0), 1H) 4.82 (d(4.1), 1H) NCH2CH3 Table 4.6 Comparison of 1H NMR data for Authentic (ref. 15) vs. Synthetic (–)talatisamine.15 Chapter 4 – Completion of Aconitine Core and Synthesis of (–)-Talatisamine, (–)-Liljestrandisine, and (–)-Liljestrandinine 181 authentic talatisamine Inoue, 2020 (ref. 15) (125 MHz, C6D6) [α]D=0 ° (c =0.6, CHCl3) (ref. 16) synthetic talatisamine this report (151 MHz, C6D6) [α]D=–3.2 ° (c =0.23, CHCl3) chemical shift difference Carbon #, multiplicity 13C (δ) ppm 13C (δ) ppm 13C (Δδ) ppm 1d 86.1 86.2 +0.1 2t 26.3 26.4 +0.1 3t 33.2 33.3 +0.1 4s 38.99/39.03 39.05/39.10 5d 46.16/46.18/46.27 46.23/46.26/46.33 +0.1 6t 25.5 25.6 +0.1 7d 46.16/46.18/46.27 46.23/46.26/46.33 +0.1 8s 72.8 72.8 0 9d 47.6 47.6 0 10 d 46.16/46.18/46.27 46.23/46.26/46.33 +0.1 11 s 48.9 49.0 +0.1 0 12 t 28.0 28.1 +0.1 13 d 38.4 38.5 +0.1 14 d 75.9 76.0 +0.1 15 t 38.99/39.03 39.05/39.10 16 d 82.7 82.8 17 d 62.8 62.8 0 18 t 79.7 79.8 +0.1 19 t 53.8 53.9 +0.1 N CH2 49.6 49.7 +0.1 NCH2CH3 13.9 13.9 0 1’ 55.7 55.7 0 16’ 56.0 56.1 +0.1 18’ 59.1 59.2 +0.1 0 +0.1 Table 4.7 Comparison of 13C NMR data for Authentic (ref. 15) vs. Synthetic (–)talatisamine. Chapter 4 – Completion of Aconitine Core and Synthesis of (–)-Talatisamine, (–)-Liljestrandisine, and (–)-Liljestrandinine 182 Preparation of Wang’s reported structure of (–)-liljestrandisine (16-epi-liljestrandisine, 2): OMe NEt MeO OMe NEt MeO H2SO4 (0.5 M) 40 °C OH MOM O HO 237 94% yield OH OH HO 16-epi-liljestrandisine (2) Wang’s reported structure 0.5 M H2SO4 (1 mL) was added to a 1-dram vial charged with diol 237 (2.0 mg, 4.4 µmol, 1.0 equiv.) and the mixture was heated to 40 °C for 16 hours. The reaction was cooled to 0 °C and quenched with 1 M NaOH (1.5 mL) and sat. NaHCO3 (2 mL). The mixture was extracted with 3:1 CHCl3:iPrOH (8 x 5 mL). The combined organic phases were dried (Na2SO4), filtered and concentrated in vacuo. The product was purified by silica gel chromatography (1-2-3-4-5-6-7 % 7N NH3 in MeOH/CH2Cl2) to give 2 (1.7 mg, 4.1 µmol, 91% yield) as a colorless oil. 1 H NMR (600 MHz, Chloroform-d) δ 4.62 – 4.49 (m, 1H), 4.16 (s, 1H), 3.54 (s, 1H), 3.30 (s, 6H), 3.11 (d, J = 8.9 Hz, 1H), 2.98 (d, J = 9.1 Hz, 1H), 2.53 (d, J = 11.3 Hz, 2H), 2.45 – 2.34 (m, 2H), 2.23 (dd, J = 14.9, 7.2 Hz, 2H), 2.20 – 2.12 (m, 2H), 2.12 – 2.01 (m, 3H), 1.97 (br, 1H), 1.90 (dd, J = 15.8, 8.5 Hz, 2H), 1.77 (d, J = 13.3 Hz, 1H), 1.71 – 1.59 (m, 3H), 1.49 (dd, J = 14.7, 7.6 Hz, 1H), 1.38 (t, J = 14.8 Hz, 2H), 1.06 (t, J = 6.9 Hz, 3H). 13 C NMR (101 MHz, CDCl3) δ 86.29, 79.31, 76.01, 66.20, 63.69, 59.50, 56.41, 53.14, 49.62, 49.39, 46.42, 46.30, 45.82, 45.56, 42.83, 41.50, 38.72, 32.79, 29.69, 25.61, 24.95, 23.60, 13.69. TLC (10% 7N NH3 MeOH/92%CH2Cl2), Rf: 0.3 (KMnO4) Chapter 4 – Completion of Aconitine Core and Synthesis of (–)-Talatisamine, (–)-Liljestrandisine, and (–)-Liljestrandinine 183 Preparation of β-methoxy ketone 239: OMe Me O LiHMDS Cl NAc t-Bu OMOM 232 OMe NAc MeO S N Ph 233 16 THF, –78 to 21 °C then MeOH, pyr O 66% yield 8 MOMO OMe O 239 Ketone 232 (11.2 mg, 25 µmol, 1 equiv.) was dried by azeotroping with PhMe (3 x 5 mL) in a 2 dram vial. The vial was put under N2, THF (1.0 mL) was added, and the mixture was cooled to – 78 °C. A solution of LiHMDS (1M in THF, 0.06 mL, 60 µmol, 2.2 equiv.) was added dropwise over 1 minute. The mixture was stirred at -78 °C for 15 minutes before a solution of N-tert-butyl-phenylsulfinyl chloride (43.2 mg. 0.20 mmol, 8.0 equiv.) in THF (0.50 mL) was added dropwise over 5 minutes. The reaction was stirred at -78 °C for 15 minutes, then warmed to 21 °C and stirred an additional 1 hour. Pyridine (0.05 mL) was added, followed by freshly-distilled MeOH (0.25 mL) and the mixture was stirred for 60 hours. The reaction was quenched with sat. aq. NaHCO3 (2 mL). The mixture was extracted with 3:1 CHCl3:iPrOH (5 x 2 mL). The combined extracts were dried (Na2SO4), filtered and concentrated. The reaction was purified by column chromatography (SiO2, 1-2-3-4-5-6-7% MeOH/CH2Cl2) to give β-methoxy ketone 239 (7.9 mg, 17 µmol, 66% yield) and β-hydroxy ketone 234 (3.5 mg, 7.5 µmol, 30% yield) 1 H NMR (400 MHz, Chloroform-d): δ 4.77 (d, J = 6.8 Hz, 1H), 4.57 (d, J = 6.8 Hz, 1H), 4.13 (t, J = 4.8 Hz, 1H), 3.91 (d, J = 14.4 Hz, 1H), 3.74 (s, 1H), 3.33 (s, 3H), 3.30 (s, 3H), 3.22 – 3.18 (m, 4H), 3.14 (dd, J = 9.8, 7.2 Hz, 1H), 3.10 (s, 3H), 3.05 – 2.96 (m, 2H), 2.78 (dd, J = 7.6, 4.3 Hz, 1H), 2.61 – 2.50 (m, 2H), 2.44 (t, J = 5.7 Hz, 1H), 2.39 (d, J = 18.3 Hz, 1H), 2.25 Chapter 4 – Completion of Aconitine Core and Synthesis of (–)-Talatisamine, (–)-Liljestrandisine, and (–)-Liljestrandinine 184 (d, J = 7.8 Hz, 1H), 2.10 – 2.02 (m, 7H), 1.88 (d, J = 7.4 Hz, 1H), 1.72 – 1.64 (m, 2H), 1.45 (tdd, J = 13.2, 4.5, 1.7 Hz, 1H), 1.37 – 1.26 (m, 1H). 13 C NMR (101 MHz, CDCl3): δ 211.3, 169.7, 95.8, 82.5, 78.2, 77.7, 76.8, 59.5, 58.3, 55.7, 55.5, 52.3, 48.7 (2 overlapping peaks), 47.6, 45.4, 45.3, 44.2, 44.1, 42.7, 37.4, 31.6, 26.8, 25.4, 24.0, 22.4. FTIR (NaCl, thin film): 2930, 2890, 2825, 1710, 1639, 1430, 1151, 1114, 1092, 1048 cm-1. HRMS: (ESI-TOF) calc’d for C26H40NO7 [M + H]+ 478.2799, found 478.2790 [𝜶]𝟐𝟓 𝐃 = –120° (c = 1.5, CHCl3). TLC (5%MeOH/95%CH2Cl2), Rf: 0.5 (UV, yellow in p-anisaldehyde) Preparation of enol triflate 247: OMe NAc MeO OMe NAc MeO LiHMDS, THF, –78 °C then Comins’ reagent MOMO OMe O 239 55% yield MOMO OMe OTf 247 A 50 mL, round-bottom flask was charged with ketone 239 (20 mg, 42 µmol, 1 equiv.), and azeotroped with PhMe (from the solvent system, 2 x 3 mL) to remove H2O and put under high vacuum for two hours. The flask was then equipped with a rubber septum, purged with N2, dissolved in THF (2 mL), and cooled to –78 °C in a dry ice/acetone bath. LiHMDS (1M in THF, 84 µL, 84 µmol, 2 equiv.) was added dropwise via syringe, causing the solution to turn yellow. The LiHMDS/239 solution was allowed to stir for 30 minutes at –78 °C, and then Comins’ reagent (36.9 mg, 94 µmol, 2.25 equiv.) solution in THF (1 mL) was added dropwise via syringe. This was allowed to stir for an additional 5 minutes at –78 °C, the bath was then removed, and this was allowed to stir for 20 minutes at room temperature. The reaction was Chapter 4 – Completion of Aconitine Core and Synthesis of (–)-Talatisamine, (–)-Liljestrandisine, and (–)-Liljestrandinine 185 quenched with 0.5 M NaOH (5 mL), and the biphasic mixture was transferred to a separatory funnel with EtOAc (10 mL). The layers were separated, and the aqueous layer was extracted EtOAc (4 x 10 mL). The combined organic extracts were dried over Na2SO4, filtered, and concentrated in vacuo. The resulting crude residue was purified by preparative thin layer silica gel chromatography (mobile phase: 100% ethyl acetate) to afford enol triflate 247 (14 mg, 23 µmol, 55% yield) as a clear oil. 1 H NMR (400 MHz, Chloroform-d): δ 5.85 (t, J = 1.6 Hz, 1H), 4.71 (d, J = 6.9 Hz, 1H), 4.60 (d, J = 6.9 Hz, 1H), 3.93 – 3.85 (m, 3H), 3.36 (s, 3H), 3.30 (s, 3H), 3.21 (s, 3H), 3.19 – 3.17 (m, 4H), 3.14 – 3.10 (m, 1H), 3.02 (d, J = 9.1 Hz, 1H), 2.86 (dd, J = 14.2, 5.3 Hz, 1H), 2.72 – 2.64 (m, 1H), 2.61 (d, J = 14.4 Hz, 1H), 2.34 (t, J = 5.5 Hz, 1H), 2.27 (d, J = 7.8 Hz, 1H), 2.09 – 2.03 (m, 6H), 1.96 – 1.90 (m, 1H), 1.83 (d, J = 7.4 Hz, 1H), 1.76 – 1.68 (m, 2H), 1.43 (tdd, J = 13.2, 4.5, 1.8 Hz, 1H), 1.33 – 1.27 (m, 1H). 13 C NMR (101 MHz, CDCl3): δ 170.5, 153.9,114.2, 96.0, 82.6, 78.4, 78.2, 77.2, 59.5, 59.1, 55.6, 55.5, 49.4, 48.0, 45.2, 45.1, 44.3, 43.9, 42.8, 42.6, 37.4, 32.0, 30.7, 25.4, 23.7, 22.5. (Note: triflate carbon was not observed). FTIR (NaCl, thin film): 2938, 2829, 1636, 1418, 1210, 1142, 1094 cm-1. HRMS: (ESI-TOF) calc’d for C27H39NF3O9S [M + H]+ 610.2292, found 610.2281 [𝜶]𝟐𝟓 𝐃 = –37.7° (c = 0.50, CHCl3). TLC (100%EtOAc), Rf: 0.4 (yellow in p-anisaldehyde) Chapter 4 – Completion of Aconitine Core and Synthesis of (–)-Talatisamine, (–)-Liljestrandisine, and (–)-Liljestrandinine 186 Preparation of alkene 240: OMe NAc MeO Pd(OAc)2, PPh3 NEt3, HCO2H OMe NAc MeO DMF, 60 °C MOMO OMe OTf 79% yield MOMO 247 OMe H 240 A 50 mL, round-bottom flask was charged with enol triflate 247 (10 mg, 16.4 µmol, 1.0 equiv.), PPh3 (3.4 mg, 12.96 µmol, 0.80 equiv.), and Pd(OAc)2 (1.5 mg, 6.68 µmol, 0.40 equiv.). The flask was equipped with a rubber septum, purged with N2, and dissolved in DMF (0.8 mL). Then, the reaction mixture was sparged with an Ar balloon for 5 minutes. NEt3 (46 µL, 328 µmol, 20 equiv.) was added followed by HCO2H (3 µL, 82.1 µmol, 5 equiv.)—a cloudy gas was observed upon addition. The reaction was lowered into a 60 °C oil bath, and allowed to continue to stir at that temperature for 20 minutes. The reaction turns black during this time indicating its completion. The reaction was cooled to room temperature, diluted with sat. NH4Cl (5 mL), transferred to a separatory funnel and diluted with EtOAc (10 mL). The layers were separated and the aqueous extracted with EtOAc (4 x 10 mL). The organic extracts were dried over Na2SO4, filtered, and concentrated in vacuo. The resulting crude residue was purified by preparative thin layer silica gel chromatography (mobile phase: 100% ethyl acetate) to afford olefin 240 (6.0 mg, 13.0 µmol, 79% yield) as a clear oil. 1 H NMR (400 MHz, Chloroform-d): δ 6.17 (ddd, J = 9.8, 6.7, 1.1 Hz, 1H), 5.85 (dd, J = 9.8, 1.9 Hz, 1H), 4.75 (d, J = 6.8 Hz, 1H), 4.59 (d, J = 6.8 Hz, 1H), 3.88 (d, J = 14.2 Hz, 1H), 3.84 (t, J = 4.5 Hz, 1H), 3.79 (s, 1H), 3.36 (s, 3H), 3.30 (s, 3H), 3.23 – 3.18 (m, 4H), 3.17 (s, 3H), 3.11 (dd, J = 10.3, 7.6 Hz, 1H), 3.04 (d, J = 9.1 Hz, 1H), 2.64 – 2.54 (m, 2H), 2.41 (dd, J = 13.9, 5.3 Hz, 1H), 2.33 (t, J = 5.5 Hz, 1H), 2.14 (d, J = 7.8 Hz, 1H), 2.08 – 1.98 (m, 6H), 1.88 – 1.79 (m, 2H), 1.74 – 1.65 (m, 2H), 1.50 – 1.36 (m, 1H), 1.34 – 1.26 (m, 1H). Chapter 4 – Completion of Aconitine Core and Synthesis of (–)-Talatisamine, (–)-Liljestrandisine, and (–)-Liljestrandinine 13 187 C NMR (101 MHz, CDCl3): δ 169.9, 134.3, 125.3, 95.5, 82.9, 78.6, 77.2, 76.6, 59.5, 59.5, 55.6, 55.5, 49.2, 48.0, 45.6, 45.2, 44.3, 44.2, 42.7, 37.5, 37.4, 32.0, 31.5, 25.5, 23.5, 22.5. FTIR (NaCl, thin film): 2928, 2886, 2822, 1634, 1428, 1149, 1114, 1091, 1044 cm-1. HRMS: (ESI-TOF) calc’d for C26H40NO6 [M + H]+ 462.2850, found 462.2835 [𝜶]𝟐𝟓 𝐃 = –78.2° (c = 0.21, CHCl3). TLC (100%EtOAc), Rf: 0.3 (grey/purple in p-anisaldehyde) Preparation of (–)-liljestrandinine (3): OMe NAc MeO 8 MOMO OMe 240 MeO 1. LiAlH4 Et2O, 40 °C OMe NEt 2. H2SO4 (aq) 110 °C 60% yield (2 steps) HO OH (–)-liljestrandinine (3) A 2-dram vial was charged with acetamide 240 (6 mg, 13 µmol, 1 equiv.), azeotroped with toluene (from the solvent system, 2 x 1 mL) and put under high vacuum for 1 hour. The vial was then equipped with a rubber septum, purged with N2, and charged with Et2O (1 mL) to dissolve the substrate, and LiAlH4 solution (1 M in THF, 52 µL, 52 µmol, 4 equiv.) was added dropwise via microsyringe. The rubber septum was replaced with a vial cap, and heated to 40 °C in a pre-heated heating block. The reaction was allowed to stir at 40 °C for 40 minutes, and then was removed from the heating block and allowed to cool to room temperature. The reaction was carefully quenched with aqueous 10% NaOH solution (4 mL), and diluted with Et2O (4 mL). The layers were separated, and the aqueous layer was extracted with 20%IPA/80%CHCl3 (5 x 5 mL). The combined organic extracts were dried over Na2SO4, Chapter 4 – Completion of Aconitine Core and Synthesis of (–)-Talatisamine, (–)-Liljestrandisine, and (–)-Liljestrandinine 188 filtered, and concentrated in vacuo. The crude residue was then transferred to a new 2-dram vial, and used in the next step without further purification. The 2-dram vial containing the crude residue from the LiAlH4 reduction was dissolved in aqueous 0.5 M H2SO4 (1.5 mL), sealed with a cap, and heated to 110 °C in a pre-heated heating block. The reaction was allowed to stir at 110 °C for 3 hours, and then was cooled to room temperature. Monitoring by LCMS indicated full conversion to the product. The reaction as then carefully quenched with aqueous 10% NaOH solution (2 mL), and diluted with 20%IPA/80%CHCl3 (4 mL). The layers were separated, and the aqueous layer was extracted with 20%IPA/80%CHCl3 (8 x 4 mL). As an additional precaution, the aqueous layer was made sure to be basic with pH paper, and an LCMS sample of the aqueous layer was taken to ensure that no product was left behind. The combined organic extracts were dried over Na2SO4, filtered, and concentrated in vacuo. The crude residue was purified by preparative thin layer chromatography (mobile phase: 10% 7 N NH3 in MeOH/ 90% CH2Cl2) to furnish (–)liljestrandinine (3) (3 mg, 7.8 µmol, 60% yield over 2 steps). Notes: (1) Due to rapid protonation and other dynamic effects, the H NMR and C NMR spectra 1 13 in CDCl are inconsistent and often display broadened and poorly resolved resonances and 3 extra peaks arising from the protonated material. Characterization data was therefore obtained in benzene-d6, and was compared with synthetic liljestrandinine that had been characterized in benzene-d6. Data collected in chloroform was only of satisfactory resolution and used only 1 to compare to the reported tabulated data for isolated liljestrandinine. 1 18 H NMR (400 MHz, Benzene-d6): δ 5.70 (ddd, J = 9.5, 6.7, 1.1 Hz, 1H), 5.61 (dd, J = 9.4, 1.8 Hz, 1H), 3.78 (q, J = 4.8 Hz, 1H), 3.12 (s, 3H), 3.10 – 3.08 (m, 1H), 3.05 (s, 3H), 2.96 (d, J = Chapter 4 – Completion of Aconitine Core and Synthesis of (–)-Talatisamine, (–)-Liljestrandisine, and (–)-Liljestrandinine 189 8.8 Hz, 1H), 2.90 – 2.82 (m, 2H), 2.60 – 2.50 (m, 3H), 2.49 – 2.43 (m, 1H), 2.43 – 2.35 (m, 1H), 2.29 (dq, J = 12.0, 7.1 Hz, 1H), 2.19 – 2.11 (m, 2H), 2.10 – 2.07 (m, 1H), 2.05 – 1.97 (m, 2H), 1.97 – 1.88 (m, 2H), 1.76 – 1.72 (m, 1H), 1.64 – 1.47 (m, 5H), 1.04 (t, J = 7.2 Hz, 3H). 13 C NMR (101 MHz, CDCl3): δ 131.9, 129.8, 85.8, 79.6, 74.6, 74.3, 63.2, 59.5, 56.3, 53.2, 49.5, 48.4, 46.4, 45.8, 45.6, 42.4, 39.0, 38.5, 33.1, 32.7, 23.5, 22.7, 13.5. 13 C NMR (101 MHz, C6D6): δ 132.5, 129.7, 85.6, 80.0, 75.0, 74.5, 63.0, 59.2, 55.7, 53.9, 49.6, 48.6, 46.8, 46.3, 46.0, 42.7, 39.6, 38.9, 33.4, 33.3, 26.7, 24.2, 13.7. TLC (10%7N NH3 in MeOH/90%CH2Cl2), Rf: 0.5 (ninhydrin) FTIR (NaCl, thin film): 3332, 2921, 2823, 1642, 1462, 1095 cm-1. HRMS: (ESI-TOF) calc’d for C23H36NO4 [M + H]+ 390.2639, found 390.2622 [𝜶]𝟐𝟓 𝐃 = –17.6° (c = 0.21, CHCl3). The following carbon numbering system is used in the line list comparisons: MeO OMe NEt 2 3 18’ MeO 18 4 5 11 10 17 12 7 8 15 6 9 14 OH OH 1’ OMe 19 NEt 1 13 16 OH OH (–)-liljestrandinine (3) Chapter 4 – Completion of Aconitine Core and Synthesis of (–)-Talatisamine, (–)-Liljestrandisine, and (–)-Liljestrandinine 190 authentic liljestrandinine Wang, 2003 (ref. 18 ) (100 MHz, CHCl3) [α]D=–10 ° (c =0.5, CHCl3) synthetic liljestrandinine this report (101 MHz, CHCl3) [α]D=–17.6 ° (c =0.21, CHCl3) chemical shift difference Carbon #, multiplicity 13C (δ) ppm 13C (δ) ppm 13C (Δδ) ppm 1d 85.7 85.8 +0.1 2t 22.6 22.7 +0.1 3t 32.6 32.7 +0.1 4s 38.5 38.5 0 5d 45.6 45.6 0 6t 23.4 23.5 +0.1 7d 42.3 42.4 +0.1 8s 74.3 74.3 0 9d 46.3 46.4 +0.1 10 d 45.8 45.8 0 11 s 48.2 48.4 +0.2 12 t 33.0 33.1 +0.1 13 d 38.9 39.0 +0.1 14 d 74.5 74.6 +0.2 15 t 131.7 131.9 +0.2 16 d 129.8 129.8 0 17 d 63.2 63.2 +0.1 18 t 79.6 79.6 0 19 t 53.1 53.2 +0.1 N CH2 49.4 49.5 +0.1 NCH2CH3 13.4 13.5 +0.1 1’ 56.2 56.3 +0.1 18’ 59.4 59.5 +0.1 Table S2.5.6. Comparison of 13C NMR data for Authentic (ref. 18) vs. synthetic (–)liljestrandinine. Chapter 4 – Completion of Aconitine Core and Synthesis of (–)-Talatisamine, (–)-Liljestrandisine, and (–)-Liljestrandinine 191 synthetic liljestrandinine Sarpong, 2015 (ref. 17 ) (151 MHz, C6D6) synthetic liljestrandinine this report (101 MHz, C6D6) [α]D=–17.6 ° (c =0.21, CHCl3) chemical shift difference 13C (δ) ppm 13C (δ) ppm 13C (Δδ) ppm 132.4 132.5 +0.1 129.7 129.7 0 85.6 85.6 0 80.0 80.0 0 74.9 75.0 +0.1 74.5 74.5 0 63.0 63.0 0 59.1 59.2 +0.1 55.6 55.7 +0.1 53.9 53.9 0 49.5 49.6 +0.1 48.6 48.6 0 46.8 46.8 0 46.3 46.3 0 46.0 46.0 0 42.7 42.7 0 39.6 39.6 0 38.9 38.9 0 33.4 33.4 0 33.2 33.3 +0.1 26.7 26.7 0 24.2 24.2 0 13.7 13.7 0 Table S2.5.7. Comparison of 13C NMR data in C6D6 of Synthetic (ref. 17) vs. Synthetic (–)-liljestrandinine (this report). Chapter 4 – Completion of Aconitine Core and Synthesis of (–)-Talatisamine, (–)-Liljestrandisine, and (–)-Liljestrandinine 192 4.6 NOTES AND REFERENCES (1) (2) (3) (4) (5) (6) (7) (8) (9) (10) (11) (12) (13) (14) (15) (16) (17) (18) Surzur, J.-M.; Stella, L.; Tordo, P. Bull. Soc. Chim. Fr. 1970, 115. Stella, L. Angew. Chem. Int. Ed. Engl. 1983, 22 (5), 337–350. Stella, L.; Raynier, B.; Surzur, J.-M. Tetrahedron Lett. 1977, 18 (31), 2721–2724. Stella, L.; Raynier, B.; Surzur, J. M. Tetrahedron 1981, 37 (16), 2843–2854. Göttlich, R. Synthesis 2000, 2000 (11), 1561–1564. Das, S.; Li, Y.; Lu, L.-Q.; Junge, K.; Beller, M. Chem. - Eur. J. 2016, 22 (21), 7050– 7053. Shi, Y.; Wilmot, J. T.; Nordstrøm, L. U.; Tan, D. S.; Gin, D. Y. J. Am. Chem. Soc. 2013, 135 (38), 14313–14320. Liu, Z.-G.; Cheng, H.; Ge, M.-J.; Xu, L.; Wang, F.-P. Tetrahedron 2013, 69 (26), 5431–5437. Mukaiyama, T.; Matsuo, J.; Kitagawa, H. Chem. Lett. 2000, 29 (11), 1250–1251. Xie, G.-B.; Wang, F.-P. Chem. J. Chin. Univ. 2004, 25 (3), 482–483. Cheng, C.; Brookhart, M. J. Am. Chem. Soc. 2012, 134 (28), 11304–11307. Steves, J. E.; Stahl, S. S. J. Am. Chem. Soc. 2013, 135 (42), 15742–15745. Harrowven, D. C.; Curran, D. P.; Kostiuk, S. L.; Wallis-Guy, I. L.; Whiting, S.; Stenning, K. J.; Tang, B.; Packard, E.; Nanson, L. Chem. Commun. 2010, 46 (34), 6335. Davis, T. A.; Chopade, P. R.; Hilmersson, G.; Flowers, R. A. Org. Lett. 2005, 7 (1), 119–122. Kamakura, D.; Todoroki, H.; Urabe, D.; Hagiwara, K.; Inoue, M. Angew. Chem. Int. Ed. 2020, 59 (1), 479–486. Deng, Li-Sheng; Chen, Yao-Zu; Wu, Feng-E; Li, Bo-Gang. Phytochemistry 1990, 29 (11), 3694–3696. Marth, C. J.; Gallego, G. M.; Lee, J. C.; Lebold, T. P.; Kulyk, S.; Kou, K. G. M.; Qin, J.; Lilien, R.; Sarpong, R. Nature 2015, 528 (7583), 493–498. Wang, F.-P.; Xie, G.-B.; Chen, Q.-H.; Chen, D.-L.; Jian, X.-X. Heterocycles 2003, 60 (3), 631. About the Author 193 ABOUT THE AUTHOR Nicholas James Fastuca was born on June 4th, 1993 in Raleigh, NC, to parents Debra Fastuca and Stephen Fastuca. He grew up in West Chester, PA, where he spent his childhood surrounded by his extended family and friends. His interest in chemistry was sparked by his high school teacher, Josh Gellner. This led him to pursue a B.S. in chemistry at The Pennsylvania State University from 2012-2016 where he received encouragement and support from his academic adviser Dr. Joseph Keiser. During his time at Penn State, he had the opportunity to join Professor Alexander Radosevich’s laboratory where he was first exposed to the world of synthetic chemistry. After graduating from Penn State, he moved to Pasadena, CA, to pursue a Ph.D. in organic chemistry at Caltech in the lab of Professor Sarah Reisman. During his time at Caltech, he had the opportunity to learn from many excellent synthetic chemists. Upon completion of his Ph.D., he will move to Princeton, NJ to work as a postdoctoral scholar at Princeton University in the lab of Professor Robert Knowles. Appendix 1 – Spectra Relevant to Chapter 2 194 Appendix 1 Spectra Relevant to Chapter 2: Convergent Approach to C19-Diterpenoid Alkaloids via 1,2Addition/Semipinacol Sequence 11.0 16 40 1.0000 5.9000 3.0000 2016-01-28T13:41:44 Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date 65536 Spectral Size 9.0 24000 Acquired Size 9.5 1H Nucleus 10.0 -1009.5 Lowest Frequency 10.5 8000.0 Spectral Width 1.84 7.5 2.83 7.0 5.0 ppm 1D Experiment 5.5 s2pul Pulse Sequence 6.0 25.0 Temperature 6.5 cdcl3 Solvent 8.0 inova Instrument 8.5 Varian Origin Spectrometer Frequency 499.67 PROTON01 Title Value 7.29 / Volumes/ nmrdata/ vmak/ vnmrsys/ data/ VWM6-033/ PROTON01.fid/ fid Parameter 6.95 Data File Name 3.5 3.0 4.61 1.00 1.0 4.06 0.97 1.5 2.56 0.91 2.0 1.95 1.01 2.5 1.70 1.03 4.0 1.31 1.23 3.08 3.02 4.5 0.5 0.0 -0.5 -1.0 Appendix 1 – Spectra Relevant to Chapter 2 195 11.0 zg30 1D 16 197.4 1.0000 11.7000 4.0894 2016-03-11T19:56:13 Experiment Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date 65536 Spectral Size 9.0 32768 Acquired Size 9.5 1H Nucleus 10.0 -1545.2 Lowest Frequency 10.5 8012.8 Spectral Width 3.15 7.0 1.00 6.5 6.0 294.9 Pulse Sequence 7.5 CDCl3 Temperature 8.0 spect Solvent 8.5 Bruker BioSpin GmbH Instrument Spectrometer Frequency 400.13 VWM6-048-flashed.1.fid Origin 7.27 Title Value 6.91 / Volumes/ nmrdata/ vmak/ nmr/ VWM6-048-flashed/ 1/ fid Parameter 6.34 Data File Name 5.5 5.0 ppm 3.5 3.0 2.5 2.0 4.52 4.33 4.12 4.03 1.02 0.97 1.01 0.93 1.5 2.20 1.01 4.0 1.65 1.33 1.25 1.01 3.07 3.16 4.5 1.0 0.5 0.0 -0.5 -1.0 Appendix 1 – Spectra Relevant to Chapter 2 196 6.16 11.0 87.8 1.0000 11.7000 4.0894 2016-04-27T13:37:19 Relaxation Delay Pulse Width Acquisition Time Acquisition Date 65536 Spectral Size 9.0 32768 Acquired Size 9.5 1H Nucleus 10.0 -1545.6 Lowest Frequency 10.5 8012.8 Spectral Width 6.0 16 Receiver Gain 6.5 1D Number of Scans 7.0 zg30 Experiment 7.5 295.0 Pulse Sequence 8.0 CDCl3 Temperature 8.5 spect Solvent Spectrometer Frequency 400.13 Bruker BioSpin GmbH Instrument 5.5 5.0 ppm 4.5 4.0 3.5 3.0 2.5 2.0 1.5 5.62 5.43 1.00 1.00 ARWIII-040_flashed.1.fid 4.48 4.31 4.05 1.00 1.04 1.11 Origin 3.25 0.99 Title Value 2.46 2.34 2.09 3.11 1.10 1.07 / Volumes/ nmrdata-1/ arwong/ nmr/ ARWIII-040_flashed/ 1/ fid Parameter 1.59 1.23 1.21 1.18 3.03 3.00 Data File Name 1.0 0.5 0.0 -0.5 -1.0 Appendix 1 – Spectra Relevant to Chapter 2 197 11.0 16 127.1 1.0000 11.7000 4.0894 2017-06-28T03:44:24 Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date 65536 Spectral Size 9.0 32768 Acquired Size 9.5 1H Nucleus 10.0 -1545.6 Lowest Frequency 10.5 8012.8 Spectral Width 5.5 5.0 ppm 3.0 2.0 1.0 5.88 5.74 0.98 1.07 1.5 4.46 0.99 2.5 3.97 3.83 3.72 3.56 1.05 1.02 3.06 1.92 3.5 2.61 2.46 1.02 1.00 4.0 1.77 1.55 2.05 2.10 4.5 1.15 1.12 3.17 3.00 6.0 1D Experiment 6.5 zg30 Pulse Sequence 7.0 295.0 Temperature 7.5 CDCl3 Solvent 8.0 spect Instrument 8.5 Bruker BioSpin GmbH Origin Spectrometer Frequency 400.13 VWM7-arw4-163.1.fid Title Value / Volumes/ nmrdata/ vmak/ nmr/ VWM7-arw4-163/ 1/ fid Parameter Data File Name 0.5 0.0 -0.5 -1.0 Appendix 1 – Spectra Relevant to Chapter 2 198 220 1D 512 78.7 1.0000 10.0000 1.2976 2017-06-28T06:00:12 Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date 65536 Spectral Size 180 32768 Acquired Size 190 13C Nucleus 200 -2569.1 Lowest Frequency 210 25252.5 Spectral Width 130 zgpg30 Experiment 140 295.0 Pulse Sequence 150 CDCl3 Temperature 160 spect Solvent 170 Bruker BioSpin GmbH Instrument Spectrometer Frequency 100.62 VWM7-arw4-162.2.fid Origin Value Title 213.93 / Volumes/ nmrdata/ vmak/ nmr/ VWM7-arw4-162/ 2/ fid Parameter 131.40 129.59 Data File Name 120 110 100 ppm 90 70 81.07 75.20 73.87 66.98 80 60 51.56 49.49 45.91 50 40 32.18 30 23.62 19.99 20 10 0 -10 Appendix 1 – Spectra Relevant to Chapter 2 199 11.0 16 127.1 1.0000 11.7000 4.0894 2017-06-28T03:44:24 Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date 65536 Spectral Size 9.0 32768 Acquired Size 9.5 1H Nucleus 10.0 -1545.6 Lowest Frequency 10.5 8012.8 Spectral Width 5.5 5.0 ppm 3.0 2.0 1.0 5.88 5.74 0.98 1.07 1.5 4.46 0.99 2.5 3.97 3.83 3.72 3.56 1.05 1.02 3.06 1.92 3.5 2.61 2.46 1.02 1.00 4.0 1.77 1.55 2.05 2.10 4.5 1.15 1.12 3.17 3.00 6.0 1D Experiment 6.5 zg30 Pulse Sequence 7.0 295.0 Temperature 7.5 CDCl3 Solvent 8.0 spect Instrument 8.5 Bruker BioSpin GmbH Origin Spectrometer Frequency 400.13 VWM7-arw4-163.1.fid Title Value / Volumes/ nmrdata/ vmak/ nmr/ VWM7-arw4-163/ 1/ fid Parameter Data File Name 0.5 0.0 -0.5 -1.0 Appendix 1 – Spectra Relevant to Chapter 2 200 220 1D 1024 78.7 1.0000 10.0000 1.2976 2017-06-28T04:25:11 Experiment Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date 65536 Spectral Size 180 32768 Acquired Size 190 13C Nucleus 200 -2569.4 Lowest Frequency 210 25252.5 Spectral Width 130 zgpg30 Pulse Sequence 140 295.0 Temperature 150 CDCl3 Solvent 160 spect Instrument 170 Bruker BioSpin GmbH Origin Spectrometer Frequency 100.62 VWM7-arw4-163.2.fid Title Value / Volumes/ nmrdata/ vmak/ nmr/ VWM7-arw4-163/ 2/ fid Parameter 129.73 126.50 Data File Name 120 110 100 ppm 90 70 78.27 75.36 72.03 71.75 68.09 80 60 50 45.49 45.42 40.81 33.85 40 30 23.43 20.11 20 10 0 -10 Appendix 1 – Spectra Relevant to Chapter 2 201 11.0 16 78.7 1.0000 11.7000 4.0894 2017-05-03T00:37:05 Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date 65536 Spectral Size 9.0 32768 Acquired Size 9.5 1H Nucleus 10.0 -1545.6 Lowest Frequency 10.5 8012.8 Spectral Width 0.98 1.00 6.0 5.5 1D Experiment 6.5 zg30 Pulse Sequence 7.0 295.0 Temperature 7.5 CDCl3 Solvent 8.0 spect Instrument 8.5 Bruker BioSpin GmbH Origin Spectrometer Frequency 400.13 VWM-ARWIV-164-flashed.1.fid Title Value / Volumes/ nmrdata/ vmak/ nmr/ VWM-ARWIV-164-flashed/ 1/ fid Parameter 5.83 5.74 Data File Name 5.0 ppm 2.5 2.0 1.5 1.0 4.59 0.99 3.0 3.98 3.75 3.39 2.00 2.01 0.98 3.5 2.78 2.72 0.98 0.98 4.0 1.82 1.71 1.59 1.51 1.16 1.12 1.05 0.96 0.98 1.10 1.10 1.02 3.04 2.98 9.20 8.84 4.5 0.5 0.0 -0.5 -1.0 Appendix 1 – Spectra Relevant to Chapter 2 202 78.7 2.0000 10.0000 1.2780 2017-05-03T01:34:44 Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date 180 65536 Spectral Size 190 32768 Acquired Size 200 13C Nucleus 210 -2762.6 Lowest Frequency 220 25641.0 Spectral Width 110 1024 Number of Scans 120 1D Experiment 130 zgpg30 Pulse Sequence 140 294.9 Temperature 150 CDCl3 Solvent 160 spect Instrument 170 Bruker BioSpin GmbH Origin Spectrometer Frequency 100.62 VWM-ARWIV-164-flashed.2.fid Title Value / Volumes/ nmrdata/ vmak/ nmr/ VWM-ARWIV-164-flashed/ 2/ fid Parameter 129.78 129.14 Data File Name 100 ppm 90 70 76.36 74.68 73.73 70.99 67.62 80 60 40 30 20 46.37 45.93 38.56 32.97 28.67 28.18 23.56 21.22 20.67 19.98 50 10 0 -10 -20 Appendix 1 – Spectra Relevant to Chapter 2 203 Bruker BioSpin GmbH spect CDCl3 294.9 zg30 1D 16 78.7 1.0000 11.7000 4.0894 2017-06-28T04:29:23 Origin Instrument Solvent Temperature Pulse Sequence Experiment Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date -1545.6 1H 32768 65536 Lowest Frequency Nucleus Acquired Size Spectral Size 1.11 1.05 0.96 3.03 8.93 8.90 1.76 1.67 1.02 1.42 2.16 3.00 2.42 1.00 2.69 3.08 3.65 1.00 3.99 3.92 1.02 1.01 4.55 1.00 0.99 1.01 8.0 7.8 7.6 7.4 7.2 7.0 6.8 6.6 6.4 6.2 6.0 5.8 5.6 5.4 5.2 5.0 4.8 4.6 4.4 4.2 4.0 3.8 3.6 3.4 3.2 3.0 2.8 2.6 2.4 2.2 2.0 1.8 1.6 1.4 1.2 1.0 0.8 0.6 0.4 ppm 8012.8 Spectral Width Spectrometer Frequency 400.13 VWM7-arw4-165.1.fid Title Value / Volumes/ nmrdata/ vmak/ nmr/ VWM7-arw4-165/ 1/ fid Parameter 5.81 5.73 Data File Name Appendix 1 – Spectra Relevant to Chapter 2 204 207.22 1024 78.7 1.0000 10.0000 1.2976 2017-06-28T05:10:11 Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date 180 65536 Spectral Size 190 32768 Acquired Size 200 13C Nucleus 210 -2568.7 Lowest Frequency 220 25252.5 Spectral Width 120 1D Experiment 130 zgpg30 Pulse Sequence 140 295.0 Temperature 150 CDCl3 Solvent 160 spect Instrument 170 Bruker BioSpin GmbH Origin Spectrometer Frequency 100.62 VWM7-arw4-165.2.fid Title Value / Volumes/ nmrdata/ vmak/ nmr/ VWM7-arw4-165/ 2/ fid Parameter 129.54 129.38 Data File Name 110 100 ppm 90 70 76.86 73.79 71.09 70.17 80 60 40 30 20 51.04 46.65 38.48 32.67 31.18 28.66 28.21 21.21 20.66 19.96 50 10 0 -10 -20 Appendix 1 – Spectra Relevant to Chapter 2 205 zg30 1D 16 98.9 1.0000 11.7000 4.0894 2017-05-06T05:53:04 Pulse Sequence Experiment Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date 65536 Spectral Size 12.5 32768 Acquired Size 13.5 1H Nucleus 14.5 -1545.6 Lowest Frequency 15.5 8012.8 Spectral Width 8.5 295.0 Temperature 9.5 CDCl3 Solvent 10.5 spect Instrument 11.5 Bruker BioSpin GmbH Origin Spectrometer Frequency 400.13 VWM-arw4-166.1.fid Title Value / Volumes/ nmrdata/ vmak/ nmr/ VWM-arw4-166/ 1/ fid Parameter Data File Name 7.5 3.5 4.06 4.01 1.04 1.01 4.78 1.00 5.82 5.77 1.03 1.02 1.5 2.76 2.63 1.02 1.01 2.5 1.82 1.70 1.04 1.04 4.5 1.07 0.98 9.18 9.14 6.5 5.5 ppm 0.5 -0.5 -1.5 -2.5 -3.5 Appendix 1 – Spectra Relevant to Chapter 2 206 1D 512 78.7 2.0000 10.0000 1.3631 2017-05-06T06:22:57 Experiment Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date 65536 Spectral Size 180 32768 Acquired Size 190 13C Nucleus 200 -1961.2 Lowest Frequency 210 24038.5 Spectral Width 130 zgpg30 Pulse Sequence 140 294.9 Temperature 150 CDCl3 Solvent 160 spect Instrument 170 Bruker BioSpin GmbH Origin Spectrometer Frequency 100.62 VWM-arw4-166.2.fid Title Value / Volumes/ nmrdata/ vmak/ nmr/ VWM-arw4-166/ 2/ fid Parameter 129.60 129.01 Data File Name 120 110 100 ppm 90 80 73.59 71.83 71.01 70 60 50.37 50 30 20 38.66 34.07 28.65 28.20 21.24 20.67 40 10 0 -10 Appendix 1 – Spectra Relevant to Chapter 2 207 1D 16 64.2 1.0000 11.7000 4.0894 2017-05-03T01:39:19 Experiment Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date 65536 Spectral Size 12.5 32768 Acquired Size 13.5 1H Nucleus 14.5 -1545.2 Lowest Frequency 15.5 8012.8 Spectral Width 7.5 zg30 Pulse Sequence 8.5 294.9 Temperature 9.5 CDCl3 Solvent 10.5 spect Instrument 11.5 Bruker BioSpin GmbH Origin Spectrometer Frequency 400.13 VWM-ARWIV-167-flashed.1.fid Title Value / Volumes/ nmrdata/ vmak/ nmr/ VWM-ARWIV-167-flashed/ 1/ fid Parameter Data File Name 6.08 5.76 0.99 0.99 1.5 4.59 4.27 0.99 0.99 2.5 3.15 3.02 0.98 1.00 3.5 2.34 2.27 1.03 1.00 4.5 1.09 0.99 9.00 9.03 6.5 5.5 ppm 0.5 -0.5 -1.5 -2.5 -3.5 Appendix 1 – Spectra Relevant to Chapter 2 208 206.68 72.0 2.0000 10.0000 1.2780 2017-05-03T02:36:58 Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date 180 65536 Spectral Size 190 32768 Acquired Size 200 13C Nucleus 210 -2763.0 Lowest Frequency 220 25641.0 Spectral Width 110 1024 Number of Scans 120 1D Experiment 130 zgpg30 Pulse Sequence 140 295.0 Temperature 150 CDCl3 Solvent 160 spect Instrument 170 Bruker BioSpin GmbH Origin Spectrometer Frequency 100.62 VWM-ARWIV-167-flashed.2.fid Title Value 131.35 / Volumes/ nmrdata/ vmak/ nmr/ VWM-ARWIV-167-flashed/ 2/ fid Parameter 124.84 Data File Name 100 ppm 90 80 71.65 69.80 70 60 50 38.49 37.56 40 20 28.60 28.18 21.12 20.80 30 10 0 -10 -20 Appendix 1 – Spectra Relevant to Chapter 2 209 55.07 295.0 zg30 1D 16 156.2 1.0000 11.7000 4.0894 2017-05-06T06:27:59 Pulse Sequence Experiment Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date 65536 Spectral Size 7.6 32768 Acquired Size 8.0 1H Nucleus 8.4 -1545.2 Lowest Frequency 8.8 8012.8 Spectral Width 6.4 CDCl3 Temperature 6.8 spect Solvent 7.2 Bruker BioSpin GmbH Instrument Spectrometer Frequency 400.13 VWM-arw4-168.1.fid Origin 6.21 Title Value 5.84 / Volumes/ nmrdata/ vmak/ nmr/ VWM-arw4-168/ 1/ fid Parameter 5.49 Data File Name 1.01 6.0 0.99 5.6 5.2 4.0 3.6 2.4 2.0 1.6 4.69 1.01 1.2 4.28 1.00 2.8 3.13 1.02 3.2 2.91 1.00 4.4 ppm 1.08 1.01 9.16 9.06 4.8 0.8 0.4 0.0 Appendix 1 – Spectra Relevant to Chapter 2 210 1.00 163.28 131.74 130.78 Bruker BioSpin GmbH spect CDCl3 294.9 zgpg30 1D 512 55.5 2.0000 10.0000 1.3631 2017-05-06T06:57:52 Origin Instrument Solvent Temperature Pulse Sequence Experiment Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date -1960.7 13C 32768 65536 Lowest Frequency Nucleus Acquired Size Spectral Size 175 170 165 160 155 150 145 140 135 130 125 120 115 110 105 100 95 24038.5 Spectral Width Spectrometer Frequency 100.62 VWM-arw4-168.2.fid Title Value 118.50 / Volumes/ nmrdata/ vmak/ nmr/ VWM-arw4-168/ 2/ fid Parameter 113.19 Data File Name 90 ppm 85 80 75.68 75 70 64.95 65 60 55 50 45.17 44.18 45 40 35 25 28.79 28.18 30 21.15 20.72 20 15 10 Appendix 1 – Spectra Relevant to Chapter 2 211 296.1 zg30 1D Z122623_0045 (CPP BBO 400S1 BB-H&F-D-05 Z) 16 142.8 1.0000 12.5000 4.0894 2019-06-20T12:03:36 2019-06-20T12:03:36 Temperature Pulse Sequence Experiment Probe Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date Modification Date 32768 65536 Acquired Size Spectral Size 13.5 1H Nucleus 14.5 -1545.4 Lowest Frequency 15.5 8012.8 Spectral Width 10.5 CDCl3 Solvent 11.5 spect Instrument 12.5 Bruker BioSpin GmbH Origin Spectrometer Frequency 400.13 NJF-3-222.1.fid Title Value / Volumes/ nmrdata/ nfastuca/ nmr/ NJF-3-222/ 1/ fid 9.5 8.5 7.26 CDCl3 7.5 4.23 6.24 5.86 5.80 1.00 0.98 1.01 4.5 4.64 1.01 1.00 6.5 5.5 ppm 3.5 3.04 2.5 1.04 1.00 Parameter Data File Name 1.07 1.01 1.5 0.5 -0.5 -1.5 -2.5 -3.5 Appendix 1 – Spectra Relevant to Chapter 2 212 zgpg30 1D Z122623_0045 (CPP BBO 400S1 BB-H&F-D-05 Z) 512 72.0 1.0000 10.0000 1.3631 2019-06-20T12:24:50 2019-06-20T12:24:50 Pulse Sequence Experiment Probe Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date Modification Date -1961.4 13C 32768 65536 190 Lowest Frequency Nucleus Acquired Size Spectral Size 210 200 24038.5 Spectral Width 150 296.1 Temperature 160 CDCl3 Solvent 170 spect Instrument 180 Bruker BioSpin GmbH Origin Spectrometer Frequency 100.62 NJF-3-222.2.fid Title Value / Volumes/ nmrdata/ nfastuca/ nmr/ NJF-3-222/ 2/ fid Parameter Data File Name 130 135.90 132.94 130.36 128.20 140 120 110 100 ppm 90 77.00 CDCl3 76.60 80 70 60 51.64 47.28 50 40 20 28.83 28.22 21.18 20.70 30 10 0 -10 Appendix 1 – Spectra Relevant to Chapter 2 213 65.12 Bruker BioSpin GmbH Avance CDCl3 298.2 hsqcedetgpsisp2.3 HSQC-EDITED Z163739_0009 (PI HR-400-S1-BBF/ H/ D-5.0-Z SP) 4 32.0 1.5000 9.7500 0.1946 2019-06-20T22:10:42 2019-06-20T22:10:42 (400.15, 100.63) (5263.2, 16604.4) (-760.3, -743.2) (1H, 13C) (1024, 256) (1024, 1024) Origin Instrument Solvent Temperature Pulse Sequence Experiment Probe Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date Modification Date Spectrometer Frequency Spectral Width Lowest Frequency Nucleus Acquired Size Spectral Size 11.0 10.5 10.0 9.5 9.0 8.5 8.0 7.5 7.0 6.5 6.0 5.5 5.0 4.5 4.0 3.5 3.0 2.5 2.0 1.5 1.0 0.5 0.0 -0.5 -1.0 -1.5 ppm NJF-3-222.2.ser Title Value / Volumes/ nmrdata/ nfastuca/ nmr/ NJF-3-222/ 2/ ser Parameter Data File Name 150 140 130 120 110 100 90 80 70 60 50 40 30 20 10 0 Appendix 1 – Spectra Relevant to Chapter 2 214 ppm 11.0 5.9000 3.0000 2016-04-28T14:43:32 Pulse Width Acquisition Time Acquisition Date 65536 Spectral Size 9.0 24000 Acquired Size 9.5 1H Nucleus 10.0 -1013.6 Lowest Frequency 10.5 8000.0 Spectral Width 4.5 1.0000 Relaxation Delay 5.0 ppm 30 Receiver Gain 5.5 8 Number of Scans 6.0 1D Experiment 6.5 s2pul Pulse Sequence 7.0 25.0 Temperature 7.5 cdcl3 Solvent 8.0 inova Instrument 8.5 Varian Origin Spectrometer Frequency 499.66 PROTON01 Title Value / Volumes/ nmrdata/ arwong/ vnmrsys/ data/ ARWIII-047_redistilled_2/ PROTON01.fid/ fid Parameter Data File Name 4.0 3.0 2.0 3.37 2.91 2.87 0.93 2.5 2.85 2.49 2.34 2.21 2.00 1.65 0.96 1.00 1.01 2.02 1.00 1.05 3.5 1.5 1.0 0.5 0.0 -0.5 -1.0 Appendix 1 – Spectra Relevant to Chapter 2 215 3.76 11.0 1D 16 98.9 1.0000 11.7000 4.0894 2017-05-05T05:03:32 Experiment Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date 65536 Spectral Size 9.0 32768 Acquired Size 9.5 1H Nucleus 10.0 -1545.2 Lowest Frequency 10.5 8012.8 Spectral Width 6.5 zg30 Pulse Sequence 7.0 294.9 Temperature 7.5 CDCl3 Solvent 8.0 spect Instrument 8.5 Bruker BioSpin GmbH Origin Spectrometer Frequency 400.13 VWM7-095.1.fid Title Value / Volumes/ nmrdata/ vmak/ nmr/ VWM7-095/ 1/ fid Parameter Data File Name 6.0 5.5 5.0 ppm 4.5 3.0 1.5 3.89 3.72 4.10 5.87 2.0 3.31 0.99 2.5 2.69 1.01 3.5 2.08 1.93 1.80 1.56 1.42 1.03 2.05 1.03 1.07 1.02 4.0 1.0 0.5 0.0 -0.5 -1.0 Appendix 1 – Spectra Relevant to Chapter 2 216 1D 1024 78.7 2.0000 10.0000 1.3631 2017-05-05T06:02:40 Experiment Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date 65536 Spectral Size 180 32768 Acquired Size 190 13C Nucleus 200 -1962.2 Lowest Frequency 210 24038.5 Spectral Width 130 zgpg30 Pulse Sequence 140 295.0 Temperature 150 CDCl3 Solvent 160 spect Instrument 170 Bruker BioSpin GmbH Origin Spectrometer Frequency 100.62 VWM7-095.2.fid Title Value / Volumes/ nmrdata/ vmak/ nmr/ VWM7-095/ 2/ fid Parameter 169.00 168.93 Data File Name 116.95 120 110 100 ppm 90 80 60 50 64.27 64.25 56.69 52.42 52.40 70 30 40.62 36.82 35.55 28.25 40 20 10 0 -10 Appendix 1 – Spectra Relevant to Chapter 2 217 11.0 1.0000 5.8000 3.0000 2017-05-08T06:02:22 Pulse Width Acquisition Time Acquisition Date 65536 Spectral Size 9.0 24000 Acquired Size 9.5 1H Nucleus 10.0 -1017.6 Lowest Frequency 10.5 8000.0 Spectral Width 5.0 ppm 28 Relaxation Delay 5.5 8 Receiver Gain 6.0 1D Number of Scans 6.5 s2pul Experiment 7.0 25.0 Pulse Sequence 7.5 cdcl3 Temperature 8.0 inova Solvent 8.5 Varian Instrument Spectrometer Frequency 499.65 PROTON01 4.5 4.0 3.5 3.0 2.5 2.0 1.5 4.83 0.94 Origin 3.89 3.71 8.13 5.73 Title Value 2.65 1.00 / Volumes/ nmrdata-1/ vmak/ vnmrsys/ data/ VWM7-097/ PROTON01.fid/ fid Parameter 2.00 1.79 1.57 2.90 4.22 3.02 Data File Name 1.0 0.5 0.0 -0.5 -1.0 Appendix 1 – Spectra Relevant to Chapter 2 218 220 30 1.0000 4.6125 1.0420 2017-05-08T06:03:12 Relaxation Delay Pulse Width Acquisition Time Acquisition Date 65536 Spectral Size 180 32768 Acquired Size 190 13C Nucleus 200 -1906.1 Lowest Frequency 210 31446.5 Spectral Width 110 100 ppm 2000 Receiver Gain 120 1D Number of Scans 130 s2pul Experiment 140 25.0 Pulse Sequence 150 cdcl3 Temperature 160 inova Solvent 170 Varian Instrument Spectrometer Frequency 125.65 CARBON01 Origin 171.24 171.13 Title Value 116.59 / Volumes/ nmrdata-1/ vmak/ vnmrsys/ data/ VWM7-097/ CARBON01.fid/ fid Parameter 103.95 Data File Name 90 80 60 50 64.89 64.89 64.21 64.12 59.81 52.15 52.12 70 30 40.39 37.99 35.34 28.97 27.94 25.36 40 20 10 0 -10 Appendix 1 – Spectra Relevant to Chapter 2 219 11.0 zg30 1D 16 87.8 1.0000 11.7000 4.0894 2017-05-25T07:17:46 Pulse Sequence Experiment Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date 65536 Spectral Size 9.0 32768 Acquired Size 9.5 1H Nucleus 10.0 -1545.2 Lowest Frequency 10.5 8012.8 Spectral Width 7.0 294.9 Temperature 7.5 CDCl3 Solvent 8.0 spect Instrument 8.5 Bruker BioSpin GmbH Origin Spectrometer Frequency 400.13 VWM7-099.1.fid Title Value / Volumes/ nmrdata/ vmak/ nmr/ VWM7-099/ 1/ fid Parameter 6.69 Data File Name 0.96 6.5 6.0 5.5 5.0 ppm 4.5 2.5 2.0 3.78 3.67 2.98 2.88 3.0 3.10 1.00 3.5 2.41 2.26 2.00 4.06 2.06 2.06 4.0 1.5 1.0 0.5 0.0 -0.5 -1.0 Appendix 1 – Spectra Relevant to Chapter 2 220 204.61 1D 1024 72.0 2.0000 10.0000 1.3631 2017-05-25T08:16:54 Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date 65536 Spectral Size 180 32768 Acquired Size 190 13C Nucleus 200 -1963.2 Lowest Frequency 210 24038.5 Spectral Width 130 zgpg30 Experiment 140 294.9 Pulse Sequence 150 CDCl3 Temperature 160 spect Solvent 170 Bruker BioSpin GmbH Instrument Spectrometer Frequency 100.62 VWM7-099.2.fid Origin 171.83 169.44 Title Value 137.08 / Volumes/ nmrdata/ vmak/ nmr/ VWM7-099/ 2/ fid Parameter 131.32 Data File Name 120 110 100 ppm 90 80 70 50 55.01 52.70 52.17 60 37.74 42.78 40 20 29.37 23.90 23.24 30 10 0 -10 Appendix 1 – Spectra Relevant to Chapter 2 221 11.0 5.9000 3.0000 2016-05-03T22:10:42 Acquisition Time Acquisition Date 65536 Spectral Size 9.0 24000 Acquired Size 9.5 1H Nucleus 10.0 -1014.1 Lowest Frequency 10.5 8000.0 Spectral Width 1.00 4.5 4.0 1.0000 Pulse Width 5.0 ppm 38 Relaxation Delay 5.5 8 Receiver Gain 6.0 1D Number of Scans 6.5 s2pul Experiment 7.0 25.0 Pulse Sequence 7.5 cdcl3 Temperature 8.0 inova Solvent 8.5 Varian Instrument Spectrometer Frequency 499.66 PROTON01 Origin 4.37 Title Value 3.5 3.0 2.5 2.0 3.77 3.74 2.87 2.87 / Volumes/ nmrdata/ arwong/ vnmrsys/ data/ ARWIII-054_recryst_crop1/ PROTON01.fid/ fid Parameter 3.10 2.81 2.75 2.51 2.37 2.24 2.02 1.06 0.92 1.11 1.09 2.15 2.21 2.17 Data File Name 1.5 1.0 0.5 0.0 -0.5 -1.0 Appendix 1 – Spectra Relevant to Chapter 2 222 1D 1024 72.0 2.0000 10.0000 1.3631 2017-05-25T10:24:24 Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date 65536 Spectral Size 180 32768 Acquired Size 190 13C Nucleus 200 -1960.9 Lowest Frequency 210 24038.5 Spectral Width 130 zgpg30 Experiment 140 295.0 Pulse Sequence 150 CDCl3 Temperature 160 spect Solvent 170 Bruker BioSpin GmbH Instrument Spectrometer Frequency 100.62 VWM7-101.2.fid Origin Value Title 211.61 / Volumes/ nmrdata/ vmak/ nmr/ VWM7-101/ 2/ fid Parameter 171.29 171.15 Data File Name 120 110 100 ppm 90 76.53 80 70 60 40 55.48 52.97 52.47 47.92 42.24 36.15 50 20 28.21 27.63 21.30 30 10 0 -10 Appendix 1 – Spectra Relevant to Chapter 2 223 11.0 1.0000 5.8000 3.0000 2017-06-03T06:33:08 Relaxation Delay Pulse Width Acquisition Time Acquisition Date 65536 Spectral Size 9.0 24000 Acquired Size 9.5 1H Nucleus 10.0 -1017.6 Lowest Frequency 10.5 8000.0 Spectral Width 5.0 ppm 40 Receiver Gain 5.5 8 Number of Scans 6.0 1D Experiment 6.5 s2pul Pulse Sequence 7.0 25.0 Temperature 7.5 cdcl3 Solvent 8.0 inova Instrument 8.5 Varian Origin Spectrometer Frequency 499.65 PROTON01 Title Value / Volumes/ nmrdata/ vmak/ vnmrsys/ data/ VWM-arw3-055/ PROTON01.fid/ fid Parameter Data File Name 4.5 2.5 2.0 1.5 3.76 3.74 3.02 3.93 3.0 2.93 2.68 2.58 2.39 2.07 1.00 1.04 1.06 4.16 1.09 3.5 1.66 1.07 4.0 1.0 0.5 0.0 -0.5 -1.0 Appendix 1 – Spectra Relevant to Chapter 2 224 220 30 1.0000 4.6125 1.0420 2017-06-03T06:33:58 Relaxation Delay Pulse Width Acquisition Time Acquisition Date 65536 Spectral Size 180 32768 Acquired Size 190 13C Nucleus 200 -1904.0 Lowest Frequency 210 31446.5 Spectral Width 110 100 ppm 1000 Receiver Gain 120 1D Number of Scans 130 s2pul Experiment 140 25.0 Pulse Sequence 150 cdcl3 Temperature 160 inova Solvent 170 Varian Instrument Spectrometer Frequency 125.65 CARBON01 Origin Value Title 211.97 / Volumes/ nmrdata/ vmak/ vnmrsys/ data/ VWM-arw3-055/ CARBON01.fid/ fid Parameter 171.38 169.75 Data File Name 90 80 70 50 63.03 56.13 55.16 53.06 52.36 60 38.40 40 20 29.33 22.98 21.74 30 10 0 -10 Appendix 1 – Spectra Relevant to Chapter 2 225 43.80 11.0 1D 16 176.8 1.0000 11.7000 4.0894 2017-05-25T08:21:50 Experiment Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date 65536 Spectral Size 9.0 32768 Acquired Size 9.5 1H Nucleus 10.0 -1545.2 Lowest Frequency 10.5 8012.8 Spectral Width 6.5 zg30 Pulse Sequence 7.0 295.0 Temperature 7.5 CDCl3 Solvent 8.0 spect Instrument 8.5 Bruker BioSpin GmbH Origin Spectrometer Frequency 400.13 VWM7-103.1.fid Title Value / Volumes/ nmrdata/ vmak/ nmr/ VWM7-103/ 1/ fid Parameter Data File Name 5.0 ppm 3.0 2.5 2.0 1.5 1.0 0.5 0.0 6.03 5.68 5.57 0.99 0.99 0.97 3.5 4.48 4.26 1.02 1.01 4.0 3.74 3.67 3.30 3.06 2.90 2.74 2.61 2.34 2.12 2.04 1.96 1.83 3.01 3.00 1.00 1.01 1.00 1.00 1.01 2.03 0.99 0.97 0.97 0.99 4.5 1.45 1.09 1.02 1.01 9.23 9.34 5.5 0.05 9.21 6.0 -0.5 -1.0 Appendix 1 – Spectra Relevant to Chapter 2 226 1D 1024 87.8 2.0000 10.0000 1.3631 2017-05-25T09:20:57 Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date 65536 Spectral Size 180 32768 Acquired Size 190 13C Nucleus 200 -1961.0 Lowest Frequency 210 24038.5 Spectral Width 130 zgpg30 Experiment 140 295.0 Pulse Sequence 150 CDCl3 Temperature 160 spect Solvent 170 Bruker BioSpin GmbH Instrument Spectrometer Frequency 100.62 VWM7-103.2.fid Origin Value Title 172.00 170.38 163.21 / Volumes/ nmrdata/ vmak/ nmr/ VWM7-103/ 2/ fid Parameter 135.08 128.98 122.51 Data File Name 120 110 100 ppm 90 77.68 77.62 80 60 50 40 30 20 10 70.08 66.13 54.99 54.04 52.78 52.05 45.82 44.92 41.40 34.45 29.74 28.91 28.31 22.24 21.62 21.24 20.72 70 2.17 0 -10 Appendix 1 – Spectra Relevant to Chapter 2 227 11.0 8 46 1.0000 5.8000 3.0000 2017-05-04T06:45:07 Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date 65536 Spectral Size 9.0 24000 Acquired Size 9.5 1H Nucleus 10.0 -1017.6 Lowest Frequency 10.5 8000.0 Spectral Width 5.5 1.00 1.01 1.00 6.0 5.0 ppm 1D Experiment 6.5 s2pul Pulse Sequence 7.0 25.0 Temperature 7.5 cdcl3 Solvent 8.0 inova Instrument 8.5 Varian Origin Spectrometer Frequency 499.65 PROTON01 Title Value / Volumes/ nmrdata-1/ vmak/ vnmrsys/ data/ VWM6-274/ PROTON01.fid/ fid Parameter 5.88 5.65 5.40 Data File Name 4.0 1.0 0.5 0.0 4.66 4.31 4.23 1.00 0.99 0.98 1.5 3.74 3.67 3.40 2.97 2.91 0.99 2.0 2.98 2.02 2.5 2.48 2.32 2.11 1.75 1.64 1.52 0.98 0.98 3.05 0.97 1.03 0.95 3.0 1.08 1.00 9.12 8.90 3.5 0.05 8.21 4.5 -0.5 -1.0 Appendix 1 – Spectra Relevant to Chapter 2 228 220 1.0000 4.6125 1.0420 2017-05-04T06:45:56 Pulse Width Acquisition Time Acquisition Date 65536 Spectral Size 180 32768 Acquired Size 190 13C Nucleus 200 -1902.9 Lowest Frequency 210 31446.5 Spectral Width 110 100 ppm 30 Relaxation Delay 120 2000 Receiver Gain 130 1D Number of Scans 140 s2pul Experiment 150 25.0 Pulse Sequence 160 cdcl3 Temperature 170 inova Solvent Spectrometer Frequency 125.65 Varian Instrument Value CARBON01 215.90 Origin 170.67 170.59 Title 158.88 / Volumes/ nmrdata-1/ vmak/ vnmrsys/ data/ VWM6-274/ CARBON01.fid/ fid Parameter 134.82 128.20 124.42 Data File Name 90 80 76.72 69.87 66.23 70 50 40 30 20 10 58.04 56.25 52.78 52.64 46.20 46.15 41.76 38.83 28.93 28.30 27.38 23.66 21.22 20.67 19.32 60 0.01 0 -10 Appendix 1 – Spectra Relevant to Chapter 2 229 Appendix 2 – Spectra Relevant to Chapter 3 230 Appendix 2 Spectra Relevant to Chapter 3: Towards C19-Diterpenoid Alkaloids: Approaches to the C4 Chiral Quaternary Center 11.0 295.1 zg30 1D 16 87.8 1.0000 11.7000 4.0894 2018-05-07T12:26:02 Temperature Pulse Sequence Experiment Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date 65536 Spectral Size 9.0 32768 Acquired Size 9.5 1H Nucleus 10.0 -1545.7 Lowest Frequency 10.5 8012.8 Spectral Width 7.5 CDCl3 Solvent 8.0 spect Instrument 8.5 Bruker BioSpin GmbH Origin Spectrometer Frequency 400.13 ARWV-251_flashed_char.1.fid Title Value / Volumes/ nmrdata/ arwong/ nmr/ ARWV-251_flashed_char/ 1/ fid Parameter Data File Name 7.0 6.5 5.5 3.5 1.5 1.0 6.10 5.79 5.75 1.00 1.00 0.98 2.0 4.93 0.96 2.5 4.35 4.21 0.98 0.95 3.0 3.83 3.02 4.0 3.10 3.03 2.00 0.97 4.5 2.52 2.35 2.18 2.04 1.95 1.87 1.76 1.04 2.11 1.13 1.05 0.91 1.18 1.08 5.0 ppm 1.05 0.99 9.03 9.22 6.0 0.5 0.0 -0.5 -1.0 Appendix 2 – Spectra Relevant to Chapter 3 231 220 1.0000 10.0000 1.2496 2018-05-07T12:54:02 Pulse Width Acquisition Time Acquisition Date 65536 Spectral Size 180 32768 Acquired Size 190 13C Nucleus 200 -2048.3 Lowest Frequency 210 26223.8 Spectral Width 110 100 ppm 72.0 Relaxation Delay 120 256 Receiver Gain 130 1D Number of Scans 140 zgpg30 Experiment 150 295.2 Pulse Sequence 160 CDCl3 Temperature 170 spect Solvent Spectrometer Frequency 100.62 Bruker BioSpin GmbH Instrument Value ARWV-251_flashed_char.4.fid 211.98 Origin 169.77 169.59 Title 156.40 / Volumes/ nmrdata/ arwong/ nmr/ ARWV-251_flashed_char/ 4/ fid Parameter 134.07 130.11 127.01 Data File Name 90 78.63 77.47 80 70 50 40 30 20 10 57.99 53.52 52.87 45.86 44.83 43.04 35.29 28.77 28.19 26.10 23.50 22.35 21.16 20.66 60 0 -10 Appendix 2 – Spectra Relevant to Chapter 3 232 65.65 11.0 295.1 zg30 1D 16 127.1 1.0000 11.7000 4.0894 2018-05-07T12:59:11 Temperature Pulse Sequence Experiment Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date 65536 Spectral Size 9.0 32768 Acquired Size 9.5 1H Nucleus 10.0 -1545.1 Lowest Frequency 10.5 8012.8 Spectral Width 7.5 CDCl3 Solvent 8.0 spect Instrument 8.5 Bruker BioSpin GmbH Origin Spectrometer Frequency 400.13 ARWV-255_flashed_char.1.fid Title Value / Volumes/ nmrdata/ arwong/ nmr/ ARWV-255_flashed_char/ 1/ fid Parameter Data File Name 7.0 6.5 5.0 ppm 3.5 2.5 2.0 1.5 1.0 6.06 5.72 5.59 1.00 2.03 0.99 3.0 4.78 4.49 4.23 1.00 1.05 1.01 4.0 3.82 2.83 4.5 3.10 2.95 2.86 2.50 2.34 2.12 1.79 1.02 1.09 2.00 1.01 1.05 2.18 1.06 5.5 1.08 1.00 8.81 9.20 6.0 0.5 0.0 -0.5 Appendix 2 – Spectra Relevant to Chapter 3 233 220 295.2 zgpg30 1D 256 72.0 1.0000 10.0000 1.2496 2018-05-07T13:27:05 Pulse Sequence Experiment Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date 65536 Spectral Size 180 32768 Acquired Size 190 13C Nucleus 200 -2047.5 Lowest Frequency 210 26223.8 Spectral Width 150 CDCl3 Temperature 160 spect Solvent 170 Bruker BioSpin GmbH Instrument Spectrometer Frequency 100.62 ARWV-255_flashed_char.4.fid Origin Value Title 169.51 168.98 / Volumes/ nmrdata/ arwong/ nmr/ ARWV-255_flashed_char/ 4/ fid Parameter 156.90 Data File Name 140 133.42 129.85 126.07 130 118.30 114.33 120 110 100 ppm 90 77.20 75.26 80 70 50 40 30 20 56.75 52.95 52.74 46.23 46.18 43.86 32.50 28.81 28.22 26.57 21.89 21.20 20.68 60 10 0 -10 Appendix 2 – Spectra Relevant to Chapter 3 234 65.17 145.17 11.0 295.1 zg30 1D 32 98.9 1.0000 11.7000 4.0894 2018-05-07T06:37:28 Temperature Pulse Sequence Experiment Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date 65536 Spectral Size 9.0 32768 Acquired Size 9.5 1H Nucleus 10.0 -1545.0 Lowest Frequency 10.5 8012.8 Spectral Width 7.5 CDCl3 Solvent 8.0 spect Instrument 8.5 Bruker BioSpin GmbH Origin Spectrometer Frequency 400.13 ARWV-282_flashed.1.fid Title Value / Volumes/ nmrdata/ arwong/ nmr/ ARWV-282_flashed/ 1/ fid Parameter Data File Name 7.0 6.5 5.0 ppm 3.5 2.5 2.0 1.5 1.0 6.06 5.72 5.51 5.46 1.00 2.03 0.99 1.01 3.0 4.61 4.46 4.22 1.00 1.01 1.01 4.0 3.80 2.96 4.5 3.04 2.90 2.79 2.48 2.27 2.17 1.99 1.85 1.02 2.02 1.01 1.04 1.01 0.99 1.03 1.02 5.5 1.07 1.00 8.84 8.81 6.0 0.5 0.0 -0.5 -1.0 Appendix 2 – Spectra Relevant to Chapter 3 235 220 295.2 zgpg30 1D 256 35.5 1.0000 10.0000 1.2496 2018-05-07T07:05:31 Pulse Sequence Experiment Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date 65536 Spectral Size 180 32768 Acquired Size 190 13C Nucleus 200 -2048.0 Lowest Frequency 210 26223.8 Spectral Width 150 CDCl3 Temperature 160 spect Solvent 170 Bruker BioSpin GmbH Instrument Spectrometer Frequency 100.62 ARWV-282_flashed.4.fid Origin Value Title 170.37 170.27 / Volumes/ nmrdata/ arwong/ nmr/ ARWV-282_flashed/ 4/ fid Parameter 161.26 Data File Name 130 134.65 132.02 131.30 129.13 140 121.77 120 110 100 ppm 90 78.68 77.32 80 70 50 40 65.89 59.84 53.38 52.71 46.41 45.73 44.38 38.24 60 20 28.81 28.23 26.78 22.38 21.19 20.66 30 10 0 -10 Appendix 2 – Spectra Relevant to Chapter 3 236 11.0 295.2 zg30 1D 32 87.8 1.0000 11.7000 4.0894 2019-01-27T04:11:16 Temperature Pulse Sequence Experiment Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date 65536 Spectral Size 9.0 32768 Acquired Size 9.5 1H Nucleus 10.0 -1545.1 Lowest Frequency 10.5 8012.8 Spectral Width 7.5 CDCl3 Solvent 8.0 spect Instrument 8.5 Bruker BioSpin GmbH Origin Spectrometer Frequency 400.13 ARWVI-201_flashed_char.1.fid Title Value / Volumes/ nmrdata/ arwong/ nmr/ ARWVI-201_flashed_char/ 1/ fid Parameter Data File Name 7.0 6.5 5.0 ppm 3.0 2.5 2.0 1.5 1.0 6.06 5.71 5.49 1.00 2.09 1.95 3.5 4.62 4.45 4.23 4.03 0.98 1.04 1.07 1.03 4.0 3.42 1.07 4.5 3.04 2.77 2.58 2.41 2.12 1.90 1.57 1.02 0.99 1.07 3.26 2.28 1.12 0.99 5.5 1.07 1.00 8.91 9.51 6.0 0.5 0.0 -0.5 -1.0 Appendix 2 – Spectra Relevant to Chapter 3 237 295.1 zgpg30 1D 1024 72.0 1.0000 10.0000 1.3631 2019-01-27T05:05:04 Pulse Sequence Experiment Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date 65536 Spectral Size 180 32768 Acquired Size 190 13C Nucleus 200 -1962.1 Lowest Frequency 210 24038.5 Spectral Width 150 CDCl3 Temperature 160 spect Solvent 170 Bruker BioSpin GmbH Instrument Spectrometer Frequency 100.62 ARWVI-201_flashed_char.3.fid Origin Value Title 176.58 / Volumes/ nmrdata/ arwong/ nmr/ ARWVI-201_flashed_char/ 3/ fid Parameter 161.37 Data File Name 130 134.70 131.87 131.25 129.04 140 121.58 120 110 100 ppm 90 78.57 80 60 50 40 30 20 65.92 64.16 60.40 53.73 46.43 46.25 45.73 44.49 34.98 28.82 28.24 24.64 22.32 21.19 20.66 70 10 0 -10 Appendix 2 – Spectra Relevant to Chapter 3 238 11.0 295.2 zg30 1D 32 87.8 1.0000 11.7000 4.0894 2019-01-25T06:21:05 Temperature Pulse Sequence Experiment Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date 65536 Spectral Size 9.0 32768 Acquired Size 9.5 1H Nucleus 10.0 -1570.7 Lowest Frequency 10.5 8012.8 Spectral Width 7.5 CDCl3 Solvent 8.0 spect Instrument 8.5 Bruker BioSpin GmbH Origin Spectrometer Frequency 400.13 ARWVI-130_flashed_char.1.fid Title Value / Volumes/ nmrdata/ arwong/ nmr/ ARWVI-130_flashed_char/ 1/ fid Parameter Data File Name 7.0 6.5 5.0 ppm 3.5 3.0 2.5 2.0 1.5 1.0 5.99 5.63 5.43 5.40 1.00 2.03 0.98 0.95 4.0 4.53 4.39 4.16 0.96 1.00 1.00 4.5 3.58 3.38 3.30 2.96 2.71 2.56 2.28 2.04 1.85 1.58 0.99 1.00 3.11 1.01 1.00 2.03 0.97 0.99 2.09 0.98 5.5 1.01 0.94 9.33 9.16 6.0 0.5 0.0 -0.5 -1.0 Appendix 2 – Spectra Relevant to Chapter 3 239 295.2 zgpg30 1D 1500 78.7 0.5000 10.0000 1.3631 2019-01-25T07:32:03 Pulse Sequence Experiment Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date 65536 Spectral Size 180 32768 Acquired Size 190 13C Nucleus 200 -1961.7 Lowest Frequency 210 24038.5 Spectral Width 150 CDCl3 Temperature 160 spect Solvent 170 Bruker BioSpin GmbH Instrument Spectrometer Frequency 100.62 ARWVI-130_flashed_char.3.fid Origin Value Title 175.13 / Volumes/ nmrdata/ arwong/ nmr/ ARWVI-130_flashed_char/ 3/ fid Parameter 161.64 Data File Name 130 134.87 132.16 131.26 128.84 140 121.44 120 110 100 ppm 90 70 60 77.92 77.32 72.80 65.99 59.84 59.53 80 40 30 20 46.44 45.75 44.83 43.23 35.03 28.83 28.25 25.53 21.93 21.20 20.66 50 10 0 -10 Appendix 2 – Spectra Relevant to Chapter 3 240 295.2 hsqcedetgpsisp2.4 HSQC-EDITED 4 197.4 1.5000 11.7000 0.2135 2019-01-25T06:22:25 Temperature Pulse Sequence Experiment Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date 8.0 7.5 (1024, 1024) Spectral Size 8.5 (1024, 180) Acquired Size 9.0 (1H, 13C) Nucleus 9.5 (-517.1, -1262.8) Lowest Frequency 10.5 10.0 (4795.4, 16611.3) Spectral Width 6.0 CDCl3 Solvent 6.5 spect Instrument 7.0 Bruker BioSpin GmbH Origin Spectrometer Frequency (400.13, 100.62) ARWVI-130_flashed_char.2.ser Title Value / Volumes/ nmrdata/ arwong/ nmr/ ARWVI-130_flashed_char/ 2/ ser Parameter Data File Name 5.5 5.0 4.5 ppm 4.0 3.5 3.0 2.5 2.0 1.5 1.0 0.5 0.0 -0.5 -1.0 150 140 130 120 110 100 90 80 70 60 50 40 30 20 10 0 -10 Appendix 2 – Spectra Relevant to Chapter 3 241 ppm 11.0 295.1 zg30 1D 32 87.8 1.0000 11.7000 4.0894 2019-01-24T21:06:12 Temperature Pulse Sequence Experiment Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date 65536 Spectral Size 9.0 32768 Acquired Size 9.5 1H Nucleus 10.0 -1545.1 Lowest Frequency 10.5 8012.8 Spectral Width 7.5 CDCl3 Solvent 8.0 spect Instrument 8.5 Bruker BioSpin GmbH Origin Spectrometer Frequency 400.13 ARWVI-168_flashed_charact.1.fid Title Value / Volumes/ nmrdata/ arwong/ nmr/ ARWVI-168_flashed_charact/ 1/ fid Parameter Data File Name 7.0 6.5 5.0 ppm 2.5 2.0 1.5 1.0 6.19 6.08 1.00 0.97 3.0 5.67 5.49 5.44 1.07 1.06 1.03 3.5 4.44 4.22 4.03 1.11 1.12 1.04 4.0 3.55 3.46 3.31 3.24 3.04 2.91 2.68 2.51 2.39 2.18 1.12 3.02 1.02 2.15 1.03 1.03 1.05 1.10 4.5 1.68 4.43 5.5 1.11 1.08 1.01 1.25 9.03 9.03 6.0 0.5 0.0 -0.5 -1.0 Appendix 2 – Spectra Relevant to Chapter 3 242 295.2 zgpg30 1D 1500 72.0 0.5000 10.0000 1.3631 2019-01-24T22:17:11 Temperature Pulse Sequence Experiment Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date 65536 Spectral Size 180 32768 Acquired Size 190 13C Nucleus 200 -1961.5 Lowest Frequency 210 24038.5 Spectral Width 150 CDCl3 Solvent 160 spect Instrument 170 Bruker BioSpin GmbH Origin Spectrometer Frequency 100.62 ARWVI-168_flashed_charact.3.fid Title Value / Volumes/ nmrdata/ arwong/ nmr/ ARWVI-168_flashed_charact/ 3/ fid Parameter 165.43 Data File Name 130 135.91 135.58 133.61 128.35 140 120.86 120 110 100 ppm 90 70 79.74 77.07 72.64 68.18 66.33 80 50 40 30 20 59.08 56.13 46.47 46.25 41.52 39.55 34.99 28.86 28.29 24.57 21.24 20.70 20.67 60 10 0 -10 Appendix 2 – Spectra Relevant to Chapter 3 243 11.0 295.2 zg30 1D 32 112.8 1.0000 11.7000 4.0894 2019-01-24T23:09:00 Temperature Pulse Sequence Experiment Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date 65536 Spectral Size 9.0 32768 Acquired Size 9.5 1H Nucleus 10.0 -1545.2 Lowest Frequency 10.5 8012.8 Spectral Width 7.5 CDCl3 Solvent 8.0 spect Instrument 8.5 Bruker BioSpin GmbH Origin Spectrometer Frequency 400.13 ARWVI-181_flashed_charact.1.fid Title Value / Volumes/ nmrdata/ arwong/ nmr/ ARWVI-181_flashed_charact/ 1/ fid Parameter Data File Name 7.0 6.5 5.0 ppm 2.5 2.0 1.5 1.0 0.5 6.18 6.07 1.00 0.97 3.0 5.66 5.44 5.40 1.00 1.00 0.96 3.5 4.49 4.21 4.03 0.97 1.01 0.98 4.0 3.49 3.27 3.25 3.19 3.06 2.87 0.99 1.98 2.93 0.99 1.01 0.98 4.5 2.37 2.22 2.02 1.00 5.5 1.80 1.71 1.46 1.31 1.09 1.01 0.95 0.58 0.98 2.11 1.01 1.11 9.22 9.25 9.24 6.36 6.0 0.0 -0.5 -1.0 Appendix 2 – Spectra Relevant to Chapter 3 244 295.1 zgpg30 1D 1500 78.7 0.5000 10.0000 1.3631 2019-01-25T00:19:58 Temperature Pulse Sequence Experiment Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date 65536 Spectral Size 180 32768 Acquired Size 190 13C Nucleus 200 -1961.0 Lowest Frequency 210 24038.5 Spectral Width 150 CDCl3 Solvent 160 spect Instrument 170 Bruker BioSpin GmbH Origin Spectrometer Frequency 100.62 ARWVI-181_flashed_charact.3.fid Title Value / Volumes/ nmrdata/ arwong/ nmr/ ARWVI-181_flashed_charact/ 3/ fid Parameter 165.51 Data File Name 130 136.14 135.73 133.88 128.24 140 120.86 120 110 100 ppm 90 70 77.08 72.29 68.07 66.30 66.11 80 58.84 55.91 60 40 30 20 46.26 46.23 43.14 40.51 34.44 28.90 28.29 24.06 21.27 20.70 18.56 50 6.85 4.31 10 0 -10 Appendix 2 – Spectra Relevant to Chapter 3 245 11.0 295.1 zg30 1D 32 72.0 1.0000 11.7000 4.0894 2019-01-27T05:11:09 Temperature Pulse Sequence Experiment Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date 65536 Spectral Size 9.0 32768 Acquired Size 9.5 1H Nucleus 10.0 -1545.2 Lowest Frequency 10.5 8012.8 Spectral Width 7.5 CDCl3 Solvent 8.0 spect Instrument 8.5 Bruker BioSpin GmbH Origin Spectrometer Frequency 400.13 ARWVI-182_flashed_char.1.fid Title Value / Volumes/ nmrdata/ arwong/ nmr/ ARWVI-182_flashed_char/ 1/ fid Parameter Data File Name 7.0 6.5 5.0 ppm 3.0 2.5 2.0 1.5 1.0 0.5 6.08 5.88 5.65 5.38 1.00 1.00 1.01 2.04 3.5 4.47 4.0 4.22 3.55 3.45 3.30 3.24 3.24 3.21 3.16 3.04 2.87 2.31 2.23 1.01 1.02 1.01 1.01 1.13 3.11 2.71 1.04 0.92 1.04 1.00 2.00 1.03 4.5 1.67 1.57 1.31 1.09 1.01 0.95 1.03 1.59 2.31 9.28 9.28 9.49 5.5 0.57 6.20 6.0 0.0 -0.5 -1.0 Appendix 2 – Spectra Relevant to Chapter 3 246 295.1 zgpg30 1D 1024 64.2 1.0000 10.0000 1.3631 2019-01-27T06:04:57 Temperature Pulse Sequence Experiment Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date 65536 Spectral Size 180 32768 Acquired Size 190 13C Nucleus 200 -1961.0 Lowest Frequency 210 24038.5 Spectral Width 150 CDCl3 Solvent 160 spect Instrument 170 Bruker BioSpin GmbH Origin Spectrometer Frequency 100.62 ARWVI-182_flashed_char.3.fid Title Value / Volumes/ nmrdata/ arwong/ nmr/ ARWVI-182_flashed_char/ 3/ fid Parameter 166.58 Data File Name 130 135.95 133.97 131.59 127.98 140 119.83 120 110 100 ppm 90 70 60 50 40 30 20 79.72 77.10 73.39 66.47 65.99 58.71 57.23 56.54 46.20 46.20 43.70 40.60 34.77 28.92 28.31 21.42 21.26 20.94 20.69 80 6.84 4.32 10 0 -10 Appendix 2 – Spectra Relevant to Chapter 3 247 CDCl3 295.2 hsqcedetgpsisp2.4 HSQC-EDITED 2 197.4 1.5000 11.7000 0.2135 2019-01-27T05:12:25 Solvent Temperature Pulse Sequence Experiment Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date 8.0 7.5 (1024, 1024) Spectral Size 8.5 (1024, 180) Acquired Size 9.0 (1H, 13C) Nucleus 9.5 (-553.5, -1277.7) Lowest Frequency 10.5 10.0 (4795.4, 16611.3) Spectral Width 6.5 spect Instrument 7.0 Bruker BioSpin GmbH Origin Spectrometer Frequency (400.13, 100.62) ARWVI-182_flashed_char.2.ser Title Value / Volumes/ nmrdata/ arwong/ nmr/ ARWVI-182_flashed_char/ 2/ ser Parameter Data File Name 6.0 5.5 5.0 4.5 ppm 4.0 3.5 3.0 2.5 2.0 1.5 1.0 0.5 0.0 -0.5 -1.0 150 140 130 120 110 100 90 80 70 60 50 40 30 20 10 0 -10 Appendix 2 – Spectra Relevant to Chapter 3 248 ppm 11.0 295.1 zg30 1D 32 78.7 1.0000 11.7000 4.0894 2019-01-25T01:11:36 Temperature Pulse Sequence Experiment Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date 65536 Spectral Size 9.0 32768 Acquired Size 9.5 1H Nucleus 10.0 -1545.4 Lowest Frequency 10.5 8012.8 Spectral Width 7.5 CDCl3 Solvent 8.0 spect Instrument 8.5 Bruker BioSpin GmbH Origin Spectrometer Frequency 400.13 ARWVI-184_flashed_charact.1.fid Title Value / Volumes/ nmrdata/ arwong/ nmr/ ARWVI-184_flashed_charact/ 1/ fid Parameter Data File Name 7.0 6.5 5.0 ppm 4.0 3.0 2.5 2.0 1.5 1.0 6.08 5.90 5.66 5.43 1.00 1.01 1.03 2.04 3.5 4.44 4.23 1.03 1.02 4.5 3.54 3.46 3.27 3.26 3.21 3.02 2.91 2.64 2.46 2.30 2.10 2.05 3.00 2.91 0.99 1.91 1.01 1.00 1.02 1.05 5.5 1.65 1.52 1.15 1.08 1.01 1.22 2.19 1.09 8.84 9.00 6.0 0.5 0.0 -0.5 -1.0 Appendix 2 – Spectra Relevant to Chapter 3 249 295.1 zgpg30 1D 1500 87.8 0.5000 10.0000 1.3631 2019-01-25T02:22:35 Temperature Pulse Sequence Experiment Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date 65536 Spectral Size 180 32768 Acquired Size 190 13C Nucleus 200 -1958.4 Lowest Frequency 210 24038.5 Spectral Width 150 CDCl3 Solvent 160 spect Instrument 170 Bruker BioSpin GmbH Origin Spectrometer Frequency 100.62 ARWVI-184_flashed_charact.3.fid Title Value / Volumes/ nmrdata/ arwong/ nmr/ ARWVI-184_flashed_charact/ 3/ fid Parameter 166.45 Data File Name 130 135.77 133.67 131.49 128.15 140 119.58 120 110 100 ppm 90 70 80.53 79.72 77.03 72.38 66.50 80 50 40 30 20 58.86 57.23 56.76 46.37 46.16 42.33 39.67 35.46 28.88 28.29 23.21 21.58 21.24 20.66 60 10 0 -10 Appendix 2 – Spectra Relevant to Chapter 3 250 11.0 Parameter 9.53 10.5 10.0 Spectral Size Acquired Size Nucleus Lowest Frequency Spectral Width 9.5 0.89 Value 9.0 65536 32768 1H -1570.9 8012.8 8.5 8.0 2019-01-27T03:10:20 4.0894 11.7000 1.0000 98.9 32 1D zg30 295.1 CDCl3 spect Bruker BioSpin GmbH 7.5 ARWVI-185_flashed_char.1.fid / Volumes/ nmrdata/ arwong/ nmr/ ARWVI-185_flashed_char/ 1/ fid Spectrometer Frequency 400.13 Acquisition Date Acquisition Time Pulse Width Relaxation Delay Receiver Gain Number of Scans Experiment Pulse Sequence Temperature Solvent Instrument Origin Title Data File Name 7.0 6.5 5.0 ppm 4.0 3.0 2.5 2.0 1.5 1.0 6.09 5.85 5.68 5.62 5.40 1.00 1.00 1.06 0.96 0.96 3.5 4.46 4.5 4.21 1.05 1.00 5.5 3.49 3.39 3.31 3.27 3.26 3.02 2.88 2.44 2.29 2.22 2.12 1.83 1.55 1.40 1.08 1.02 1.01 1.14 1.26 2.81 2.93 1.03 0.99 0.91 0.97 0.95 0.95 0.94 1.05 0.88 9.09 9.08 6.0 0.5 0.0 -0.5 -1.0 Appendix 2 – Spectra Relevant to Chapter 3 251 203.99 295.2 zgpg30 1D 1024 72.0 1.0000 10.0000 1.3631 2019-01-27T04:04:10 Temperature Pulse Sequence Experiment Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date 65536 Spectral Size 180 32768 Acquired Size 190 13C Nucleus 200 -1958.4 Lowest Frequency 210 24038.5 Spectral Width 150 CDCl3 Solvent 160 spect Instrument 170 Bruker BioSpin GmbH Origin Spectrometer Frequency 100.62 ARWVI-185_flashed_char.3.fid Title Value / Volumes/ nmrdata/ arwong/ nmr/ ARWVI-185_flashed_char/ 3/ fid Parameter 164.49 Data File Name 130 120 135.22 133.95 131.97 128.49 121.77 140 110 100 ppm 90 79.54 77.32 76.13 80 60 50 40 30 20 66.29 59.26 58.11 56.95 50.83 46.16 45.99 44.10 34.53 28.90 28.28 21.49 21.23 21.14 20.69 70 10 0 -10 Appendix 2 – Spectra Relevant to Chapter 3 252 CDCl3 295.1 hsqcedetgpsisp2.4 HSQC-EDITED 2 197.4 1.5000 11.7000 0.2135 2019-01-27T03:11:36 Solvent Temperature Pulse Sequence Experiment Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date 8.0 7.5 (1024, 1024) Spectral Size 8.5 (1024, 180) Acquired Size 9.0 (1H, 13C) Nucleus 9.5 (-521.8, -1271.5) Lowest Frequency 10.5 10.0 (4795.4, 16611.3) Spectral Width 6.5 spect Instrument 7.0 Bruker BioSpin GmbH Origin Spectrometer Frequency (400.13, 100.62) ARWVI-185_flashed_char.2.ser Title Value / Volumes/ nmrdata/ arwong/ nmr/ ARWVI-185_flashed_char/ 2/ ser Parameter Data File Name 6.0 5.5 5.0 4.5 ppm 4.0 3.5 3.0 2.5 2.0 1.5 1.0 0.5 0.0 -0.5 -1.0 150 140 130 120 110 100 90 80 70 60 50 40 30 20 10 0 -10 Appendix 2 – Spectra Relevant to Chapter 3 253 ppm 11.0 296.2 zg30 1D 32 30.3 1.0000 11.7000 4.0894 2019-04-23T14:18:57 Temperature Pulse Sequence Experiment Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date 65536 Spectral Size 9.0 32768 Acquired Size 9.5 1H Nucleus 10.0 -1545.5 Lowest Frequency 10.5 8012.8 Spectral Width 7.5 CDCl3 Solvent 8.0 spect Instrument 8.5 Bruker BioSpin GmbH Origin Spectrometer Frequency 400.13 ARWVII-030_flashed.1.fid Title Value / Volumes/ nmrdata/ arwong/ nmr/ ARWVII-030_flashed/ 1/ fid Parameter Data File Name 7.0 6.5 4.0 3.0 2.5 2.0 1.5 1.0 6.06 5.87 5.64 5.40 5.16 5.08 0.96 1.98 1.00 1.92 1.00 0.92 3.5 4.44 4.22 1.00 0.99 4.5 3.55 3.24 3.02 2.88 0.99 10.24 1.00 0.94 5.0 ppm 2.49 2.41 2.30 2.01 1.01 2.07 5.5 1.66 1.54 1.36 1.07 1.00 1.18 2.10 1.01 9.26 9.29 6.0 0.5 0.0 -0.5 -1.0 Appendix 2 – Spectra Relevant to Chapter 3 254 296.2 zgpg30 1D 1024 64.2 1.0000 10.0000 1.3631 2019-04-23T15:01:56 Temperature Pulse Sequence Experiment Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date 65536 Spectral Size 180 32768 Acquired Size 190 13C Nucleus 200 -1961.8 Lowest Frequency 210 24038.5 Spectral Width 150 CDCl3 Solvent 160 spect Instrument 170 Bruker BioSpin GmbH Origin Spectrometer Frequency 100.62 ARWVII-030_flashed.2.fid Title Value / Volumes/ nmrdata/ arwong/ nmr/ ARWVII-030_flashed/ 2/ fid Parameter 166.48 Data File Name 130 136.80 135.84 133.98 131.28 128.05 140 119.65 116.08 120 110 100 ppm 90 79.63 77.00 76.54 80 60 50 40 30 20 10 66.49 58.75 57.16 56.81 56.29 53.03 46.26 46.16 44.50 39.28 35.36 28.89 28.29 24.35 21.73 21.23 20.66 70 0 -10 Appendix 2 – Spectra Relevant to Chapter 3 255 11.0 295.1 zg30 1D 32 64.2 1.0000 11.7000 4.0894 2018-12-11T01:00:34 Temperature Pulse Sequence Experiment Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date 65536 Spectral Size 9.0 32768 Acquired Size 9.5 1H Nucleus 10.0 -1545.4 Lowest Frequency 10.5 8012.8 Spectral Width 7.5 CDCl3 Solvent 8.0 spect Instrument 8.5 Bruker BioSpin GmbH Origin Spectrometer Frequency 400.13 ARWVI-270_flashed_char.1.fid Title Value / Volumes/ nmrdata/ arwong/ nmr/ ARWVI-270_flashed_char/ 1/ fid Parameter Data File Name 7.0 6.5 5.0 ppm 4.0 3.0 2.5 2.0 1.5 1.0 6.06 5.87 5.65 5.41 1.00 1.04 1.10 2.10 3.5 4.46 4.23 1.05 1.01 4.5 3.56 3.24 3.22 3.18 3.03 2.88 2.63 2.58 2.39 2.29 1.02 6.10 1.15 1.05 1.04 1.00 1.11 1.07 1.08 2.09 5.5 1.68 1.50 1.29 1.08 1.00 1.15 4.17 1.32 9.38 9.31 6.0 0.5 0.0 -0.5 -1.0 Appendix 2 – Spectra Relevant to Chapter 3 256 295.1 zgpg30 1D 1024 78.7 1.0000 10.0000 1.3631 2018-12-11T03:29:33 Temperature Pulse Sequence Experiment Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date 65536 Spectral Size 180 32768 Acquired Size 190 13C Nucleus 200 -1962.2 Lowest Frequency 210 24038.5 Spectral Width 150 CDCl3 Solvent 160 spect Instrument 170 Bruker BioSpin GmbH Origin Spectrometer Frequency 100.62 ARWVI-270_flashed_char.6.fid Title Value / Volumes/ nmrdata/ arwong/ nmr/ ARWVI-270_flashed_char/ 6/ fid Parameter 166.41 Data File Name 130 135.78 134.09 131.14 128.10 140 119.67 120 110 100 ppm 90 79.67 77.00 76.11 80 60 50 40 30 20 10 66.47 58.80 57.20 56.93 49.47 46.29 46.16 44.04 39.74 35.21 28.88 28.29 23.89 21.77 21.23 20.66 70 0 -10 Appendix 2 – Spectra Relevant to Chapter 3 257 spect CDCl3 295.1 hsqcedetgpsisp2.4 HSQC-EDITED 2 197.4 1.5000 11.7000 0.2135 2018-12-11T01:01:53 Instrument Solvent Temperature Pulse Sequence Experiment Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date 7.5 (1024, 1024) Spectral Size 8.0 (1024, 180) Acquired Size 8.5 (1H, 13C) Nucleus 9.0 (-524.6, -1262.8) Lowest Frequency 10.5 10.0 9.5 (4795.4, 16611.3) Spectral Width 6.5 Bruker BioSpin GmbH Origin 7.0 ARWVI-270_flashed_char.2.ser Title Spectrometer Frequency (400.13, 100.62) / Volumes/ nmrdata/ arwong/ nmr/ ARWVI-270_flashed_char/ 2/ ser Value Data File Name Parameter 6.0 5.5 5.0 4.5 ppm 4.0 3.5 3.0 2.5 2.0 1.5 1.0 0.5 0.0 -0.5 -1.0 150 140 130 120 110 100 90 80 70 60 50 40 30 20 10 0 -10 Appendix 2 – Spectra Relevant to Chapter 3 258 ppm 11.0 32 50 1.0000 5.8000 3.0000 2019-04-19T16:11:29 Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date 65536 Spectral Size 9.0 24000 Acquired Size 9.5 1H Nucleus 10.0 -1029.7 Lowest Frequency 10.5 8000.0 Spectral Width 6.5 1D Number of Scans 7.0 s2pul Experiment 7.5 3.0 Pulse Sequence 8.0 cdcl3 Temperature 8.5 inova Solvent Spectrometer Frequency 499.63 Varian Instrument 6.0 5.5 5.0 ppm 4.5 4.0 3.5 3.0 2.5 2.0 1.5 1.0 6.33 6.13 6.10 5.78 5.66 5.51 5.34 5.14 5.11 0.99 1.01 1.04 1.13 1.09 1.05 1.05 1.01 0.97 PROTON01 4.54 4.18 3.92 3.83 3.70 1.00 0.99 1.03 2.30 2.95 Origin 3.17 3.00 2.90 1.07 1.03 1.05 Title Value 2.34 2.09 1.78 3.11 1.08 3.17 / Volumes/ nmrdata-1/ arwong/ vnmrsys/ data/ ARWVII-020_flashed_char/ PROTON01.fid/ fid Parameter 1.08 1.01 9.06 8.95 Data File Name 0.5 0.0 -0.5 -1.0 Appendix 2 – Spectra Relevant to Chapter 3 259 220 s2pul 1D 512 30 1.0000 4.8375 1.0420 2019-04-19T16:13:56 Experiment Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date 65536 Spectral Size 180 32768 Acquired Size 190 13C Nucleus 200 -1907.9 Lowest Frequency 210 31446.5 Spectral Width 140 3.0 Pulse Sequence 150 cdcl3 Temperature 160 inova Solvent 170 Varian Instrument Spectrometer Frequency 125.64 CARBON01 Origin Value Title 174.86 168.61 163.50 / Volumes/ nmrdata-1/ arwong/ vnmrsys/ data/ ARWVII-020_flashed_char/ CARBON01.fid/ fid Parameter 130 120 135.62 135.18 133.67 132.84 128.48 122.05 116.69 Data File Name 110 100 ppm 90 76.60 80 60 50 40 30 20 66.94 66.24 56.95 55.61 52.95 46.36 46.30 46.23 42.13 35.32 28.89 28.26 25.55 21.24 20.67 18.09 70 10 0 -10 Appendix 2 – Spectra Relevant to Chapter 3 260 CDCl3 296.1 zg30 1D 32 176.8 1.0000 11.7000 4.0894 2019-04-22T19:07:00 Solvent Temperature Pulse Sequence Experiment Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date 32768 65536 Acquired Size Spectral Size 13.5 1H Nucleus 14.5 -1545.3 Lowest Frequency 15.5 8012.8 Spectral Width 11.5 spect Instrument 12.5 Bruker BioSpin GmbH Origin Spectrometer Frequency 400.13 ARWVII-025_flashed.1.fid Title Value / Volumes/ nmrdata-1/ arwong/ nmr/ ARWVII-025_flashed/ 1/ fid 10.5 9.5 8.5 6.19 6.13 6.06 5.82 5.66 5.44 5.17 4.53 4.21 4.02 3.94 3.84 3.76 3.55 3.05 2.95 2.86 2.25 1.85 1.43 1.08 1.00 7.5 6.5 5.5 ppm 4.5 3.5 2.5 1.5 0.5 1.00 0.90 0.99 1.08 1.00 0.93 0.97 2.13 1.01 0.99 1.03 1.10 1.18 1.14 1.00 1.77 0.94 0.95 3.14 2.09 1.95 9.07 9.17 Parameter Data File Name -0.5 -1.5 -2.5 -3.5 Appendix 2 – Spectra Relevant to Chapter 3 261 296.2 zgpg30 1D 1024 72.0 1.0000 10.0000 1.3631 2019-04-22T19:48:55 Temperature Pulse Sequence Experiment Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date 32768 65536 Acquired Size Spectral Size 190 13C Nucleus 200 -1960.8 Lowest Frequency 210 24038.5 Spectral Width 160 CDCl3 Solvent 170 spect Instrument 180 Bruker BioSpin GmbH Origin Spectrometer Frequency 100.62 ARWVII-025_flashed.2.fid Title Value 176.16 / Volumes/ nmrdata-1/ arwong/ nmr/ ARWVII-025_flashed/ 2/ fid Parameter 164.39 Data File Name 150 130 120 136.30 135.55 133.86 133.79 128.51 121.76 116.77 140 110 100 ppm 90 80 60 50 40 30 20 10 67.16 66.23 65.47 55.97 47.57 46.46 46.25 41.76 41.57 29.69 28.90 28.30 24.08 21.21 20.70 17.99 70 0 -10 Appendix 2 – Spectra Relevant to Chapter 3 262 11.0 296.2 zg30 1D 32 127.1 1.0000 11.7000 4.0894 2019-04-23T19:17:24 Temperature Pulse Sequence Experiment Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date 65536 Spectral Size 9.0 32768 Acquired Size 9.5 1H Nucleus 10.0 -1545.4 Lowest Frequency 10.5 8012.8 Spectral Width 7.5 CDCl3 Solvent 8.0 spect Instrument 8.5 Bruker BioSpin GmbH Origin Spectrometer Frequency 400.13 ARWVII-031_flash2.2.fid Title Value / Volumes/ nmrdata/ arwong/ nmr/ ARWVII-031_flash2/ 2/ fid Parameter Data File Name 6.5 4.0 3.5 3.0 2.5 2.0 1.5 1.0 6.80 0.91 4.5 6.06 5.83 5.67 5.44 5.39 5.13 1.00 2.18 1.06 0.93 1.00 2.22 5.0 ppm 4.49 0.98 5.5 4.24 3.86 3.60 3.42 3.28 3.26 3.21 3.04 2.87 2.69 2.59 2.30 2.20 1.73 1.64 1.02 2.17 0.98 1.21 2.82 2.93 1.10 1.07 1.06 1.01 0.98 1.25 1.15 1.30 0.99 6.0 1.17 1.08 1.01 1.26 9.12 9.49 7.0 0.5 0.0 -0.5 -1.0 Appendix 2 – Spectra Relevant to Chapter 3 263 296.1 zgpg30 1D 1024 87.8 1.0000 10.0000 1.3631 2019-04-23T19:59:55 Pulse Sequence Experiment Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date 65536 Spectral Size 180 32768 Acquired Size 190 13C Nucleus 200 -1961.0 Lowest Frequency 210 24038.5 Spectral Width 150 CDCl3 Temperature 160 spect Solvent 170 Bruker BioSpin GmbH Instrument Spectrometer Frequency 100.62 ARWVII-031_flash2.3.fid Origin Value Title 174.87 / Volumes/ nmrdata/ arwong/ nmr/ ARWVII-031_flash2/ 3/ fid Parameter 165.96 Data File Name 130 135.64 134.57 133.36 131.59 128.21 140 119.42 115.84 120 110 100 ppm 90 78.88 76.83 75.96 80 66.45 70 50 40 30 20 10 58.42 57.17 56.89 46.97 46.38 46.20 43.35 41.82 36.52 28.88 28.30 21.48 21.25 21.06 20.67 60 0 -10 Appendix 2 – Spectra Relevant to Chapter 3 264 spect CDCl3 297.2 zg30 1D Z122623_0045 (CPP BBO 400S1 BB-H&F-D-05 Z) 16 98.9 1.0000 12.5000 2.8224 2020-07-13T15:24:24 2020-07-13T15:24:24 Instrument Solvent Temperature Pulse Sequence Experiment Probe Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date Modification Date -1613.4 1H 32768 131072 Lowest Frequency Nucleus Acquired Size Spectral Size 12.0 11.5 11.0 10.5 10.0 9.5 11609.9 Spectral Width 9.0 Bruker BioSpin GmbH Origin Spectrometer Frequency 400.13 NJF-4-228-spot2.1.fid Title Value / Volumes/ nmrdata/ nfastuca/ nmr/ NJF-4-228-spot2/ 1/ fid Parameter Data File Name 8.5 8.0 7.5 7.0 5.0 4.5 ppm 4.0 3.5 3.0 2.5 2.0 1.5 1.0 6.30 6.07 5.84 5.65 5.46 5.39 5.19 5.14 0.81 1.00 1.91 1.04 0.82 1.02 0.99 0.93 5.5 4.50 4.22 3.88 3.73 3.54 3.24 3.03 2.92 2.70 0.96 1.01 2.08 0.93 1.74 2.89 1.20 0.90 0.83 6.0 2.23 2.06 1.75 1.40 1.08 1.00 2.90 0.87 2.21 0.90 8.95 8.99 6.5 0.5 0.0 -0.5 -1.0 -1.5 -2.0 Appendix 2 – Spectra Relevant to Chapter 3 265 297.1 hsqcedetgpsisp2.4 HSQC-EDITED Z122623_0045 (CPP BBO 400S1 BBH&F-D-05 Z) 2 197.4 1.5000 12.5000 0.2135 2020-07-13T15:31:25 2020-07-13T15:41:53 Temperature Pulse Sequence Experiment Probe Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date Modification Date (1024, 180) (1024, 1024) Acquired Size Spectral Size 8.5 (1H, 13C) Nucleus 9.0 (-527.2, -1229.4) Lowest Frequency 10.5 10.0 9.5 (4795.4, 16611.3) Spectral Width 7.0 CDCl3 Solvent 7.5 spect Instrument 8.0 Bruker BioSpin GmbH Origin Spectrometer Frequency (400.13, 100.62) NJF-4-228-spot2.3.ser Title Value / Volumes/ nmrdata/ nfastuca/ nmr/ NJF-4-228-spot2/ 3/ ser Parameter Data File Name 6.5 6.0 5.5 5.0 4.5 ppm 4.0 3.5 3.0 2.5 2.0 1.5 1.0 0.5 0.0 -0.5 -1.0 150 140 130 120 110 100 90 80 70 60 50 40 30 20 10 0 -10 Appendix 2 – Spectra Relevant to Chapter 3 266 ppm CDCl3 297.2 zgpg30 1D Z122623_0045 (CPP BBO 400S1 BB-H&F-D-05 Z) 512 55.5 1.0000 10.0000 1.3631 2020-07-13T16:03:14 2020-07-13T16:03:14 Solvent Temperature Pulse Sequence Experiment Probe Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date Modification Date 32768 65536 Acquired Size Spectral Size 190 13C Nucleus 200 -1961.8 Lowest Frequency 210 24038.5 Spectral Width 170 spect Instrument 180 Bruker BioSpin GmbH Origin Spectrometer Frequency 100.62 NJF-4-228-spot2.4.fid Title Value 176.13 / Volumes/ nmrdata/ nfastuca/ nmr/ NJF-4-228-spot2/ 4/ fid Parameter 165.83 Data File Name 160 150 130 120 135.72 134.05 133.79 131.80 128.25 120.76 116.61 140 110 100 ppm 90 78.62 80 66.30 66.06 70 50 40 30 20 10 57.77 56.33 47.94 46.38 46.19 42.81 41.84 36.16 28.90 28.29 21.72 21.20 20.68 19.48 60 0 -10 Appendix 2 – Spectra Relevant to Chapter 3 267 Bruker BioSpin GmbH spect CDCl3 297.1 cosygpppqf COSY Z122623_0045 (CPP BBO 400S1 BBH&F-D-05 Z) 1 50.3 2.0004 12.5000 0.1966 2020-07-13T15:25:27 2020-07-13T15:30:12 Origin Instrument Solvent Temperature Pulse Sequence Experiment Probe Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date Modification Date (-1810.9, -1811.1) (1H, 1H) (1024, 128) (1024, 1024) Lowest Frequency Nucleus Acquired Size Spectral Size 8.0 7.5 7.0 6.5 6.0 5.5 5.0 4.5 4.0 3.5 3.0 2.5 2.0 1.5 f2 (ppm) (5208.3, 5208.3) Spectral Width Spectrometer Frequency (400.13, 400.13) NJF-4-228-spot2.2.ser Title Value / Volumes/ nmrdata/ nfastuca/ nmr/ NJF-4-228-spot2/ 2/ ser Data File Name Parameter NJF-4-228-spot2.2.ser 1.0 0.5 0.0 -0.5 -1.0 -1.5 -2.0 8 7 6 5 4 3 2 1 0 -1 -2 Appendix 2 – Spectra Relevant to Chapter 3 268 f1 (ppm) CDCl3 297.1 zg30 1D Z122623_0045 (CPP BBO 400S1 BB-H&F-D-05 Z) 16 55.5 1.0000 12.5000 2.8224 2020-07-13T16:08:13 2020-07-13T16:08:13 Solvent Temperature Pulse Sequence Experiment Probe Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date Modification Date -1613.3 1H 32768 131072 Lowest Frequency Nucleus Acquired Size Spectral Size 12.0 11.5 11.0 10.5 10.0 11609.9 Spectral Width 9.0 spect Instrument 9.5 Bruker BioSpin GmbH Origin Spectrometer Frequency 400.13 NJF-4-228-spot3.1.fid Title Value / Volumes/ nmrdata/ nfastuca/ nmr/ NJF-4-228-spot3/ 1/ fid 8.5 8.0 7.5 7.0 5.5 5.0 4.5 ppm 4.0 3.5 3.0 2.5 6.62 6.11 6.06 5.82 5.66 5.50 5.41 5.16 5.10 0.80 0.95 0.96 0.96 1.00 0.92 0.96 1.11 0.84 6.0 4.46 4.22 4.04 3.86 3.49 3.29 3.04 3.01 2.89 2.66 2.33 0.99 0.97 0.96 1.89 0.97 2.97 1.04 0.94 0.95 3.13 6.5 1.79 2.0 1.35 1.08 1.00 1.5 1.0 2.37 0.97 8.98 8.90 Parameter Data File Name 0.5 0.0 -0.5 -1.0 -1.5 -2.0 Appendix 2 – Spectra Relevant to Chapter 3 269 spect CDCl3 297.1 hsqcedetgpsisp2.4 HSQC-EDITED Z122623_0045 (CPP BBO 400S1 BBH&F-D-05 Z) 2 197.4 1.5000 12.5000 0.2135 2020-07-13T16:15:13 2020-07-13T16:25:41 Instrument Solvent Temperature Pulse Sequence Experiment Probe Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date Modification Date (1024, 1024) Spectral Size 8.0 (1024, 180) Acquired Size 8.5 (1H, 13C) Nucleus 9.0 (-527.1, -1232.4) Lowest Frequency 10.5 10.0 9.5 (4795.4, 16611.3) Spectral Width 7.0 Bruker BioSpin GmbH Origin 7.5 NJF-4-228-spot3.3.ser Spectrometer Frequency (400.13, 100.62) / Volumes/ nmrdata/ nfastuca/ nmr/ NJF-4-228-spot3/ 3/ ser Title Value Data File Name Parameter 6.5 6.0 5.5 5.0 4.5 ppm 4.0 3.5 3.0 2.5 2.0 1.5 1.0 0.5 0.0 -0.5 -1.0 150 140 130 120 110 100 90 80 70 60 50 40 30 20 10 0 -10 Appendix 2 – Spectra Relevant to Chapter 3 270 ppm CDCl3 297.2 zgpg30 1D Z122623_0045 (CPP BBO 400S1 BB-H&F-D-05 Z) 512 78.7 1.0000 10.0000 1.3631 2020-07-13T16:47:01 2020-07-13T16:47:01 Solvent Temperature Pulse Sequence Experiment Probe Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date Modification Date 32768 65536 Acquired Size Spectral Size 190 13C Nucleus 200 -1962.6 Lowest Frequency 210 24038.5 Spectral Width 170 spect Instrument 180 Bruker BioSpin GmbH Origin Spectrometer Frequency 100.62 NJF-4-228-spot3.4.fid Title Value 174.91 / Volumes/ nmrdata/ nfastuca/ nmr/ NJF-4-228-spot3/ 4/ fid Parameter 164.93 Data File Name 160 150 130 115.91 120 135.69 135.49 134.47 133.37 128.42 120.97 140 110 100 ppm 90 76.88 75.21 80 67.53 66.25 70 50 40 30 20 10 58.75 56.34 47.06 46.43 46.23 42.49 41.71 36.11 28.85 28.27 24.60 21.23 20.65 18.54 60 0 -10 Appendix 2 – Spectra Relevant to Chapter 3 271 CDCl3 297.1 cosygpppqf COSY Z122623_0045 (CPP BBO 400S1 BBH&F-D-05 Z) 1 21.4 1.9410 12.5000 0.2560 2020-07-13T16:09:15 2020-07-13T16:14:00 Solvent Temperature Pulse Sequence Experiment Probe Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date Modification Date (1024, 1024) Spectral Size 6.5 (1024, 128) Acquired Size 7.0 (1H, 1H) Nucleus 7.5 (-726.3, -726.3) Lowest Frequency 8.0 (4000.0, 4000.0) Spectral Width 5.5 spect Instrument 6.0 Bruker BioSpin GmbH Origin Spectrometer Frequency (400.13, 400.13) NJF-4-228-spot3.2.ser Title Value / Volumes/ nmrdata/ nfastuca/ nmr/ NJF-4-228-spot3/ 2/ ser Data File Name Parameter NJF-4-228-spot3.2.ser 5.0 4.5 4.0 3.5 3.0 2.5 f2 (ppm) 2.0 1.5 1.0 0.5 0.0 -0.5 -1.0 -1.5 8 7 6 5 4 3 2 1 0 -1 Appendix 2 – Spectra Relevant to Chapter 3 272 f1 (ppm) 11.0 72.0 1.0000 11.7000 4.0894 2018-05-07T13:32:13 Relaxation Delay Pulse Width Acquisition Time Acquisition Date 65536 Spectral Size 9.0 32768 Acquired Size 9.5 1H Nucleus 10.0 -1545.0 Lowest Frequency 10.5 8012.8 Spectral Width 0.92 2.04 6.0 5.5 16 Receiver Gain 6.5 1D Number of Scans 7.0 zg30 Experiment 7.5 295.1 Pulse Sequence 8.0 CDCl3 Temperature 8.5 spect Solvent Spectrometer Frequency 400.13 Bruker BioSpin GmbH 6.22 6.11 Instrument 5.0 ppm 4.5 4.0 3.5 3.0 2.5 2.0 1.5 1.0 5.65 5.50 5.33 1.04 1.03 0.99 ARWV-258_flashed_char.1.fid 4.54 4.17 3.91 3.68 1.00 1.00 1.01 3.12 Origin 3.28 3.16 2.98 2.90 1.09 2.12 1.02 1.01 Title Value 2.32 2.06 1.75 3.15 1.02 3.79 / Volumes/ nmrdata/ arwong/ nmr/ ARWV-258_flashed_char/ 1/ fid Parameter 1.09 1.00 12.01 9.08 Data File Name 0.5 0.0 -0.5 -1.0 Appendix 2 – Spectra Relevant to Chapter 3 273 220 55.5 1.0000 10.0000 1.2496 2018-05-07T14:00:13 Relaxation Delay Pulse Width Acquisition Time Acquisition Date 65536 Spectral Size 180 32768 Acquired Size 190 13C Nucleus 200 -2048.3 Lowest Frequency 210 26223.8 Spectral Width 110 100 ppm 256 Receiver Gain 120 1D Number of Scans 130 zgpg30 Experiment 140 295.2 Pulse Sequence 150 CDCl3 Temperature 160 spect Solvent 170 Bruker BioSpin GmbH Instrument Spectrometer Frequency 100.62 ARWV-258_flashed_char.4.fid Origin Value Title 174.93 168.57 163.56 / Volumes/ nmrdata/ arwong/ nmr/ ARWV-258_flashed_char/ 4/ fid Parameter 135.63 135.14 132.84 128.45 121.96 Data File Name 90 76.59 80 60 50 40 30 20 10 66.94 66.24 56.93 55.48 52.89 46.36 46.31 46.21 35.24 34.73 28.88 28.26 25.57 21.23 20.66 18.04 14.62 70 0 -10 Appendix 2 – Spectra Relevant to Chapter 3 274 16 zg30 1D 16 78.7 1.0000 12.5000 Pulse Sequence Experiment Number of Scans Receiver Gain Relaxation Delay Pulse Width 1H 32768 65536 Nucleus Acquired Size Spectral Size 14 -1545.2 Lowest Frequency 15 8012.8 Spectral Width 10 297.2 Temperature 11 CDCl3 Solvent 12 spect Instrument 13 Bruker BioSpin GmbH Origin Spectrometer Frequency 400.13 NJF-4-079-char.1.fid Title Value / Volumes/ nmrdata/ nfastuca/ nmr/ NJF-4-079-char/ 1/ fid Parameter Data File Name 9 8 7 6.17 6.04 5.65 5.55 5.44 4.54 4.46 4.20 4.01 3.75 3.52 3.27 2.95 2.86 2.22 1.96 1.84 1.40 1.25 1.13 1.07 0.99 6 f1 (ppm) 5 4 3 2 1 0 1.00 2.09 1.19 0.69 1.22 1.07 1.23 1.01 0.93 0.93 3.26 1.21 0.96 0.95 3.23 0.98 2.03 0.83 0.91 3.22 9.25 10.15 NJF-4-079-char.1.fid -1 -2 -3 Appendix 2 – Spectra Relevant to Chapter 3 275 7.26 CDCl3 Bruker BioSpin GmbH spect CDCl3 297.2 zgpg30 1D 512 55.5 1.0000 10.0000 Instrument Solvent Temperature Pulse Sequence Experiment Number of Scans Receiver Gain Relaxation Delay Pulse Width -1945.7 13C 32768 65536 Lowest Frequency Nucleus Acquired Size Spectral Size 136.42 135.74 133.95 128.66 121.89 210 200 190 180 170 160 150 140 130 120 110 100 90 f1 (ppm) 24038.5 Spectral Width Spectrometer Frequency 100.62 NJF-4-079-char.2.fid Origin Value Title 176.44 / Volumes/ nmrdata/ nfastuca/ nmr/ NJF-4-079-char/ 2/ fid 164.58 Data File Name Parameter NJF-4-079-char.2.fid 77.16 CDCl3 77.04 80 50 40 30 20 10 56.13 47.38 46.64 46.40 41.63 36.17 34.45 29.06 28.45 24.23 21.35 20.85 18.20 14.89 60 67.34 66.40 65.63 70 0 -10 Appendix 2 – Spectra Relevant to Chapter 3 276 16 1D Z122623_0045 (CPP BBO 400S1 BB-H&F-D-05 Z) 16 72.0 1 12 4 2019-11-08T00:56:35 Experiment Probe Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date 32768 65536 Acquired Size Spectral Size 13 1H Nucleus 14 -1545.3 Lowest Frequency 15 8012.8 Spectral Width 9 zg30 Pulse Sequence 10 297.1 Temperature 11 CDCl3 Solvent 12 spect Instrument Spectrometer Frequency 400.13 NJF-4-081-spot2-repur.1.fid Title Value / Volumes/ nmrdata/ nfastuca/ nmr/ NJF-4-081-spot2-repur/ 1/ fid Parameter Data File Name 8 7 6.16 6.07 5.87 5.65 5.46 5.39 4.51 4.22 3.71 3.52 3.32 3.24 3.03 2.93 2.71 2.18 1.74 1.38 1.13 1.07 1.00 6 f1 (ppm) 5 4 3 2 1 0.94 1.00 1.00 1.03 1.00 0.96 1.00 0.99 1.02 1.99 1.18 4.13 1.90 0.98 0.97 3.37 2.02 1.05 3.33 9.11 9.09 NJF-4-081-spot2-repur.1.fid 0 -1 -2 -3 Appendix 2 – Spectra Relevant to Chapter 3 277 Bruker BioSpin GmbH spect CDCl3 297.2 zgpg30 1D Z122623_0045 (CPP BBO 400S1 BB-H&F-D-05 Z) 512 55.5 1.0000 10.0000 1.3631 2019-11-08T02:19:23 2019-11-08T02:19:23 Instrument Solvent Temperature Pulse Sequence Experiment Probe Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date Modification Date -1946.1 13C 32768 65536 Lowest Frequency Nucleus Acquired Size Spectral Size 120.89 210 200 190 180 170 160 150 140 130 120 110 100 90 f1 (ppm) 24038.5 Spectral Width Spectrometer Frequency 100.62 NJF-4-081-spot2-repur.5.fid Origin 176.34 Title Value 165.99 / Volumes/ nmrdata/ nfastuca/ nmr/ NJF-4-081-spot2-repur/ 5/ fid 135.90 133.95 131.96 128.40 Data File Name Parameter NJF-4-081-spot2-repur.5.fid 78.80 80 66.47 66.23 70 50 40 30 20 10 57.96 56.46 47.74 46.56 46.35 42.82 36.22 34.46 29.06 28.45 21.91 21.35 20.84 19.66 14.89 60 0 -10 Appendix 2 – Spectra Relevant to Chapter 3 278 CDCl3 297.2 hsqcedetgpsisp2.4 HSQC-EDITED Z122623_0045 (CPP BBO 400S1 BB-H&F-D-05 Z) 2 197.4 1.5000 12.5000 0.2135 2019-11-08T00:58:06 2019-11-08T01:08:34 Solvent Temperature Pulse Sequence Experiment Probe Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date Modification Date (1024, 180) (1024, 1024) Acquired Size Spectral Size 8.0 (1H, 13C) Nucleus 9.0 (-527.0, -1265.2) Lowest Frequency 10.0 (4795.4, 16611.3) Spectral Width 5.0 4.0 f2 (ppm) spect Instrument 6.0 Bruker BioSpin GmbH Origin 7.0 NJF-4-081-spot2-repur.2.ser Spectrometer Frequency (400.13, 100.62) / Volumes/ nmrdata/ nfastuca/ nmr/ NJF-4-081-spot2-repur/ 2/ ser Title Value Data File Name Parameter NJF-4-081-spot2-repur.2.ser 3.0 2.0 1.0 0.0 -1.0 150 140 130 120 110 100 90 80 70 60 50 40 30 20 10 0 -10 Appendix 2 – Spectra Relevant to Chapter 3 279 f1 (ppm) 7.0 cosygpppqf COSY Z122623_0045 (CPP BBO 400S1 BB-H&F-D-05 Z) 1 28.2 1.8796 12.5000 0.3174 2019-11-08T01:09:30 2019-11-08T01:14:16 Pulse Sequence Experiment Probe Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date Modification Date (1024, 128) (1024, 1024) Acquired Size Spectral Size 5.5 (1H, 1H) Nucleus 6.0 (-399.0, -399.0) Lowest Frequency 6.5 (3225.8, 3225.8) Spectral Width 3.5 297.2 Temperature 4.0 CDCl3 Solvent 4.5 spect Instrument 5.0 Bruker BioSpin GmbH Origin Spectrometer Frequency (400.13, 400.13) NJF-4-081-spot2-repur.3.ser Title Value / Volumes/ nmrdata/ nfastuca/ nmr/ NJF-4-081-spot2-repur/ 3/ ser Data File Name Parameter NJF-4-081-spot2-repur.3.ser 3.0 2.5 f2 (ppm) 2.0 1.5 1.0 0.5 0.0 -0.5 7.0 6.5 6.0 5.5 5.0 4.5 4.0 3.5 3.0 2.5 2.0 1.5 1.0 0.5 0.0 -0.5 Appendix 2 – Spectra Relevant to Chapter 3 280 f1 (ppm) CDCl3 297.1 hmbcetgpl3nd HMBC Z122623_0045 (CPP BBO 400S1 BB-H&F-D-05 Z) 4 197.4 2.0000 12.5000 0.4260 2019-11-08T01:15:21 2019-11-08T01:58:02 Solvent Temperature Pulse Sequence Experiment Probe Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date Modification Date (2048, 256) (2048, 1024) Acquired Size Spectral Size 9.0 (1H, 13C) Nucleus 10.0 (-173.0, -1118.9) Lowest Frequency 11.0 (4807.7, 22321.4) Spectral Width 7.0 spect Instrument 8.0 Bruker BioSpin GmbH Origin Spectrometer Frequency (400.13, 100.62) NJF-4-081-spot2-repur.4.ser Title Value / Volumes/ nmrdata/ nfastuca/ nmr/ NJF-4-081-spot2-repur/ 4/ ser Data File Name Parameter NJF-4-081-spot2-repur.4.ser 6.0 5.0 f2 (ppm) 4.0 3.0 2.0 1.0 0.0 -10 0 10 20 30 40 50 60 70 80 90 100 110 120 130 140 150 160 170 180 190 200 210 Appendix 2 – Spectra Relevant to Chapter 3 281 f1 (ppm) 14 cdcl3 25.0 s2pul 1D penta 4 18 1.0000 2.6500 1.7046 2019-11-08T00:50:35 2019-11-08T00:51:13 Solvent Temperature Pulse Sequence Experiment Probe Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date Modification Date 1H 16384 65536 Nucleus Acquired Size Spectral Size 12 -1196.2 Lowest Frequency 13 9611.9 Spectral Width 11 inova Instrument Spectrometer Frequency 599.59 Varian Origin Value NJF-4-081-spot3 Title Parameter NJF-4-081-spot3 STANDARD 1H OBSERVE - profile 10 9 8 7 5 3 2 1 6.48 6.11 6.05 5.66 5.49 5.40 0.91 1.00 1.05 1.05 0.98 0.95 4 4.46 4.21 4.03 3.48 3.27 3.04 2.89 2.63 2.40 2.33 2.24 1.74 1.33 1.10 1.08 1.00 1.02 1.01 0.95 0.99 6.30 1.15 0.95 1.01 1.02 1.12 1.16 2.01 1.05 3.24 9.09 9.37 6 f1 (ppm) 0 -1 Appendix 2 – Spectra Relevant to Chapter 3 282 spect CDCl3 297.1 hsqcedetgpsisp2.4 HSQC-EDITED Z122623_0045 (CPP BBO 400S1 BB-H&F-D-05 Z) 2 197.4 1.5000 12.5000 0.2135 2019-11-08T02:24:59 2019-11-08T02:35:28 Instrument Solvent Temperature Pulse Sequence Experiment Probe Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date Modification Date (1024, 180) (1024, 1024) Acquired Size Spectral Size 8.0 (1H, 13C) Nucleus 9.0 (-530.2, -1262.8) Lowest Frequency 10.0 (4795.4, 16611.3) Spectral Width 6.0 Bruker BioSpin GmbH Origin 7.0 NJF-4-081-spot3.2.ser Spectrometer Frequency (400.13, 100.62) / Volumes/ nmrdata-1/ nfastuca/ nmr/ NJF-4-081-spot3/ 2/ ser Title Value Data File Name Parameter NJF-4-081-spot3.2.ser 5.0 4.0 f2 (ppm) 3.0 2.0 1.0 0.0 -1.0 150 140 130 120 110 100 90 80 70 60 50 40 30 20 10 0 -10 Appendix 2 – Spectra Relevant to Chapter 3 283 f1 (ppm) 297.2 cosygpppqf COSY Z122623_0045 (CPP BBO 400S1 BB-H&F-D-05 Z) 1 18.8 1.8755 12.5000 0.3215 2019-11-08T02:36:23 2019-11-08T02:41:09 Temperature Pulse Sequence Experiment Probe Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date Modification Date (1024, 128) (1024, 1024) Acquired Size Spectral Size 6.0 (1H, 1H) Nucleus 6.5 (-266.4, -266.4) Lowest Frequency 7.0 (3184.7, 3184.7) Spectral Width 4.0 CDCl3 Solvent 4.5 spect Instrument 5.0 Bruker BioSpin GmbH 5.5 NJF-4-081-spot3.3.ser Origin Spectrometer Frequency (400.13, 400.13) / Volumes/ nmrdata-1/ nfastuca/ nmr/ NJF-4-081-spot3/ 3/ ser Title Value Data File Name Parameter NJF-4-081-spot3.3.ser 3.5 3.0 f2 (ppm) 2.5 2.0 1.5 1.0 0.5 0.0 -0.5 7.0 6.5 6.0 5.5 5.0 4.5 4.0 3.5 3.0 2.5 2.0 1.5 1.0 0.5 0.0 -0.5 Appendix 2 – Spectra Relevant to Chapter 3 284 f1 (ppm) CDCl3 297.1 hmbcetgpl3nd HMBC Z122623_0045 (CPP BBO 400S1 BB-H&F-D-05 Z) 4 197.4 2.0000 12.5000 0.4260 2019-11-08T02:42:14 2019-11-08T03:24:55 Solvent Temperature Pulse Sequence Experiment Probe Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date Modification Date (2048, 256) (2048, 1024) Acquired Size Spectral Size 9.0 (1H, 13C) Nucleus 10.0 (-163.1, -1116.4) Lowest Frequency 11.0 (4807.7, 22321.4) Spectral Width 7.0 spect Instrument 8.0 Bruker BioSpin GmbH Origin Spectrometer Frequency (400.13, 100.62) NJF-4-081-spot3.4.ser Title Value / Volumes/ nmrdata-1/ nfastuca/ nmr/ NJF-4-081-spot3/ 4/ ser Data File Name Parameter NJF-4-081-spot3.4.ser 6.0 5.0 f2 (ppm) 4.0 3.0 2.0 1.0 0.0 -10 0 10 20 30 40 50 60 70 80 90 100 110 120 130 140 150 160 170 180 190 200 210 Appendix 2 – Spectra Relevant to Chapter 3 285 f1 (ppm) Bruker BioSpin GmbH spect CDCl3 297.2 zgpg30 1D Z122623_0045 (CPP BBO 400S1 BB-H&F-D-05 Z) 512 55.5 1.0000 10.0000 1.3631 2019-11-08T03:46:13 2019-11-08T03:46:13 Origin Instrument Solvent Temperature Pulse Sequence Experiment Probe Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date Modification Date -1946.9 13C 32768 65536 Lowest Frequency Nucleus Acquired Size Spectral Size 121.07 135.84 135.66 133.52 128.56 210 200 190 180 170 160 150 140 130 120 110 100 90 f1 (ppm) 24038.5 Spectral Width Spectrometer Frequency 100.62 NJF-4-081-spot3.5.fid Title 175.01 / Volumes/ nmrdata-1/ nfastuca/ nmr/ NJF-4-081-spot3/ 5/ fid Value 165.12 Data File Name Parameter NJF-4-081-spot3.5.fid 70 77.16 CDCl3 77.03 75.39 67.69 66.41 80 50 40 30 20 10 58.88 56.48 47.00 46.58 46.38 42.75 36.18 34.34 29.01 28.43 24.78 21.38 20.81 18.64 15.00 60 0 -10 Appendix 2 – Spectra Relevant to Chapter 3 286 16 297.2 zg30 1D Z122623_0045 (CPP BBO 400S1 BB-H&F-D-05 Z) 16 78.7 1.0000 12.5000 4.0894 2019-11-01T19:22:10 2019-11-01T19:22:10 Temperature Pulse Sequence Experiment Probe Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date Modification Date 1H 32768 65536 Nucleus Acquired Size Spectral Size 14 -1545.2 Lowest Frequency 15 8012.8 Spectral Width 11 CDCl3 Solvent 12 spect Instrument 13 Bruker BioSpin GmbH Origin Spectrometer Frequency 400.13 NJF-4-084.1.fid Title Value / Volumes/ nmrdata/ nfastuca/ nmr/ NJF-4-084/ 1/ fid Parameter Data File Name 10 9 8 7 5 4 3 2 1 0 6.06 5.88 5.65 5.41 1.00 0.99 1.01 1.95 6 f1 (ppm) 4.46 4.23 3.56 3.25 3.24 3.19 3.03 2.89 2.59 2.46 2.40 2.31 1.67 1.53 1.35 1.07 1.01 1.00 0.98 0.96 3.13 2.85 1.98 0.99 0.96 2.03 1.90 1.04 1.98 1.05 2.15 1.15 12.18 9.18 NJF-4-084.1.fid -1 -2 -3 Appendix 2 – Spectra Relevant to Chapter 3 287 7.26 CDCl3 8.0 CDCl3 297.2 cosygpppqf COSY Z122623_0045 (CPP BBO 400S1 BB-H&F-D-05 Z) 1 28.2 1.9083 12.5000 0.2888 2019-11-01T19:23:08 2019-11-01T19:27:54 Solvent Temperature Pulse Sequence Experiment Probe Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date Modification Date (1024, 1024) Spectral Size 6.0 (1024, 128) Acquired Size 6.5 (1H, 1H) Nucleus 7.0 (-280.4, -280.4) Lowest Frequency 7.5 (3546.1, 3546.1) Spectral Width 5.0 spect Instrument 5.5 Bruker BioSpin GmbH Origin Spectrometer Frequency (400.13, 400.13) NJF-4-084.2.ser Title Value / Volumes/ nmrdata/ nfastuca/ nmr/ NJF-4-084/ 2/ ser Data File Name Parameter NJF-4-084.2.ser 4.5 4.0 3.5 f2 (ppm) 3.0 2.5 2.0 1.5 1.0 0.5 0.0 -0.5 8 7 6 5 4 3 2 1 0 Appendix 2 – Spectra Relevant to Chapter 3 288 f1 (ppm) spect CDCl3 297.1 hsqcedetgpsisp2.4 HSQC-EDITED Z122623_0045 (CPP BBO 400S1 BB-H&F-D-05 Z) 2 197.4 1.5000 12.5000 0.2135 2019-11-01T19:29:47 2019-11-01T19:40:16 Instrument Solvent Temperature Pulse Sequence Experiment Probe Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date Modification Date (1024, 180) (1024, 1024) Acquired Size Spectral Size 8.0 (1H, 13C) Nucleus 9.0 (-527.1, -1250.3) Lowest Frequency 10.0 (4795.4, 16611.3) Spectral Width 6.0 Bruker BioSpin GmbH Origin 7.0 NJF-4-084.3.ser Spectrometer Frequency (400.13, 100.62) / Volumes/ nmrdata/ nfastuca/ nmr/ NJF-4-084/ 3/ ser Title Value Data File Name Parameter NJF-4-084.3.ser 5.0 4.0 f2 (ppm) 3.0 2.0 1.0 0.0 -1.0 150 140 130 120 110 100 90 80 70 60 50 40 30 20 10 0 -10 Appendix 2 – Spectra Relevant to Chapter 3 289 f1 (ppm) NJF-4-084.4.fid Bruker BioSpin GmbH spect CDCl3 297.1 zgpg30 1D Z122623_0045 (CPP BBO 400S1 BB-H&F-D-05 Z) 512 55.5 1.0000 10.0000 1.3631 2019-11-01T20:01:37 2019-11-01T20:01:37 Title Origin Instrument Solvent Temperature Pulse Sequence Experiment Probe Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date Modification Date -1945.2 13C 32768 65536 Lowest Frequency Nucleus Acquired Size Spectral Size 119.72 136.06 134.18 131.52 128.20 210 200 190 180 170 160 150 140 130 120 110 100 90 f1 (ppm) 24038.5 Spectral Width Spectrometer Frequency 100.62 / Volumes/ nmrdata/ nfastuca/ nmr/ NJF-4-084/ 4/ fid Value 166.80 Data File Name Parameter NJF-4-084.4.fid 79.92 77.16 CDCl3 76.52 80 60 50 40 30 20 10 66.69 58.93 57.31 56.98 56.89 46.43 46.35 45.06 44.81 39.48 35.51 29.06 28.46 24.58 21.94 21.40 20.83 15.57 70 0 -10 Appendix 2 – Spectra Relevant to Chapter 3 290 14 inova cdcl3 25.0 s2pul 1D 8 30 2.0000 2.6500 Instrument Solvent Temperature Pulse Sequence Experiment Number of Scans Receiver Gain Relaxation Delay Pulse Width 1H 16384 65536 Nucleus Acquired Size Spectral Size 12 -1190.2 Lowest Frequency 13 9611.9 Spectral Width Spectrometer Frequency 599.60 Varian Origin Value NJF-2-253 Parameter Title 11 10 9 8 7 6 f1 (ppm) 3 2 4.86 1.00 4 3.98 3.91 3.88 3.69 3.60 2.11 2.14 4.17 2.34 2.20 5 2.59 2.14 1.85 1.71 1.60 1.52 1.99 1.19 2.26 5.15 3.18 2.21 NJF-2-253 new experiment 1 0 -1 Appendix 2 – Spectra Relevant to Chapter 3 291 7.26 CDCl3 Bruker BioSpin GmbH spect CDCl3 295.2 zgpg30 1D 512 72.0 1.0000 10.0000 Origin Instrument Solvent Temperature Pulse Sequence Experiment Number of Scans Receiver Gain Relaxation Delay Pulse Width -1946.0 13C 32768 65536 Lowest Frequency Nucleus Acquired Size Spectral Size 104.95 117.35 210 200 190 180 170 160 150 140 130 120 110 100 90 f1 (ppm) 24038.5 Spectral Width Spectrometer Frequency 100.62 NJF-2-253.4.fid Title Value / Volumes/ nmrdata-1/ nfastuca/ nmr/ NJF-2-253/ 4/ fid Data File Name Parameter NJF-2-253.4.fid 77.16 CDCl3 80 60 67.20 65.11 64.49 64.34 70 50 30 20 42.10 39.59 36.75 35.82 27.33 24.46 23.90 40 10 0 -10 Appendix 2 – Spectra Relevant to Chapter 3 292 14 inova cdcl3 25.0 s2pul 1D 8 42 2.0000 5.8000 Instrument Solvent Temperature Pulse Sequence Experiment Number of Scans Receiver Gain Relaxation Delay Pulse Width 1H 24000 65536 Nucleus Acquired Size Spectral Size 12 -1002.2 Lowest Frequency 13 8000.0 Spectral Width Spectrometer Frequency 499.63 Varian Origin Value PROTON01 Title Parameter PROTON01 NJF-3-063-diol 11 10 9 8 6 f1 (ppm) 5 3 3.96 3.82 3.71 3.54 0.99 1.11 1.09 1.00 6.82 0.91 2 2.59 2.36 2.20 0.99 2.33 6.95 4 1.55 1.31 1.96 1.07 7 1 0 -1 -2 Appendix 2 – Spectra Relevant to Chapter 3 293 NJF-3-084-diol.2.fid 205.86 Bruker BioSpin GmbH spect CDCl3 297.2 zgpg30 1D Z122623_0045 (CPP BBO 400S1 BB-H&F-D-05 Z) 512 64.2 1 10 1 2022-05-10T19:54:14 Origin Instrument Solvent Temperature Pulse Sequence Experiment Probe Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date -1945.6 13C 32768 65536 Lowest Frequency Nucleus Acquired Size Spectral Size 210 200 190 180 170 160 150 140 130 120 110 100 90 f1 (ppm) 24038.5 Spectral Width Spectrometer Frequency 100.62 NJF-3-084-diol.2.fid Title Value 138.24 / Volumes/ nmrdata/ nfastuca/ nmr/ NJF-3-084-diol/ 2/ fid Parameter 132.79 Data File Name 80 71.63 70 63.51 60 50 43.61 39.43 38.33 40 20 27.04 23.11 22.61 30 10 0 -10 Appendix 2 – Spectra Relevant to Chapter 3 294 inova cdcl3 25.0 s2pul 1D 8 50 2.0000 5.8000 Instrument Solvent Temperature Pulse Sequence Experiment Number of Scans Receiver Gain Relaxation Delay Pulse Width 8000.0 -1014.6 1H 24000 65536 12 Spectral Width Lowest Frequency Acquired Size Spectral Size 13 Nucleus Spectrometer Frequency 499.63 Varian Origin Value PROTON01 Parameter Title 11 10 9 8 7 6 f1 (ppm) 5 0.94 0.99 1.00 0.98 0.95 4 3 2 1 4.16 3.77 3.62 3.48 3.43 2.61 2.41 2.33 2.24 2.09 1.97 1.78 1.69 1.52 1.10 1.95 0.99 1.98 1.03 1.14 1.84 1.19 PROTON01 NJF-2-286-fr2 0 -1 -2 Appendix 2 – Spectra Relevant to Chapter 3 295 7.26 C6D6 Bruker BioSpin GmbH spect CDCl3 297.2 zgpg30 1D Z122623_0045 (CPP BBO 400S1 BB-H&F-D-05 Z) 1024 72.0 1 10 1 2022-04-23T04:27:27 Origin Instrument Solvent Temperature Pulse Sequence Experiment Probe Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date -1950.2 13C 32768 65536 Lowest Frequency Nucleus Acquired Size Spectral Size 210 200 190 180 170 160 150 140 130 120 110 100 90 f1 (ppm) 24038.5 Spectral Width Spectrometer Frequency 100.62 NJF-3-036-oxy_michael.1.fid Title Value / Volumes/ nmrdata/ nfastuca/ nmr/ NJF-3-036-oxy_michael/ 1/ fid 211.14 Data File Name Parameter NJF-3-036-oxy_michael.1.fid 80 60 67.96 66.34 65.66 70 50.37 50 20 27.66 25.66 21.32 30 38.85 37.69 36.18 40 10 0 -10 Appendix 2 – Spectra Relevant to Chapter 3 296 inova cdcl3 25.0 HSQCAD HSQC-EDITED 2 50 1.0000 11.6000 Instrument Solvent Temperature Pulse Sequence Experiment Number of Scans Receiver Gain Relaxation Delay Pulse Width (1200, 96) (2048, 1024) Acquired Size Spectral Size 11 (1H, 13C) Nucleus 12 (-1011.2, -1256.1) Lowest Frequency 13 (8000.0, 25125.6) Spectral Width Spectrometer Frequency (499.63, 125.64) Varian Origin Value HSQCAD01 Title HSQCAD01 NJF-2-286-fr2 Parameter 10 9 8 7 6 5 f2 (ppm) 4 3 2 1 0 -1 -2 180 170 160 150 140 130 120 110 100 90 80 70 60 50 40 30 20 10 0 Appendix 2 – Spectra Relevant to Chapter 3 297 f1 (ppm) Varian inova cdcl3 25.0 gCOSY COSY 1 38 1.0000 11.6000 Instrument Solvent Temperature Pulse Sequence Experiment Number of Scans Receiver Gain Relaxation Delay Pulse Width (1200, 128) (2048, 2048) Acquired Size Spectral Size 4.5 (1H, 1H) Nucleus 5.0 (-1014.6, -1014.6) Lowest Frequency 5.5 (8000.0, 8000.0) Spectral Width Spectrometer Frequency (499.63, 499.63) gCOSY01 Origin Value Title Parameter gCOSY01 NJF-2-286-fr2-1 4.0 3.5 3.0 f2 (ppm) 2.5 2.0 1.5 1.0 0.5 5.5 5.0 4.5 4.0 3.5 3.0 2.5 2.0 1.5 1.0 0.5 0.0 Appendix 2 – Spectra Relevant to Chapter 3 298 f1 (ppm) inova cdcl3 25.0 gHMBCAD HMBC 4 38 1.0000 11.6000 Instrument Solvent Temperature Pulse Sequence Experiment Number of Scans Receiver Gain Relaxation Delay Pulse Width (2048, 2048) Spectral Size 10 (1200, 200) Acquired Size 11 (1H, 13C) Nucleus 12 (-1014.6, -1889.2) Lowest Frequency 13 (8000.0, 30154.5) Spectral Width Spectrometer Frequency (499.63, 125.64) Varian Origin Value gHMBCAD01 Title Parameter gHMBCAD01 NJF-2-286-fr2-1 9 8 7 6 5 f2 (ppm) 4 3 2 1 0 -1 -2 220 200 180 160 140 120 100 80 60 40 20 0 Appendix 2 – Spectra Relevant to Chapter 3 299 f1 (ppm) 14 inova cdcl3 25.0 s2pul 1D 8 44 2.0000 5.8000 Instrument Solvent Temperature Pulse Sequence Experiment Number of Scans Receiver Gain Relaxation Delay Pulse Width 1H 24000 65536 Nucleus Acquired Size Spectral Size 12 -1002.2 Lowest Frequency 13 8000.0 Spectral Width Spectrometer Frequency 499.63 Varian Origin Value PROTON01 Parameter Title 11 10 9 8 7 6 f1 (ppm) 5 4.17 3.80 3.65 3.34 3.19 3.15 2.60 2.44 2.43 2.35 2.24 2.09 2.01 1.82 1.68 1.52 4 3 2 1 1.00 1.13 1.09 3.18 1.12 1.04 1.08 2.11 0.94 2.03 1.01 1.13 1.19 1.07 PROTON01 NJF-2-296 0 -1 -2 Appendix 2 – Spectra Relevant to Chapter 3 300 Bruker BioSpin GmbH spect CDCl3 297.1 zgpg30 1D Z122623_0045 (CPP BBO 400S1 BB-H&F-D-05 Z) 512 78.7 1 10 1 2022-05-10T23:12:56 Origin Instrument Solvent Temperature Pulse Sequence Experiment Probe Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date -1442.1 13C 32768 65536 Lowest Frequency Nucleus Acquired Size Spectral Size 220 210 200 190 180 170 160 150 140 130 120 110 100 f1 (ppm) 24038.5 Spectral Width Spectrometer Frequency 100.62 NJF-5-214.4.fid Title Value / Volumes/ nmrdata/ nfastuca/ nmr/ NJF-5-214/ 4/ fid 220.43 Data File Name Parameter NJF-5-214.4.fid 90 76.24 80 60 67.99 66.57 59.67 70 50.48 50 30 20 38.89 38.36 35.57 28.32 25.76 21.53 40 10 0 -10 Appendix 2 – Spectra Relevant to Chapter 3 301 spect CDCl3 295.2 hsqcedetgpsisp2.4 HSQC-EDITED 2 197.4 1.5000 11.7000 Instrument Solvent Temperature Pulse Sequence Experiment Number of Scans Receiver Gain Relaxation Delay Pulse Width (1024, 180) (1024, 1024) Acquired Size Spectral Size 8.0 (1H, 13C) Nucleus 9.0 (-526.9, -1265.2) Lowest Frequency 10.0 (4795.4, 16611.3) Spectral Width 6.0 Bruker BioSpin GmbH Origin 7.0 NJF-2-296.3.ser Spectrometer Frequency (400.13, 100.62) / Volumes/ nmrdata-1/ nfastuca/ nmr/ NJF-2-296/ 3/ ser Title Value Data File Name Parameter NJF-2-296.3.ser 5.0 4.0 f2 (ppm) 3.0 2.0 1.0 0.0 -1.0 150 140 130 120 110 100 90 80 70 60 50 40 30 20 10 0 -10 Appendix 2 – Spectra Relevant to Chapter 3 302 f1 (ppm) 14 inova cdcl3 25.0 s2pul 1D 24 40 2.0000 2.6500 Instrument Solvent Temperature Pulse Sequence Experiment Number of Scans Receiver Gain Relaxation Delay Pulse Width 1H 16384 65536 Nucleus Acquired Size Spectral Size 12 -1194.7 Lowest Frequency 13 9611.9 Spectral Width Spectrometer Frequency 599.60 Varian Origin Value NJF-3-075 Parameter Title 11 10 9 8 6 f1 (ppm) 5 1 0 6.80 0.92 2 3.89 3.74 3.57 3.42 1.00 1.03 1.02 1.00 3 2.65 2.48 2.31 2.17 2.06 0.98 0.96 2.97 2.46 1.05 4 1.47 1.20 0.93 1.16 1.02 9.01 7 0.11 5.80 NJF-3-075 new experiment -1 Appendix 2 – Spectra Relevant to Chapter 3 303 7.26 C6D6 NJF-5-213-major.2.fid 205.77 Bruker BioSpin GmbH spect CDCl3 297.2 zgpg30 1D Z122623_0045 (CPP BBO 400S1 BB-H&F-D-05 Z) 512 72.0 1 10 1 2022-05-12T03:08:27 Origin Instrument Solvent Temperature Pulse Sequence Experiment Probe Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date -1945.3 13C 32768 65536 Lowest Frequency Nucleus Acquired Size Spectral Size 210 200 190 180 170 160 150 140 130 120 110 100 90 f1 (ppm) 24038.5 Spectral Width Spectrometer Frequency 100.62 NJF-5-213-major.2.fid Title Value 138.04 / Volumes/ nmrdata/ nfastuca/ nmr/ NJF-5-213-major/ 2/ fid Parameter 133.18 Data File Name 80 72.30 70 63.43 60 50 43.33 39.30 38.30 40 20 27.09 26.00 23.27 22.10 18.32 30 10 0 -5.45 -5.53 -10 Appendix 2 – Spectra Relevant to Chapter 3 304 16 zg30 1D Z122623_0045 (CPP BBO 400S1 BB-H&F-D-05 Z) 16 112.8 1 12 4 2022-05-18T08:56:19 Pulse Sequence Experiment Probe Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date 32768 65536 Acquired Size Spectral Size 13 1H Nucleus 14 -1545.2 Lowest Frequency 15 8012.8 Spectral Width 9 297.2 Temperature 10 CDCl3 Solvent 11 spect Instrument 12 Bruker BioSpin GmbH Origin Spectrometer Frequency 400.13 NJF-5-219-major.1.fid Title Value / Volumes/ nmrdata-1/ nfastuca/ nmr/ NJF-5-219-major/ 1/ fid Data File Name Parameter NJF-5-219-major.1.fid 8 7 5 2 1 0 5.84 1.00 3 4.43 3.85 3.68 3.45 2.88 2.33 2.14 2.02 1.81 1.44 1.20 0.91 1.00 1.03 1.03 2.06 0.83 1.07 3.21 1.25 1.14 1.33 3.04 9.05 4 0.09 0.09 2.99 2.85 6 f1 (ppm) -1 -2 -3 Appendix 2 – Spectra Relevant to Chapter 3 305 Bruker BioSpin GmbH spect CDCl3 297.2 zgpg30 1D Z122623_0045 (CPP BBO 400S1 BB-H&F-D-05 Z) 1024 87.8 1 10 1 2022-05-18T09:50:16 Origin Instrument Solvent Temperature Pulse Sequence Experiment Probe Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date -1945.4 13C 32768 65536 Lowest Frequency Nucleus Acquired Size Spectral Size 122.53 210 200 190 180 170 160 150 140 130 120 110 100 90 f1 (ppm) 24038.5 Spectral Width Spectrometer Frequency 100.62 NJF-5-219-major.3.fid Title Value / Volumes/ nmrdata-1/ nfastuca/ nmr/ NJF-5-219-major/ 3/ fid 144.24 Data File Name Parameter NJF-5-219-major.3.fid 70 74.08 73.39 80 60 50 30 20 43.11 39.14 34.23 27.22 26.01 24.01 22.74 18.31 40 10 0 -5.48 -5.55 -10 Appendix 2 – Spectra Relevant to Chapter 3 306 64.08 297.2 hsqcedetgpsisp2.4 HSQC-EDITED Z122623_0045 (CPP BBO 400S1 BB-H&F-D-05 Z) 2 197.4 2 12 0 2022-05-18T09:08:04 Temperature Pulse Sequence Experiment Probe Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date (1024, 180) (1024, 1024) Acquired Size Spectral Size 8.0 (1H, 13C) Nucleus 9.0 (-526.9, -1265.2) Lowest Frequency 10.0 (4795.4, 16611.3) Spectral Width 5.0 4.0 f2 (ppm) CDCl3 Solvent 6.0 spect Instrument 7.0 Bruker BioSpin GmbH Origin Spectrometer Frequency (400.13, 100.62) NJF-5-219-major.2.ser Title Value / Volumes/ nmrdata-1/ nfastuca/ nmr/ NJF-5-219-major/ 2/ ser Data File Name Parameter NJF-5-219-major.2.ser 3.0 2.0 1.0 0.0 -1.0 150 140 130 120 110 100 90 80 70 60 50 40 30 20 10 0 -10 Appendix 2 – Spectra Relevant to Chapter 3 307 f1 (ppm) cosygpppqf COSY Z122623_0045 (CPP BBO 400S1 BB-H&F-D-05 Z) 1 64.2 2 12 0 2022-05-18T09:55:56 Pulse Sequence Experiment Probe Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date 6.0 (1024, 1024) Spectral Size 6.5 (1024, 128) Acquired Size 7.0 (1H, 1H) Nucleus 7.5 (-436.2, -436.2) Lowest Frequency 8.0 (3731.3, 3731.3) Spectral Width 4.0 3.5 3.0 f2 (ppm) 297.2 Temperature 4.5 CDCl3 Solvent 5.0 spect Instrument 5.5 Bruker BioSpin GmbH Origin Spectrometer Frequency (400.13, 400.13) NJF-5-219-major.4.ser Title Value / Volumes/ nmrdata-1/ nfastuca/ nmr/ NJF-5-219-major/ 4/ ser Data File Name Parameter NJF-5-219-major.4.ser 2.5 2.0 1.5 1.0 0.5 0.0 -0.5 -1.0 8 7 6 5 4 3 2 1 0 -1 Appendix 2 – Spectra Relevant to Chapter 3 308 f1 (ppm) 16 zg30 1D Z122623_0045 (CPP BBO 400S1 BB-H&F-D-05 Z) 16 127.1 1 12 4 2022-05-21T05:40:34 Pulse Sequence Experiment Probe Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date 32768 65536 Acquired Size Spectral Size 13 1H Nucleus 14 -1545.4 Lowest Frequency 15 8012.8 Spectral Width 9 297.1 Temperature 10 CDCl3 Solvent 11 spect Instrument 12 Bruker BioSpin GmbH Origin Spectrometer Frequency 400.13 NJF-5-222.1.fid Title Value / Volumes/ nmrdata/ dcharbon/ nmr/ NJF-5-222/ 1/ fid Data File Name Parameter NJF-5-222.1.fid 8 7 6 f1 (ppm) 5 0 4.26 4.17 3.65 3.55 3.31 1.00 0.96 1.95 1.99 0.92 1 2.58 2.26 1.87 1.65 1.51 1.34 1.04 2.32 3.37 2.06 2 0.87 9.09 3 0.02 0.02 2.58 3.13 4 -1 -2 -3 Appendix 2 – Spectra Relevant to Chapter 3 309 Bruker BioSpin GmbH spect CDCl3 297.1 zgpg30 1D Z122623_0045 (CPP BBO 400S1 BB-H&F-D-05 Z) 1024 78.7 1 10 1 2022-05-21T06:23:41 Origin Instrument Solvent Temperature Pulse Sequence Experiment Probe Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date -1945.2 13C 32768 65536 Lowest Frequency Nucleus Acquired Size Spectral Size 94.54 210 200 190 180 170 160 150 140 130 120 110 100 90 f1 (ppm) 24038.5 Spectral Width Spectrometer Frequency 100.62 NJF-5-222.2.fid Title Value / Volumes/ nmrdata/ dcharbon/ nmr/ NJF-5-222/ 2/ fid Data File Name Parameter NJF-5-222.2.fid 70 79.43 72.14 70.62 65.94 80 60 50.62 46.46 50 30 20 35.88 29.89 27.79 25.99 22.57 18.38 40 10 0 -5.41 -5.44 -10 Appendix 2 – Spectra Relevant to Chapter 3 310 297.2 hsqcedetgpsisp2.4 HSQC-EDITED Z122623_0045 (CPP BBO 400S1 BB-H&F-D-05 Z) 2 197.4 2 12 0 2022-05-21T06:35:25 Temperature Pulse Sequence Experiment Probe Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date (1024, 180) (1024, 1024) Acquired Size Spectral Size 8.0 (1H, 13C) Nucleus 9.0 (-527.0, -1254.0) Lowest Frequency 10.0 (4795.4, 16611.3) Spectral Width 5.0 4.0 f2 (ppm) CDCl3 Solvent 6.0 spect Instrument 7.0 Bruker BioSpin GmbH Origin Spectrometer Frequency (400.13, 100.62) NJF-5-222.3.ser Title Value / Volumes/ nmrdata/ dcharbon/ nmr/ NJF-5-222/ 3/ ser Data File Name Parameter NJF-5-222.3.ser 3.0 2.0 1.0 0.0 -1.0 150 140 130 120 110 100 90 80 70 60 50 40 30 20 10 0 -10 Appendix 2 – Spectra Relevant to Chapter 3 311 f1 (ppm) 297.1 cosygpppqf COSY Z122623_0045 (CPP BBO 400S1 BB-H&F-D-05 Z) 1 64.2 2 12 0 2022-05-21T08:10:44 Temperature Pulse Sequence Experiment Probe Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date 6.0 (1024, 1024) Spectral Size 6.5 (1024, 128) Acquired Size 7.0 (1H, 1H) Nucleus 7.5 (-428.1, -424.5) Lowest Frequency 8.0 (3703.7, 3703.7) Spectral Width 4.0 3.5 3.0 f2 (ppm) CDCl3 Solvent 4.5 spect Instrument 5.0 Bruker BioSpin GmbH 5.5 NJF-5-222.5.ser Origin Spectrometer Frequency (400.13, 400.13) / Volumes/ nmrdata/ dcharbon/ nmr/ NJF-5-222/ 5/ ser Title Value Data File Name Parameter NJF-5-222.5.ser 2.5 2.0 1.5 1.0 0.5 0.0 -0.5 -1.0 8 7 6 5 4 3 2 1 0 -1 Appendix 2 – Spectra Relevant to Chapter 3 312 f1 (ppm) inova cdcl3 3.0 s2pul 1D 16 56 1.0000 5.8000 Instrument Solvent Temperature Pulse Sequence Experiment Number of Scans Receiver Gain Relaxation Delay Pulse Width 1H 24000 65536 Nucleus Acquired Size Spectral Size 12 -1029.5 Lowest Frequency 13 8000.0 Spectral Width Spectrometer Frequency 499.63 Varian Origin Value PROTON01 Parameter Title 11 10 9 8 7 6 f1 (ppm) 3 2 4.90 0.97 4 3.97 3.87 2.00 1.99 5 2.67 2.47 2.22 2.09 1.94 1.68 1.54 1.00 2.00 2.03 1.05 4.04 1.10 8.65 PROTON01 NJF-3-130-fr1 1 0 -1 -2 Appendix 2 – Spectra Relevant to Chapter 3 313 7.26 CDCl3 Bruker BioSpin GmbH spect CDCl3 297.2 zgpg30 1D Z122623_0045 (CPP BBO 400S1 BB-H&F-D-05 Z) 512 64.2 1 10 1 2022-04-24T19:54:00 Origin Instrument Solvent Temperature Pulse Sequence Experiment Probe Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date -1945.9 13C 32768 65536 Lowest Frequency Nucleus Acquired Size Spectral Size 103.02 117.63 210 200 190 180 170 160 150 140 130 120 110 100 90 f1 (ppm) 24038.5 Spectral Width Spectrometer Frequency 100.62 NJF-3-168-minor.2.fid Title Value / Volumes/ nmrdata/ nfastuca/ nmr/ NJF-3-168-minor/ 2/ fid 166.95 Data File Name Parameter NJF-3-168-minor.2.fid 80 60 65.19 65.17 70 50 30 20 43.32 40.64 38.27 30.83 29.89 28.01 27.94 25.84 40 10 0 -10 Appendix 2 – Spectra Relevant to Chapter 3 314 54.57 85.01 inova cdcl3 3.0 s2pul 1D 64 58 1.0000 5.8000 Instrument Solvent Temperature Pulse Sequence Experiment Number of Scans Receiver Gain Relaxation Delay Pulse Width 1H 24000 65536 Nucleus Acquired Size Spectral Size 12 -1029.5 Lowest Frequency 13 8000.0 Spectral Width Spectrometer Frequency 499.63 Varian Origin Value PROTON01 Parameter Title 11 10 9 8 7 6 f1 (ppm) 4.92 0.93 5 1.97 1.92 4 3 2 1 0 3.98 3.88 3.15 3.15 3.14 3.13 3.13 3.12 3.11 3.10 2.65 2.47 2.30 2.23 2.00 1.89 1.70 1.52 1.00 1.08 2.85 1.13 3.00 0.97 1.13 9.00 PROTON01 NJF-3-130-fr2 -1 -2 Appendix 2 – Spectra Relevant to Chapter 3 315 7.26 CDCl3 Bruker BioSpin GmbH spect CDCl3 297.1 zgpg30 1D Z122623_0045 (CPP BBO 400S1 BB-H&F-D-05 Z) 2048 78.7 1 10 1 2022-04-22T02:47:59 Origin Instrument Solvent Temperature Pulse Sequence Experiment Probe Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date -1945.1 13C 32768 65536 Lowest Frequency Nucleus Acquired Size Spectral Size 103.11 117.69 210 200 190 180 170 160 150 140 130 120 110 100 90 f1 (ppm) 24038.5 Spectral Width Spectrometer Frequency 100.62 NJF-3-168-major.1.fid Title Value / Volumes/ nmrdata/ nfastuca/ nmr/ NJF-3-168-major/ 1/ fid 167.03 Data File Name Parameter NJF-3-168-major.1.fid 80 60 65.21 65.19 70 50 30 20 43.03 41.27 38.22 29.89 29.55 27.95 25.19 40 10 0 -10 Appendix 2 – Spectra Relevant to Chapter 3 316 54.21 85.04 14 cdcl3 25.0 s2pul 1D 8 48 2.0000 2.6500 1.7046 Solvent Temperature Pulse Sequence Experiment Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time 1H 16384 65536 Nucleus Acquired Size Spectral Size 12 -1195.0 Lowest Frequency 13 9611.9 Spectral Width 11 inova Instrument Spectrometer Frequency 599.60 Varian Origin Value NJF-3-099-repurified Title Parameter NJF-3-099-repurified new experiment 10 9 5 4 7.78 7.32 7.30 7.26 C6D6 7.14 2.00 2.04 1.95 4.04 6 f1 (ppm) 5.88 1.91 7 4.10 6.21 8 3 2 1 0 -1 Appendix 2 – Spectra Relevant to Chapter 3 317 cdcl3 3.0 s2pul 1D 8 54 1.0000 5.8000 3.0000 2019-02-17T11:45:15 Solvent Temperature Pulse Sequence Experiment Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date 1H 24000 65536 Nucleus Acquired Size Spectral Size 12 -1014.5 Lowest Frequency 13 8000.0 Spectral Width 11 inova Instrument Spectrometer Frequency 499.63 Varian Origin Value PROTON01 Title Parameter PROTON01 NJF-3-118 10 9 6 f1 (ppm) 5 4 7.87 7.52 7.49 7.26 CDCl3 7.24 7.13 2.00 2.04 1.94 2.11 1.93 7 4.11 6.14 8 3 2 1 0 -1 -2 Appendix 2 – Spectra Relevant to Chapter 3 318 s2pul 1D 4 60 5.0000 5.8000 3.0000 2019-02-21T15:03:51 Experiment Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date 1H 24000 65536 Nucleus Acquired Size Spectral Size 12 -1029.5 Lowest Frequency 13 8000.0 Spectral Width 8 3.0 Pulse Sequence 9 cdcl3 Temperature 10 inova Solvent 11 Varian Instrument Spectrometer Frequency 499.63 PROTON01 Origin 7 6 f1 (ppm) 5 4 3 2 7.79 7.35 7.26 CDCl3 7.23 7.06 6.96 2.00 1.97 1.96 2.03 1.86 Title Value 4.04 5.93 / Volumes/ nmrdata-1/ nfastuca/ vnmrsys/ data/ NJF-3-125/ PROTON01.fid/ fid 1.93 5.65 Data File Name Parameter PROTON01 NJF-3-125 1 0 -1 -2 Appendix 2 – Spectra Relevant to Chapter 3 319 3.0 s2pul 1D 8 60 1.0000 5.8000 3.0000 2019-03-01T12:10:17 Temperature Pulse Sequence Experiment Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date 1H 24000 65536 Nucleus Acquired Size Spectral Size 12 -1029.6 Lowest Frequency 13 8000.0 Spectral Width 9 cdcl3 Solvent 10 inova Instrument 11 Varian Origin Spectrometer Frequency 499.63 PROTON01 Title Value / Volumes/ nmrdata/ nfastuca/ vnmrsys/ data/ NJF-3-135/ PROTON01.fid/ fid Parameter 7.95 7.53 7.33 7.26 CDCl3 6.98 Data File Name 7 4.01 2.00 2.03 1.89 8 6 f1 (ppm) 5 4 3 2.05 2 5.74 PROTON01 NJF-3-135 1 0 -1 -2 Appendix 2 – Spectra Relevant to Chapter 3 320 3.0 s2pul 1D 8 60 1.0000 5.8000 3.0000 2019-03-01T12:18:19 Temperature Pulse Sequence Experiment Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date 1H 24000 65536 Nucleus Acquired Size Spectral Size 12 -1029.6 Lowest Frequency 13 8000.0 Spectral Width 9 cdcl3 Solvent 10 inova Instrument 11 Varian Origin Spectrometer Frequency 499.63 PROTON01 Value / Volumes/ nmrdata/ nfastuca/ vnmrsys/ data/ NJF-3-137/ PROTON01.fid/ fid Title Parameter Data File Name 7.76 7.68 7.26 CDCl3 7.15 7.08 2.08 2.00 8 6 f1 (ppm) 6.43 2.06 7 5.94 2.00 0.90 1.87 PROTON01 NJF-3-137 5 4 3 2 1 0 -1 -2 Appendix 2 – Spectra Relevant to Chapter 3 321 3.0 s2pul 1D 8 46 1.0000 5.8000 3.0000 2019-02-22T22:32:49 Temperature Pulse Sequence Experiment Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date 1H 24000 65536 Nucleus Acquired Size Spectral Size 12 -1029.4 Lowest Frequency 13 8000.0 Spectral Width 9 cdcl3 Solvent 10 inova Instrument 11 Varian Origin Spectrometer Frequency 499.63 PROTON01 Title Value / Volumes/ nmrdata/ nfastuca/ vnmrsys/ data/ NJF-3-126/ PROTON01.fid/ fid 8.24 7.95 Data File Name Parameter PROTON01 NJF-3-126 2.00 1.00 8 7 6 f1 (ppm) 5 4 3 2 1 0 -1 -2 Appendix 2 – Spectra Relevant to Chapter 3 322 12.29 7.26 CDCl3 s2pul 1D 8 50 1.0000 5.8000 3.0000 2019-03-05T13:47:37 Pulse Sequence Experiment Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date 1H 24000 65536 Nucleus Acquired Size Spectral Size 12 -1014.6 Lowest Frequency 13 8000.0 Spectral Width 9 3.0 Temperature 10 cdcl3 Solvent 11 inova Instrument Spectrometer Frequency 499.63 Varian Origin Value PROTON01 4.00 1.96 1.99 1.06 8 7 6 f1 (ppm) 8.10 7.95 7.90 7.83 7.26 CDCl3 7.18 7.11 2.11 1.93 / Volumes/ nmrdata/ nfastuca/ vnmrsys/ data/ NJF-3-138-column3-fr2-3/ PROTON01.fid/ fid 6.51 1.98 Title 6.13 2.01 Data File Name Parameter PROTON01 NJF-3-138-column3-fr2-3 5 4 3 2 1 0 -1 -2 Appendix 2 – Spectra Relevant to Chapter 3 323 3.0 s2pul 1D 32 60 1.0000 5.8000 3.0000 2019-03-09T17:27:09 Temperature Pulse Sequence Experiment Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date 1H 24000 65536 Nucleus Acquired Size Spectral Size 12 -1029.5 Lowest Frequency 13 8000.0 Spectral Width 9 cdcl3 Solvent 10 inova Instrument 11 Varian Origin Spectrometer Frequency 499.63 PROTON01 Title Value / Volumes/ nmrdata/ nfastuca/ vnmrsys/ data/ NJF-3-144/ PROTON01.fid/ fid Parameter 8.11 7.97 7.93 7.77 7.49 7.33 7.26 CDCl3 7.18 Data File Name 7 8.16 4.00 4.12 5.84 2.02 2.01 3.01 8 6 f1 (ppm) 5 4 3 2.36 2.07 2 5.65 PROTON01 NJF-3-144 1 0 -1 -2 Appendix 2 – Spectra Relevant to Chapter 3 324 3.0 s2pul 1D 8 60 1.0000 5.8000 3.0000 2019-03-11T12:43:32 Pulse Sequence Experiment Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date 1H 24000 65536 Nucleus Acquired Size Spectral Size 12 -1029.6 Lowest Frequency 13 8000.0 Spectral Width 9 cdcl3 Temperature 10 inova Solvent 11 Varian Instrument Spectrometer Frequency 499.63 PROTON01 Origin 8 7 6 f1 (ppm) 5 8.34 2.00 6.12 4.46 1.84 2.11 2.07 2.04 Title Value 8.16 8.12 8.11 8.06 7.99 7.94 7.87 7.60 7.38 7.26 CDCl3 7.24 7.23 7.18 / Volumes/ nmrdata/ nfastuca/ vnmrsys/ data/ NJF-3-141-fr3/ PROTON01.fid/ fid 4.37 4.32 1.80 1.84 Data File Name Parameter PROTON01 NJF-3-141-fr3 4 3 2 1 0 -1 -2 Appendix 2 – Spectra Relevant to Chapter 3 325 2.36 s2pul 1D 16 50 5.0000 5.8000 3.0000 2019-03-12T21:51:43 Pulse Sequence Experiment Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date 1H 24000 65536 Nucleus Acquired Size Spectral Size 12 -1029.7 Lowest Frequency 13 8000.0 Spectral Width 9 3.0 Temperature 10 cdcl3 Solvent 11 inova Instrument Spectrometer Frequency 499.63 Varian Origin Value PROTON01 8 7 6 f1 (ppm) 5 4 3 8.21 8.13 7.98 7.94 7.90 7.89 7.71 7.55 7.48 7.26 CDCl3 2.07 9.71 1.94 2.01 2.14 1.82 3.94 2.00 2.00 / Volumes/ nmrdata/ nfastuca/ vnmrsys/ data/ NJF-3-145/ PROTON01.fid/ fid 5.27 5.24 0.99 0.93 Title 4.04 3.88 3.50 3.11 3.00 1.93 1.91 1.98 2.00 1.82 Data File Name Parameter PROTON01 NJF-3-145 2 1 0 -1 -2 Appendix 2 – Spectra Relevant to Chapter 3 326 Appendix 3 – Spectra Relevant to Chapter 4 327 Appendix 3 Spectra Relevant to Chapter 4: Completion of Aconitine Core and Synthesis of (–)-Talatisamine, (–)-Liljestrandisine, and (–)-Liljestrandinine 11.0 1.0000 11.7000 4.0894 2018-04-17T03:48:29 Pulse Width Acquisition Time Acquisition Date 65536 Spectral Size 9.0 32768 Acquired Size 9.5 1H Nucleus 10.0 -1545.2 Lowest Frequency 10.5 8012.8 Spectral Width 5.5 127.1 Relaxation Delay 6.0 32 Receiver Gain 6.5 1D Number of Scans 7.0 zg30 Experiment 7.5 295.2 Pulse Sequence 8.0 CDCl3 Temperature 8.5 spect Solvent Spectrometer Frequency 400.13 Bruker BioSpin GmbH 5.0 ppm 4.5 4.0 3.5 3.0 2.5 2.0 1.5 1.0 6.05 5.89 5.64 5.46 5.27 1.01 1.01 1.04 1.01 0.97 Instrument 4.41 4.21 0.97 1.03 ARWV-267_flashed.1.fid 3.58 3.54 3.23 2.97 2.77 2.56 2.43 2.31 2.98 1.00 3.09 1.01 3.00 3.06 1.05 2.05 Origin 1.84 1.01 Title Value 1.48 3.29 / Volumes/ nmrdata-2/ arwong/ nmr/ ARWV-267_flashed/ 1/ fid Parameter 1.07 1.01 9.06 12.11 Data File Name 0.5 0.0 -0.5 Appendix 3 – Spectra Relevant to Chapter 4 328 220 1024 78.7 1.0000 10.0000 1.2496 2018-04-17T06:16:34 Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date 65536 Spectral Size 180 32768 Acquired Size 190 13C Nucleus 200 -2047.2 Lowest Frequency 210 26223.8 Spectral Width 120 1D Number of Scans 130 zgpg30 Experiment 140 295.1 Pulse Sequence 150 CDCl3 Temperature 160 spect Solvent 170 Bruker BioSpin GmbH Instrument Spectrometer Frequency 100.62 ARWV-267_flashed.6.fid 176.94 Origin 164.65 Title Value 135.60 133.35 131.08 128.19 / Volumes/ nmrdata-2/ arwong/ nmr/ ARWV-267_flashed/ 6/ fid Parameter 119.81 Data File Name 110 100 ppm 90 77.69 76.74 80 60 50 40 30 20 10 66.56 58.67 57.67 57.07 51.53 47.90 46.22 46.13 46.02 44.50 35.42 28.89 28.28 23.71 21.83 21.23 20.65 15.14 70 0 -10 Appendix 3 – Spectra Relevant to Chapter 4 329 11.0 zg30 1D 64 197.4 1.0000 11.7000 4.0894 2018-05-07T15:09:23 Pulse Sequence Experiment Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date 65536 Spectral Size 9.0 32768 Acquired Size 9.5 1H Nucleus 10.0 -1545.7 Lowest Frequency 10.5 8012.8 Spectral Width 1.00 7.0 6.5 295.2 Temperature 7.5 CDCl3 Solvent 8.0 spect Instrument 8.5 Bruker BioSpin GmbH Origin Spectrometer Frequency 400.13 ARWV-294_prepTLC_redo.1.fid Title Value 6.94 / Volumes/ nmrdata/ arwong/ nmr/ ARWV-294_prepTLC_redo/ 1/ fid Parameter 5.96 5.76 5.50 5.37 Data File Name 5.5 1.02 1.01 1.03 1.00 6.0 5.0 ppm 4.0 2.0 1.5 1.0 0.5 4.65 1.00 2.5 3.61 3.46 3.19 3.08 2.92 2.77 2.58 2.43 2.33 2.20 3.00 1.01 3.02 2.04 1.08 1.04 3.30 1.05 1.15 1.00 3.0 1.78 1.59 1.18 2.99 3.5 1.03 0.93 0.58 3.36 9.34 6.37 4.5 0.0 -0.5 -1.0 Appendix 3 – Spectra Relevant to Chapter 4 330 220 196.96 850 35.5 1.0000 10.0000 1.2496 2018-05-07T06:31:22 Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date 65536 Spectral Size 180 32768 Acquired Size 190 13C Nucleus 200 -2046.9 Lowest Frequency 210 26223.8 Spectral Width 120 1D Number of Scans 130 zgpg30 Experiment 140 295.1 Pulse Sequence 150 CDCl3 Temperature 160 spect Solvent 170 Bruker BioSpin GmbH Instrument Spectrometer Frequency 100.62 ARWV-294_prepTLC.4.fid 176.83 Origin 161.39 Title Value 149.16 / Volumes/ nmrdata/ arwong/ nmr/ ARWV-294_prepTLC/ 4/ fid Parameter 132.77 132.24 126.60 120.60 Data File Name 110 100 ppm 90 78.18 80 60 50 40 62.07 58.52 57.37 57.16 51.71 48.16 47.91 45.59 44.54 34.91 70 30 22.92 22.51 20 6.71 4.68 10 0 -10 Appendix 3 – Spectra Relevant to Chapter 4 331 15.14 84.60 11.0 1.0000 11.7000 4.0894 2018-05-05T19:42:25 Pulse Width Acquisition Time Acquisition Date 65536 Spectral Size 9.0 32768 Acquired Size 9.5 1H Nucleus 10.0 -1545.3 Lowest Frequency 10.5 8012.8 Spectral Width 1.15 7.0 5.5 197.4 Relaxation Delay 6.0 32 Receiver Gain 6.5 1D Number of Scans 7.5 zg30 Experiment 8.0 295.1 Pulse Sequence 8.5 CDCl3 Temperature Spectrometer Frequency 400.13 spect Solvent 6.93 Bruker BioSpin GmbH 5.0 ppm 4.5 4.0 3.5 3.0 2.5 2.0 1.5 1.0 0.5 5.94 5.76 5.51 5.37 1.13 1.11 0.95 1.10 Instrument 4.68 1.00 ARWV-295_prepTLC.1.fid 3.60 3.49 3.28 3.19 3.07 2.93 2.92 1.06 1.06 3.29 3.37 3.13 Origin 2.36 3.20 Title Value 1.80 1.70 1.58 1.03 1.16 3.56 / Volumes/ nmrdata/ arwong/ nmr/ ARWV-295_prepTLC/ 1/ fid Parameter 1.16 0.93 0.58 3.14 9.33 6.22 Data File Name 0.0 -0.5 -1.0 Appendix 3 – Spectra Relevant to Chapter 4 332 220 196.92 1000 72.0 1.0000 10.0000 1.2496 2018-05-05T20:43:37 Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date 65536 Spectral Size 180 32768 Acquired Size 190 13C Nucleus 200 -2047.1 Lowest Frequency 210 26223.8 Spectral Width 120 1D Number of Scans 130 zgpg30 Experiment 140 295.1 Pulse Sequence 150 CDCl3 Temperature 160 spect Solvent 170 Bruker BioSpin GmbH Instrument Spectrometer Frequency 100.62 ARWV-295_prepTLC.3.fid 175.93 Origin 161.20 Title Value 149.09 / Volumes/ nmrdata/ arwong/ nmr/ ARWV-295_prepTLC/ 3/ fid Parameter 133.03 131.76 126.62 120.58 Data File Name 110 100 ppm 90 84.59 77.91 72.30 80 60 50 40 62.04 60.36 57.34 57.30 51.70 48.42 47.92 45.86 70 30 23.10 22.30 20 12.80 6.70 4.69 10 0 -10 Appendix 3 – Spectra Relevant to Chapter 4 333 35.27 14 cdcl3 25.0 s2pul 1D penta 8 40 1.0000 2.6500 1.7078 2021-04-02T00:11:09 2021-04-02T00:11:35 Solvent Temperature Pulse Sequence Experiment Probe Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date Modification Date 1H 16384 65536 Nucleus Acquired Size Spectral Size 12 -1193.2 Lowest Frequency 13 9593.5 Spectral Width 11 inova Instrument Spectrometer Frequency 599.58 Varian Origin Value PROTON_01 Parameter Title 10 9 8 7 5.75 5.70 1.16 1.18 6 f1 (ppm) 1.40 1.06 0.94 0.59 4.89 4.78 3.76 3.71 3.54 3.28 3.20 3.15 2.90 2.61 2.51 2.41 2.24 2.13 2.03 5 4 3 2 1 0 1.00 0.92 0.97 2.88 0.99 1.11 1.03 3.16 1.09 0.96 1.23 2.33 1.26 1.13 1.05 1.07 1.25 1.47 3.29 9.13 6.09 PROTON_01 NJF-5-073 -1 Appendix 3 – Spectra Relevant to Chapter 4 334 7.26 CDCl3 Bruker BioSpin GmbH spect CDCl3 297.2 zgpg30 1D Z122623_0045 (CPP BBO 400S1 BB-H&F-D-05 Z) 13117 87.8 1.0000 10.0000 1.3631 2021-04-01T08:08:41 2021-04-01T08:08:41 Instrument Solvent Temperature Pulse Sequence Experiment Probe Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date Modification Date -1961.0 13C 32768 65536 Lowest Frequency Nucleus Acquired Size Spectral Size 114.69 135.18 131.10 210 200 190 180 170 160 150 140 130 120 110 100 90 f1 (ppm) 24038.5 Spectral Width Spectrometer Frequency 100.62 NJF-5-073.1.fid Origin Value Title 177.41 / Volumes/ nmrdata/ nfastuca/ nmr/ NJF-5-073/ 1/ fid 156.74 Data File Name Parameter 205.83 NJF-5-073.1.fid 70 60 50 40 30 77.00 CDCl3 74.65 71.69 62.98 61.05 56.10 54.88 53.65 52.10 48.67 47.99 44.52 41.90 33.91 80 25.23 21.32 15.01 20 6.75 4.66 10 0 -10 Appendix 3 – Spectra Relevant to Chapter 4 335 14 Varian inova cdcl3 25.0 gHMBCAD HMBC penta 16 40 1.0000 5.3000 0.1500 2021-04-02T17:29:00 2021-04-02T19:41:38 Instrument Solvent Temperature Pulse Sequence Experiment Probe Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date Modification Date (1439, 200) (2048, 2048) Acquired Size Spectral Size 11 (1H, 13C) Nucleus 12 (-1199.3, -2261.1) Lowest Frequency 13 (9593.5, 36182.7) Spectral Width Spectrometer Frequency (599.58, 150.78) gHMBCAD_01 Origin Value Title Parameter gHMBCAD_01 NJF-5-073-1 10 9 8 7 6 f2 (ppm) 5 4 3 2 1 0 -1 220 200 180 160 140 120 100 80 60 40 20 0 Appendix 3 – Spectra Relevant to Chapter 4 336 f1 (ppm) Varian inova cdcl3 25.0 gCOSY COSY penta 4 40 1.0000 5.3000 0.1501 2021-04-02T19:41:40 2021-04-02T19:52:33 Instrument Solvent Temperature Pulse Sequence Experiment Probe Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date Modification Date (1024, 1024) Spectral Size 6.5 (810, 128) Acquired Size 7.0 (1H, 1H) Nucleus 7.5 (-465.1, -465.1) Lowest Frequency 8.0 (5398.1, 5398.1) Spectral Width Spectrometer Frequency (599.58, 599.58) gCOSY_01 Origin Value Title Parameter gCOSY_01 NJF-5-073-1 6.0 5.5 5.0 4.5 4.0 3.5 f2 (ppm) 3.0 2.5 2.0 1.5 1.0 0.5 0.0 -0.5 8 7 6 5 4 3 2 1 0 Appendix 3 – Spectra Relevant to Chapter 4 337 f1 (ppm) inova cdcl3 25.0 s2pul 1D 22 36 1.0000 2.6500 Instrument Solvent Temperature Pulse Sequence Experiment Number of Scans Receiver Gain Relaxation Delay Pulse Width -1208.4 1H 16384 65536 Lowest Frequency Nucleus Acquired Size Spectral Size 1.27 2.92 2.73 2.45 2.34 2.14 2.00 1.90 1.04 2.05 6.52 1.30 1.18 4.05 3.75 3.70 3.34 3.12 3.02 1.03 5.67 1.00 14.0 13.5 13.0 12.5 12.0 11.5 11.0 10.5 10.0 9.5 9.0 8.5 8.0 7.5 7.0 6.5 6.0 5.5 5.0 4.5 4.0 3.5 3.0 2.5 2.0 1.5 1.0 0.5 0.0 -0.5 -1.0 -1.5 -2.0 ppm 9611.9 Spectral Width Spectrometer Frequency 599.60 Varian Origin Value ARWV-178_flashed Parameter Title Appendix 3 – Spectra Relevant to Chapter 4 338 cdcl3 25.0 s2pul 1D 48 24 1.0000 2.6500 Solvent Temperature Pulse Sequence Experiment Number of Scans Receiver Gain Relaxation Delay Pulse Width 16384 65536 Acquired Size Spectral Size 7.2 1H Nucleus 7.4 -1202.4 Lowest Frequency 7.6 9611.9 Spectral Width 7.0 inova Instrument Spectrometer Frequency 599.60 Varian Origin Value ARWV-179_flashed Title Parameter 6.8 6.6 ARWV-179_flashed STANDARD 1H OBSERVE - profile 6.4 6.2 6.0 5.8 5.6 5.0 4.8 4.6 4.2 4.0 5.37 1.00 3.4 4.32 0.99 3.6 3.73 3.02 3.8 3.64 3.00 4.4 3.36 0.98 5.4 5.2 f1 (ppm) 3.2 Appendix 3 – Spectra Relevant to Chapter 4 339 inova cdcl3 25.0 s2pul 1D 62 40 1.0000 2.6500 Instrument Solvent Temperature Pulse Sequence Experiment Number of Scans Receiver Gain Relaxation Delay Pulse Width -1208.4 1H 16384 65536 Lowest Frequency Nucleus Acquired Size Spectral Size 2.58 2.44 2.34 2.17 1.90 1.70 1.07 2.12 2.90 2.08 5.18 1.05 3.23 1.02 3.83 2.81 5.06 0.94 5.78 1.00 14.0 13.5 13.0 12.5 12.0 11.5 11.0 10.5 10.0 9.5 9.0 8.5 8.0 7.5 7.0 6.5 6.0 5.5 5.0 4.5 4.0 3.5 3.0 2.5 2.0 1.5 1.0 0.5 0.0 -0.5 -1.0 -1.5 -2.0 f1 (ppm) 9611.9 Spectral Width Spectrometer Frequency 599.60 Varian Origin Value ARWV-180_flashed Title Parameter ARWV-180_flashed STANDARD 1H OBSERVE - profile Appendix 3 – Spectra Relevant to Chapter 4 340 inova cdcl3 25.0 s2pul 1D 34 40 1.0000 2.6500 Instrument Solvent Temperature Pulse Sequence Experiment Number of Scans Receiver Gain Relaxation Delay Pulse Width -1202.4 1H 16384 65536 Lowest Frequency Nucleus Acquired Size Spectral Size 3.81 2.91 4.70 1.00 5.69 5.59 5.51 0.92 1.03 0.93 7.26 CDCl3 14.0 13.5 13.0 12.5 12.0 11.5 11.0 10.5 10.0 9.5 9.0 8.5 8.0 7.5 7.0 6.5 6.0 5.5 5.0 4.5 4.0 3.5 3.0 2.5 2.0 1.5 1.0 0.5 0.0 -0.5 -1.0 -1.5 -2.0 f1 (ppm) 9611.9 Spectral Width Spectrometer Frequency 599.60 Varian Origin Value ARWV-185_flashed Parameter Title 3.08 2.91 2.45 2.36 2.26 1.99 1.91 1.03 1.08 1.08 2.89 1.99 2.16 3.27 ARWV-185_flashed STANDARD 1H OBSERVE - profile Appendix 3 – Spectra Relevant to Chapter 4 341 Bruker BioSpin GmbH spect CDCl3 297.2 zgpg30 1D Z122623_0045 (CPP BBO 400S1 BB-H&F-D-05 Z) 512 38.0 1 10 1 2022-04-24T20:19:48 Instrument Solvent Temperature Pulse Sequence Experiment Probe Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date -1946.1 13C 32768 65536 Lowest Frequency Nucleus Acquired Size Spectral Size 116.90 116.34 129.91 210 200 190 180 170 160 150 140 130 120 110 100 90 f1 (ppm) 24038.5 Spectral Width Spectrometer Frequency 100.62 NJF-5-207.2.fid Origin 169.98 169.79 Title Value 146.02 / Volumes/ nmrdata/ nfastuca/ nmr/ NJF-5-207/ 2/ fid 139.86 Data File Name Parameter NJF-5-207.2.fid 76.26 80 70 50 56.45 53.11 53.03 60 30 20 33.14 33.05 32.35 26.65 22.83 21.89 43.60 40 10 0 -10 Appendix 3 – Spectra Relevant to Chapter 4 342 zg30 1D Z163739_0009 (PI HR-400-S1-BBF/ H/ D-5.0-Z SP) 16 101.0 1 10 4 2022-04-21T18:11:56 Pulse Sequence Experiment Probe Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date 65536 Spectral Size 13 32768 Acquired Size 14 1H Nucleus 15 -1637.1 Lowest Frequency 16 8196.7 Spectral Width 9 298.2 Temperature 10 CDCl3 Solvent 11 Avance Instrument 12 Bruker BioSpin GmbH Origin Spectrometer Frequency 400.15 NJF-5-202-arwV-185.1.fid Title Value / Volumes/ nmrdata/ nfastuca/ nmr/ NJF-5-202-arwV-185/ 1/ fid Parameter Data File Name 8 7 5 4.70 0.98 5.68 5.59 5.50 1.00 0.92 0.95 6 f1 (ppm) 3.77 4 3.07 2.91 2.35 3 2 0.99 1.03 6.91 6.17 NJF-5-202-arwV-185.1.fid 1 0 -1 -2 -3 -4 Appendix 3 – Spectra Relevant to Chapter 4 343 Bruker BioSpin GmbH spect CDCl3 297.2 zgpg30 1D Z122623_0045 (CPP BBO 400S1 BB-H&F-D-05 Z) 2048 72.0 1 10 1 2022-04-21T23:54:25 Instrument Solvent Temperature Pulse Sequence Experiment Probe Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date -1946.7 13C 32768 65536 Lowest Frequency Nucleus Acquired Size Spectral Size 132.39 130.90 125.13 210 200 190 180 170 160 150 140 130 120 110 100 90 f1 (ppm) 24038.5 Spectral Width Spectrometer Frequency 100.62 NJF-5-202.1.fid Origin Value Title 170.86 170.78 / Volumes/ nmrdata/ nfastuca/ nmr/ NJF-5-202/ 1/ fid 145.07 Data File Name Parameter NJF-5-202.1.fid 79.50 80 70 50 59.31 54.02 52.79 60 30 20 44.87 38.25 32.92 32.45 26.62 23.65 22.50 40 10 0 -10 Appendix 3 – Spectra Relevant to Chapter 4 344 zg30 1D Z163739_0009 (PI HR-400-S1-BBF/ H/ D-5.0-Z SP) 16 101.0 1 10 4 2022-04-21T18:24:20 Pulse Sequence Experiment Probe Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date 65536 Spectral Size 13 32768 Acquired Size 14 1H Nucleus 15 -1637.2 Lowest Frequency 16 8196.7 Spectral Width 9 298.2 Temperature 10 CDCl3 Solvent 11 Avance Instrument 12 Bruker BioSpin GmbH Origin Spectrometer Frequency 400.15 NJF-5-203-arwV-186.1.fid Title Value / Volumes/ nmrdata-1/ nfastuca/ nmr/ NJF-5-203-arwV-186/ 1/ fid Data File Name Parameter NJF-5-203-arwV-186.1.fid 8 7 5 3 6.22 6.08 5.49 5.40 0.96 0.99 0.97 1.02 1 3.95 3.67 3.25 0.98 3.06 3.07 2 2.46 2.29 2.08 1.83 1.05 6.12 1.08 5.08 4 1.09 3.00 6 f1 (ppm) 0 -1 -2 -3 -4 Appendix 3 – Spectra Relevant to Chapter 4 345 Bruker BioSpin GmbH spect CDCl3 297.2 zgpg30 1D Z122623_0045 (CPP BBO 400S1 BB-H&F-D-05 Z) 2048 72.0 1 10 1 2022-04-22T01:21:12 Instrument Solvent Temperature Pulse Sequence Experiment Probe Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date -1945.8 13C 32768 65536 Lowest Frequency Nucleus Acquired Size Spectral Size 125.44 210 200 190 180 170 160 150 140 130 120 110 100 90 f1 (ppm) 24038.5 Spectral Width Spectrometer Frequency 100.62 NJF-5-203.1.fid 174.64 Origin 169.11 Title Value 146.86 / Volumes/ nmrdata/ nfastuca/ nmr/ NJF-5-203/ 1/ fid 134.74 134.38 Data File Name Parameter NJF-5-203.1.fid 80 67.89 70 50 57.00 55.75 52.87 46.45 60 30 20 10 35.46 34.85 33.24 33.14 25.75 23.38 18.60 14.81 40 0 -10 Appendix 3 – Spectra Relevant to Chapter 4 346 16 297.2 zg30 1D Z122623_0045 (CPP BBO 400S1 BB-H&F-D-05 Z) 16 50.3 1 12 4 2022-04-24T18:21:43 Temperature Pulse Sequence Experiment Probe Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date 32768 65536 Acquired Size Spectral Size 13 1H Nucleus 14 -1545.1 Lowest Frequency 15 8012.8 Spectral Width 10 CDCl3 Solvent 11 spect Instrument 12 Bruker BioSpin GmbH Origin Spectrometer Frequency 400.13 NJF-5-208.1.fid Title Value / Volumes/ nmrdata/ nfastuca/ nmr/ NJF-5-208/ 1/ fid Data File Name Parameter NJF-5-208.1.fid 9 8 7 5 4 2 6.10 5.83 5.47 5.33 0.83 1.00 1.08 1.11 3 3.68 3.43 3.22 2.84 2.28 1.84 1.56 1.08 3.02 1.04 6.94 0.56 8.45 3.04 1.21 3.21 6 f1 (ppm) 1 0 -1 -2 -3 Appendix 3 – Spectra Relevant to Chapter 4 347 Bruker BioSpin GmbH spect CDCl3 297.2 zgpg30 1D Z122623_0045 (CPP BBO 400S1 BB-H&F-D-05 Z) 512 72.0 1 10 1 2022-04-24T20:43:52 Instrument Solvent Temperature Pulse Sequence Experiment Probe Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date -1947.6 13C 32768 65536 Lowest Frequency Nucleus Acquired Size Spectral Size 134.36 130.81 124.10 210 200 190 180 170 160 150 140 130 120 110 100 90 f1 (ppm) 24038.5 Spectral Width Spectrometer Frequency 100.62 NJF-5-208.2.fid Value Origin 174.77 Title 169.33 / Volumes/ nmrdata/ nfastuca/ nmr/ NJF-5-208/ 2/ fid 148.58 Data File Name Parameter NJF-5-208.2.fid 78.47 80 70 50 57.46 57.06 55.98 52.82 47.32 60 30 20 10 35.72 34.87 32.84 32.53 23.52 22.12 19.95 14.75 40 0 -10 Appendix 3 – Spectra Relevant to Chapter 4 348 16 297.1 zg30 1D Z122623_0045 (CPP BBO 400S1 BB-H&F-D-05 Z) 16 176.8 1 12 4 2022-05-26T04:50:56 Temperature Pulse Sequence Experiment Probe Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date 32768 65536 Acquired Size Spectral Size 13 1H Nucleus 14 -1535.4 Lowest Frequency 15 8012.8 Spectral Width 10 C6D6 Solvent 11 spect Instrument 12 Bruker BioSpin GmbH Origin Spectrometer Frequency 400.13 NJF-5-220.1.fid Title Value / Volumes/ nmrdata/ nfastuca/ nmr/ NJF-5-220/ 1/ fid Data File Name Parameter NJF-5-220.1.fid 9 8 7 5 3 2 1 5.83 5.65 5.45 1.00 0.90 0.88 4 3.69 3.36 3.11 3.08 2.77 2.51 2.36 2.23 2.05 1.81 1.66 1.55 0.91 0.97 2.81 2.95 0.94 1.01 2.40 8.02 1.24 2.11 1.29 1.11 3.45 6 f1 (ppm) 0 -1 -2 -3 Appendix 3 – Spectra Relevant to Chapter 4 349 Bruker BioSpin GmbH spect C6D6 297.1 zgpg30 1D Z122623_0045 (CPP BBO 400S1 BB-H&F-D-05 Z) 2048 43.7 1 10 1 2022-05-26T06:14:53 Instrument Solvent Temperature Pulse Sequence Experiment Probe Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date -1921.8 13C 32768 65536 Lowest Frequency Nucleus Acquired Size Spectral Size 210 200 190 180 170 160 150 140 130 120 110 100 90 f1 (ppm) 24038.5 Spectral Width Spectrometer Frequency 100.62 NJF-5-220.2.fid Origin 176.56 Title Value 148.73 / Volumes/ nmrdata/ nfastuca/ nmr/ NJF-5-220/ 2/ fid 136.23 130.36 123.68 Data File Name Parameter NJF-5-220.2.fid 79.90 80 70 50 40 30 20 59.16 57.52 57.16 51.16 48.39 46.74 44.91 35.40 33.43 33.35 23.93 23.72 23.06 15.52 60 10 0 -10 Appendix 3 – Spectra Relevant to Chapter 4 350 8.0 297.1 cosygpppqf COSY Z122623_0045 (CPP BBO 400S1 BB-H&F-D-05 Z) 1 112.8 2 12 0 2022-05-26T06:20:36 Temperature Pulse Sequence Experiment Probe Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date (1024, 1024) Spectral Size 6.0 (1024, 128) Acquired Size 6.5 (1H, 1H) Nucleus 7.0 (-310.8, -310.8) Lowest Frequency 7.5 (3623.2, 3623.2) Spectral Width 4.5 C6D6 Solvent 5.0 spect Instrument 5.5 Bruker BioSpin GmbH Origin Spectrometer Frequency (400.13, 400.13) NJF-5-220.3.ser Title Value / Volumes/ nmrdata/ nfastuca/ nmr/ NJF-5-220/ 3/ ser Data File Name Parameter NJF-5-220.3.ser 4.0 3.5 f2 (ppm) 3.0 2.5 2.0 1.5 1.0 0.5 0.0 -0.5 8 7 6 5 4 3 2 1 0 Appendix 3 – Spectra Relevant to Chapter 4 351 f1 (ppm) spect C6D6 297.1 hsqcedetgpsisp2.4 HSQC-EDITED Z122623_0045 (CPP BBO 400S1 BB-H&F-D-05 Z) 4 197.4 2 12 0 2022-05-26T06:42:47 Instrument Solvent Temperature Pulse Sequence Experiment Probe Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date (1024, 180) (1024, 1024) Acquired Size Spectral Size 8.0 (1H, 13C) Nucleus 9.0 (-517.1, -1236.9) Lowest Frequency 10.0 (4795.4, 16611.3) Spectral Width 6.0 Bruker BioSpin GmbH Origin 7.0 NJF-5-220.4.ser Spectrometer Frequency (400.13, 100.62) / Volumes/ nmrdata/ nfastuca/ nmr/ NJF-5-220/ 4/ ser Title Value Data File Name Parameter NJF-5-220.4.ser 5.0 4.0 f2 (ppm) 3.0 2.0 1.0 0.0 -1.0 150 140 130 120 110 100 90 80 70 60 50 40 30 20 10 0 -10 Appendix 3 – Spectra Relevant to Chapter 4 352 f1 (ppm) inova cdcl3 3.0 s2pul 1D 16 38 2.0000 5.6500 Instrument Solvent Temperature Pulse Sequence Experiment Number of Scans Receiver Gain Relaxation Delay Pulse Width 1H 24000 65536 Nucleus Acquired Size Spectral Size 12 -1030.4 Lowest Frequency 13 8000.0 Spectral Width Spectrometer Frequency 499.61 Varian Origin Value PROTON01 Parameter Title 11 10 9 8 7 5 4 3 2 1 5.85 5.44 5.39 1.00 0.97 1.01 6 f1 (ppm) 3.70 3.57 3.33 3.28 3.00 2.94 2.86 2.54 2.34 2.15 1.94 1.78 1.50 1.15 1.06 2.92 1.05 2.93 1.03 1.91 0.99 1.05 5.02 1.10 1.11 3.14 1.00 2.93 PROTON01 NJF-4-070 0 -1 -2 Appendix 3 – Spectra Relevant to Chapter 4 353 7.26 CDCl3 16 zg30 1D 32 197.4 1.0000 12.5000 Pulse Sequence Experiment Number of Scans Receiver Gain Relaxation Delay Pulse Width 1H 32768 65536 Nucleus Acquired Size Spectral Size 14 -1532.3 Lowest Frequency 15 8012.8 Spectral Width 10 297.1 Temperature 11 C6D6 Solvent 12 spect Instrument 13 Bruker BioSpin GmbH Origin Spectrometer Frequency 400.13 NJF-4-077-spot1.1.fid Title Value / Volumes/ nmrdata/ nfastuca/ nmr/ NJF-4-077-spot1/ 1/ fid 9 8 7.16 CDCl3 7 6 f1 (ppm) 5 4 3 2 1 6.29 5.77 5.51 1.00 1.03 1.08 Parameter Data File Name 4.31 3.66 3.41 3.38 3.29 3.12 2.63 2.45 2.31 1.90 1.70 1.54 1.38 1.03 1.01 0.97 0.99 3.84 1.09 3.37 1.13 5.18 3.09 3.95 1.44 1.42 1.95 3.20 NJF-4-077-spot1.1.fid 0 -1 -2 -3 Appendix 3 – Spectra Relevant to Chapter 4 354 1.0 0.0 -1.0 197.4 Receiver Gain 2.0 4 Number of Scans 3.0 Z122623_0045 (CPP BBO 400S1 BB-H&F-D-05 Z) Probe 5.0 4.0 f2 (ppm) HSQC-EDITED Experiment (1024, 180) (1024, 1024) Acquired Size Spectral Size 8.0 (1H, 13C) Nucleus 9.0 (-514.0, -1261.0) Lowest Frequency 10.0 (4795.4, 16611.3) Spectral Width 6.0 hsqcedetgpsisp2.4 Pulse Sequence 7.0 297.1 Temperature Spectrometer Frequency (400.13, 100.62) C6D6 Solvent 150 140 130 120 110 100 90 80 70 60 50 40 30 spect Instrument 10 20 NJF-4-077-mixed.2.ser 0 -10 Site / Volumes/ nmrdata/ nfastuca/ nmr/ NJF-4-077-mixed/ 2/ ser Title Value Data File Name Parameter NJF-4-077-mixed.2.ser Appendix 3 – Spectra Relevant to Chapter 4 355 f1 (ppm) NJF-4-077-spot1.2.ser C6D6 297.2 cosygpppqf COSY Z122623_0045 (CPP BBO 400S1 BB-H&F-D-05 Z) 4 142.8 Solvent Temperature Pulse Sequence Experiment Probe Number of Scans Receiver Gain (-280.6, -280.6) (1H, 1H) (1024, 128) (1024, 1024) Lowest Frequency Nucleus Acquired Size Spectral Size 5.4 5.2 5.0 4.8 4.6 4.4 4.2 4.0 3.8 3.6 3.4 3.2 3.0 2.8 2.6 2.4 2.2 2.0 1.8 1.6 1.4 1.2 1.0 0.8 0.6 0.4 0.2 f2 (ppm) (3676.5, 3676.5) Spectral Width Spectrometer Frequency (400.13, 400.13) spect Instrument Site Title Value / Volumes/ nmrdata/ nfastuca/ nmr/ NJF-4-077-spot1/ 2/ ser Data File Name Parameter NJF-4-077-spot1.2.ser 7.5 7.0 6.5 6.0 5.5 5.0 4.5 4.0 3.5 3.0 2.5 2.0 1.5 1.0 0.5 0.0 -0.5 Appendix 3 – Spectra Relevant to Chapter 4 356 f1 (ppm) 16 NJF-4-085.1.fid 1D Z122623_0045 (CPP BBO 400S1 BB-H&F-D-05 Z) 16 142.8 Experiment Probe Number of Scans Receiver Gain 1H 32768 65536 Nucleus Acquired Size Spectral Size 14 -1545.2 Lowest Frequency 15 8012.8 Spectral Width 11 zg30 Pulse Sequence 12 297.1 Temperature 13 CDCl3 Solvent Spectrometer Frequency 400.13 spect Instrument Site Title Value / Volumes/ nmrdata/ nfastuca/ nmr/ NJF-4-085/ 1/ fid Data File Name Parameter NJF-4-085.1.fid 10 9 8 7 5 3 2 1 6.07 5.87 5.65 5.42 1.00 1.00 1.01 1.97 4 4.47 4.23 3.57 3.25 3.22 2.97 2.89 2.47 2.30 1.60 1.44 1.19 1.09 1.01 1.01 1.00 1.01 4.90 2.97 5.11 1.02 1.02 2.08 3.26 0.97 3.18 9.22 9.21 6 f1 (ppm) 0 -1 -2 -3 Appendix 3 – Spectra Relevant to Chapter 4 357 hsqcedetgpsisp2.4 HSQC-EDITED Z122623_0045 (CPP BBO 400S1 BB-H&F-D-05 Z) 2 197.4 Pulse Sequence Experiment Probe Number of Scans Receiver Gain (1024, 180) (1024, 1024) Acquired Size Spectral Size 8.0 (1H, 13C) Nucleus 9.0 (-526.9, -1265.2) Lowest Frequency 10.0 (4795.4, 16611.3) Spectral Width 6.0 297.2 Temperature 7.0 CDCl3 Solvent Spectrometer Frequency (400.13, 100.62) spect Instrument Site NJF-4-085.2.ser Title Value / Volumes/ nmrdata/ nfastuca/ nmr/ NJF-4-085/ 2/ ser Data File Name Parameter NJF-4-085.2.ser 5.0 4.0 f2 (ppm) 3.0 2.0 1.0 0.0 -1.0 150 140 130 120 110 100 90 80 70 60 50 40 30 20 10 0 -10 Appendix 3 – Spectra Relevant to Chapter 4 358 f1 (ppm) NJF-4-085.3.ser cosygpppqf COSY Z122623_0045 (CPP BBO 400S1 BB-H&F-D-05 Z) 1 64.2 Pulse Sequence Experiment Probe Number of Scans Receiver Gain (1024, 128) (1024, 1024) Acquired Size Spectral Size 5.5 (1H, 1H) Nucleus 6.0 (-13.0, -13.0) Lowest Frequency 6.5 (2762.4, 2762.4) Spectral Width 4.5 297.1 Temperature 5.0 CDCl3 Solvent Spectrometer Frequency (400.13, 400.13) spect Instrument Site Title Value / Volumes/ nmrdata/ nfastuca/ nmr/ NJF-4-085/ 3/ ser Data File Name Parameter NJF-4-085.3.ser 4.0 3.5 3.0 f2 (ppm) 2.5 2.0 1.5 1.0 0.5 0.0 6.5 6.0 5.5 5.0 4.5 4.0 3.5 3.0 2.5 2.0 1.5 1.0 0.5 0.0 Appendix 3 – Spectra Relevant to Chapter 4 359 f1 (ppm) NJF-4-085.4.fid Title CDCl3 297.2 zgpg30 1D Z122623_0045 (CPP BBO 400S1 BB-H&F-D-05 Z) 512 55.5 Solvent Temperature Pulse Sequence Experiment Probe Number of Scans Receiver Gain -1944.9 13C 32768 65536 Lowest Frequency Nucleus Acquired Size Spectral Size 119.94 136.04 134.27 131.37 128.25 210 200 190 180 170 160 150 140 130 120 110 100 90 f1 (ppm) 24038.5 Spectral Width Spectrometer Frequency 100.62 spect Instrument Site / Volumes/ nmrdata/ nfastuca/ nmr/ NJF-4-085/ 4/ fid Value 166.56 Data File Name Parameter NJF-4-085.4.fid 70 60 50 40 30 20 10 79.64 77.36 74.37 69.55 66.68 60.90 58.33 57.36 57.26 46.45 46.36 44.59 40.82 35.61 29.07 28.48 24.01 21.86 21.41 20.84 13.25 80 0 -10 Appendix 3 – Spectra Relevant to Chapter 4 360 16 15 14 NJF-4-120.1.fid 13 12 11 10 9 8 7 6 f1 (ppm) 5 4 3 2 1 0 1.00 0.97 0.99 0.96 1.00 1.00 1.29 3.25 1.00 2.96 2.91 3.19 1.13 1.08 1.06 1.16 2.04 1.67 3.12 9.23 8.99 6.95 6.07 5.74 5.64 5.38 5.37 5.36 5.35 5.34 5.34 5.33 5.33 5.30 -1 -2 -3 4.56 4.47 4.25 3.78 3.68 3.68 3.66 3.66 3.64 3.38 3.35 3.32 3.28 3.22 3.08 2.83 2.26 2.25 2.24 2.23 2.22 2.21 2.20 2.01 2.00 1.39 1.08 1.00 Appendix 3 – Spectra Relevant to Chapter 4 361 CDCl3 297.1 cosygpppqf COSY Z122623_0045 (CPP BBO 400S1 BB-H&F-D-05 Z) 4 176.8 1.8878 12.5000 0.3092 2019-12-28T14:30:39 2019-12-28T14:49:46 Solvent Temperature Pulse Sequence Experiment Probe Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date Modification Date (1024, 1024) Spectral Size 6.5 (1024, 128) Acquired Size 7.0 (1H, 1H) Nucleus 7.5 (-54.0, -54.0) Lowest Frequency 8.0 (3311.3, 3311.3) Spectral Width 5.0 spect Instrument 5.5 Bruker BioSpin GmbH 6.0 NJF-4-120.3.ser Origin Spectrometer Frequency (400.13, 400.13) / Volumes/ nmrdata/ nfastuca/ nmr/ NJF-4-120/ 3/ ser Title Value Data File Name NJF-4-120.3.ser Parameter 4.5 4.0 3.5 f2 (ppm) 3.0 2.5 2.0 1.5 1.0 0.5 0.0 8.0 7.5 7.0 6.5 6.0 5.5 5.0 4.5 4.0 3.5 3.0 2.5 2.0 1.5 1.0 0.5 0.0 Appendix 3 – Spectra Relevant to Chapter 4 362 f1 (ppm) 11.0 10.0 NJF-4-120.7.ser 9.0 8.0 7.0 6.0 5.0 f2 (ppm) 4.0 3.0 2.0 1.0 0.0 -10 0 10 20 30 40 50 60 70 80 90 100 110 120 130 140 150 160 170 180 190 200 210 Appendix 3 – Spectra Relevant to Chapter 4 363 f1 (ppm) 16 297.2 zg30 1D Z122623_0045 (CPP BBO 400S1 BB-H&F-D-05 Z) 16 112.8 1.0000 12.5000 4.0894 2019-11-14T20:35:50 2019-11-14T20:35:50 Temperature Pulse Sequence Experiment Probe Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date Modification Date 1H 32768 65536 Nucleus Acquired Size Spectral Size 14 -1545.1 Lowest Frequency 15 8012.8 Spectral Width 11 CDCl3 Solvent 12 spect Instrument 13 Bruker BioSpin GmbH Origin Spectrometer Frequency 400.13 NJF-4-095.2.fid Title Value / Volumes/ nmrdata/ nfastuca/ nmr/ NJF-4-095/ 2/ fid Parameter Data File Name 10 9 8 7 5.97 5.88 5.76 5.47 5.37 4.38 3.92 3.54 3.25 3.24 3.19 2.81 2.71 2.60 2.47 2.39 2.31 1.57 1.34 1.07 6 f1 (ppm) 5 4 3 2 1 0.98 1.00 1.01 0.99 0.97 1.02 1.00 1.02 3.31 2.89 1.96 1.12 1.11 2.10 2.01 1.08 2.06 3.38 1.11 3.07 NJF-4-095.2.fid 0 -1 -2 -3 Appendix 3 – Spectra Relevant to Chapter 4 364 Bruker BioSpin GmbH spect CDCl3 297.2 zgpg30 1D Z122623_0045 (CPP BBO 400S1 BB-H&F-D-05 Z) 512 55.5 1.0000 10.0000 1.3631 2019-11-14T20:58:05 2019-11-14T20:58:05 Origin Instrument Solvent Temperature Pulse Sequence Experiment Probe Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date Modification Date -1945.6 13C 32768 65536 Lowest Frequency Nucleus Acquired Size Spectral Size 80 70 60 50 40 79.80 77.36 77.16 CDCl3 76.51 74.85 66.16 59.05 57.37 56.97 56.72 47.78 44.99 44.74 39.38 35.45 120.14 134.31 132.42 131.58 129.34 210 200 190 180 170 160 150 140 130 120 110 100 90 f1 (ppm) 24038.5 Spectral Width Spectrometer Frequency 100.62 NJF-4-095.3.fid Title Value / Volumes/ nmrdata/ nfastuca/ nmr/ NJF-4-095/ 3/ fid 165.03 Data File Name Parameter NJF-4-095.3.fid 30 24.47 21.91 15.35 20 10 0 -10 Appendix 3 – Spectra Relevant to Chapter 4 365 14 inova cdcl3 25.0 s2pul 1D 8 36 2.0000 2.6500 Instrument Solvent Temperature Pulse Sequence Experiment Number of Scans Receiver Gain Relaxation Delay Pulse Width 1H 16384 65536 Nucleus Acquired Size Spectral Size 12 -1193.2 Lowest Frequency 13 9611.9 Spectral Width Spectrometer Frequency 599.59 Varian Origin Value NJF-4-171 Parameter Title 11 10 9 8 5 2 0.57 0.92 1 6.94 1.00 3 5.94 5.78 5.66 5.34 1.06 0.95 1.03 1.09 4 4.66 1.09 6 f1 (ppm) 3.51 3.24 3.22 3.18 3.12 2.61 2.50 2.36 1.11 3.18 3.27 2.22 1.10 2.11 3.25 2.03 7 1.68 1.57 1.36 3.24 1.19 3.27 8.98 6.09 NJF-4-171 STANDARD 1H OBSERVE - profile 0 -1 Appendix 3 – Spectra Relevant to Chapter 4 366 7.26 CDCl3 9.57 14 cdcl3 25.0 s2pul 1D penta 6 40 2.0000 2.6500 1.7046 2020-02-13T02:28:47 2020-02-13T02:29:40 Solvent Temperature Pulse Sequence Experiment Probe Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date Modification Date 1H 16384 65536 Nucleus Acquired Size Spectral Size 12 -1192.8 Lowest Frequency 13 9611.9 Spectral Width 11 inova Instrument Spectrometer Frequency 599.59 Varian Origin Value NJF-4-174 Parameter Title 10 9 8 5 3 2 1 6.94 1.00 4 5.93 5.78 5.66 5.34 0.95 0.91 1.01 1.00 6 f1 (ppm) 4.66 0.98 7 3.53 3.29 3.22 3.18 3.11 3.08 2.98 2.54 2.31 1.71 1.60 1.20 0.92 0.57 1.01 2.01 6.06 1.02 1.03 1.02 3.11 0.99 2.07 2.01 1.01 0.99 3.17 8.98 5.89 NJF-4-174 STANDARD 1H OBSERVE - profile 0 -1 Appendix 3 – Spectra Relevant to Chapter 4 367 7.26 CDCl3 14 cdcl3 3.0 s2pul 1D OneNMR 200 30 1.0000 5.0000 2.5559 2020-02-17T15:27:10 2020-02-17T15:27:27 Solvent Temperature Pulse Sequence Experiment Probe Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date Modification Date 1H 16384 65536 Nucleus Acquired Size Spectral Size 12 -806.4 Lowest Frequency 13 6410.3 Spectral Width 11 vnmrs Instrument Spectrometer Frequency 399.80 Varian Origin Value NJF-4-176-spot2 Parameter Title 10 9 8 7.06 1.04 6 f1 (ppm) 5.96 5.92 5.78 5.76 5.47 0.90 0.83 0.89 0.78 7 5 4.64 4.14 3.65 3.37 3.33 3.17 3.11 3.04 2.65 2.47 2.38 2.14 1.87 1.83 1.10 0.92 0.57 3 2 1.00 1.00 2.35 3.75 3.79 1.82 1.07 1.09 0.99 2.04 1.02 1.90 4 1 4.17 9.35 6.12 NJF-4-176-spot2 z0gmap_calib Selective band center: 4.56 (ppm); width: 39.4 (Hz) Selective band center: 4.26 (ppm); width: 38.3 (Hz) 0 -1 -2 Appendix 3 – Spectra Relevant to Chapter 4 368 11.0 1D 16 64.2 1.0000 11.7000 4.0894 2018-12-10T20:31:59 Experiment Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date 65536 Spectral Size 9.0 32768 Acquired Size 9.5 1H Nucleus 10.0 -1545.3 Lowest Frequency 10.5 8012.8 Spectral Width 6.5 zg30 Pulse Sequence 7.0 295.1 Temperature 7.5 CDCl3 Solvent 8.0 spect Instrument 8.5 Bruker BioSpin GmbH Origin Spectrometer Frequency 400.13 ARWVI-271_flashed_char.1.fid Title Value 6.11 / Volumes/ nmrdata/ arwong/ nmr/ ARWVI-271_flashed_char/ 1/ fid Parameter 5.67 5.45 Data File Name 1.00 6.0 1.03 0.98 5.5 5.0 ppm 3.0 2.5 2.0 1.5 1.0 4.51 1.11 3.5 4.20 1.04 4.0 3.35 3.27 3.09 3.03 2.95 2.88 2.78 2.59 2.47 2.13 1.82 1.63 1.46 1.30 1.07 1.01 4.05 3.03 1.05 1.06 2.10 1.01 1.00 0.92 0.97 1.04 0.97 2.08 2.06 1.15 9.08 9.15 4.5 0.5 0.0 -0.5 -1.0 Appendix 3 – Spectra Relevant to Chapter 4 369 50.3 1.0000 10.0000 1.3631 2018-12-10T23:36:00 Relaxation Delay Pulse Width Acquisition Time Acquisition Date 65536 Spectral Size 180 32768 Acquired Size 190 13C Nucleus 200 -1958.4 Lowest Frequency 210 24038.5 Spectral Width 110 100 ppm 1024 Receiver Gain 120 1D Number of Scans 130 zgpg30 Experiment 140 295.1 Pulse Sequence 150 CDCl3 Temperature 160 spect Solvent 170 Bruker BioSpin GmbH Instrument Spectrometer Frequency 100.62 ARWVI-271_flashed_char.7.fid Origin Value Title 162.71 / Volumes/ nmrdata/ arwong/ nmr/ ARWVI-271_flashed_char/ 7/ fid Parameter 134.95 128.84 123.89 Data File Name 90 70 60 50 40 30 20 10 78.93 77.32 66.29 59.32 56.77 50.18 50.04 46.03 45.07 38.13 35.92 35.65 33.70 32.86 28.89 28.26 23.96 22.62 21.19 20.68 80 0 -10 Appendix 3 – Spectra Relevant to Chapter 4 370 2 197.4 1.5000 11.7000 0.2135 2018-12-10T21:08:18 Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date 8.0 7.5 (1024, 1024) Spectral Size 8.5 (1024, 180) Acquired Size 9.0 (1H, 13C) Nucleus 9.5 (-517.1, -1262.8) Lowest Frequency 10.5 10.0 (4795.4, 16611.3) Spectral Width 4.0 HSQC-EDITED Experiment 5.0 4.5 ppm hsqcedetgpsisp2.4 Pulse Sequence 5.5 295.1 Temperature 6.0 CDCl3 Solvent 6.5 spect Instrument 7.0 Bruker BioSpin GmbH Origin Spectrometer Frequency (400.13, 100.62) ARWVI-271_flashed_char.3.ser Title Value / Volumes/ nmrdata/ arwong/ nmr/ ARWVI-271_flashed_char/ 3/ ser Parameter Data File Name 3.5 3.0 2.5 2.0 1.5 1.0 0.5 0.0 -0.5 -1.0 150 140 130 120 110 100 90 80 70 60 50 40 30 20 10 0 -10 Appendix 3 – Spectra Relevant to Chapter 4 371 ppm 11.0 16 72.0 1.0000 11.7000 4.0894 2018-12-11T14:02:19 Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date 65536 Spectral Size 9.0 32768 Acquired Size 9.5 1H Nucleus 10.0 -1545.0 Lowest Frequency 10.5 8012.8 Spectral Width 6.5 1D Number of Scans 7.0 zg30 Experiment 7.5 295.1 Pulse Sequence 8.0 CDCl3 Temperature 8.5 spect Solvent Spectrometer Frequency 400.13 Bruker BioSpin GmbH Instrument 6.17 ARWVI-272_flashed.2.fid 1.00 6.0 5.5 5.0 ppm 4.5 4.0 3.5 3.0 2.5 2.0 1.5 1.0 5.71 1.96 Origin 4.74 4.65 0.84 1.00 Title Value 4.21 4.06 1.03 1.92 / Volumes/ nmrdata/ arwong/ nmr/ ARWVI-272_flashed/ 2/ fid Parameter 3.29 3.23 3.14 3.07 3.02 2.51 2.38 2.29 2.18 2.03 1.71 1.41 1.12 1.08 1.03 3.01 3.30 1.07 1.15 2.02 0.98 1.01 2.04 3.03 1.21 1.05 2.14 0.77 9.20 9.16 Data File Name 0.5 0.0 -0.5 -1.0 Appendix 3 – Spectra Relevant to Chapter 4 372 78.7 1.0000 10.0000 1.3631 2018-12-11T23:16:52 Relaxation Delay Pulse Width Acquisition Time Acquisition Date 65536 Spectral Size 180 32768 Acquired Size 190 13C Nucleus 200 -1961.9 Lowest Frequency 210 24038.5 Spectral Width 110 100 ppm 2000 Receiver Gain 120 1D Number of Scans 130 zgpg30 Experiment 140 295.2 Pulse Sequence 150 CDCl3 Temperature 160 spect Solvent 170 Bruker BioSpin GmbH Instrument Spectrometer Frequency 100.62 ARWVI-272_flashed_char.1.fid Origin 169.04 Title Value 135.91 / Volumes/ nmrdata/ arwong/ nmr/ ARWVI-272_flashed_char/ 1/ fid Parameter 128.20 124.88 Data File Name 90 70 60 50 40 30 20 10 80.60 78.08 77.64 66.65 66.22 59.45 55.68 53.13 47.30 46.26 45.95 45.66 42.80 35.52 35.40 32.19 29.68 28.97 28.33 24.58 21.81 21.26 20.75 80 0 -10 Appendix 3 – Spectra Relevant to Chapter 4 373 11.0 16 112.8 1.0000 11.7000 4.0894 2018-12-13T02:10:44 Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date 65536 Spectral Size 9.0 32768 Acquired Size 9.5 1H Nucleus 10.0 -1545.4 Lowest Frequency 10.5 8012.8 Spectral Width 2.00 1.00 6.0 5.5 1D Experiment 6.5 zg30 Pulse Sequence 7.0 295.2 Temperature 7.5 CDCl3 Solvent 8.0 spect Instrument 8.5 Bruker BioSpin GmbH Origin Spectrometer Frequency 400.13 ARWVI-274_flashed.1.fid Title Value 6.10 / Volumes/ nmrdata-1/ arwong/ nmr/ ARWVI-274_flashed/ 1/ fid Parameter 5.80 Data File Name 5.0 ppm 3.5 3.0 2.5 2.0 1.5 1.0 4.66 1.93 4.0 4.05 3.96 3.29 3.21 3.18 3.13 3.03 2.92 2.81 2.75 2.51 2.38 2.28 2.19 2.01 1.70 1.46 1.33 1.95 1.03 3.00 3.01 0.91 1.18 1.28 1.06 0.73 1.10 0.97 1.01 1.96 3.02 1.00 1.88 1.09 0.99 4.5 0.5 0.0 -0.5 -1.0 Appendix 3 – Spectra Relevant to Chapter 4 374 2000 78.7 1.0000 10.0000 1.3631 2018-12-13T05:19:26 Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date 65536 Spectral Size 180 32768 Acquired Size 190 13C Nucleus 200 -1962.6 Lowest Frequency 210 24038.5 Spectral Width 120 1D Number of Scans 130 zgpg30 Experiment 140 295.1 Pulse Sequence 150 CDCl3 Temperature 160 spect Solvent 170 Bruker BioSpin GmbH Instrument Spectrometer Frequency 100.62 ARWVI-274_flashed.6.fid Origin Value Title 169.10 / Volumes/ nmrdata-1/ arwong/ nmr/ ARWVI-274_flashed/ 6/ fid Parameter 132.43 129.04 125.13 Data File Name 110 100 ppm 90 70 60 50 40 30 80.85 78.04 75.10 66.64 65.61 59.45 55.80 52.94 47.44 47.43 45.49 44.54 42.80 35.47 35.44 32.16 80 24.54 21.85 20 10 0 -10 Appendix 3 – Spectra Relevant to Chapter 4 375 197.4 2.0000 11.7000 0.4260 2018-12-13T02:29:40 Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date (2048, 256) (2048, 1024) Acquired Size Spectral Size 8.5 (1H, 13C) Nucleus 9.0 (-163.1, -1116.4) Lowest Frequency 11.5 11.0 10.5 10.0 9.5 (4807.7, 22321.4) Spectral Width 5.0 4 Number of Scans 5.5 ppm HMBC Experiment 6.0 hmbcetgpl3nd Pulse Sequence 6.5 295.1 Temperature 7.0 CDCl3 Solvent 7.5 spect Instrument 8.0 Bruker BioSpin GmbH Origin Spectrometer Frequency (400.13, 100.62) ARWVI-274_flashed.4.ser Title Value / Volumes/ nmrdata-1/ arwong/ nmr/ ARWVI-274_flashed/ 4/ ser Parameter Data File Name 4.5 4.0 3.5 3.0 2.5 2.0 1.5 1.0 0.5 0.0 210 200 190 180 170 160 150 140 130 120 110 100 90 80 70 60 50 40 30 20 10 0 -10 Appendix 3 – Spectra Relevant to Chapter 4 376 ppm 11.0 zg30 1D 16 142.8 1.0000 11.7000 4.0894 2018-12-15T20:12:13 Pulse Sequence Experiment Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date 65536 Spectral Size 9.0 32768 Acquired Size 9.5 1H Nucleus 10.0 -1545.6 Lowest Frequency 10.5 8012.8 Spectral Width 1.00 7.0 6.5 295.2 Temperature 7.5 CDCl3 Solvent 8.0 spect Instrument 8.5 Bruker BioSpin GmbH Origin Spectrometer Frequency 400.13 ARWVI-275_repurify.1.fid Title Value 7.19 / Volumes/ nmrdata/ arwong/ nmr/ ARWVI-275_repurify/ 1/ fid Parameter 5.93 Data File Name 2.04 6.0 5.5 4.5 2.5 2.0 1.5 5.14 1.04 3.0 4.71 1.03 3.5 4.09 3.46 3.33 3.29 3.24 3.21 3.14 3.04 2.10 1.00 1.15 2.89 1.10 3.03 1.10 1.06 4.0 2.60 2.41 2.36 2.28 2.21 2.01 1.72 1.41 2.10 1.11 1.01 1.20 3.21 1.12 1.04 2.35 5.0 ppm 1.0 0.5 0.0 -0.5 -1.0 Appendix 3 – Spectra Relevant to Chapter 4 377 2048 72.0 1.0000 10.0000 1.3631 2018-12-15T22:31:19 Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date 65536 Spectral Size 180 32768 Acquired Size 190 13C Nucleus 200 -1961.9 Lowest Frequency 210 24038.5 Spectral Width 120 1D Number of Scans 130 zgpg30 Experiment 140 295.1 Pulse Sequence 150 CDCl3 Temperature 160 spect Solvent 170 Bruker BioSpin GmbH Instrument Spectrometer Frequency 100.62 ARWVI-275_repurify.3.fid 168.98 Origin 160.15 Title Value 150.91 / Volumes/ nmrdata/ arwong/ nmr/ ARWVI-275_repurify/ 3/ fid 110 100 ppm 90 85.05 82.00 77.94 80 70 66.77 60.87 59.45 55.14 52.93 47.75 47.49 45.40 42.72 35.49 35.29 31.97 24.50 21.87 60 50 40 30 20 10 162.1 161.2 160.3 159.4 158.5 157.6 ppm 160.15 195.12 Parameter 126.33 124.06 Data File Name 0 -10 Appendix 3 – Spectra Relevant to Chapter 4 378 11.0 zg30 1D 16 30.3 1.0000 11.7000 4.0894 2018-12-17T13:29:46 Pulse Sequence Experiment Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date 65536 Spectral Size 9.0 32768 Acquired Size 9.5 1H Nucleus 10.0 -1545.0 Lowest Frequency 10.5 8012.8 Spectral Width 1.00 7.0 6.5 295.2 Temperature 7.5 CDCl3 Solvent 8.0 spect Instrument 8.5 Bruker BioSpin GmbH Origin Spectrometer Frequency 400.13 ARWVI-279_flashed.1.fid Title Value 7.11 / Volumes/ nmrdata/ arwong/ nmr/ ARWVI-279_flashed/ 1/ fid Parameter 5.95 5.81 Data File Name 1.00 0.97 6.0 5.5 3.5 3.0 2.5 2.0 1.5 4.92 4.70 4.65 0.86 1.04 1.92 4.0 4.08 3.48 3.39 3.32 3.27 3.23 3.19 3.13 3.03 2.05 1.01 1.21 3.02 3.14 1.08 3.10 1.11 1.08 4.5 2.58 2.37 2.26 2.21 2.00 1.71 1.39 1.17 2.17 1.11 3.00 1.11 1.09 2.16 5.0 ppm 1.0 0.5 0.0 -0.5 -1.0 Appendix 3 – Spectra Relevant to Chapter 4 379 195.62 1024 64.2 1.0000 10.0000 1.3631 2018-12-17T15:59:23 Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date 65536 Spectral Size 180 32768 Acquired Size 190 13C Nucleus 200 -1965.5 Lowest Frequency 210 24038.5 Spectral Width 120 1D Number of Scans 130 zgpg30 Experiment 140 295.1 Pulse Sequence 150 CDCl3 Temperature 160 spect Solvent 170 Bruker BioSpin GmbH Instrument Spectrometer Frequency 100.62 ARWVI-279_flashed.6.fid 168.94 Origin 160.03 Title Value 150.60 / Volumes/ nmrdata/ arwong/ nmr/ ARWVI-279_flashed/ 6/ fid Parameter 96.10 110 100 ppm 90.57 90 81.97 77.92 80 70 60 161.4 50 160.9 40 160.4 66.68 59.40 59.33 55.92 55.12 52.89 47.57 46.16 45.46 42.66 35.43 35.26 31.94 30 159.9 ppm 160.03 125.42 123.94 Data File Name 24.46 21.92 20 159.4 10 158.9 0 158.4 -10 Appendix 3 – Spectra Relevant to Chapter 4 380 11.0 1D 32 64.2 1.0000 11.7000 4.0894 2018-12-18T14:24:51 Experiment Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date 65536 Spectral Size 9.0 32768 Acquired Size 9.5 1H Nucleus 10.0 -1545.1 Lowest Frequency 10.5 8012.8 Spectral Width 6.5 zg30 Pulse Sequence 7.0 295.1 Temperature 7.5 CDCl3 Solvent 8.0 spect Instrument 8.5 Bruker BioSpin GmbH Origin Spectrometer Frequency 400.13 ARWVI-280_flashed.1.fid Title Value / Volumes/ nmrdata/ arwong/ nmr/ ARWVI-280_flashed/ 1/ fid Parameter 5.83 Data File Name 1.00 6.0 5.5 5.0 ppm 3.0 2.5 2.0 1.5 4.66 4.48 2.29 1.01 3.5 4.07 3.74 3.58 3.34 3.30 3.29 3.21 3.03 2.91 2.68 1.00 0.99 1.02 2.94 3.05 2.97 2.17 1.03 0.98 2.02 4.0 2.28 2.11 1.97 1.73 1.44 1.32 3.25 5.29 1.12 2.53 1.26 1.28 4.5 1.0 0.5 0.0 -0.5 -1.0 Appendix 3 – Spectra Relevant to Chapter 4 381 204.64 1024 64.2 0.5000 10.0000 1.2496 2018-12-18T16:43:16 Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date 65536 Spectral Size 180 32768 Acquired Size 190 13C Nucleus 200 -2049.6 Lowest Frequency 210 26223.8 Spectral Width 120 1D Number of Scans 130 zgpg30 Experiment 140 295.1 Pulse Sequence 150 CDCl3 Temperature 160 spect Solvent 170 Bruker BioSpin GmbH Instrument Spectrometer Frequency 100.62 ARWVI-280_flashed.6.fid Origin 169.80 Title Value 153.37 / Volumes/ nmrdata/ arwong/ nmr/ ARWVI-280_flashed/ 6/ fid Parameter 121.42 Data File Name 96.31 110 100 ppm 90 70 60 50 40 30 20 80.01 79.00 78.18 65.03 59.67 59.45 55.90 55.09 51.55 46.25 45.39 42.50 42.20 39.23 37.67 33.82 31.88 30.53 24.69 22.49 80 10 0 -10 Appendix 3 – Spectra Relevant to Chapter 4 382 HSQC-EDITED 2 197.4 1.5000 11.7000 0.2135 2018-12-18T14:26:09 Experiment Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date 8.0 7.5 (1024, 1024) Spectral Size 8.5 (1024, 180) Acquired Size 9.0 (1H, 13C) Nucleus 9.5 (-526.5, -1262.8) Lowest Frequency 10.5 10.0 (4795.4, 16611.3) Spectral Width 5.0 4.5 ppm hsqcedetgpsisp2.4 Pulse Sequence 5.5 295.1 Temperature 6.0 CDCl3 Solvent 6.5 spect Instrument 7.0 Bruker BioSpin GmbH Origin Spectrometer Frequency (400.13, 100.62) ARWVI-280_flashed.2.ser Title Value / Volumes/ nmrdata/ arwong/ nmr/ ARWVI-280_flashed/ 2/ ser Parameter Data File Name 4.0 3.5 3.0 2.5 2.0 1.5 1.0 0.5 0.0 -0.5 -1.0 150 140 130 120 110 100 90 80 70 60 50 40 30 20 10 0 -10 Appendix 3 – Spectra Relevant to Chapter 4 383 ppm 4 197.4 2.0000 11.7000 0.4260 2018-12-18T14:37:41 Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date (2048, 256) (2048, 1024) Acquired Size Spectral Size 8.5 (1H, 13C) Nucleus 9.0 (-173.0, -1118.1) Lowest Frequency 11.5 11.0 10.5 10.0 9.5 (4807.7, 22321.4) Spectral Width 5.0 HMBC Experiment 5.5 ppm hmbcetgpl3nd Pulse Sequence 6.0 295.2 Temperature 6.5 CDCl3 Solvent 7.0 spect Instrument 7.5 Bruker BioSpin GmbH Origin 8.0 ARWVI-280_flashed.3.ser Spectrometer Frequency (400.13, 100.62) / Volumes/ nmrdata/ arwong/ nmr/ ARWVI-280_flashed/ 3/ ser Title Value Data File Name Parameter 4.5 4.0 3.5 3.0 2.5 2.0 1.5 1.0 0.5 0.0 210 200 190 180 170 160 150 140 130 120 110 100 90 80 70 60 50 40 30 20 10 0 -10 Appendix 3 – Spectra Relevant to Chapter 4 384 ppm NOESY 4 28.2 1.8000 11.7000 0.2560 2018-12-18T15:27:55 Experiment Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date 7.6 7.2 6.8 (1024, 1024) Spectral Size 8.0 (1024, 256) Acquired Size 8.4 (1H, 1H) Nucleus 8.8 (-168.5, -166.8) Lowest Frequency 9.2 (4000.0, 4000.0) Spectral Width 4.8 4.4 ppm noesygpphzs Pulse Sequence 5.2 295.2 Temperature 5.6 CDCl3 Solvent 6.0 spect Instrument 6.4 Bruker BioSpin GmbH Origin Spectrometer Frequency (400.13, 400.13) ARWVI-280_flashed.5.ser Title Value / Volumes/ nmrdata/ arwong/ nmr/ ARWVI-280_flashed/ 5/ ser Parameter Data File Name 4.0 3.6 3.2 2.8 2.4 2.0 1.6 1.2 0.8 0.4 0.0 -0.4 9 8 7 6 5 4 3 2 1 0 Appendix 3 – Spectra Relevant to Chapter 4 385 ppm 1 30.3 2.0000 11.7000 0.1970 2018-12-18T15:21:19 Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date 8.0 7.6 7.2 (1024, 1024) Spectral Size 8.4 (1024, 128) Acquired Size 8.8 (1H, 1H) Nucleus 9.2 (-208.0, -204.1) Lowest Frequency 10.0 9.6 (5197.5, 5197.5) Spectral Width 4.8 4.4 ppm COSY Experiment 5.2 cosygpppqf Pulse Sequence 5.6 295.1 Temperature 6.0 CDCl3 Solvent 6.4 spect Instrument 6.8 Bruker BioSpin GmbH Origin Spectrometer Frequency (400.13, 400.13) ARWVI-280_flashed.4.ser Title Value / Volumes/ nmrdata/ arwong/ nmr/ ARWVI-280_flashed/ 4/ ser Parameter Data File Name 4.0 3.6 3.2 2.8 2.4 2.0 1.6 1.2 0.8 0.4 0.0 -0.4 10 9 8 7 6 5 4 3 2 1 0 Appendix 3 – Spectra Relevant to Chapter 4 386 ppm 11.0 32 30.3 1.0000 11.7000 4.0894 2018-12-19T14:12:10 Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date 65536 Spectral Size 9.0 32768 Acquired Size 9.5 1H Nucleus 10.0 -1545.0 Lowest Frequency 10.5 8012.8 Spectral Width 6.0 1D Experiment 6.5 zg30 Pulse Sequence 7.0 295.1 Temperature 7.5 CDCl3 Solvent 8.0 spect Instrument 8.5 Bruker BioSpin GmbH Origin Spectrometer Frequency 400.13 ARWVI-282_flashed.1.fid Title Value / Volumes/ nmrdata/ arwong/ nmr/ ARWVI-282_flashed/ 1/ fid Parameter Data File Name 5.5 5.0 ppm 3.5 3.0 2.5 2.0 1.5 1.0 4.60 1.91 4.0 4.11 3.99 3.86 3.31 3.28 3.20 3.09 2.98 2.76 2.70 2.58 2.52 2.41 2.33 2.06 1.89 1.68 1.43 1.29 0.97 1.01 1.00 2.80 3.40 5.08 1.32 1.03 1.12 1.17 1.01 1.22 1.19 1.12 7.22 2.22 2.20 1.28 1.09 4.5 0.5 0.0 -0.5 -1.0 Appendix 3 – Spectra Relevant to Chapter 4 387 220 1024 55.5 0.5000 10.0000 1.2496 2018-12-19T16:31:04 Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date 65536 Spectral Size 180 32768 Acquired Size 190 13C Nucleus 200 -2051.1 Lowest Frequency 210 26223.8 Spectral Width 120 1D Number of Scans 130 zgpg30 Experiment 140 295.1 Pulse Sequence 150 CDCl3 Temperature 160 spect Solvent 170 Bruker BioSpin GmbH Instrument Spectrometer Frequency 100.62 ARWVI-282_flashed.6.fid Origin Value Title 212.90 / Volumes/ nmrdata/ arwong/ nmr/ ARWVI-282_flashed/ 6/ fid Parameter 169.74 Data File Name 95.97 110 100 ppm 90 70 60 50 40 30 20 10 84.00 79.69 78.18 59.42 56.78 55.77 55.60 52.28 48.28 46.78 46.11 44.50 43.85 39.11 37.42 36.65 33.80 32.04 28.63 27.28 25.18 22.48 80 0 -10 Appendix 3 – Spectra Relevant to Chapter 4 388 14 cdcl3 25.0 s2pul 1D penta 10 36 1.0000 2.6500 1.7078 2020-09-29T16:11:18 2020-09-29T16:12:08 Solvent Temperature Pulse Sequence Experiment Probe Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date Modification Date 1H 16384 65536 Nucleus Acquired Size Spectral Size 12 -1193.3 Lowest Frequency 13 9593.5 Spectral Width 11 inova Instrument Spectrometer Frequency 599.58 Varian Origin Value NJF-4-288 Parameter Title 10 9 8 7 6 f1 (ppm) 5 4.67 4.27 3.98 3.90 3.35 3.31 3.18 3.02 2.87 2.79 2.63 2.48 2.13 2.02 1.92 1.80 1.71 1.46 1.30 4 3 2 1 0 2.11 1.00 1.00 0.96 3.16 3.06 6.18 0.92 1.01 1.79 0.98 2.06 8.18 1.18 0.92 0.97 1.20 1.15 1.32 NJF-4-288 -1 Appendix 3 – Spectra Relevant to Chapter 4 389 7.26 CDCl3 220 1500 72.0 0.5000 10.0000 1.2496 2019-01-04T23:54:05 Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date 65536 Spectral Size 180 32768 Acquired Size 190 13C Nucleus 200 -2048.9 Lowest Frequency 210 26223.8 Spectral Width 120 1D Number of Scans 130 zgpg30 Experiment 140 295.1 Pulse Sequence 150 CDCl3 Temperature 160 spect Solvent 170 Bruker BioSpin GmbH Instrument Spectrometer Frequency 100.62 ARWVI-285_flashed.6.fid Origin Value Title 210.36 / Volumes/ nmrdata/ arwong/ nmr/ ARWVI-285_flashed/ 6/ fid Parameter 169.71 Data File Name 96.26 110 100 ppm 90 70 60 50 40 30 20 10 82.74 80.55 78.04 73.41 59.47 58.52 56.05 55.55 52.18 51.70 50.64 48.30 45.59 45.29 44.43 44.32 37.44 31.94 26.98 25.34 25.15 22.52 80 0 -10 Appendix 3 – Spectra Relevant to Chapter 4 390 cdcl3 25.0 s2pul 1D penta 8 30 5.0000 2.6500 1.7078 2020-12-19T20:24:18 2020-12-19T20:25:33 Solvent Temperature Pulse Sequence Experiment Probe Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date Modification Date 1H 16384 65536 Nucleus Acquired Size Spectral Size 12 -1221.7 Lowest Frequency 13 9593.5 Spectral Width 11 inova Instrument Spectrometer Frequency 599.58 Varian Origin Value NJF-5-035-C6D6 Parameter Title 10 9 8 7 6 f1 (ppm) 5 4.33 3.91 3.79 3.21 3.18 3.11 3.07 3.07 2.97 2.87 2.80 2.68 2.51 2.42 2.24 2.08 1.96 1.88 1.56 1.34 1.03 4 3 2 1 0 2.00 0.95 0.97 0.99 1.00 2.97 2.83 2.88 1.02 1.00 1.99 0.98 1.01 2.09 5.01 2.05 2.04 1.08 4.13 1.07 2.96 NJF-5-035-C6D6 -1 -2 Appendix 3 – Spectra Relevant to Chapter 4 391 Bruker BioSpin GmbH spect CDCl3 297.2 zgpg30 1D Z122623_0045 (CPP BBO 400S1 BB-H&F-D-05 Z) 2048 78.7 1.0000 10.0000 1.3631 2020-12-19T23:19:14 2020-12-19T23:19:14 Origin Instrument Solvent Temperature Pulse Sequence Experiment Probe Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date Modification Date -1930.1 13C 32768 65536 Lowest Frequency Nucleus Acquired Size Spectral Size 80 70 60 50 40 30 20 10 0 85.81 81.40 79.68 73.16 72.72 62.46 59.15 55.69 55.53 53.77 49.53 48.97 46.67 46.21 46.01 45.56 44.89 41.49 38.93 33.26 28.59 26.55 25.53 13.83 96.17 210 200 190 180 170 160 150 140 130 120 110 100 90 f1 (ppm) 24038.5 Spectral Width Spectrometer Frequency 100.62 NJF-5-035-C6D6.1.fid Title Value / Volumes/ nmrdata/ nfastuca/ nmr/ NJF-5-035-C6D6/ 1/ fid Data File Name Parameter NJF-5-035-C6D6.1.fid -10 Appendix 3 – Spectra Relevant to Chapter 4 392 14 inova c6d6 25.0 HSQCAD HSQC-EDITED penta 2 32 1.0000 5.3000 0.1500 2020-12-19T20:31:18 2020-12-19T20:39:46 Instrument Solvent Temperature Pulse Sequence Experiment Probe Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date Modification Date (1439, 96) (2048, 1024) Acquired Size Spectral Size 11 (1H, 13C) Nucleus 12 (-1166.0, -1500.6) Lowest Frequency 13 (9593.5, 30154.5) Spectral Width 10 Varian Origin Spectrometer Frequency (599.58, 150.78) HSQCAD_01 Value Title Parameter HSQCAD_01 NJF-5-035-C6D6 9 8 {7.16,127.99}C6D6 7 6 f2 (ppm) 5 4 3 2 1 0 -1 190 180 170 160 150 140 130 120 110 100 90 80 70 60 50 40 30 20 10 0 Appendix 3 – Spectra Relevant to Chapter 4 393 f1 (ppm) 8.0 7.5 7.0 inova c6d6 25.0 gCOSY COSY penta 1 32 1.0000 5.3000 0.1500 2020-12-19T20:27:58 2020-12-19T20:31:14 Instrument Solvent Temperature Pulse Sequence Experiment Probe Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date Modification Date (742, 128) (1024, 1024) Acquired Size Spectral Size 5.5 (1H, 1H) Nucleus 6.0 (-136.7, -136.7) Lowest Frequency 6.5 (4947.7, 4947.7) Spectral Width 5.0 Varian Spectrometer Frequency (599.58, 599.58) gCOSY_01 Value Origin Parameter Title gCOSY_01 NJF-5-035-C6D6 4.5 4.0 3.5 f2 (ppm) 3.0 2.5 2.0 1.5 1.0 0.5 0.0 8.0 7.5 7.0 6.5 6.0 5.5 5.0 4.5 4.0 3.5 3.0 2.5 2.0 1.5 1.0 0.5 0.0 Appendix 3 – Spectra Relevant to Chapter 4 394 f1 (ppm) CDCl3 297.1 hmbcetgpl3nd HMBC Z122623_0045 (CPP BBO 400S1 BB-H&F-D-05 Z) 4 197.4 2.0000 12.5000 0.4260 2020-12-21T20:34:26 2020-12-21T21:17:06 Solvent Temperature Pulse Sequence Experiment Probe Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date Modification Date (2048, 256) (2048, 1024) Acquired Size Spectral Size 9.0 (1H, 13C) Nucleus 10.0 (-195.1, -1091.4) Lowest Frequency 11.0 (4807.7, 22321.4) Spectral Width 7.0 spect Instrument 8.0 Bruker BioSpin GmbH Origin Spectrometer Frequency (400.13, 100.62) NJF-5-035-C6D6.3.ser Title Value / Volumes/ nmrdata/ nfastuca/ nmr/ NJF-5-035-C6D6/ 3/ ser Data File Name Parameter NJF-5-035-C6D6.3.ser 6.0 5.0 f2 (ppm) 4.0 3.0 2.0 1.0 0.0 -10 0 10 20 30 40 50 60 70 80 90 100 110 120 130 140 150 160 170 180 190 200 210 Appendix 3 – Spectra Relevant to Chapter 4 395 f1 (ppm) 14 cdcl3 25.0 s2pul 1D penta 8 40 1.0000 2.6500 1.7078 2021-03-08T23:21:57 2021-03-08T23:22:29 Solvent Temperature Pulse Sequence Experiment Probe Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date Modification Date 1H 16384 65536 Nucleus Acquired Size Spectral Size 12 -1193.2 Lowest Frequency 13 9593.5 Spectral Width 11 inova Instrument Spectrometer Frequency 599.58 Varian Origin Value PROTON_01 Parameter Title 10 9 8 7 6 f1 (ppm) 5 4.25 4.16 3.87 3.30 3.26 3.20 3.18 3.11 3.07 2.99 2.89 2.64 2.52 2.39 2.26 2.08 1.96 1.83 1.77 1.72 1.66 1.49 1.39 1.06 4 3 2 1 0 -1 1.00 0.91 0.97 3.06 2.98 0.78 1.05 1.03 1.01 1.01 0.89 1.00 2.00 1.04 3.06 1.05 4.14 1.94 1.05 1.05 1.06 1.08 1.07 2.94 PROTON_01 NJF-5-066 Appendix 3 – Spectra Relevant to Chapter 4 396 7.26 CDCl3 inova cdcl3 25.0 s2pul 1D penta 10000 30 1.0000 6.4875 0.8696 2021-03-09T02:48:32 2021-03-09T08:00:26 Instrument Solvent Temperature Pulse Sequence Experiment Probe Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date Modification Date -2255.2 13C 32768 65536 Lowest Frequency Nucleus Acquired Size Spectral Size 80 70 60 50 40 30 20 10 0 86.22 79.44 77.00 CDCl3 76.05 73.59 72.63 63.09 59.50 56.34 53.10 49.55 48.63 46.57 46.47 46.08 45.74 42.52 40.89 38.68 32.77 27.92 25.83 24.62 13.69 230 220 210 200 190 180 170 160 150 140 130 120 110 100 90 f1 (ppm) 37682.5 Spectral Width Spectrometer Frequency 150.78 Varian Origin Value CARBON_01 Title Parameter CARBON_01 NJF-5-066 -10 Appendix 3 – Spectra Relevant to Chapter 4 397 14 cdcl3 25.0 HSQCAD HSQC-EDITED penta 16 40 1.0000 5.3000 0.1500 2021-03-08T23:22:32 2021-03-09T00:24:41 Solvent Temperature Pulse Sequence Experiment Probe Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date Modification Date (1439, 96) (2048, 1024) Acquired Size Spectral Size 11 (1H, 13C) Nucleus 12 (-1193.2, -1507.0) Lowest Frequency 13 (9593.5, 30154.5) Spectral Width 10 inova Instrument Spectrometer Frequency (599.58, 150.78) Varian Origin Value HSQCAD_01 Title Parameter HSQCAD_01 NJF-5-066 9 8 7 6 5 f2 (ppm) 4 3 2 1 0 -1 190 180 170 160 150 140 130 120 110 100 90 80 70 60 50 40 30 20 10 0 Appendix 3 – Spectra Relevant to Chapter 4 398 f1 (ppm) inova cdcl3 25.0 gCOSY COSY penta 4 40 1.0000 5.3000 0.1501 2021-03-09T02:37:25 2021-03-09T02:48:17 Solvent Temperature Pulse Sequence Experiment Probe Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date Modification Date (1024, 1024) Spectral Size 6.5 (807, 128) Acquired Size 7.0 (1H, 1H) Nucleus 7.5 (-442.6, -442.6) Lowest Frequency 8.0 (5377.1, 5377.1) Spectral Width 6.0 Varian Instrument Spectrometer Frequency (599.58, 599.58) gCOSY_01 Origin Value Title Parameter gCOSY_01 NJF-5-066 5.5 5.0 4.5 4.0 3.5 f2 (ppm) 3.0 2.5 2.0 1.5 1.0 0.5 0.0 -0.5 8 7 6 5 4 3 2 1 0 Appendix 3 – Spectra Relevant to Chapter 4 399 f1 (ppm) 14 inova cdcl3 25.0 gHMBCAD HMBC penta 16 40 1.0000 5.3000 0.1500 2021-03-09T00:24:44 2021-03-09T02:37:22 Solvent Temperature Pulse Sequence Experiment Probe Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date Modification Date (1439, 200) (2048, 2048) Acquired Size Spectral Size 11 (1H, 13C) Nucleus 12 (-1193.2, -2259.6) Lowest Frequency 13 (9593.5, 36182.7) Spectral Width 10 Varian Instrument Spectrometer Frequency (599.58, 150.78) gHMBCAD_01 Origin Value Title Parameter gHMBCAD_01 NJF-5-066 9 8 7 6 5 f2 (ppm) 4 3 2 1 0 -1 220 200 180 160 140 120 100 80 60 40 20 0 Appendix 3 – Spectra Relevant to Chapter 4 400 f1 (ppm) 14 13 PROTON_01 NJF-5-066 12 11 10 9 8 NOESY1D_01 NJF-5-066 Selective band center: 3.86 (ppm); width: 32.3 (Hz) NOESY1D_01 NJF-5-066 Selective band center: 3.86 (ppm); width: 32.3 (Hz) 7 6 f1 (ppm) 5 4 1.95 3 2 1.90 1 0 1.85 1.80 f1 (ppm) -1 1.75 1 2 3 1 2 3 Appendix 3 – Spectra Relevant to Chapter 4 401 14 c6d6 25.0 s2pul 1D penta 18 46 1.0000 2.6500 1.7078 2021-03-19T23:25:14 2021-03-19T23:26:28 Solvent Temperature Pulse Sequence Experiment Probe Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date Modification Date 1H 16384 65536 Nucleus Acquired Size Spectral Size 12 -1167.6 Lowest Frequency 13 9593.5 Spectral Width 11 inova Instrument Spectrometer Frequency 599.58 Varian Origin Value NJF-5-074-fr111-114 Parameter Title 10 9 8 7.16 C6D6 7 6 f1 (ppm) 5 3.22 3.13 3.11 3.07 2.95 2.81 2.50 2.31 2.16 2.07 1.97 1.90 1.62 1.51 1.35 1.05 4 3 2 1 0 3.27 0.97 1.00 0.98 2.89 6.04 0.93 2.27 1.30 4.47 3.33 2.12 3.27 1.36 1.22 1.14 3.71 2.14 NJF-5-074-fr111-114 NJF-5-066 -1 Appendix 3 – Spectra Relevant to Chapter 4 402 Bruker BioSpin GmbH spect C6D6 297.1 zgpg30 1D Z122623_0045 (CPP BBO 400S1 BB-H&F-D-05 Z) 17500 43.7 1.0000 10.0000 1.3631 2021-03-28T08:51:36 2021-03-28T08:51:36 Origin Instrument Solvent Temperature Pulse Sequence Experiment Probe Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date Modification Date -1946.3 13C 32768 65536 Lowest Frequency Nucleus Acquired Size Spectral Size 80 70 60 50 40 30 20 10 0 86.00 80.79 79.36 75.48 66.01 63.14 58.90 55.59 55.19 53.59 49.56 46.21 45.84 45.59 45.31 42.50 40.93 38.88 33.12 29.98 25.96 25.59 23.55 13.70 95.59 210 200 190 180 170 160 150 140 130 120 110 100 90 f1 (ppm) 24038.5 Spectral Width Spectrometer Frequency 100.62 NJF-5-074.6.fid Title Value / Volumes/ nmrdata/ nfastuca/ nmr/ NJF-5-074/ 6/ fid Data File Name Parameter NJF-5-074.6.fid -10 Appendix 3 – Spectra Relevant to Chapter 4 403 spect C6D6 297.1 hsqcedetgpsisp2.4 HSQC-EDITED Z122623_0045 (CPP BBO 400S1 BB-H&F-D-05 Z) 6 197.4 1.5000 12.5000 0.2135 2021-03-26T02:03:09 2021-03-26T02:03:09 Instrument Solvent Temperature Pulse Sequence Experiment Probe Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date Modification Date (1H, 13C) (1024, 180) (1024, 1024) Nucleus Acquired Size Spectral Size 10.0 (-513.0, -1262.8) Lowest Frequency 9.0 8.0 {7.16,128.12}CDCl3 (4795.4, 16611.3) Spectral Width 6.0 Bruker BioSpin GmbH Origin 7.0 NJF-5-074.3.ser Spectrometer Frequency (400.13, 100.62) / Volumes/ nmrdata/ nfastuca/ nmr/ NJF-5-074/ 3/ ser Title Value Data File Name Parameter NJF-5-074.3.ser 5.0 4.0 f2 (ppm) 3.0 2.0 1.0 0.0 -1.0 150 140 130 120 110 100 90 80 70 60 50 40 30 20 10 0 -10 Appendix 3 – Spectra Relevant to Chapter 4 404 f1 (ppm) Bruker BioSpin GmbH spect C6D6 297.1 hmbcetgpl3nd HMBC Z122623_0045 (CPP BBO 400S1 BB-H&F-D-05 Z) 12 197.4 2.0000 12.5000 0.4260 2021-03-26T04:12:32 2021-03-26T04:12:34 Instrument Solvent Temperature Pulse Sequence Experiment Probe Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date Modification Date (2048, 256) (2048, 1024) Acquired Size Spectral Size 9.0 (1H, 13C) Nucleus 10.0 (-163.1, -1116.4) Lowest Frequency 11.0 (4807.7, 22321.4) Spectral Width 8.0 NJF-5-074.4.ser Origin Spectrometer Frequency (400.13, 100.62) / Volumes/ nmrdata/ nfastuca/ nmr/ NJF-5-074/ 4/ ser Title Value Data File Name Parameter NJF-5-074.4.ser 7.0 6.0 5.0 f2 (ppm) 4.0 3.0 2.0 1.0 0.0 -10 0 10 20 30 40 50 60 70 80 90 100 110 120 130 140 150 160 170 180 190 200 210 Appendix 3 – Spectra Relevant to Chapter 4 405 f1 (ppm) 297.2 cosygpppqf COSY Z122623_0045 (CPP BBO 400S1 BB-H&F-D-05 Z) 2 156.2 1.9062 12.5000 0.2908 2021-03-26T01:30:07 2021-03-26T01:30:07 Temperature Pulse Sequence Experiment Probe Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date Modification Date (1024, 1024) Spectral Size 6.5 (1024, 128) Acquired Size 7.0 (1H, 1H) Nucleus 7.5 (-275.6, -275.6) Lowest Frequency 8.0 (3521.1, 3521.1) Spectral Width 5.0 C6D6 Solvent 5.5 spect Instrument 6.0 Bruker BioSpin GmbH Origin Spectrometer Frequency (400.13, 400.13) NJF-5-074.2.ser Title Value / Volumes/ nmrdata/ nfastuca/ nmr/ NJF-5-074/ 2/ ser Data File Name Parameter NJF-5-074.2.ser 4.5 4.0 3.5 f2 (ppm) 3.0 2.5 2.0 1.5 1.0 0.5 0.0 -0.5 8 7 6 5 4 3 2 1 0 Appendix 3 – Spectra Relevant to Chapter 4 406 f1 (ppm) 14 cdcl3 25.0 s2pul 1D penta 64 46 2.0000 2.6500 1.7078 2021-04-05T21:24:45 2021-04-05T21:29:06 Solvent Temperature Pulse Sequence Experiment Probe Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date Modification Date 1H 16384 65536 Nucleus Acquired Size Spectral Size 12 -1193.3 Lowest Frequency 13 9593.5 Spectral Width 11 inova Instrument Spectrometer Frequency 599.58 Varian Origin Value NJF-5-078 Parameter Title 10 9 8 7 6 f1 (ppm) 5 3 2 1 4.55 1.00 4 4.17 3.54 3.30 3.11 2.98 2.53 2.40 2.23 2.16 2.07 1.97 1.90 1.77 1.65 1.49 1.38 1.06 0.97 0.94 5.97 1.17 0.95 2.20 1.89 1.91 2.08 2.98 1.05 2.02 1.01 3.12 1.13 1.98 2.79 NJF-5-078 NJF-5-073 0 -1 Appendix 3 – Spectra Relevant to Chapter 4 407 7.26 CDCl3 Bruker BioSpin GmbH spect CDCl3 297.2 zgpg30 1D Z122623_0045 (CPP BBO 400S1 BB-H&F-D-05 Z) 16500 72.0 1.0000 10.0000 1.3631 2021-04-07T09:00:21 2021-04-07T09:00:21 Origin Instrument Solvent Temperature Pulse Sequence Experiment Probe Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date Modification Date -1960.9 13C 32768 65536 Lowest Frequency Nucleus Acquired Size Spectral Size 80 70 60 50 40 30 20 10 0 86.29 79.31 77.00 CDCl3 76.01 66.20 63.69 59.50 56.41 53.14 49.62 49.39 46.42 46.30 45.82 45.56 42.83 41.50 38.72 32.79 29.69 25.61 24.95 23.60 13.69 210 200 190 180 170 160 150 140 130 120 110 100 90 f1 (ppm) 24038.5 Spectral Width Spectrometer Frequency 100.62 NJF-5-077.5.fid Title Value / Volumes/ nmrdata-1/ nfastuca/ nmr/ NJF-5-077/ 5/ fid Data File Name Parameter NJF-5-077.5.fid -10 Appendix 3 – Spectra Relevant to Chapter 4 408 inova c6d6 25.0 s2pul 1D penta 8 46 5.0000 2.6500 1.7078 2021-01-15T01:22:12 2021-01-15T01:24:01 Instrument Solvent Temperature Pulse Sequence Experiment Probe Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date Modification Date 16384 65536 Acquired Size Spectral Size 12 1H Nucleus 13 -1167.0 Lowest Frequency 14 9593.5 Spectral Width Spectrometer Frequency 599.58 Varian Origin Value NJF-5-039-C6D6 Title Parameter NJF-5-039-C6D6 NJF-5-035-C6D6 11 10 9 8 7.16 C6D6 7 6 f1 (ppm) 3 2 1 0 -1 4.82 0.99 4 4.15 3.99 3.26 3.12 3.08 3.04 2.94 2.87 2.85 2.84 2.56 2.46 2.38 2.32 2.27 2.23 2.17 2.04 1.94 1.85 1.59 1.55 1.48 1.37 1.06 0.97 0.94 0.95 3.14 2.96 1.03 1.03 0.50 3.03 1.00 2.11 1.02 1.98 1.09 1.05 2.03 1.05 2.12 1.04 1.08 1.13 2.20 1.15 3.02 5 Appendix 3 – Spectra Relevant to Chapter 4 409 150.78 37682.5 -2195.0 13C 32768 65536 Value CARBON_01 Varian inova c6d6 25.0 s2pul 1D penta 10000 30 1.0000 6.4875 0.8696 80 70 60 50 40 30 20 10 0 86.15 82.75 79.80 75.96 72.84 62.83 59.18 56.08 55.74 53.89 49.68 48.99 47.62 46.33 46.26 46.23 39.10 39.05 38.51 33.25 28.10 26.35 25.56 13.93 128.06 C6D6 230 220 210 200 190 180 170 160 150 140 130 120 110 100 90 f1 (ppm) Parameter Title Origin Instrument Solvent Temperature Pulse Sequence Experiment Probe Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date 2021-01-25T21 45 35 Modification Date 2021-01-26T02 57 29 Spectrometer Frequency Spectral Width Lowest Frequency Nucleus Acquired Size Spectral Size CARBON_01 NJF-5-039 -10 Appendix 3 – Spectra Relevant to Chapter 4 410 14 c6d6 25.0 HSQCAD HSQC-EDITED penta 8 38 1.0000 5.3000 0.1500 2021-01-25T17:26:21 2021-01-25T17:57:49 Solvent Temperature Pulse Sequence Experiment Probe Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date Modification Date (1439, 96) (2048, 1024) Acquired Size Spectral Size 11 (1H, 13C) Nucleus 12 (-1167.0, -1451.5) Lowest Frequency 13 (9593.5, 30154.5) Spectral Width 10 inova Instrument Spectrometer Frequency (599.58, 150.78) Varian Origin Value HSQCAD_01 Title Parameter HSQCAD_01 NJF-5-039 9 8 7 6 f2 (ppm) 5 4 3 2 1 0 -1 190 180 170 160 150 140 130 120 110 100 90 80 70 60 50 40 30 20 10 0 Appendix 3 – Spectra Relevant to Chapter 4 411 f1 (ppm) 8.0 c6d6 25.0 gCOSY COSY penta 4 38 1.0000 5.3000 0.1500 2021-01-25T17:15:24 2021-01-25T17:26:17 Solvent Temperature Pulse Sequence Experiment Probe Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date Modification Date (768, 128) (1024, 1024) Acquired Size Spectral Size 6.5 (1H, 1H) Nucleus 7.0 (-288.5, -288.5) Lowest Frequency 7.5 (5120.7, 5120.7) Spectral Width 6.0 inova Instrument Spectrometer Frequency (599.58, 599.58) Varian Origin Value gCOSY_01 Title Parameter gCOSY_01 NJF-5-039 5.5 5.0 4.5 4.0 3.5 f2 (ppm) 3.0 2.5 2.0 1.5 1.0 0.5 0.0 8 7 6 5 4 3 2 1 0 Appendix 3 – Spectra Relevant to Chapter 4 412 f1 (ppm) 14 c6d6 25.0 gHMBCAD HMBC penta 16 38 1.0000 5.3000 0.1500 2021-01-25T17:57:52 2021-01-25T20:10:30 Solvent Temperature Pulse Sequence Experiment Probe Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date Modification Date (1439, 200) (2048, 2048) Acquired Size Spectral Size 11 (1H, 13C) Nucleus 12 (-1167.0, -2251.6) Lowest Frequency 13 (9593.5, 36182.7) Spectral Width 10 inova Instrument Spectrometer Frequency (599.58, 150.78) Varian Origin Value gHMBCAD_01 Title Parameter gHMBCAD_01 NJF-5-039 9 8 7 6 f2 (ppm) 5 4 3 2 1 0 -1 220 200 180 160 140 120 100 80 60 40 20 0 Appendix 3 – Spectra Relevant to Chapter 4 413 f1 (ppm) 11.0 32 98.9 1.0000 11.7000 4.0894 2019-01-10T14:52:36 Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date 65536 Spectral Size 9.0 32768 Acquired Size 9.5 1H Nucleus 10.0 -1545.4 Lowest Frequency 10.5 8012.8 Spectral Width 6.0 1D Experiment 6.5 zg30 Pulse Sequence 7.0 295.2 Temperature 7.5 CDCl3 Solvent 8.0 spect Instrument 8.5 Bruker BioSpin GmbH Origin Spectrometer Frequency 400.13 ARWVI-191_flashed.1.fid Title Value / Volumes/ nmrdata/ arwong/ nmr/ ARWVI-191_flashed/ 1/ fid Parameter Data File Name 5.5 4.0 3.5 3.0 2.5 2.0 1.5 1.0 4.77 4.57 0.99 0.90 4.5 4.13 3.91 3.74 3.33 3.30 3.20 3.14 3.10 3.00 2.78 2.56 2.44 2.39 2.25 2.06 1.88 1.69 1.45 1.32 1.00 0.93 0.92 3.17 3.01 4.02 1.35 2.87 2.23 1.13 2.07 1.09 1.23 1.06 7.08 1.05 2.02 1.13 1.22 5.0 ppm 0.5 0.0 -0.5 -1.0 Appendix 3 – Spectra Relevant to Chapter 4 414 1300 78.7 0.5000 10.0000 1.3631 2019-01-10T15:34:47 Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date 65536 Spectral Size 180 32768 Acquired Size 190 13C Nucleus 200 -1962.5 Lowest Frequency 210 24038.5 Spectral Width 120 1D Number of Scans 130 zgpg30 Experiment 140 295.2 Pulse Sequence 150 CDCl3 Temperature 160 spect Solvent 170 Bruker BioSpin GmbH Instrument Spectrometer Frequency 100.62 ARWVI-191_flashed.2.fid Origin Value Title 211.27 / Volumes/ nmrdata/ arwong/ nmr/ ARWVI-191_flashed/ 2/ fid Parameter 169.75 Data File Name 95.76 110 100 ppm 90 70 60 50 40 30 20 10 82.50 78.24 77.68 76.78 59.46 58.32 55.68 55.52 52.34 48.66 47.58 45.43 45.27 44.24 44.08 42.65 37.39 31.64 26.79 25.35 24.03 22.39 80 0 -10 Appendix 3 – Spectra Relevant to Chapter 4 415 11.0 32 142.8 1.0000 11.7000 4.0894 2019-01-18T00:41:35 Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date 65536 Spectral Size 9.0 32768 Acquired Size 9.5 1H Nucleus 10.0 -1545.4 Lowest Frequency 10.5 8012.8 Spectral Width 1.00 6.0 5.5 1D Experiment 6.5 zg30 Pulse Sequence 7.0 295.1 Temperature 7.5 CDCl3 Solvent 8.0 spect Instrument 8.5 Bruker BioSpin GmbH Origin Spectrometer Frequency 400.13 ARWVI-296_prepTLC_B.1.fid Title Value / Volumes/ nmrdata-2/ arwong/ nmr/ ARWVI-296_prepTLC_B/ 1/ fid Parameter 5.85 Data File Name 4.0 3.5 3.0 2.5 2.0 1.5 1.0 4.71 4.60 1.11 1.12 4.5 3.89 3.36 3.30 3.21 3.18 3.12 3.02 2.86 2.68 2.61 2.34 2.27 2.06 1.93 1.83 1.72 1.43 1.30 3.13 3.10 3.22 3.15 4.09 1.24 1.09 1.22 1.22 1.07 1.16 1.14 6.06 1.03 1.10 2.26 1.30 0.96 5.0 ppm 0.5 0.0 -0.5 -1.0 Appendix 3 – Spectra Relevant to Chapter 4 416 87.8 0.5000 10.0000 1.3631 2019-01-18T04:15:36 Relaxation Delay Pulse Width Acquisition Time Acquisition Date 65536 Spectral Size 180 32768 Acquired Size 190 13C Nucleus 200 -1961.5 Lowest Frequency 210 24038.5 Spectral Width 110 3000 Receiver Gain 120 1D Number of Scans 130 zgpg30 Experiment 140 295.1 Pulse Sequence 150 CDCl3 Temperature 160 spect Solvent 170 Bruker BioSpin GmbH Instrument Spectrometer Frequency 100.62 ARWVI-296_prepTLC_B.6.fid Value Origin 170.55 Title 153.85 / Volumes/ nmrdata-2/ arwong/ nmr/ ARWVI-296_prepTLC_B/ 6/ fid Parameter 114.16 Data File Name 95.97 100 ppm 90 70 60 50 40 30 20 10 82.58 78.36 78.23 77.20 59.47 59.09 55.62 55.49 49.36 48.05 45.22 45.09 44.34 43.90 42.83 42.62 37.39 31.95 30.74 25.38 23.74 22.53 80 0 -10 Appendix 3 – Spectra Relevant to Chapter 4 417 11.0 1D 32 197.4 1.0000 11.7000 4.0894 2019-01-18T22:07:10 Number of Scans Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date 65536 Spectral Size 9.0 32768 Acquired Size 9.5 1H Nucleus 10.0 -1545.3 Lowest Frequency 10.5 8012.8 Spectral Width 6.5 zg30 Experiment 7.0 295.1 Pulse Sequence 7.5 CDCl3 Temperature 8.0 spect Solvent 8.5 Bruker BioSpin GmbH Instrument Spectrometer Frequency 400.13 ARWVI-297_prepTLC.1.fid 6.17 Origin 1.00 6.0 5.5 5.0 ppm 4.5 4.0 3.5 3.0 2.5 2.0 1.5 5.85 0.93 Title Value 4.75 4.59 1.10 1.01 / Volumes/ nmrdata/ arwong/ nmr/ ARWVI-297_prepTLC/ 1/ fid Parameter 3.88 3.84 3.79 3.36 3.30 3.21 3.17 3.11 3.04 2.58 2.41 2.33 2.14 2.03 1.83 1.69 1.43 1.30 1.01 0.99 0.97 2.98 3.02 4.14 2.91 1.29 0.98 2.03 1.01 1.02 1.05 5.99 2.10 2.10 1.07 1.06 Data File Name 1.0 0.5 0.0 -0.5 Appendix 3 – Spectra Relevant to Chapter 4 418 3000 72.0 0.5000 10.0000 1.3631 2019-01-19T01:41:17 Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date 65536 Spectral Size 180 32768 Acquired Size 190 13C Nucleus 200 -1961.2 Lowest Frequency 210 24038.5 Spectral Width 120 1D Number of Scans 130 zgpg30 Experiment 140 295.2 Pulse Sequence 150 CDCl3 Temperature 160 spect Solvent 170 Bruker BioSpin GmbH Instrument Spectrometer Frequency 100.62 ARWVI-297_prepTLC.6.fid Origin 169.88 Title Value 134.32 / Volumes/ nmrdata/ arwong/ nmr/ ARWVI-297_prepTLC/ 6/ fid 95.51 110 100 ppm 90 80 70 60 59.63 59.52 59.41 59.30 59.19 ppm 59.51 59.49 50 40 30 20 10 77.40 77.15 76.90 76.65 76.40 ppm 77.20 Parameter 82.90 78.56 77.20 76.57 59.51 59.49 55.59 55.50 49.22 48.00 45.59 45.16 44.29 44.22 42.67 37.54 37.38 32.00 31.52 25.50 23.48 22.53 76.57 125.27 Data File Name 0 -10 Appendix 3 – Spectra Relevant to Chapter 4 419 11.0 197.4 1.0000 11.7000 4.0894 2019-01-29T00:14:13 Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date 65536 Spectral Size 9.0 32768 Acquired Size 9.5 1H Nucleus 10.0 -1532.2 Lowest Frequency 10.5 8012.8 Spectral Width 1.00 1.00 5.5 5.0 ppm 64 Number of Scans 6.0 1D Experiment 6.5 zg30 Pulse Sequence 7.0 295.1 Temperature 7.5 C6D6 Solvent 8.0 spect Instrument 8.5 Bruker BioSpin GmbH Origin Spectrometer Frequency 400.13 ARWVI-299_prepTLC2_C6D6.1.fid Title Value / Volumes/ nmrdata/ arwong/ nmr/ ARWVI-299_prepTLC2_C6D6/ 1/ fid Parameter 5.70 5.61 Data File Name 4.5 3.78 3.12 3.09 3.05 2.96 2.86 2.55 2.46 2.39 2.29 2.15 2.09 2.01 1.93 1.74 1.56 1.04 3.5 3.0 2.5 2.0 1.5 1.0 1.02 2.98 1.03 2.98 1.02 2.10 3.03 0.83 1.10 1.04 2.09 1.18 2.02 2.06 0.97 5.26 3.08 4.0 0.5 0.0 -0.5 -1.0 Appendix 3 – Spectra Relevant to Chapter 4 420 87.8 0.5000 10.0000 1.3631 2019-01-29T07:45:26 Receiver Gain Relaxation Delay Pulse Width Acquisition Time Acquisition Date 65536 Spectral Size 180 32768 Acquired Size 190 13C Nucleus 200 -1921.2 Lowest Frequency 210 24038.5 Spectral Width 110 100 ppm 13000 Number of Scans 120 1D Experiment 130 zgpg30 Pulse Sequence 140 295.1 Temperature 150 C6D6 Solvent 160 spect Instrument 170 Bruker BioSpin GmbH Origin Spectrometer Frequency 100.62 ARWVI-299_prepTLC2_C6D6.3.fid Title Value / Volumes/ nmrdata/ arwong/ nmr/ ARWVI-299_prepTLC2_C6D6/ 3/ fid Parameter 132.53 129.69 Data File Name 90 70 60 50 40 30 20 85.63 80.00 75.00 74.45 62.99 59.19 55.68 53.91 49.56 48.63 46.77 46.33 46.03 42.70 39.59 38.93 33.44 33.28 26.74 24.23 13.74 80 10 0 -10 Appendix 3 – Spectra Relevant to Chapter 4 421