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CHAMNI-DISSERTATION.Pdf NEW DIAZO REAGENTS AND APPLICATIONS OF β-LACTONES FOR SYNTHESIS AND BIOLOGICAL EVALUATION OF NATURAL PRODUCTS A Dissertation by SUPAKARN CHAMNI Submitted to the Office of Graduate Studies of Texas A&M University in partial fulfillment of the requirements for the degree of DOCTOR OF PHILOSOPHY December 2011 Major Subject: Chemistry New Diazo Reagents and Applications of β-Lactones for Synthesis and Biological Evaluation of Natural Products Copyright 2011 Supakarn Chamni NEW DIAZO REAGENTS AND APPLICATIONS OF β-LACTONES FOR SYNTHESIS AND BIOLOGICAL EVALUATION OF NATURAL PRODUCTS A Dissertation by SUPAKARN CHAMNI Submitted to the Office of Graduate Studies of Texas A&M University in partial fulfillment of the requirements for the degree of DOCTOR OF PHILOSOPHY Approved by: Chair of Committee, Daniel Romo Committee Members, David E. Bergbreiter Coran M. H. Watanabe Thomas McKnight Head of Department, David H. Russell December 2011 Major Subject: Chemistry iii ABSTRACT New Diazo Reagents and Applications of β-Lactones for Synthesis and Biological Evaluation of Natural Products. (December 2011) Supakarn Chamni, B.S., Thammasat University Chair of Advisory Committee: Prof. Daniel Romo Natural products are essential tools for basic cellular studies leading to the identification of medically relevant protein targets and the discovery of potential therapeutic agents. We have developed a set of second-generation diazo reagents with small steric footprints, namely an α-trifluoroethyl (HTFB) diazo reagent, for simultaneous arming and SAR studies of bioactive natural products. The Rh(II)- catalyzed O−H insertions of several alcohol-containing natural products, including the potent translation inhibitor lactimidomycin, are investigated and useful reactivity and both chemo- and site- (chemosite) selectivities are observed. The α-trifluoroethyl diazo reagents (HTFB) shows clear differences in the IL-2 reporter assay with FK506 derivatives and provides greater retention of biological activity in a hMetAP2 proliferation assay of fumagillol derivatives compared to the first generation p- bromophenyl diazo reagent (HBPA). The synthetic utilities of the new α-trifluoroethyl diazo reagent (HTFB) provide a great new tool for basic cellular studies facilitating the discovery of new drug candidates for human disease. iv Furthermore, we are interested in methodologies for β-lactone synthesis and transformations. In this study, we demonstrated synthetic versatilities of β-lactones for the synthesis of β-lactam congeners of orlistat as fatty acid synthase inhibitors via SnCl4- promoted tandem Mukaiyama aldol-lactonization (TMAL) reaction and a one-pot, mild conversion of β-lactones to β-lactams. The inhibitory activities of the derived β-lactam derivatives are determined in a biochemical fluorogenic assay using recombinant FAS- TE, and the micro-molar range FAS-TE inhibitory activities were observed. Additionally, we pursued synthetic studies toward the total synthesis of spongiolactone, which is a unique β-lactone-containing marine diterpenoid, isolated from the marine sponge Spongionella gracilis. This natural product bears a unique tricyclic β-lactone core possessing four contiguous stereogenic centers and an additional stereogenic quaternary carbon on a cyclohexyl appendage. We completed the total synthesis of 6,15-bis-epi-spongiolactone by employing an intramolecular nucleophile- catalyzed aldol-lactonization (NCAL) process as the key step to construct the fused tricyclic β-lactone core. Importantly, we developed a double diastereoselective and, for the first time, a kinetic resolution via the NCAL process that enables an enantioselective strategy to the tricyclic β-lactone core of (+)-spongiolactone. v DEDICATION To my beloved parents, Worranart and Sirin Chamni and sister, Nuanphan Chamni. vi ACKNOWLEDGEMENTS I would like to sincerely thank Prof. Daniel Romo, my advisor, for his guidance and support. It has been an excellent experience to work under his supervision. I really appreciate his time in teaching me not only an intellectual research, but also a life-lesson throughout my time at Texas A&M. Thank you for giving me such a great opportunity to expand my knowledge into several research areas during my Ph.D. studies. This opportunity has given me the wonderful chance to learn so much chemistry and increase my research skill and has given me the vision to support my future career as an academician. I also would like to thank my committee members, Prof. David Bergbreiter, Prof. Coran Watanabe and Prof. Thomas McKnight, for their guidance and assistance throughout the course of this research. Many thanks also to Prof. Daniel Singleton, Prof. Brian Connell and, once again, Prof. David Bergbreiter for the excellent classes to advance my chemistry knowledge and assist my research. Thanks also to both former and current Romo group members for their friendship, laughs, helps and discussions. It has been a great memorable moment to share with all of you. I would like to thank Dr. Wei Zhang, my research mentor who taught me excellent laboratory tricks. Much appreciation goes to Dr. Sung Wook Cho and Dr. Satyamaheshwar Peddibhotla for their initial studies toward my research. Thank you to Dr. Ke Kong, Gang Liu and Dr. Yonggang Wang for being great chemist-role models. Thanks to Dr. Henry Nguyen, as known as my uncle, for sincere friendship and vii for keeping me entertained while working under pressure for several years. Thank you to Dr. Carolyn Leverett for warm kindness, sweet-treats, and for always being my research partner and personal supporter. Thanks to JC Reyes for sharing the creative lifestyle, laughable conversations and for being my birthday buddy for a couple of years. Special thanks for all the girls in Romo group and 315-labmate for making the lab an enjoyable place. Thank to Dr. Kay Morris, Morgan Shirley, Natalie Harvey and Rae Lynn McFarlin for moral support and fun activities. Thanks also go to Alfred Tuley and Mikail Abbasov for making me laugh. Thank you to Dr. Omar Robbles, Dr. Sreekumar Vellalath and Dr. Chun-Xiao Xu for research advice and discussion. I also want to thank my mentees, Natalie Harvey and Sandra Fiorentini, for their excellent hard work toward the total synthesis of spongiolactone. I also would like to thanks the former and current Thai chemistry students, especially Dr. Chayanant Hongfa and Dr. Osit Karroonniran, for encouragement, lasting friendship, entertainment and research assistance. I also want to extend my gratitude to the Royal Thai Government Scholarship for financial support throughout my Ph.D. career, and for a job position at the Department of Pharmacognosy and Pharmaceutical Botany, Faulty of Pharmaceutical Sciences, Chulalongkorn University, Thailand. Finally, I would like to thank my parents and my sister for their encouragement throughout five years of my graduated study. I truly appreciate their love and support, which has made me a better person and a successful chemist. viii TABLE OF CONTENTS Page ABSTRACT .......................................................................................................... iii DEDICATION ...................................................................................................... v ACKNOWLEDGEMENTS ................................................................................... vi TABLE OF CONTENTS....................................................................................... viii LIST OF FIGURES............................................................................................... xi LIST OF TABLES................................................................................................. xiv CHAPTER I INTRODUCTION TO METHODS FOR NATURAL PRODUCT DERIVATIZATION ......................................................................... 1 1.1. Basic Technology of Cellular Probe...................................... 2 1.2. Methods for Natural Product Derivatization Leading to Cellular Probe Synthesis ....................................................... 3 II NEW DIAZO REAGENTS WITH SMALL STERIC FOOTPRINTS FOR SIMULTANEOUS ARMING AND STRUCTURE-ACTIVITY RELATIONSHIP STUDIES OF ALCOHOL-CONTAINING NATURAL PRODUCTS .................................................................. 13 2.1. Introduction .......................................................................... 13 2.1.1. Simultaneous Arming and Structure-Activity Relationship (SAR) Studies of Natural Products for Chemical Proteomics ...................................... 14 2.1.2. Derivatization of Alcohol-Containing Natural Products via Chemo- and Site Selective Rh(II)- Catalyzed O−H Insertions..................................... 16 2.1.3. Rh(II)-Mediated O−H Insertion: Mechanism and Carbene Type ....................................................... 17 2.2. Results and Discussion.......................................................... 20 2.2.1. Alteration of Chemosite Selectivity Using Chiral Rh(II) Catalysts..................................................... 21 ix CHAPTER Page 2.2.2. Development of Second Generation Diazo Reagents ............................................................... 27 2.2.3. Proof of Principle Protein Assays and Affinity Experiments.......................................................... 36 2.3. Conclusion............................................................................ 45 III SYNTHESIS OF β-LACTAM CONGENERS OF ORLISTAT AS FATTY ACID SYNTHASE INHIBITORS ....................................... 47 3.1. Introduction
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