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Development of Specific Natural Bacterial Beta-Glucuronidase Inhibitors for Reducing Irinotecan-Associated Diarrhea

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Development of Specific Natural Bacterial Beta-Glucuronidase Inhibitors for Reducing Irinotecan-Associated Diarrhea Wayne State University Wayne State University Dissertations January 2020 Development Of Specific Natural Bacterial Beta-Glucuronidase Inhibitors For Reducing Irinotecan-Associated Diarrhea Fei Yang Wayne State University Follow this and additional works at: https://digitalcommons.wayne.edu/oa_dissertations Part of the Food Science Commons Recommended Citation Yang, Fei, "Development Of Specific Natural Bacterial Beta-Glucuronidase Inhibitors For Reducing Irinotecan-Associated Diarrhea" (2020). Wayne State University Dissertations. 2515. https://digitalcommons.wayne.edu/oa_dissertations/2515 This Open Access Dissertation is brought to you for free and open access by DigitalCommons@WayneState. It has been accepted for inclusion in Wayne State University Dissertations by an authorized administrator of DigitalCommons@WayneState. DEVELOPMENT OF SPECIFIC NATURAL BACTERIAL BETA-GLUCURONIDASE INHIBITORS FOR REDUCING IRINOTECAN-ASSOCIATED DIARRHEA by FEI YANG DISSERTATION Submitted to the Graduate School of Wayne State University, Detroit, Michigan in partial fulfillment of the requirements for the degree of DOCTOR OF PHILOSOPHY 2020 MAJOR: NUTRITION AND FOOD SCIENCE Approved By: Advisor Date DEDICATION To my beloved father and mother. To my darling wife and daughter. ii ACKNOWLEDGMENTS My deepest and sincere gratitude goes first to my supervisor, Prof. Dr. Kequan Zhou. He opened a window for me to the amazing world of gut microbiota by bacterial β-glucuronidases and new probiotics, which gave me a chance to improve professional skills, and an insight into the state-of-art of my research fields. During these wonderful years with him, his competent guidance and instruction gave me the power to move forward, his constant trust and support provided me the confidence to overcome any difficulties faced. More importantly, his scientific spirit of suspicion, innovation and pursing truth have greatly enlightened me and will benefit my endeavors in the future. Never enough words to say thank you for his help and supports. I am very grateful to Dr. K-L Catherine Jen, Dr. Smiti V. Gupta, and Dr. Weilong Hao, the honorable members of my dissertation committee. From the meetings we had, their valuable advices clarified my direction, and their brilliant ideas enabled me to settle specific problems. I am so lucky to have them on my committee. I extend my gratitude to the department of Nutrition and Food Science and the Graduate School of Wayne State University for offering me the scholarships and awards. My special thanks are addressed to all the faculty and staff of our department for their professionalism and considerate supports. How to miss this unique chance to thank the present and past members in Dr. Zhou’s lab: Wenjun Zhu, Qing Ai, Paba Edirisuriya, Dr. Shi Sun, Dr. Jiangqi Tang (Kiki), Dr. Kai Nie, Lingjing Liu, Jiarun Cui, Jun Ma, Sampurna Guhathakurta, Maria Elena Hakim and Ninghui Zhou et al. We did research together, learn together, and play together. We share the joys when someone succeeds, we give a hand when someone is in difficulty. What a big and sweet family. iii I would also like to acknowledge Dr. Nicholas Peraino and Mr. Dennis Anderson in Lumigen Instrument Center of Wayne State University for their expertise training and patient instruction in the HRESIMS and NMR, respectively. My great gratitude is also devoted to my dearest family: dad Xiaoliu Yang, mom Qiaoxiang Wang, wife Yueyuan Chen and daughter Erin Enxin Yang. I could not have made it without their warm company and selfless love. Finally, I am infinitely thankful to all of you who have contributed to my achievement of this project by any mean. iv TABLE OF CONTENTS DEDICATION .............................................................................................................................................. ii ACKNOWLEDGMENTS ........................................................................................................................... iii LIST OF TABLES ..................................................................................................................................... viii LIST OF FIGURES ..................................................................................................................................... ix LIST OF ABBREVIATIONS ...................................................................................................................... xi CHAPTER 1 Introduction ............................................................................................................................. 1 1.1. Irinotecan and Delayed Diarrhea .................................................................................................. 1 1.2. Bacterial β-Glucuronidases and Inhibitors .................................................................................... 4 1.3. Specific Aims ................................................................................................................................ 7 CHAPTER 2 Isolation of Bioactive Compounds from Noni Fruits ............................................................. 9 2.1. Introduction of Noni (Morinda citrifolia) ................................................................................... 10 2.1.1. Nutritional content and chemical constituents of noni ............................................................ 11 2.1.2. Biological activities and safety of noni ................................................................................... 12 2.2. Isolation and Purification ............................................................................................................ 14 2.2.1. Equipment and materials ......................................................................................................... 15 2.2.2. Isolation procedures ................................................................................................................ 16 CHAPTER 3 Elucidation of Chemical Structures ...................................................................................... 19 3.1. Spectroscopic Data Acquisition .................................................................................................. 22 3.2. ECD Computation ....................................................................................................................... 25 3.2.1. Computational chemistry and its application in CD calculation ............................................. 25 v 3.2.2. ECD calculation of compounds 1-4 ........................................................................................ 27 3.3. Structural Elucidation ................................................................................................................. 31 3.3.1. (7S,8S,7'R,8'R)-Isoamericanol B ............................................................................................. 32 3.3.2. Americanol B .......................................................................................................................... 34 3.3.3. Moricitrin A ............................................................................................................................ 37 3.3.4. Moricitrin B ............................................................................................................................ 39 CHAPTER 4 Evaluation of Inhibitory Activities ....................................................................................... 43 4.1. Inhibitory Assays on Enzymes .................................................................................................... 43 4.1.1. Enzymes and Reagents ............................................................................................................ 43 4.1.2. Bacterial β-Glucuronidase Assay ............................................................................................ 44 4.1.3. α-Amylase Assay .................................................................................................................... 44 4.1.4. α-Glucosidase Assay ............................................................................................................... 45 4.1.5. Pancreatic Lipase Assay .......................................................................................................... 45 4.1.6. Data Analysis .......................................................................................................................... 45 4.2. Specific Inhibition Against Bacterial β-Glucuronidase .............................................................. 45 4.3. Inhibition on Digestive Enzymes ................................................................................................ 47 CHAPTER 5 Discussion ............................................................................................................................. 49 5.1. Bioassay-guided Isolation ........................................................................................................... 49 5.2. Structure Elucidations ................................................................................................................. 50 5.3. In vitro Study .............................................................................................................................. 51 vi 5.4. In vivo Study ............................................................................................................................... 53 APPENDIX ................................................................................................................................................
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