Pharmacognostic Investigations and in Vitro Biological Profile of Illicium Angustisepalum
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Pharmacognostic Investigations and in vitro Biological Profile of Illicium angustisepalum By KARINA MARIANNA SZYMULANSKA-RAMAMURTHY Magister Farmacji, Jagiellonian University, Poland, 2009 DISSERTATION Submitted as partial fulfillment of the requirements for the degree of Doctor of Philosophy in Pharmacognosy in the Graduate College of the University of Illinois at Chicago, 2015 Chicago, Illinois Doctoral Committee: Dr. Chun-Tao Che, Chair/Advisor Dr. Birgit U. Jaki Dr. Hyun-Young Jeong, Department of Pharmacy Practice Dr. Jeremy J. Johnson Dr. Djaja D. Soejarto DEDICATION I dedicate this dissertation to my husband, Saikrishnan Ramamurthy, who has given me the motivation and strength to persevere beyond all the challenges that I have faced during the past five years. Without his presence, support and love, I could not have completed this dissertation. I love you forever. II ACKNOWLEDGEMENTS I would like to express my sincere gratitude to Professor Chun-Tao Che, my advisor, for the opportunity to conduct my research work under his guidance, for providing essential environment to grow as a scientist and also for his generous support and inspiration throughout the past four years. I extend my gratitude to the members of my dissertation committee: Dr. Birgit U. Jaki for the thoughtful review of my work and for insightful comments that helped me to enrich my dissertation, and also for the encouragement and for the support throughout my tenure in the program. Dr. Djaja D. Soejarto for providing me with opportunities to broaden my knowledge of medicinal plants and for teaching me about the plant taxonomy, and also for the insightful review of this work and for the encouragement while in the program. Dr. Hyun-Young Jeong and Dr. Jeremy J. Johnson for the review of my dissertation and for their valuable comments and also for good advice. Many thanks to Dr. Guido F. Pauli for providing me with the opportunity to participate in this program and to Dr. Jimmy Orjala for his guidance. And also, thanks to Dr. Brian T. Murphy and Dr. Birgit Dietz for their help and support. Special thanks to Dr. Ming Zhao for training me in various research techniques and for his continuous support during my tenure as a graduate student. III ACKNOWLEDGEMENTS (continued) I would also like to express my appreciation to the following people, without whose assistance my work would have not move forward: Dr. Aleksjej Krunic and Dr. Benjamin Ramirez for providing me with the NMR training. Dr. Jerry White, Caleb Nienow and Monika Lysakowska for providing mass spectra of the compounds isolated in the present study. Dr. Arthur Anderson for help with GC-MS analysis used to establish the metabolite fingerprinting. Wei-Lun Chen, Dr. Steven Swanson and Dr. Joanna Burdette for testing the compounds isolated in the present study in the cytotoxic assay. Dr. Hyunwoo Lee, Joy Barranis and Dr. Tamiko Oguri for testing the compounds isolated in the present study in the antimicrobial assay. Dr. Bernard Santarsiero for collecting x-ray diffraction crystallographic data. Dr. Shi Lei and Dr. Paul Ip for testing compounds and fractions in the neural cell protection assay and in the promotion of neural growth assay. Also thanks to all past and present members of Professor’s Che laboratory for their help. Many thanks go to the faculty, staff and graduate students of the Department of Medicinal Chemistry and Pharmacognosy for assistance and conversations. IV ACKNOWLEDGEMENTS (continued) I am very grateful to the Department of Medicinal Chemistry and Pharmacognosy for the teaching assistantships provided during my tenure as a graduate student. I thank my friends, Dr. Edyta Grzelak, Rasika Phansalkar, Dr. Yu Zhang, Joy Barranis, Wei- Lun Chen, Yang Liu, Sunaina Premkumar, Prabha Venkat, Meera Ranganathan, Dr. Sai Hari Gandham, Vaishnav Vijayakumar, Naren Babu, Prashanthi Gandham and Amruth Rao for their friendship and support. Last, and the most important, I thank my parents, Jozefa and Michal, who instilled in me the perseverance and passion for science and to my brothers, Michal, Daniel and Wojciech, who continuously supported me in my efforts despite the geographical distance between us. This work would not be possible without their love and caring support. I also thank my uncle, Janusz Borek, who inspired me to pursue my career goals. I express my gratitude to my parents-in-law, Uma Maheswari and Ramamurthy, sisters in law, Uthara Prashant, Magdalena Szymulanska, and Meng-Yi Wu for their encouragement and love. - KSR V TABLE OF CONTENTS CHAPTER 1 - LITERATURE INFORMATION ........................................................................1 INTRODUCTION ................................................................................................1 LITERATURE REVIEW ON THE GENUS ILLICIUM .......................................4 Taxonomy of the genus Illicium ..............................................................4 Ethnomedical and other uses of species of Illicium ...............................7 Brief history of scientific investigations on the genus Illicium ................9 Biological activities of Illicium plants .................................................... 12 1.2.4.1 In vitro biological activities of species of Illicium ...................... 12 1.2.4.1.1 Acetylcholinesterase inhibiting activity ...................................... 12 1.2.4.1.2 α-Glucosidase inhibiting activity ................................................ 16 1.2.4.1.3 Anti-inflammatory activity ........................................................... 17 1.2.4.1.4 Antimicrobial activity .................................................................. 20 1.2.4.1.5 Antioxidant activity ..................................................................... 21 1.2.4.1.6 Antiviral activity .......................................................................... 24 1.2.4.1.7 Chemopreventive activity........................................................... 25 1.2.4.1.8 Cytotoxic activity ........................................................................ 26 1.2.4.1.9 Estrogenic activity ...................................................................... 29 1.2.4.1.10 Lipase inhibition activity ............................................................. 30 1.2.4.1.11 Neural cell protection activity ..................................................... 30 1.2.4.1.12 Promotion of neural growth activity ........................................... 31 1.2.4.1.13 Promotion of ChAT activity ........................................................ 33 1.2.4.2 In vivo biological activities of Illicium plants ............................. 34 1.2.4.2.1 Anti-depressant .......................................................................... 34 1.2.4.2.2 Anti-inflammatory ....................................................................... 34 1.2.4.2.3 Chemopreventive ....................................................................... 35 1.2.4.2.4 Fumigant .................................................................................... 36 Morphological description of I. angustisepalum .................................. 36 Chemical constituents of I. angustisepalum ....................................... 37 OBJECTIVES OF THE STUDY ...................................................................... 40 CHAPTER 2 - METABOLITE FINGERPRINTING OF I. ANGUSTISEPALUM ................. 42 RATIONALE .................................................................................................... 42 TABLE OF CONTENTS (continued) EXPERIMENTAL PROCEDURES ................................................................. 43 Plant material collection ....................................................................... 43 Plant material extraction ...................................................................... 44 2.2.3 GC-MS analysis, data processing and compound identification ........ 44 RESULTS ........................................................................................................ 46 Identification of volatile secondary metabolites .................................. 46 Metabolite fingerprint of I. angustisepalum ......................................... 51 Comparative profiles between I. angustisepalum, I. lanceolatum and I. verum ................................................................................................ 52 DISCUSSION .................................................................................................. 54 CONCLUSIONS .............................................................................................. 56 CHAPTER 3 - PHYTOCHEMICAL STUDY OF I. ANGUSTISEPALUM ............................ 57 RATIONALE .................................................................................................... 57 GENERAL EXPERIMENTAL PROCEDURES .............................................. 57 PLANT MATERIAL .......................................................................................... 58 EXTRACTION OF THE PLANT MATERIAL .................................................. 58 FRACTIONATION OF THE ETHANOL EXTRACT ....................................... 58 ISOLATION AND STRUCTURE ELUCIDATION .......................................... 59 Thymol (1) ............................................................................................ 68 (-)-T-Muurolol (2) .................................................................................. 68 2-Hydroxy-2-methyl-6-methyleneoct-7-en-3-yl benzoate (3) ............. 70