Nghiên cứu thành phần hóa học của hai loài địa y lobaria orientalis and dendriscosticta plantyphylloides (lobariaceae)

VIETNAM NATIONAL UNIVERSITY – HO CHI MINH CITY UNIVERSITY OF SCIENCE - NGUYEN THI MY DUNG STUDY ON CHEMICAL CONSTITUENTS OF TWO LICHEN SPECIES LOBARIA ORIENTALIS AND DENDRISCOSTICTA PLATYPHYLLOIDES (LOBARIACEAE) PhD THESIS IN CHEMISTRY HO CHI MINH CITY – 2018 VIETNAM NATIONAL UNIVERSITY – HO CHI MINH CITY UNIVERSITY OF SCIENCE NGUYEN THI MY DUNG STUDY ON CHEMICAL CONSTITUENTS OF TWO LICHEN SPECIES LOBARIA ORIENTALIS AND DENDRISCOSTICTA PLATYPHYLLOIDES (LOBARIACEAE) Major: Organic Chemistry Major Code: 62 44 01 14 Reviewer 1: Assoc Prof Dr Trần Hùng Reviewer 2: Assoc Prof Dr Lê Tiến Dũng Reviewer 3: Assoc Prof Dr Hà Diệu Ly Independent reviewer 1: Assoc Prof Dr Nguyễn Thị Hoàng Anh Independent reviewer 2: Dr Võ Thị Ngà SUPERVISORS: PROF DR NGUYEN KIM PHI PHUNG PROF DR JACQUES MORTIER HO CHI MINH CITY – 2018 SOCIALIST REPUBLIC OF VIETNAM INDEPENDENCE-FREEDOM-HAPPINESS DECLARATION The work presented in this thesis was completed in the period of November 2013 to November 2016 under the co-supervision of Professor Nguyen Kim Phi Phung of the University of Science, Vietnam National University, Ho Chi Minh City, Vietnam and Professor Jacques Mortier from Institute for Molecules and Materials, University of Maine, France In compliance with the university regulations, I declare that: Except where due acknowledgement has been made, the work is that of the author alone; The work has not been submitted previously, in whole or in part, to qualify for any other academic award; The content of the thesis is the result of work which has been carried out since the official commencement date of the approved doctoral research program; Ethics procedures and guidelines have been followed PhD student NGUYỄN THỊ MỸ DUNG i ACKNOWLEDGEMENTS There are many individuals without whom the work described in this thesis might not have been possible, and to whom I am greatly indebted Firstly, I wish to thank my supervisor, Prof Dr Nguyen Kim Phi Phung for her knowledge, support, and guidance, hundreds of meetings/emails and for always keeping me on my toes, from the very beginning to the end of my PhD I would also like to acknowledge my second supervisor, Prof Dr Jacques Mortier for his guidance, patience and who has taught me the true spirit of research I am deeply indebted to Prof Dr Warinthorn Chavasiri of Chulalongkorn University, Thailand for his teachings, kindness, helpful suggestion and valuable advice in this research I would also like to express my sincere thanks to Dr Robert Lücking in the Department of Botany, the Field Museum, Illinois USA for his expertise in the authentification of lichen I am very grateful to thank PhD Do Thi My Lien for giving up her precious time to help me with CD spectra Similarly, I would also like to thank my lecturers, friends and students in the Department of Organic Chemistry, Faculty of Chemistry, University of Science, Vietnam National University-Ho Chi Minh City Finally, I would like to thank my parents for believing in me and for being proud of me Their unconditional love and support have given me the strength and courage while I am far from home ii CONTENTS DECLARATION ACKNOWLEDGEMENTS CONTENTS LIST OF ABBREVIATIONS LIST OF TABLES LIST OF FIGURES PREFACE CHAPTER 1: LITERATURE REVIEW 1.1 GENERAL INFORMATION 1.1.1 The lichen 1.1.2 Identification of lichens substances by chemical methods 1.1.2.1 Spot test 1.1.2.2 Thin layer chromatography method (TLC) 1.1.2.3 Microcrystallographic test 1.1.3 Biosynthetic pathways to lichen secondary metabolites 1.2 GENERAL DESCRIPTION OF LOBARIA ORIENTALIS AND DENDRISCOSTICTA PLATYPHYLLOIDES (LOBARIACEAE FAMILY).6 1.2.1 Lobaria orientalis (Asahina) Yoshim 1.2.2 Dendriscosticta platyphylloides (Nyl.) B Moncada & Lücking comb nov.10 1.3 CHEMICAL STUDIES ON THE GENUS LOBARIA 10 1.3.1 N-Containing compounds 10 1.3.2 Carbohydrate compounds 10 1.3.3 Monoaromatic compounds 11 1.3.4 Terphenylquinone compounds 11 1.3.5 Dibenzofuran compounds 11 1.3.6 Depside compounds 11 1.3.7 Depsidone compounds 12 1.3.8 Sesterterpenoid compounds 13 iii 1.3.9 Triterpenoid compounds 13 1.3.10 Steroid compounds 14 1.4 CHEMICAL STUDIES ON THE GENUS DENDRISCOSTICTA 14 1.4.1 N-Containing compounds 14 1.4.2 Triterpenoid compounds 15 1.4.3 Steroid compounds 15 1.5 PHARMACOLOGICAL AND CLINIC PROPERTY 15 1.5.1 Lobaria genus 15 1.5.1.1 Traditional medical use 15 1.5.1.2 Biological activity of Lobaria genus 16 1.5.2 Sticta genus 16 CHAPTER 2: EXPERIMENTS 19 2.1 MATERIALS AND EQUIPMENTS 19 2.2 LICHEN MATERIAL 20 2.3 EXTRACTION AND ISOLATION PROCEDURES 20 2.3.1 Extraction and isolation of compounds from Lobaria orientalis (Asahina) Yoshim 20 2.3.1.1 Isolating compounds from the ethyl acetate EA1 fraction 21 2.3.1.2 Isolating compounds from the ethyl acetate EA2 fraction 21 2.3.1.3 Isolating compounds from the ethyl acetate EA3 fraction 22 2.3.2 Extraction and isolation of compounds from Dendriscosticta platyphylloides (Nyl.) 22 2.3.2.1 Isolating compounds from the ethyl acetate EAS1 fraction 23 2.3.2.2 Isolating compounds from the ethyl acetate EAS2 fraction 23 2.3.2.3 Isolating compounds from the ethyl acetate EAS3 fraction 23 2.4 CYTOTOXIC ASSAY 25 CHAPTER 3: RESULTS AND DISCUSSIONS 29 3.1 MONOCYCLIC COMPOUNDS 29 3.1.1 Methyl orselinate (LOBA 1) 30 iv 3.1.2 Methyl β-orcinolcarboxylate (LOBA 9) 31 3.1.3 Orsellinic acid (LOBA 29) 31 3.1.4 Methyl 2,4-dimethoxy-5-formyl-6-methylbenzoate (LOBA 31) 32 3.1.5 Methyl 2,4-dihydroxy-3,5,6-trimethylbenzoate (STIC 9) 33 3.1.6 Orcinol (LOBA 13) 34 3.1.7 5-(Hydroxymethyl)-2-methoxy-1,3-benzenediol (LOBA 33) 35 3.1.8 Resorcinol (LOBA 34) 36 3.1.9 Methyl caffeate (STIC 5) 37 3.1.10 (+)-D-Montagnetol (STIC 14) 37 3.2 DEPSIDE COMPOUNDS 39 3.2.1 Lecanorin (LOBA 14) 39 3.2.2 Isolecanoric acid (LOBA 21) 40 3.3 DEPSIDONE COMPOUNDS 42 3.3.1 1,10-Di-O-methylstictic acid (LOBA 7) 42 3.3.2 Lobarientalone A (LOBA 8) 43 3.3.3 Lobarientalone B (LOBA 28) 46 3.4 DIPHENYL ETHER COMPOUNDS 48 3.4.1 Lobariether A (LOBA 2) 49 3.4.2 Lobariether B (LOBA 12) 51 3.4.3 Lobariether C (LOBA 6) 52 3.4.4 Lobariether D (LOBA 23) 54 3.4.5 Lobariether E (LOBA 26) 55 3.5 STEROID COMPOUNDS 56 3.5.1 Ergosterol-5α,8α-peroxide (LOBA 5) 57 3.5.2 Cerevisterol (LOBA 11) 58 3.5.3 Sitosterol 3-O-β-D-glucopyranoside (STIC 21) 60 3.6 TRITERPENOID COMPOUNDS 61 3.6.1 Zeorin (LOBA 19) 61 3.6.2 16β-Acetoxyhopane-6α,22-diol (LOBA 4) 62 v 3.6.3 15α-Acetoxyhopan-22-ol (STIC 22) 63 3.6.4 Hopane-15α, 22-diol (LOBA 15) 64 3.6.5 Retigeric acid A (LOBA 22) 65 3.6.6 Retigeric acid B (LOBA 25) 67 3.7 N-CONTAINING COMPOUNDS 68 3.7.1 Evodiamine (STIC 15) 68 3.7.2 Rutaecarpine (STIC 29) 69 3.8 BIOLOGICAL ACTIVITY OF ISOLATED COMPOUNDS 70 CHAPTER 4: CONCLUSIONS 73 4.1 CHEMICAL CONSTITUENTS 73 4.2 BIOLOGICAL ASSAY 77 FUTURE OUTLOOK LIST OF PUBLICATIONS REFERENCES APPENDICES vi LIST OF ABBREVIATIONS 1D One dimensional 2D Two dimensional Ac Acetone AcOH Acetic acid br Broad C Chloroform calcd Calculated CC Column chromatography CD Circular dichroism COSY Correlation spectroscopy d Doublet dd Doublet of doublets DMSO Dimethyl sulfoxide EA Ethyl acetate ECD Electronic circular dichroism EI-MS Electron-impact ionization mass spectrum EtOH Ethanol EtOAc Ethyl acetate H n-Hexane HMBC Heteronuclear multiple bond correlation spectroscopy HR-ESI-MS High resolution electrospray ionization mass spectrometry HSQC Heteronuclear single quantum correlation spectroscopy IR Infrared spectrophotometry IC50 The half maximal inhibitory concentration m Multiplet MeOH Methanol vii Minutes MS Mass spectrum NMR Nuclear magnetic resonance NOESY Nuclear overhauser enhancement spectroscopy ppm Parts per million (chemical shift value) RP Reversed phase Prep-TLC Preparative thin-layer chromatography q Quartet s Singlet t Triplet TLC Thin-layer chromatography TMS Tetramethylsilane UV Ultraviolet viii Appendix 23a: 1H-NMR spectrum of STIC 21 (DMSO–d6) 21 20 19 OH O HOHO O 10 11 13 22 18 23 24 15 OH Appendix 23b: 1H-NMR spectrum of STIC 21 (DMSO–d6) (expanded) 26 25 17 29 28 27 21 20 19 OH O HOHO O 10 18 23 24 26 25 17 13 11 29 28 22 27 15 OH Appendix 23c: 13C-NMR spectrum of STIC 21 (DMSO–d6) 19 28 23 24 OH 30 22 16 10 17 14 18 13 26 25 29 21 12 11 20 15 OCOCH3 27 OH Appendix 24a: 1H-NMR spectrum of LOBA (Chloroform–d) 19 28 23 17 24 OH 16 10 22 30 14 18 13 26 25 29 21 12 11 20 15 OCOCH3 27 OH Appendix 24b: 13C-NMR spectrum of LOBA (Chloroform–d) Appendix 25a: 1H-NMR spectrum of LOBA 19 (DMSO–d6) 19 20 28 11 26 25 10 23 OH 22 17 13 30 16 18 14 29 21 12 24 15 27 OH Appendix 25b: 13C-NMR spectrum of LOBA 19 (DMSO–d6) 20 28 25 26 10 23 18 14 16 29 21 12 22 OH 30 27 OH 24 Appendix 26a: 1H-NMR spectrum of LOBA 15 (Chloroform–d) 20 28 26 25 23 18 14 16 10 29 21 12 22 OH 30 27 OH 24 Appendix 26b: 13C-NMR spectrum of LOBA 15 (Chloroform–d) 28 25 19 21 13 11 22 29 OH 17 26 30 15 10 23 27 OCOCH3 24 Appendix 27a: 1H-NMR spectrum of STIC 22 (Chloroform–d) 28 25 19 21 13 11 29 22 OH 17 26 30 15 10 23 27 OCOCH3 24 Appendix 27b: 13C-NMR spectrum of STIC 22 (Chloroform–d) 28 25 19 21 13 11 22 29 OH 17 26 30 15 10 23 27 OCOCH3 24 Appendix 27c: HMBC spectrum of STIC 22 (Chloroform–d) Appendix 28a: 1H-NMR spectrum of LOBA 22 (Chloroform–d) Appendix 28b: 1H-NMR spectrum of LOBA 22 (DMSO – d6) (expanded) Appendix 28c: 13C-NMR spectrum of LOBA 22 (DMSO – d6) 20 19 29 26 12 25 HO 10 HO 17 14 21 22 18 13 11 HOOC 28 30 16 15 27 23 24 COOH Appendix 29a: 1H-NMR spectrum of LOBA 25 (DMSO – d6) 20 19 29 26 12 25 11 HOOC HO 10 HO 21 22 18 13 17 14 16 30 15 27 28 23 COOH 24 Appendix 29b: 1H-NMR spectrum of LOBA 25 (DMSO – d6) (expanded) 20 19 29 26 12 25 HO 10 HO 17 14 21 22 18 13 11 HOOC 28 30 16 15 27 23 24 COOH Appendix 29c: 13C-NMR spectrum of LOBA 25 (DMSO – d6) Appendix 30a: HR-ESI-MS spectrometry of STIC 15 Appendix 30b: 1H-NMR spectrum of STIC 15 (DMSO – d6) O 4a 1a 14 N N 8a CH 13a 13b 8b 13 HN 15 CH3 10 12a 12 11 Appendix 30c: 13C-NMR spectrum of STIC 15 (DMSO – d6) O 4a 1a 14 N N 8a CH 13a 13b 8b 13 15 CH3 HN 12a 12 10 11 Appendix 30d: COSY spectrum of STIC 15 (DMSO – d6) O 1a N 4a 8a CH 13a 14 13b N 8b 13 HN 15 CH3 10 12a 11 12 Appendix 30e: HSQC spectrum of STIC 15 (DMSO – d6) O 4a 1a 14 N N 8a CH 13a 13b 8b 13 15 CH3 HN 12a 12 10 11 Appendix 30f: HMBC spectrum of STIC 15 (DMSO – d6) O 4a 1a 14 N N 8a CH 13a 13b 8b 13 15 CH3 HN 12a 12 10 11 Appendix 30g: HMBC spectrum of STIC 15 (DMSO – d6) (expanded) Appendix 31a: ESI MS spectrometry of STIC 29 O 4a 1a 14 N N C 13b 8a 13a 8b 13 HN 12a 12 Appendix 31b: 1H-NMR spectrum of STIC 29 (DMSO – d6) O 4a N 1a C 14 N 13b 8a 13a 8b 13 HN 12a 12 10 11 Appendix 31c: 13C-NMR spectrum of STIC 29 (DMSO – d6) 10 11 O 4a N 1a C 14 N 13b 8a 13a 8b 13 HN 10 12a 12 11 Appendix 31d: HSQC spectrum of STIC 29 (DMSO – d6) O 4a 1a 14 N N C 13b 8a 13a 8b 13 HN 12a 12 10 11 Appendix 31e: HMBC spectrum of STIC 29 (DMSO – d6) ... NATIONAL UNIVERSITY – HO CHI MINH CITY UNIVERSITY OF SCIENCE NGUYEN THI MY DUNG STUDY ON CHEMICAL CONSTITUENTS OF TWO LICHEN SPECIES LOBARIA ORIENTALIS AND DENDRISCOSTICTA PLATYPHYLLOIDES (LOBARIACEAE). .. secondary metabolites 1.2 GENERAL DESCRIPTION OF LOBARIA ORIENTALIS AND DENDRISCOSTICTA PLATYPHYLLOIDES (LOBARIACEAE FAMILY).6 1.2.1 Lobaria orientalis (Asahina) Yoshim 1.2.2 Dendriscosticta. .. pathways: acetate–polymalonate pathway, shikimic acid pathway and mevalonic acid pathway [68] (Figure 1.4) 1.2 GENERAL DESCRIPTION OF LOBARIA ORIENTALIS AND DENDRISCOSTICTA PLATYPHYLLOIDES (LOBARIACEAE