CYCLOPEPTIDE ALKALOIDS FROM SOME ZIZIPHUS PLANTS A THESIS BY NATTHAKALN LOMCHOEY Presented in Partial Fulfillment of the Requirements for the Master of Science Degree in Chemistry at Srinakharinwirot University May 2011 CYCLOPEPTIDE ALKALOIDS FROM SOME ZIZIPHUS PLANTS A THESIS BY NATTHAKALN LOMCHOEY Presented in Partial Fulfillment of the Requirements for the Master of Science Degree in Chemistry at Srinakharinwirot University May 2011 Copyright 2011 by Srinakharinwirot University CYCLOPEPTIDE ALKALOIDS FROM SOME ZIZIPHUS PLANTS AN ABSTRACT BY NATTHAKALN LOMCHOEY Presented in Partial Fulfillment of the Requirements for the Master of Science Degree in Chemistry at Srinakharinwirot University May 2011 Natthakaln Lomchoey. (2011). Cyclopeptide alkaloids from some ziziphus plants. Thesis, M.Sc. (Chemistry). Bangkok: Graduate School, Srinakharinwirot University. Advisor Committee: Assoc. Prof. Dr. Sunit Suksamrarn, Dr. Prasert Pattanaprateeb. Phytochemical studies have established the genus Ziziphus (Rhamnaceae family) to be a rich source of cyclopeptides. Investigation of selected Thai Ziziphus plants (Z. cambodiana root bark and Z. mauritiana stem bark), led to isolation of two new cyclopeptide alkaloids as 5(14)-scutianine A-type cyclopeptide alkaloid (cambodine) and 5(13)-zizyphine A-type cyclopeptide alkaloid (mauritine M), from Z. cambodiana and Z. mauritiana, respectively. Moreover, three known cyclopeptides, frangufoline and lotusanine B, as 4(14)-membered cyclopeptides were isolated from Z. cambodiana, and nummularine H as 5(13)-cyclopeptide was isolated from Z. mauritiana. Their structures and relative stereochemistry of the new compounds were elucidated mainly on the basis of extensive NMR and mass spectroscopic analysis. The stereochemical assignments were established by their CD spectra analysis 2D NMR experiments and by comparison with other related compounds of known stereochemistry. ไซโคลเปปไทดอัลคาลอยดจากพืช ZIZIPHUS บางชนิด บทคัดยอ ของ ณัฏฐกัลย ลมเชย เสนอตอบัณฑิตวทยาลิ ัย มหาวิทยาลัยศรีนครินทรวิโรฒ เพื่อเปนสวนหนึ่งของการศึกษา ตามหลักสูตรวิทยาศาสตรมหาบัณฑิต สาขาวิชาเคม ี พฤษภาคม 2554 ณัฏฐกัลย ลมเชย. (2554). ไซโคลเปปไทดอัลคาลอยดจากพืช ZIZIPHUS บางชนิด. ปริญญานิพนธ วท.ม. (เคมี). กรุงเทพฯ: บัณฑิตวิทยาลัย มหาวิทยาลัยศรีนครินทรวิโรฒ. คณะกรรมการควบคุม: รองศาสตราจารย ดร.สุนิตย สุขสําราญ, ดร. ประเสริฐ พัฒนา- ประทีป. จากการศึกษาองคประกอบทางเคมีจากเปลือกรากตะครองและเปลือกตนพุทรา สามารถ แยกสารไซโคลเปบไทดอัลคาลอยดชนิดใหมได 2 สาร คือ cambodine จากเปลือกรากตะครอง และ mauritine M จากเปลือกตนพุทรา นอกจากนี้ยังพบสารประกอบไซโคลเปบไทดอัลคาลอยดที่เคย พบแลว 3 สาร ไดแก frangufoline และ lotusanine B จากตะครอง และ nummularine H จากพุทรา โครงสรางของสารทราบไดจากการวิเคราะหขอมูลสเปกโทรสโคป โดยใชเทคนิค NMR และ แมสสเปกโทรสโคปเปนสวนใหญ สําหรับการหาสเตอริโอเคมีของสารใชเทคนิค CD, 2D NMR และ ทําการเปรียบเทียบขอมูลกับสารประกอบอื่นที่มีโครงสรางใกลเคียงกัน The thesis title “Cyclopeptide alkaloids from some Ziziphus plants” by Natthakaln Lomchoey has been approved by the Graduate School as partial fulfillment of the requirements for the Master of Science Degree in Chemistry of Srinakharinwirot University. …………………………………………… Dean of Graduate School (Assoc. Prof. Dr. Somchai Santiwattanakul) May 27, 2011 Thesis Committee Oral Defense Committee .................................................. Major advisor ................................................. Chair (Assoc. Prof. Dr. Sunit Suksamrarn) (Assoc. Prof. Dr. Thanattkhun Mongkolaussavaratana) .................................................. Co-advisor ................................................. Committee (Dr. Prasert Pattanaprateeb) (Assoc. Prof. Dr. Sunit Suksamrarn) ................................................. Committee (Asst. Prof. Dr. Siritron Samosorn) ................................................. Committee (Dr. Prasert Pattanaprateeb) Acknowledgements The first and foremost, I would like to express my sincere gratitude to my supervisor, Assoc. Prof. Dr. Sunit Suksamrarn, for her kind and helpful supervision, hearty encouragement and research assistantship support throughout this work. I would like to thank all my committee members, Dr. Prasert Pattanaprateeb my co-advisors, Asst. Prof. Dr. Siritron Samosorn, for their endless kindness, thoughtful advice, valuable time, patient reading and warm encouragement. In addition, I feel grateful to Assoc. Prof. Dr. Thanattkhun Mongkolaussavaratana, Chulabhorn Graduate Institute, for his useful comments and encouragement. This work was supported by the Center of Excellence for Innovation in Chemistry (PERCH-CIC). I am indebted to Mr. Wicharn Wisetsri and Mr. Jessada Netsawangwicha for collections of the plant materials. I am grateful to Mr. Pichit Sudta and Mr. Payung Jiarawapi, Department of Chemistry, Faculty of Science, Srinakharinwirot University for recording the nuclear magnetic resonance spectra, Mr. Nitirat Chimnoi, Chulabhorn Research Institute for recording the HRMS, Mr. Samran Prabpai and Assoc. Prof. Dr. Palangpon Kongsaeree, Department of Chemistry, Faculty of Science, Mahidol University for X-ray diffraction data, and to Asst. Prof. Dr. Boon-Ek Yingyongnarongkul, Mr. Widchaya Radchatawedchakoon and Miss Ratchanaporn Chikchaisiri, Department of Chemistry, Faculty of Science, Ramkhamhaeng University for recording the ESIMS and optical rotations. Many special thanks also go to my teachers, friends, colleagues and staff of the Department of Chemistry, Faculty of Science, Srinakharinwirot University for their friendship, kind support and encouragement. Finally, I wish to express my profound gratitude to my parents and family for their love, unconditional support and encouragement throughout my hold life. Natthakaln Lomchoey TABLE OF CONTENTS Chapter Page 1 INTRODUCTION …………………………………………………………………….. 1 Background ...................................................................................................... 1 Ethnopharmacological uses ............................................................................. 1 Objectives of the study ..................................................................................... 2 2 REVIEW OF LITERATURE ………………………………………………………… 3 Sources of cyclopeptide alkaloids .................................................................... 3 Classification of cyclopeptide alkaloids .......................................................... 3 Nomenclature of cyclopeptide alkaloids .......................................................... 4 Types of cyclopeptide alkaloids ....................................................................... 5 Literature reviews of Ziziphus cambodiana Pierre ........................................... 12 Literature reviews of cyclopeptides from Ziziphus mauritiana Lam. …….…… 17 3 EXPERIMENTAL …………………………………………………………………….. 19 Sources of plant materials ............................................................................... 19 General techniques .......................................................................................... 19 Physical properties ........................................................................................... 20 Spectroscopy .................................................................................................... 21 Extraction and isolation procedures ................................................................. 21 Physical and spectral data of the isolated compounds from the root bark of Z. cambodiana .………………………………………………………..……….. 28 Physical and spectral data of the isolated compounds from the stem bark of Z. mauritiana ……………………………………………………………............ 29 TABLE OF CONTENTS (continued) Chapter Page 4 RESULTS AND DISCUSSION ……………………………………………………… 31 Cyclopeptide alkaloids from the MeOH extract of Z. cambodiana root bark ... 31 Structure determination of compound ZC-M1 (frangufoline, sss4449) .…. 32 Structure determination of compound ZC-M2 (lotusanine B, sss4635) .... 39 Structure determination of compound ZC-M3 (Cambodine, sss4402) .….. 46 Cyclopeptide alkaloids from the MeOH extract of Z. mauritiana stem bark .... 51 Structure determination of compound ZM-M1 (Nummularine H, sss4971).. 52 Structure determination of compound ZM-M2 (Mauritine M, sss4973) .… 57 5 CONCLUSION .………………………………………………………………………. 64 BIBLIOGRAPHY ............................................................................................................. 65 APPENDIX ..................................................................................................................... 70 GLOSSARY ................................................................................................................... 77 CURRICULUM VITAE ................................................................................................... 83 LIST OF TABLES Table Page 1 Nomenclature of cyclopeptide alkaloids .…….…………………………………… 5 2 IC50 values for antiplasmodial and MIC values for antimycobacterial activities of triterpenes 1-39 ............................................................................................. 13 3 IC50 values for neuraminidase inhibitory activity of flavonoid glycosides 40-42 .……………………………………............................................................ 14 4 IC50 values (µM) of triterpenes 34, 36-37, 39 and 43 with GLI1-mediated transcriptional inhibitory activity, and cytotoxicity against PANC1, DU145, and C3H10T1/2 cells .……………..…………………………………..…………………. 15 5 1H, 13C NMR and 2D NMR data of compound ZC-M1 (sss4449) in CDCl3 .…………………………...…………………………………………………… 34 6 Comparison of 1H and 13C NMR data of compound ZC-M1