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Purification and structure analysis of mammosphere-forming inhibitors derived from Cynanchum auriculatum

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Author(s)
등홍원
Issued Date
2017
URI
http://dcoll.jejunu.ac.kr/jsp/common/DcLoOrgPer.jsp?sItemId=000000007919
Abstract
Many studies have supported the existence of cancer stem cells (CSCs) as a subpopulation of cancer cells. CSCs are drug-resistant and radiation-resistant cancer cells to be responsible for tumor progress, maintenance and recurrence of cancer, and metastasis. The breast CSCs inhibitor that inhibits cancer stem cell derived from MCF-7 cells was isolated from the root parts of C. auriculatum. The ethyl acetate fraction obtained from methanol extracts exhibited the outstanding inhibitory effect on mammospohere formation. From the ethyl acetate fraction, a new compound named DH was purified through Silica gel column chromatography, ODS open column chromatography, Sephadex LH-20 column chromatography, and preparative thin-layer chromatography. DH-1 and DH-2 were purified using reversed-phase high performance liquid chromatography. Compound DH-1 and DH-2 are steroidal glycoside called kidjoranin-3-O-β-D-cymaropyranoside and Caudatin-3-O-β-D-cymaropyranoside. We purified CSCs inhibitor from C. auriculatum. Thus, the isolated compounds exhibited a strong anti-CSCs activity, which may contribute to the interpretation of the pre-pharmacological and clinical effects of C. auriculatum.
Department
대학원 차세대융복합과학기술협동과정
Advisor
이동선
Awarded Date
2017. 2
Table Of Contents
CONTENTS i
LIST OF ABBREVIATIONS iii
LIST OF FIGURES iv
LIST OF TABLES vi
ABSTRACT 1
INTRODUCTION 2
MATERIALS AND METHODS 5
1. General Experimental Procedures 5
2. Plant Material 6
3. Extraction and characterization of DH-1 and DH-2 from C. auriculatum 6
3.1. Preparation of mehanolic crude extracts 8
3.2. Fractionation of the methanolic extracts by means of liquid-liquid partitioning 9
3.3. Isolation and purification of active substances in the bioactive ethyl acetate fraction by bioassay-guided fractionation 9
3.4. Identification of active components by Preparative TLC using UV254 nm fluorescence detection 11
3.5. HPLC analysis 12
4. Mammosphere formation assay 12
5. Structure analysis of purified DH-1 and DH-2 13
RESULTS 16
1. Purification of active substances from C. auriculatum 16
1.1. The anti-mammosphere-forming activity of solvent-partitioning fractions 16
1.2. The anti-mammosphere-forming activity of fractions obtained from Ethyl acetate aqueous fraction 16
1.3. Open ODS column chromatography of the anti-mammosphere-forming activity using the F2 compound 20
1.4. Separation of the anti-mammosphere-forming activity compound from 30% and 50% acetonitrile elutes using Sephadex LH-20 column 22
1.5. Isolation of the anti- mammosphere-forming activity compound from compounds separted using Preparative TLC 24
1.6. Purification of active substance in partial purified sample using Preparative HPLC 27
2. Structure analysis of DH-1 and DH-2 using 1-,2-NMR, and Mass spectrophotomter 44
DISCUSSION 45
CONCLUSION 47
REFERENCS 49
ACKNOWLEDGEMENTS 53
Degree
Master
Publisher
제주대학교 일반대학원
Citation
등홍원. (2017). Purification and structure analysis of mammosphere-forming inhibitors derived from Cynanchum auriculatum
Type
Dissertation
Appears in Collections:
General Graduate School > Interdisciplinary Graduate Program in Advanced Convergence Technology and Science
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