MINISTRY OF EDUCATION AND TRAINING MINISTRY OF HEALTH
INSTITUTE OF MEDICINAL MATERIALS
Maybe you are interested!
-
Study on botanical characteristics, chemical composition and acetylcholinesterase enzyme inhibitory effects of two species Piper thomsonii (C. DC.) Hook. f. var. thomsonii and Piper hymenophyllum Miq., Piperaceae family - 24 -
Study on chemical composition, toxicity and some biological effects supporting the treatment of gastric and duodenal ulcers of Sanchezia nobilis Hook.F. leaves - 2 -
Study on botanical characteristics, chemical composition and acetylcholinesterase enzyme inhibitory effects of two species Piper thomsonii (C. DC.) Hook. f. var. thomsonii and Piper hymenophyllum Miq., Piperaceae family - 23 -
Study on botanical characteristics, chemical composition and acetylcholinesterase enzyme inhibitory effects of two species Piper thomsonii (C. DC.) Hook. f. var. thomsonii and Piper hymenophyllum Miq., Piperaceae family - 21 -
Study on botanical characteristics, chemical composition and acetylcholinesterase enzyme inhibitory effects of two species Piper thomsonii (C. DC.) Hook. f. var. thomsonii and Piper hymenophyllum Miq., Piperaceae family - 27
TRAN THI THU HIEN
RESEARCH ON CHEMICAL COMPOSITION AND EVALUATION OF ANTI-CANCER EFFECTS OF THE STEMS AND LEAVES OF THE CƯƠM PLANT
( Stephania dielsiana YC Wu)
DOCTORAL THESIS IN PHARMACY
HANOI, 2022
MINISTRY OF EDUCATION AND TRAINING MINISTRY OF HEALTH
INSTITUTE OF MEDICINAL MATERIALS
TRAN THI THU HIEN
RESEARCH ON CHEMICAL COMPOSITION AND EVALUATION OF ANTI-CANCER EFFECTS OF THE STEMS AND LEAVES OF THE CƯƠM PLANT
( Stephania dielsiana YC Wu)
DOCTORAL THESIS IN PHARMACY
MAJORITY: MEDICINAL HERBS - TRADITIONAL PHARMACY CODE: 9720206
Scientific supervisor: 1. Dr. Le Thi Kim Van
2. Associate Professor, Dr. Nguyen Quoc Huy
HANOI, 2022
COMMITMENT
I hereby declare that this is my own research work under the guidance of Dr. Le Thi Kim Van and Associate Professor Dr. Nguyen Quoc Huy. The data and results presented in this thesis are honest and have never been published in any other research work.
Author
Tran Thi Thu Hien
ACKNOWLEDGEMENTS
During the process of researching and completing this thesis, I received a lot of valuable help from teachers, scientists in many fields, colleagues, friends and family.
First of all, I would like to express my respect and deep gratitude to Dr. Le Thi Kim Van and Associate Professor Dr. Nguyen Quoc Huy for their direct guidance and wholehearted instruction throughout my study and scientific research.
I would like to sincerely thank the Board of Directors, Faculties, Departments, teachers and colleagues at the Institute of Medicinal Materials; the Experimental Oncology Research Group, Department of Cell Biology, Faculty of Biology, University of Science, Vietnam National University, Hanoi; Vinmec Stem Cell and Gene Technology Research Institute, Institute of Marine Biochemistry under the Vietnam Academy of Science and Technology for helping and creating favorable conditions to help me throughout the research process.
I would like to express my sincere thanks to Associate Professor Dr. Nguyen Thuong Dong, Associate Professor Dr. Phan Minh Giang, Associate Professor Dr. Hoang Viet Dung, Associate Professor Dr. Bui Thanh Tung, Dr. Bui Huu Tai, Dr. Nguyen Van Tai, Dr. Le Thanh Nghi, Dr. Le Thi Xoan, Dr. Nguyen Thi Ha, Dr. Nguyen Tuan Hiep for their valuable contributions to help me in the process of experimental research and thesis completion.
I would like to sincerely thank the leaders of the Vietnam Academy of Traditional Medicine and my colleagues at the Vietnam Academy of Traditional Medicine, where I work, for their encouragement and favorable conditions for me to complete this thesis.
Finally, I would like to express my deepest gratitude to my beloved family members; thank you to my close friends who have given me their love, encouragement and help over the past time.
This thesis is a part of the research of the Ministry of Health-level science and technology task managed by the Department of Science, Technology and Training - Ministry of Health (according to Decision No. 2721/QD-BYT signed on June 28, 2019 and Contract No. 09/HD-K2DT signed on September 18, 2019).
Thank you very much for all this valuable help!
INDEX
LIST OF ABBREVIATIONS LIST OF TABLES
LIST OF IMAGES
PROBLEM STATEMENT 1
CHAPTER 1. OVERVIEW 3
1.1. OVERVIEW OF PLANTS 3
1.1.1. Classification position 3
1.1.2. Botanical characteristics of the tuber species 3
1.1.3. Distribution of the species of tuber 4
1.2. CHEMICAL COMPOSITION OF THE PLANT CÚDÔM 6
1.2.1. Alkaloid 6
1.2.2. Other groups of compounds 11
1.3. BIOLOGICAL EFFECTS, USES AND TOXICITY OF DOM 13
1.3.1. Biological effects 13
1.3.2. Toxicity of the tuber of the plant 21
1.3.3. Uses 22
1.4. MOLECULAR TARGETS IN CANCER DRUG DEVELOPMENT
................................................................ ................................................................ ................................ 23
1.4.1. Overview of some molecular targets in cancer drug research and development 23
1.4.2. Aurora kinase and its role in cancer 26
CHAPTER 2. RESEARCH SUBJECTS AND METHODS 37
2.1. SUBJECT OF RESEARCH 37
2.1.1. Research materials 37
2.1.2. Some experimental cancer cell lines 37
2.1.3. Chemicals, solvents 38
2.1.4. Research machinery, equipment and tools 39
2.2. RESEARCH LOCATION 40
2.3. RESEARCH CONTENT 41
2.4. RESEARCH METHODS 41
2.4.1. Extraction, isolation and structure determination of some compounds from the stem and leaves of the plant 41
2.4.2. Initial research on the development of isolation methods and quantitative methods to monitor oxostephanin content in medicinal herbs according to harvesting time 44
2.4.3. Evaluation of cytotoxic effects of some isolated compounds and initial study of anticancer mechanism of oxostephanin 48
CHAPTER 3. RESEARCH RESULTS 61
3.1. EXTRACTION, ISOLATION AND STRUCTURAL DETERMINATION OF SOME COMPOUNDS FROM THE STEMS AND LEAVES OF THE PLANT CƯƠM 61
3.1.1. Extraction and isolation of some compounds from the stem and leaves of the plant 61
3.1.2. Determination of chemical structures of isolated compounds 62
3.2. INITIAL RESEARCH ON DEVELOPING ISOLATION METHODS AND QUANTITATIVE METHODS TO MONITOR OXOSTEPHANIN CONTENT IN MEDICINAL HERBS ACCORDING TO HARVESTING TIME 84
3.2.1. Isolation and preliminary evaluation of the purity of oxostephanin 84
3.2.2. Development and validation of a method for quantifying oxostephanin in the stems and leaves of the plant Dom 88
3.2.3. Evaluation of changes in oxostephanin content according to harvesting time. 98
3.3. EVALUATION OF THE CELLULAR TOXIC EFFECTS OF SOME ISOLATED COMPOUNDS AND INITIAL RESEARCH ON THE ANTI-CANCER MECHANISM OF OXOSTEPHANIN 99
3.3.1. Evaluation of cytotoxic effects of some isolated compounds 99
3.3.2. Study on the cytotoxic mechanism of oxostephanin 105
CHAPTER 4. DISCUSSION 123
4.1. ON EXTRACTION, ISOLATION AND STRUCTURAL DETERMINATION OF COMPOUNDS FROM THE STEMS AND LEAVES OF THE PLANT CƯƠM 123
4.2. ON THE INITIAL RESEARCH ON THE DEVELOPMENT OF ISOLATION METHODS AND QUANTITATIVE METHODS TO MONITOR OXOSTEPHANIN CONTENT IN MEDICINAL HERBS ACCORDING TO HARVESTING TIME 130
4.2.1. On the isolation and preliminary assessment of the purity of oxostephanin 130
4.2.2. On the construction and evaluation of quantitative methods 133
4.2.3. On the change in oxostephanin content according to harvesting time 135
4.3. ON THE EVALUATION OF THE CELLULAR TOXIC EFFECTS OF SOME ISOLATED COMPOUNDS AND THE INITIAL RESEARCH ON THE ANTI-CANCER MECHANISM OF OXOSTEPHANIN 138
4.3.1. Cytotoxic effects of some isolated compounds 138
4.3.2. Mechanism of cytotoxic action of oxostephanin 142
4.4. ON NEW CONTRIBUTIONS OF THE THESIS 146
CONCLUSION AND RECOMMENDATIONS 148
LIST OF PUBLISHED ARTICLES REFERENCES
LIST OF ABBREVIATIONS
Abbreviations English Vietnamese
1 H-NMR Proton Nuclear Magnetic Resonance Spectroscopy
13 C-NMR Carbon-13 Nuclear Magnetic
Resonance Spectroscopy
Proton nuclear magnetic resonance spectroscopy
Carbon 13 nuclear magnetic resonance spectroscopy
[α] D Specific rotation angle
AchE Acetycholinesterase
BchE Butyrylcholinesterase
BuOH Butanol
cDNA Complementary Deoxyribonucleic Acide
Deoxyribonucleic acid supplement
CFU Colony‐forming units CFU-EC Colony Units of Endothelial Cells
endothelial cells
CFU-F Colony Units of Fibroblasts
fibroblast
CI Cell Index Cell Index
COSY 1H–1H Correlation Spectroscopy 1 H- 1 H Correlation Spectroscopy DD Solution
Vietnamese Pharmacopoeia
DEPT Distortionless Enhancement by Polarization Transfer
DMEM Dulbecco's Modified Eagle Medium
DMSO Dimethyl sulfoxide (CH₃)₂SO
EBM Eagle's Basal Medium
EDTA Ethylene Diamine Tetraacetic Acide
ESI-MS Electrospray Ionization - Mass Spectrometry
EtOAc Ethyl acetate
DEPT spectrum
Electron spray ionization mass spectrometry
EtOH Ethanol
FBS Fetal Bovine Serum
FGF-2 Fibroblast Growth Factor‐2 Fibroblast Growth Factor
fiber-2
H358 Epithelial Cancer Cell Line
alveolar bronchial
HeLa Human cervical carcinoma Cervical cancer cells
People
HepG2 Human hepatocellular carcinoma hFBs Human dermal fibroblasts
People
HGF Hepatocyte growth factor Hepatocyte growth factor
HMBC Heteronuclear Multiple Bond Connectivity
HPLC High Performance Liquid Chromatography
HR-ESI-MS High-Resolution Electron Spray
Ionization Mass Spectrometry
HSQC Heteronuclear Single Quantum Correlation Spectroscopy
hUVECs Human Umbilical Vein Endothelial Cells
Multiple bond heteronuclear correlation spectrum
High performance liquid chromatography
High resolution electrospray ionization mass spectrometry
Heteronuclear interaction spectrum across a bond
Human umbilical vein endothelial cells
KHV Microscope
Molecular Weight
IC 50 Half-maximal inhibitory concentration
Maximum inhibitory concentration 50%
J Coherence constant (unit is Hz)
LD 50 Median Lethal Dose 50% Lethal Dose
MCF7 Human breast carcinoma Adenocarcinoma cells
multidrug resistant breast
MDA-MB-231 Hormone-independent breast
cancer cell line
MDA Hormone-independent breast cancer
Hormone-independent breast cancer cell lines
Hormone-independent breast cancer