臺灣博碩士論文加值系統

摘要
Combretastatin A-4 (簡稱CA-4)由南非一種柳樹Combretum caffrum的樹皮中分離出的，其結構屬stilbene類天然物，能夠有效抑制微管蛋白的聚合，及對抗人類不同器官之癌細胞顯示有很強的細胞毒殺活性，同時也具有破壞新生血管的活性。
由先前的文獻報導，CA-4結構上兩個芳香環為順式構型對於顯示較強的抗癌活性是很重要的，因此有許多研究人員在連接兩芳香環之碳碳雙鍵的地方設計一環狀結構來取代其碳碳雙鍵部分，避免順式構型異構化成反式構型而喪失其抗癌活性。
在本論文研究中利用isoxazole環及pyrazole環來替代CA-4的碳碳雙鍵部分，並以鹵素做為取代基，合成出一系列CA-4類似物(TYC526, TYC516, TYC506, TYC517, TYC507, TYC518, TYC508, TYC519, TYC509)，並測試對人類子宮上皮癌細胞株(HeLa)、肝臟癌細胞株(HepG2)、卵巢腺癌細胞株(OVCAR-3)的活性。其中化合物TYC526的活性最強，IC50為3.64 – 3.96 nM。

Abstract

Combretastatin A-4 (CA-4), a naturally occurring stilbene, was isolated from Combretum caffrum (African bush willow). CA-4 strongly inhibited the polymerization of tubulin by binding to the colchicine site and showed potent cytotoxicity against a variety of human cancer cell lines including multiple drug resistant cancer cell lines. CA-4 also showed strong anti-vascular activity.
In the previously comparative studies of the combretastatins it appears that the cis orientation of the two aromatic rings plays an important role of cytotoxicity. Therefore, a number of cis-restricted analogues of CA-4 were prepared using 1,2-substituted five memberd heterocycles to avoid decreasing cytotoxic activity.
In this study, we utilized isoxazole ring and pyrazole ring to mimic the cis double bond in CA-4. A series of halogen substituted 4,5-diarylisoxazoles and 4,5-diarylpyrazoles (TYC526, TYC516, TYC506, TYC517, TYC507, TYC518, TYC508, TYC519, TYC509) were prepared and their cytotoxic activity were also evaluated against human cancer cell lines including human cervical epitheloid carcinoma (HeLa), human hepatocellular carcinoma (HepG2), human ovarian adenocarcinoma (OVCAR-3), and human colon carcinoma (HT29). Among the synthesized compounds, TYC526 was the most potent compound in this series with an IC50 value of 3.64–3.96 nM.

目 錄
中文摘要 ------------------------------------------------------------------- i
英文摘要 ------------------------------------------------------------------- ii
目錄 ------------------------------------------------------------------------- iii
圖表目錄 ------------------------------------------------------------------- iv
壹、緒論 ------------------------------------------------------------------- 1
貳、實驗儀器與藥品試劑----------------------------------------------- 7
參、研究方法-------------------------------------------------------------- 10
肆、實驗步驟 ------------------------------------------------------------ 13
伍、結果與討論 ---------------------------------------------------------- 33
陸、參考文獻 ------------------------------------------------------------- 44
柒、圖表附錄 ------------------------------------------------------------- 48







圖 表 目 錄
Fig. 1 細胞週期圖 --------------------------------------------------------- 2
Fig. 2 利用五環來固定cis的結構 -------------------------------------- 5
Fig. 3 化合物TYC501之1H NMR圖譜 ----------------------------- 49
Fig. 4 化合物TYC501之13C NMR圖譜 ---------------------------- 50
Fig. 5 化合物TYC501之FTIR圖譜 --------------------------------- 51
Fig. 6 化合物TYC501之EI-MS圖譜 --------------------------------- 52
Fig. 7 化合物TYC502之EI-MS圖譜 --------------------------------- 53
Fig. 8 化合物TYC524之1H NMR圖譜 ---------------------------- 54
Fig. 9 化合物TYC524之13C NMR圖譜 --------------------------- 55
Fig. 10 化合物TYC524之FTIR圖譜 -------------------------------- 56
Fig. 11 化合物TYC524之EI-MS圖譜 ------------------------------- 57
Fig. 12 化合物TYC514之1H NMR圖譜 ---------------------------- 58
Fig. 13 化合物TYC514之13C NMR圖譜 --------------------------- 59
Fig. 14 化合物TYC514之FTIR圖譜 -------------------------------- 60
Fig. 15 化合物TYC514之EI-MS圖譜 ------------------------------ 61
Fig. 16 化合物TYC504之1H NMR圖譜 ---------------------------- 62
Fig. 17 化合物TYC504之13C NMR圖譜 --------------------------- 63
Fig. 18 化合物TYC504之FTIR圖譜 -------------------------------- 64
Fig. 19 化合物TYC504之EI-MS圖譜 ------------------------------- 65
Fig. 20 化合物TYC515之1H NMR圖譜 ---------------------------- 66
Fig. 21 化合物TYC515之13C NMR圖譜 --------------------------- 67
Fig. 22 化合物TYC515之FTIR圖譜 --------------------------------- 68
Fig. 23 化合物TYC515之EI-MS圖譜 ------------------------------- 69
Fig. 24 化合物TYC505之1H NMR圖譜 ---------------------------- 70
Fig. 25 化合物TYC505之13C NMR圖譜 --------------------------- 71
Fig. 26 化合物TYC505之FTIR圖譜 --------------------------------- 72
Fig. 27 化合物TYC505之EI-MS圖譜 -------------------------------- 73
Fig. 28 化合物TYC526之1H NMR圖譜 --------------------------- 74
Fig. 29 化合物TYC526之13C NMR圖譜 ---------------------------- 75
Fig. 30 化合物TYC526之COSY圖譜 -------------------------------- 76
Fig. 31 化合物TYC526之HMQC圖譜 ------------------------------- 77
Fig. 32 化合物TYC526之HMBC圖譜 ------------------------------- 78
Fig. 33 化合物TYC526之FTIR圖譜 --------------------------------- 79
Fig. 34 化合物TYC526之EI-MS圖譜 --------------------------------- 80
Fig. 35 化合物TYC516之1H NMR圖譜 ---------------------------- 81
Fig. 36 化合物TYC516之13C NMR圖譜 -------------------------- 82
Fig. 37 化合物TYC516之COSY圖譜 ------------------------------- 83
Fig. 38 化合物TYC516之HMQC圖譜------------------------------ 84
Fig. 39 化合物TYC516之HMBC圖譜 ------------------------------ 85
Fig. 40 化合物TYC516之FTIR圖譜--------------------------------- 86
Fig. 41 化合物TYC516之EI-MS圖譜 ------------------------------- 87
Fig. 42 化合物TYC506之1H NMR圖譜 ---------------------------- 88
Fig. 43 化合物TYC506之13C NMR圖譜 --------------------------- 89
Fig. 44 化合物TYC506之COSY圖譜 ------------------------------- 90
Fig. 45 化合物TYC506之HMQC圖譜 ------------------------------ 91
Fig. 46 化合物TYC506之HMBC圖譜 ------------------------------ 92
Fig. 47 化合物TYC506之FTIR圖譜 -------------------------------- 93
Fig. 48 化合物TYC506之EI-MS圖譜 ------------------------------- 94
Fig. 49 化合物TYC517之1H NMR圖譜 ----------------------------- 95
Fig. 50 化合物TYC517之13C NMR圖譜 ---------------------------- 96
Fig. 51 化合物TYC517之COSY圖譜 ------------------------------ 97
Fig. 52 化合物TYC517之HMQC圖譜 ----------------------------- 98
Fig. 53 化合物TYC517之HMBC圖譜------------------------------ 99
Fig. 54 化合物TYC517之FTIR圖譜 ------------------------------ 100
Fig. 55 化合物TYC517之EI-MS圖譜 ---------------------------- 101
Fig. 56 化合物TYC507之1H NMR圖譜 -------------------------- 102
Fig. 57 化合物TYC507之13CNMR圖譜 --------------------------- 103
Fig. 58 化合物TYC507之COSY圖譜 ----------------------------- 104
Fig. 59 化合物TYC507之HMQC圖譜 ----------------------------- 105
Fig. 60 化合物TYC507之HMBC圖譜 ----------------------------- 106
Fig. 61 化合物TYC507之FTIR圖譜 -------------------------------- 107
Fig. 62 化合物TYC507之EI-MS圖譜 ------------------------------- 108
Fig. 63 化合物TYC518之1H NMR圖譜 ---------------------------- 109
Fig. 64 化合物TYC518之13C NMR圖譜 --------------------------- 110
Fig. 65 化合物TYC518之COSY圖譜 ------------------------------- 111
Fig. 66 化合物TYC518之HMQC圖譜 ------------------------------ 112
Fig. 67 化合物TYC518之HMBC圖譜 ------------------------------ 113
Fig. 68 化合物TYC518之FTIR圖譜 -------------------------------- 114
Fig. 69 化合物TYC518之EI-MS圖譜 ------------------------------- 115
Fig. 70 化合物TYC519之1H NMR圖譜 ---------------------------- 116
Fig. 71 化合物TYC519之13C NMR圖譜 --------------------------- 117
Fig. 72 化合物TYC519之COSY圖譜 ------------------------------- 118
Fig. 73 化合物TYC519之HMQC圖譜 ------------------------------ 119
Fig. 74 化合物TYC519之HMBC圖譜 ----------------------------- 120
Fig. 75 化合物TYC519之FTIR圖譜 -------------------------------- 121
Fig. 76 化合物TYC519之EI-MS圖譜 -------------------------------- 122
Fig. 77 化合物TYC508之1H NMR圖譜 ---------------------------- 123
Fig. 78 化合物TYC508之13C NMR圖譜 --------------------------- 124
Fig. 79 化合物TYC508之COSY圖譜 -------------------------------- 125
Fig. 80 化合物TYC508之HMQC圖譜 ----------------------------- 126
Fig. 81 化合物TYC508之HMBC圖譜 ----------------------------- 127
Fig. 82 化合物TYC508之FTIR圖譜 -------------------------------- 128
Fig. 83 化合物TYC508之EI-MS圖譜 ------------------------------- 129
Fig. 84 化合物TYC509之1H NMR圖譜 ---------------------------- 130
Fig. 85 化合物TYC509之13C NMR圖譜 --------------------------- 131
Fig. 86 化合物TYC509之COSY圖譜 -------------------------------- 132
Fig. 87 化合物TYC509之HMQC圖譜 ----------------------------- 133
Fig. 88 化合物TYC509之HMBC圖譜 ----------------------------- 134
Fig. 89 化合物TYC509之FTIR圖譜 -------------------------------- 135
Fig. 90 化合物TYC509之EI-MS圖譜 ------------------------------ 136
表一 化合物TYC506、TYC516之1H和13C NMR數據 --------- 36
表二 化合物TYC526之1H和13C NMR數據 ----------------------- 37
表三 化合物TYC507、TYC517之1H和13C NMR數據 --------- 38
表四 化合物TYC508、TYC518之1H和13C NMR數據 --------- 39
表五 化合物TYC509、TYC519之1H和13C NMR數據 --------- 40
表六 Cytotoxicity of synthesized compounds in tumor cell lines ----- 42

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