至今為止已有許多具活性的二次代謝物從海洋無脊椎動物中被發掘出來。從過去的研究中，發現Clavularia屬之軟珊瑚有許多不同骨架且具有活性的二次代謝物，本論文以採集自綠島的踕形羽軟珊瑚Clavularia viridis中所含的二次代謝物進行分析及研究。本研究自軟珊瑚Clavularia viridis之丙酮萃取物中，分離出十個化合物，包含四個新的固醇類化合物4–7，三個已知的固醇類化合物1–3，以及三個已知的前列腺素類化合物8–10。這些二次代謝物的結構鑑定是以有機光譜的分析(包含一維及二維NMR、紅外線光譜、質譜及比旋光)以及與已知化合物的文獻比對光譜資料而確定。將化合物2–10對人類乳癌細胞(MDA-MB-231)；化合物8–10對老鼠黑色素瘤細胞(B16F10)進行細胞毒殺活性測試，發現化合物6和8–10對MDA-MB-231具有毒殺活性(IC50 分別為4.9、6.4、6.2、4.5 µM)；化合物8–10對B16F10具有良好的毒殺活性(IC50 分別為0.7、2.1、1.1 µM)。

Various bioactive secondary metabolites have been found from marine invertebrates. Soft corals of the genus Clavularia have been reported to be a rich source of secondary metabolites with different skeletons and biological activities. Therefore this thesis aim to investigate the metabolites of Clavularia viridis collected from Green Island off, Taiwan. In this study, we isolated ten compounds from acetone-soluble extract including four new steroids (4–7), three known steroids (1–3) and three known prostanoids (8–10). The structure of these compounds were determined on the basis of their spectroscopic analyses (1D NMR, 2D NMR, IR, ESIMS and optical rotations) and by comparison of the physical and spectral data with those of the related known compounds. The cytotoxicity against MDA-MB-231 and B16F10 were evaluated in vitro. Metabolites 2–10 were test against MDA-MB-231 (human breast adenocarcinoma), and metabolites 8–10 were test against B16F10 (murine skin melanoma). Metabolites 6 and 8–10 showed cytotoxic activity against MDA-MB-231 cell (IC50 = 4.9、6.4、6.2、4.5 µM, respectively ), and metabolites 8–10 exhibited potent cytotoxic activity against B16F10 cell (IC50 = 0.7、2.1、1.1 µM, respectively).

目 錄

論文審定書 i
謝辭 ii
中文摘要 iii
英文摘要 iv
化合物1-10之化學結構 v
目錄 vi
表次 viii
圖次 x
縮寫對照表 xvii


壹、緒論 1
一、前言及研究動機與目的 1
二、踕形羽軟珊瑚Clavularia viridis二次代謝物研究回顧 3
1、Prostanoid 類二次代謝物 3
2、Steroid類二次代謝物 16
3、Terpenoid類二次代謝物 24
貳、材料及研究方法 27
一、Clavularia viridis樣品採集時間、地點與鑑定 27
二、Clavularia viridis之分類地位 27
三、Clavularia viridis萃取與分離流程 28
四、Clavularia viridis之分離純化 30
1、化合物1–3之純化分離 30
2、化合物4–7之純化分離 31
3、化合物8–10之純化分離 32
参、化合物之結構解析 34
一、化合物1之結構鑑定(CDCl3) 34
二、化合物2之結構鑑定(CDCl3) 51
三、化合物3之結構鑑定(CDCl3) 64
四、化合物4之結構解析(C6D6) 77
五、化合物5之結構解析(C6D6) 104
六、化合物6之結構解析(C6D6) 130
七、化合物7之結構解析(C6D6) 157
八、化合物8之結構鑑定(CDCl3) 182
九、化合物9之結構鑑定(CDCl3) 205
十、化合物10之結構鑑定(CDCl3) 222
肆、生物活性之測試 239
一、抗乳癌活性測試方法 239
二、WST-1分析原理 240
三、活性測試結果 241
伍、結論 243
陸、參考文獻 244
柒、附錄 249
一、實驗設備儀器 249
二、實驗材料 250

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