最近我們在探索海綿Hippospongia sp.化學成份的過程中，從海綿Hippospongia sp.中分離出5個二倍半萜類化合物 (2-6)，其中發現一些化合物具有細胞毒性。本次實驗持續針對相同海綿Hippospongia sp.的後續研究，從中分離出一個新的二倍半萜類化合物hippospongide D (1)。另外，從北極收集的海綿Dendoricella flabelliformis中分離出一個新的天然物sucrose octaacetate (7)。
　　在生物活性方面，化合物2-7分別對人類膽管癌細胞 (HuCCT1) 和人類結腸癌細胞 (SW620) 進行細胞毒殺活性測試，結果顯示2和5對上述兩種細胞株具有良好的細胞毒殺活性，且兩種化合物會隨著給予細胞濃度的增加而造成細胞存活率和細胞凋亡的現象隨之增加。此外，實驗結果也顯示出化合物2和5造成細胞凋亡是經由調節caspase蛋白酶中的caspase-3、caspase-7和PARP的蛋白質表現，藉由增加其活化態，進而誘導細胞凋亡。

　　Our group recent study of the chemical constituents of the sponge Hippospongia sp. has yielded five scalarane-type sesterterpenes (2-6). Some of these have been found to possess cytotoxic activity. In continuation of our search for bioactive metabolites from the above sponge, Hippospongia sp., we have further isolated one new sesterterpene, hippospongide D (1). Moreover, our chemical investigation on the sponge Dendoricella flabelliformis which was collected off the North Pole, has led to the isolation of one new natural product sucrose octaacetate (7).
　　The cytotoxic activity of 2-7 was evaluated against two cancer cell lines including HuCCT1 and SW620. The result showed that compounds 2 and 5 exhibited significant cytotoxic activity against HuCCT1 and SW620 cancer cell lines and they had significantly suppressed cell viability and increase cell apoptosis in dose-manner. Compounds 2 and 5 administration increased protein expression of apoptosis by regulates active-form of caspase-3, caspase-7 and PARP.

第一章、緒論 1
第一節、前言 1
第二節、文獻回顧 4
第二章、生物材料與研究方法 17
第一節、生物樣品的採集與分類 17
第二節、生物樣品的萃取與分離 19
第三節、實驗器材與材料 21
第三章、結果與討論 25
第一節、Hippospongia sp.所含化合物之結構解析 25
(一)、Hippospongide D (1) 化合物結構之解析 26
第二節、Dendoricella flabelliformis所含化合物之結構解析 35
(一)、sucrose octaacetate (7) 化合物構造之解析 35
第三節、新化合物實驗數據整理 43
第四章、生物活性試驗 45
第一節、細胞毒殺試驗 45
(一)、細胞培養 45
(二)、細胞冷凍保存 45
(三)、冷凍細胞活化 45
(四)、細胞毒殺活性試驗 46
(五)、Annexin Ⅴ/ PI 雙重染色檢測細胞凋亡程度 46
(六)、西方墨點法 (Western blotting) 47
第二節、生物活性試驗結果 49
(一)、細胞毒殺活性試驗結果 49
(二)、化合物2和5誘導細胞凋亡 51
(三)、藥物增加對細胞株的蛋白質表現量之影響 52
第五章、結論 55
參考文獻 57

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