針對野外型和養殖型軟珊瑚Sinularia flexibilis 所具有的生物活性成份研究中，共分離出15個天然化合物，其中從野外型S. flexibilis中分離出4個新化合物(1–4)，flexibilisolide A (1)、flexibilisin A (2)、quercifomolide D (3)、quercifomolide B (4)和8個已知的化合物(5–12)和從養殖型S. flexibilis中也分離出3個新化合物(13–15)，flexibilisolide B (13)、flexibilisin B (14)、flexibilisquinone (15)和4個已知的化合物 (9–12)。化合物1–15的結構是經由光譜資料分析和比對相關文獻化合物而建立，化合物4的結構是經由X-ray單晶繞射解析加以鑑定。同時利用Mosher’s method 來加以確定其絶對立體位向。
於細胞毒殺活性測試方面，化合物7、9和11對四種人類癌細胞 Daoy (髓母細胞瘤)、Hep 2 (喉癌細胞)、MCF-7(乳癌細胞)和Hela (子宮頸癌細胞)具有顯著的抑制效果，此外，化合物1–15進行抗發炎活性試驗，結果發現化合物1、6、8、9、11–13和15在濃度為 10 μM下對脂多醣(lipopolysaccharide，LPS)誘發的小鼠巨噬細胞株 RAW 264.7所產生發炎性蛋白質-一氧化氮合成酶(iNOS)之蛋白質表現情形具有抑制效果，在相同濃度下化合物11和12對發炎性蛋白質環氧酶之蛋白質表現情形也具有抑制效果。

We have searched the bioactive metabolites from the wild-type and cultured soft corals Sinularia flexibilis. This study led to the isolation of fifteen natural products (1–15), including four new metabolites (1–4), flexibilisolide A (1), flexibilisin A (2), quercifomolide D (3) and quercifomolide B (4) along with eight known compounds (5–12) from the wild-type soft coral S. flexibilis, and three new metabolites (13–15), flexibilisolide B (13), flexibilisin B (14), flexibilisquinone (15) along with four known compounds (9–12) from the cultured soft coral S. flexibilis. The structures of metabolites 1–15, including their stereochemistries, have been established by detailed spectroscopic analyses and by comparison with the related physical and spectral data from other known compounds. The relative configuration of metabolite 4 was further confirmed by X-ray single-crystal diffraction analysis. Furthermore, the absolute configuration of 4 was determined by a modified Mosher’s method.
The cytotoxicity of 1–15 against the growth of human tumor cell lines, including cervical epitheloid (Hela), laryngeal (Hep 2), medulloblastoma (Daoy) and breast (MCF-7) carcinoma cells was studied. The results showed that only compounds 7, 9 and 11 exhibited moderate cytotoxicities against the tested cell lines. Furthermore, compounds 1, 6, 8, 9, 11–13 and 15 were found to significantly inhibit the accumulation of the pro-inflammatory iNOS protein in the LPS-stimulated RAW264.7 macrophage cells at 10 μM. At the same concentrations, compounds 11 and 12 also could significantly inhibit the accumulation of the pro-inflammatory COX-2 protein.

中文摘要 1
英文摘要 2
壹、緒論 4
一：前言 4
二：研究背景與動機 5
三：文獻回顧 6
貳、實驗方法與程序 14
一：生物樣品採集時間與地點 14
二：生物樣品的分類地位 14
三：萃取與分離純化步驟 15
四：實驗設備儀器與材料 19
參、實驗結果與化學構造解析 21
一、指形軟珊瑚Sinularia flexibilis 所分離化合物之結構解析 21
(一)、Flexibilisolide A (1)之結構解析 21
(二)、Flexibilisin A (2)之結構解析 30
(三)、Quercifomolide D (3)之結構解析 38
(四)、Quercifomolide B (4)之結構解析 47
（五）、Sinulariolide (5)之結構解析 57
（六）、Sinulariolone (6)之結構解析 62
（七）、Sinuflexolide (7)之結構解析 66
（八）、Dihydrosinuflexolide (8)之結構解析 71
（九）、Sinularin (9)之結構解析 75
（十）、Dihydrosinularin (10)之結構解析 79
（十一）、Dehydrosinulariolide (11)之結構解析 83
(十二)、11–Epi–sinulariolide acetate (12)之結構解析 87
(十三)、Flexibilisolide B (13)之結構解析 91
(十四)、Flexibilisin B (14)之結構解析 101
(十五)、Flexibilisquinone (15)之結構解析 110
二、化學反應步驟 118
三、實驗數據整理 121
肆、實驗結果與化學構造解析 124
一、生物活性試驗方法 124
(一)、細胞毒殺活性試驗方法 124
(二)、抗發炎活性試驗方法 126
二、生物活性試驗結果 129
(一)、細胞毒殺活性試驗結果 129
(二)、抗發炎活性試驗結果 130
伍、結論 133
陸、參考文獻 135
附錄、化合物4之X-ray實驗數據 140

1. 江昆逹、林全信；“豐富的海洋資源”；台灣書店，1997.
2. Kijjoa, A.; Sawangwong, P. “Drugs and cosmetics from the sea” Mar. Durgs 2004, 2, 73–82.
3. 戴昌鳳；“墾丁國家公園−珊瑚之美”；墾丁國家公園管理處員工消費合作社出版，中華民國，1987.
4. Newman, D. J.; Cragg, G. M. “Marine natural products and related compounds in clinical and advanced preclinical trials” J. Nat. Prod. 2004, 67, 1216–1238.
5. Simmons, T. L.; Andrianasolo, E.; McPhail, K.; Flatt, P.; Gerwick, W. H. “Marine natural products as anticancer drugs” Molecular Cancer Therapeutics 2005, 4, 333–342.
6. 趙子華 “台灣產軟珊瑚Lobophytum crassum與Dendronephthyagriffini
所含天然物及Lobohedleolide化合物之化學修飾之研究” 國立中山大學海洋資源學系博士論文，中華民國96年。
7. Hsieh, P.-W.; Chang, F.-R.; Mcphail, A. T.; Lee, K.-H.; Wu, Y.-C. “New cembranolide analogues from the Formosan soft coral Sinularia flexibilis and their cytotoxicity” Nat. Prod. Res. 2003, 17, 409–418.
8. Tursch, B.; Braekman, J. C.; Daloze, D.; Herin, M.; Karlsson, R.; Losman, D. “Chemical studies of marine invertebrates–XI. Sinulariolide, a new cembranolide diterpene from the soft coral Sinularia flexibilis” Tetrahedron 1975, 31, 129–133.
9. Herin, M.; Colin, M.; Tursch, B. “Chemical studies of marine invertebrates. XXV. Flexibilene, an unprecedented fifteen-membered ring diterpene hydrocarbon from the soft coral Sinularia flexibilis (Coelenterata, Octocorallia, Alcyonacea)” Bulletin des Societes Chimiques Belges 1976, 85, 801–803. from CAPLUS (Copyright (C)
2008 ACS on SciFinder (R)).
10. Herin, M.; Tursch, B. “Chemical studies of marine invertebrates. XXIV. Minor cembrane diterpenes from the soft coral Sinularia flexibilis (Coelenterata, Octocorallia)” Bulletin des Societes Chimiques Belges 1976, 85, 707–719. from CAPLUS (Copyright (C) 2008 ACS on SciFinder (R)).
11. Weinheimer, A. J., Matson, J. A.; Hossain, M. B.; Vander, Helm D. “Marine anticancer agents: Sinularin and dihydrosinularin, new cembranolides from the soft coral Sinularia flexibilis” Tetrahedron Lett. 1977, 34, 2923–2926.
12. Kazlauskao, R.; Murphy, P. I.; Wells, R. J.; Schonholzer, P.; Coll, J. C. “Cembranoid constituents from an Australian collection of the soft coral Sinularia flexibilis” Aust. J. Chem. 1978, 31, 1817–1824.
13. Kazlauskas, R.; Marwood, J.F.; Wells, R. J., “2- Phenylethylamides of a novel lipid acid; atrial stimulants from the soft coral Sinularia flexibilis” Aust. J. Chem. 1980, 33, 1799–1803.
14. Mori, K.; Suzuki, S.; Iguchi, K.; Yamada, Y. “8,11-Epoxy bridged cembranolide diterpene from the soft coral Sinularia flexibilis” Chem. Lett. 1983, 1515–1516.
15. Kusumi, T.; Uchida, H.; Ishitsuka, M. O.; Yamamoto, H,; Kakisawa, H. “Alcyonin, A new cladiellane diterpene from the soft coral Sinularia flexibilis” Chem .Lett. 1988, 1077–1078.
16. Hamada, T.; Kusumi, T.; Ishitsuka, M. O.; Kakisawa, H. “ Structures and absolute configurations of new lobane diterpenoids from the Okinawan soft coral Sinularia flexibilis” Chem. Lett. 1992, 333–336.
17. Guerrero, P. P.; Read, R. W.; Batley, M.; Janairo, G. C. “The structure of a novel cembranoid diterpene from a Philipine collection of the soft coral Sinularia flexibilis” J. Nat. Prod. 1995, 58, 1185–1191.
18. Anjaneyulu. A. S. R.; Sagar, K. S. “Flexibiolide and dihydroflexibiolide, the first trihydroxycembranolide lactones from the soft coral Sinularia flexibilis of the Indian Ocean.” Nat. Prod. Lett. 1996, 9, 127–135.
19. Anjaneyulu, A. S. R.; Sagar, K. S.; Rao, G. V. “New cembranoid lactones from the Indian Ocean soft coral Sinularia flexibilis” J. Nat. Prod. 1997, 60, 9–12.
20. Anjaneyulu, A. S. R.; Sagar, K. S.; Rao, G. V. “A novel cinnamide dimer from the Indian Ocean soft coral Sinularia flexibilis” Nat. Prod. Lett. 1997, 11, 5–11.
21. Duh, C.-Y.; Wang, S.-K.; Tseng, H.-K.; Sheu, J.-H.; Chiang, M. Y. “Novel cytotoxic cembranoids from the Formosan soft coral Sinularia flexibilis” J. Nat. Prod. 1998, 61, 844–847.
22. Duh, C.-Y.; Wang, S.-K.; Tseng, H.-K.; Sheu, J.-H. “A novel cytotoxic biscembranoid from the Formosan soft coral Sinularia flexibilis” Tetrahedron Lett. 1998, 39, 7121–7122.
23. Yu, S.; Deng, Z.; Ofwegen, L.-V.; Peter P., Lin, W. “5, 8-Epidioxysterols and related derivatives from a Chinese soft coral Sinularia flexibilis” Steroids 2006, 71, 955–959.
24. Su, J.; Yang, R.; Kuang, Y.; Zeng, L. “A new cembranoids from the soft coral Sinularia capillpsa” J. Nat. Prod. 2000, 63, 1543–1545.
25. Yang, R.-L.; Zeng, L.-M.; Su, J.-Y.; Lin, H. “A new furanocembranoid trihydroxylated cembranolide diterpene” Acta Chim. Sinica 2000, 58, 1186–1187.
26. Lin, C.-W.; Su, J.-Y.; Zeng, L.-M.; Qi, S.-J., Xu, A.-L. “Studies on the cembranolide of Sinularia tenella Li” Chin. J. Org. Chem. 2001, 21, 56–59.
27. Kusumi, T.; Fujita, Y.; Ohtani, I.; Kakisawa, H. “Anomaly in the modified Mosher’s method: absolute configurations of some marine cembranolides” Tetrahedron Lett. 1991, 32, 2923–2926.
28. Ohtani, I.; Kusumi, T.; Kashman, Y.; Kakisawa, H. “High-Field FT NMR application of Mosher’s method. The absolute configurations of Marine terpenoids” J. Am. Chem. Soc. 1991, 113, 4092–4096.
29. Ohtani, I.; Kusumi, T.; Kashman, Y.; Kakisawa, H. “A new aspect of the High-Field NMR application of Mosher’s Method. The absolute configuration of marine triterpene Sipholenol-A” J. Org. Chem. 1991, 56, 1296–1298.
30. Aceret, T. L.; Brown, L.; Miller, J.; Coll, J. C.; Sammarco, P.W. “Cardiac and vascular responses of isolated rat tissues treated with diterpenes form Sinularia flexibilis (Coelenterata: Octocorallia) ” Toxicon 1996, 34, 1165–1171.
31. Aceret, T. L.; John C. C.; Uchio Y.; Sammarco, P.W. “Antimicrobial activity of the diterpenes flexibilide and sinulariolide derived from Sinularia flexibilis Quoy and Gaimard 1833(Coelenterata: Alcyonacea, Octocorallia)” Comparative Biochemistry and physiology Part C 1998, 120, 121–126.
32. 盧詣 “台灣產指形軟珊瑚 Sinularia querciformis 所含 Cembranoids
之天然化合物研究” 國立中山大學海洋資源學系碩士論文，中華民國97年。
33. Kashman, Y.; Bodner. M.; Loya, Y.; Benayahu, Y. “Cembranolides from marine origin (Red Sea), survey, and isolation of new sinulariolide derivatives” Isr. J. Chem. 1977, 6, 1–3.
34. Findlay, J. A.; Patil, A. D. “Novel sterols from the finger spong
Haliclona oculata” Can. J. Chem., 1985, 63, 2406–2410.
35. Ho, F.-M.; Lai, C.-C.; Huang, L.-J.; Kuo, T.-C.; Chao, C.-M.; Lin, W.-
W. “The anti-inflammatory carbazole, LCY-2-CHO, inhibits lipopolysaccharide-induced inflammatory mediator expression through inhibition of the p38 mitogen-activated protein kinase signaling pathway in macrophages” Br. J. Pharmacol. 2004, 141, 1037–1047.
36. Alley, M. C.; Scudiero, D. A.; Monks, A.; Hursey, M. L.; Czerwinski, M. J.; Fine, D. L.; Abbott, B. J.; Mayo, J. G.; Shoemaker, R. H.; Boyd, M. R. “Feasibility of drug screening with panels of human tumor cell lines using a microculture tetraxolium assay” Cancer Res. 1988, 48, 589–601.
37. Scudiero, D. A.; Shoemaker, R. H.; Paull, K. D.; Monks, A.; Tierney,
S.; Nofziger, T. H.; Currens, M. J.; Seniff, D.; Boyd, M. R.
“Evalution of a soluble with tetrazolium/formazan assay for cell growth and drug sensitivity in culture using human and other tumor cell lines” Cancer Res. 1988, 48, 4827–4833.