本研究主要是從台灣產指型軟珊瑚 Sinularia flexibilis的有機萃取物中尋找具有生物活性的二次代謝物。從這株珊瑚中共分離得到十八個天然化合物1–18，其中分離出七個新的化合物，為cembrane-type diterpenoids類化合物flexibilisolides C–G (1, 2, 3, 6, 7)、flexiblilisin C (5)、11,12-secoflexibillin (4)及十一個已知化合物 8–18。所有化合物的化學構造均由光譜數據的分析(IR, MS, 1D、2D NMR)和比對文獻上已知化合物的光譜資料而決定。
本研究中將所獲得的化合物1–6, 8–14, 16–18進行癌細胞株人類子宮頸癌細胞(Hela)、人類肝癌細胞(SK-Hep1)與黑色素癌細胞(B16)的細胞毒殺活性篩選。結果顯示化合物1,9, 12及13對人類子宮頸癌細胞(Hela)與人類肝癌細胞(SK-Hep1)具有中等的細胞毒殺活性；而化合物12同時也對人類黑色素癌細胞(B16)具有中等的細胞毒殺活性；此外化合物14對人類子宮頸癌細胞(Hela)也具有細胞毒殺活性。另一方面，在抗發炎活性試驗中，化合物1, 9, 13及17對於在濃度為10μM下可有效抑制蛋白質iNOS與COX-2的表現；化合物2–4, 7, 8, 10, 14, 16對於iNOS有抑制效果。

In order to discover bioactive secondary metabolites, we have studied the chemical constituents from the organic extracts of soft coral Sinularia flexibilis. This study had led to the isolation of eighteen natural compounds 1–18, including seven new cembrane-type diterpenoids, flexibilisolides C–G (1, 2, 3, 6, 7), flexiblilisin C (5), 11,12-secoflexibillin (4) along with eleven know compounds. The structure of compounds 1–18 were established by detailed spectral data analysis (IR, MS, 1D, 2D NMR) and by comparison of the spectral data with those of the related known compounds.
The cytotoxicity of compounds 1–6, 8–14, 16–18 against the Hela (human cervical epitheloid carcinoma), SK-Hep1 (human liver carcinoma), and B16 (human melanin carcinoma) tumor cell lines were screened. Compounds 1, 9, 12, and 13 showed moderate activity toward the Hela and SK-Hep1 tumor cells. Compound 12 also showed moderate activity toward the B16 cells, and compound 14 showed activity toward the Hela cells. On the other hand, compounds 1, 9, 13, and 17 were found to show significant activity against the accumulation of the pro-inflammatory iNOS and COX-2 protein at 10 μM. Compounds 2–4, 7, 8, 10, 14, 16 were found to show activity against the accumulation of the pro-inflammatory iNOS protein at 10 μM.

中文摘要 IX
英文摘要 X
化合物1~9化學結構 XI
化合物10~18化學結構 XII
第一章、緒論 1
第一節、前言 1
第二節、研究背景及目的 2
第三節、文獻回顧 3
第二章、生物材料與研究方法 30
第一節、研究流程 30
第二節、Sinularia flexibilis 樣品採集時間、地點、分類地位及
分離流程 32
2.2.1 Sinularia flexibilis 樣品採集時間、地點、分類地位 32
2.2.2 Sinularia flexibilis 樣品的萃取分離 33
第三節、實驗設備儀器及材料 35
2.3.1實驗設備儀器 35
2.3.2實驗材料 36
第三章、化合物之結構證明 30
第一節：軟珊瑚Sinularia flexibilis 所分離之化合物結構解析 37
(一)、Flexibilisolide C (1) 化合物構造之解析 37
(二)、Flexibilisolide D (2) 化合物構造之解析 52
(三)、Flexibilisolide E (3) 化合物構造之解析 66
(四)、11,12-Secoflexibillin (4) 化合物構造之解析 80
(五)、Flexibilisin C (5) 化合物構造之解析 94
(六)、Flexibilisolide F (6) 化合物構造之解析 108
(七)、Flexibilisolide G (7) 化合物構造之解析 122
(八)、Flexilarin C (8) 化合物構造之解析 137
(九)、11-Dehydrosinulariolide (9) 化合物構造之解析 142
(十)、Flexilarin D (10) 化合物構造之解析 147
(十一)、Querciformilin C (11) 化合物構造之解析 152
(十二)、14-Deoxycrassin (12) 化合物構造之解析 157
(十三)、11-Epi-sinulariolide acetate (13) 化合物構造之解析 162
(十四)、3,4:8,11-Bisepoxy-7-acetoxycembra-15(17)-en-1,12-olide (14) 化合物構造之解析 167
(十五)、Flexibilisin B (15) 化合物構造之解析 173
(十六)、Sinulariolide (16) 化合物構造之解析 178
(十七)、11-Epi-sinulariolide (17) 化合物構造之解析 183
(十八)、Dihydrosinularin (18) 化合物構造之解析 188
第二節:化合物物理性質及圖譜數據整理 192
第四章、生物活性試驗 197
第一節、生物活性試驗方法 197
(一)、細胞毒殺活性試驗方法 197
(二)、抗發炎活性試驗方法 198
第二節、生物活性試驗結果 201
(一)、細胞毒殺活性試驗結果 201
(二)、抗發炎活性試驗結果 202
第五章、結論 205
第六章、參考文獻 210

1. 王寶貫，天與地，牛頓出版股份有限公司，1996。
2. 江昆達、林全信，豐富的海洋資源，臺灣書店，1998。
3. 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.
4. Kijjoa, A.; Sawangwong, P. “Drugs and cosmetics from the sea” Mar. Durgs 2004, 2, 73–82.
5. Tursch, B.; Braekman, J. C.; Daloze, D.; Herin, M.; Karlsson, R.;Losman, D. “Chemical studies of the marine invertebrates XI.Sinulariolide, a new cembranolide diterpene from the soft coralSinularia flexibilis” Tetrahedron 1975, 31, 129–133.
6. Kashman, Y.; Bodner, M.; Loya, Y.; Benayahu, Y. “Cembranolidsfrom marine origin (Red Sea), survey, and isolation of a newsinulariolide derivative” Israel J. Chem. 1977, 16, 1–3.
7. Su, J.; Yang, J. S. R.; Kuang, Y.; Zeng, L. “A new cembranoids from the soft coral Sinularia capillpsa” J. Nat. Prod. 2000, 63, 1543–1545.
8. Lin, C.-W.; Su, J.-Y.; Zeng, L.-M.; Qi, S.-J.; Xu, A.-L. “Studies on the Cembranoid Diterpenes of Sinularia Tenella Li ” Chin. J. Org. Chem. 2001, 21, 56–59.
9. 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.
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.
11. 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.
12. 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.
13. 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.
14. Yang, R.-L.; Zeng, L.-M.; Su, J.-Y.; Lin, H. “A new furanocembranoid trihydroxylated cembranolide diterpene” Acta Chim. Sinica 2000, 58, 1186–1187.
15. 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.
16. Buckle, P. J.; Baldo, B. A.; Taylor, K. M. “The anti-inflammatory activity of marine natural products – 6-n-tridecylsalicylic acid, Flexibilide and Dendalone 3-hydroxybutyrate” Inflammation Research 1980, 10, 361–367.
17. QI, S.-J.; Chen, Q.; Peng, W.-L.; Xu, A.-L. “Cytotoxic properties of cembranolides diterpenoids from soft coral Sinularia tenella Li” J. Trop. Ocean 2002, 21, 87–91.
18. 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.
19. Mori, K.; Suzuki, S.; Iguchi, K.; Yamada, Y. “8,11-Epoxy bridged cembranolide diterpene from the soft coral Sinularia flexibilis” Chem. Lett. 1983, 10, 1515–1516.
20. 林彥佑，海洋天然物11-epi-sinulariolide acetate 對於佐劑誘發類風濕性關節炎治療之影響，國立中山大學海洋生物科技暨資源學系碩士論文，中華民國98年。
21. 盧詣，台灣產指形軟珊瑚 Sinularia querciformis 所含 Cembranoids之天然化合物研究，國立中山大學海洋生物科技暨資源學系碩士論文，中華民國97年。
22. 林妤芳，野外型與養殖型柔軟指形軟珊瑚所含天然物之研究，國立中山大學海洋生物科技暨資源學系碩士論文，中華民國97年。
23. 黃瓊瑤，指型軟珊瑚 Sinularia granosa 與 Sinularia leptoclados化學成分及生物活性之研究，國立中山大學海洋生物科技暨資源學系碩士論文，中華民國98年。
24. 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, 17, 1077–1078.
25. 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, 21, 33–36.
26. Aceret, T. L.; Brown, L.; Miller, J.; Coll, J. C.; Sammarco, P. W. “Cardiac and vascular responses of isolated rat tissues treated with diterpenes from sinularia flexibilis (coelenterata: octocorallia)” Toxicon 1996, 34, 1165–1171.
27. 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.
28. 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.
29. 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.
30. 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.
31. 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.
32. 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.
33. Yu, S.; Deng, Z.; Ofwegen, L.-V.; Peter P., Lin, W.-H. “5, 8-Epidioxysterols and related derivatives from a Chinese soft coral Sinularia flexibilis” Steroids 2006, 71, 955–959.
34. Wen, T.; Ding, Y.; Deng, Z.-W.; Ofwegen, L.-V.; Proksch, P.; Lin, W.-H. “Sinulaflexiolides A—K, cembrane-type diterpenoids from the Chinese soft coral Sinularia flexibilis” J. Nat. Prod. 2008, 71, 1113–1140.
35. Lin, Y.-S.; Chen, C.-H.; Liaw, C.-C.; Chen, Y.-C.; Kuo, Y.-H.; Shen, Y.-C. “Cembrane diterpenoids from the Taiwanese soft coral Sinularia flexibilis” Tetrahedron 2009, 65, 9157–9164.
36. Lo, K.-L.; Khalil, A. T.; Chen, M.-H.; Shen, Y.-C. “New Cembrane diterpenes from Taiwanese soft coral Sinularia flexibilis” Helvetica Chimica Acta 2010, 93, 1329–1335.
37. Chen, B.-W.; Chao, C.-H.; Su, J.-H.; Huang, C.-Y.; Dai, C.-F.; Wen, Z.-H.; Sheu, J.-H. “A Novel Symmetric Sulfur-Containing Biscembranoid from the Formosan Soft Coral Sinularia flexibilis” Tetrahedron Lett. 2010, 51, 5764–5766.
38. Su, J.; Yang, J. S. R.; Kuang, Y.; Zeng, L. “A new cembranoids from the soft coral Sinularia capillpsa” J. Nat. Prod. 2000, 63, 1543–1545.
39. Ma, A.; Deng, Z.-W.; Ofwegen, L.-V.; Bayer, M; Proksch, P.; Lin W. “Dendronpholides A—R, Cembranoid diterpenes from the Chinese soft corals Dendronephthya sp.” J. Nat. Prod. 2008, 71, 1152–1160.
40. Lo, K.-L.; Khalil, A. T.; Chen, M.-H.; Shen, Y.-C. “Sinuladiterpenes A—F, new cembrane diterpenes from Sinularia flexibilis” Chemistry & Biodiversity 2009, 6, 2227–2234.
41. Li, Y.; Gao, A.-H.; Huang, H.; Li, J.; Mollo, E.; Gavagnin, M.; Cimino, G.; Gu, Y.-C.; Guo, Y.-W. “Diterpenoids from the Hainan soft coral Sinularia parva” Helvetica Chimica Acta 2009, 92, 1341–1348.
42. Lu, Y.; Huang, C.-Y.; Lin, Y.-F.; Wen, Z.-H.; Su, J.-H.; Chiang, M. Y.; Sheu, J.-H. “Anti-inflammatory cembranoids from the soft corals Sinularia querciformis and Sinularia granosa” J. Nat. Prod. 2008, 71, 1754–1759.
43. Rodriguez, A. D.; Martinez, N. “Marine antitumor agents: 14-deoxycrassin and pseudoplexaurol, new cembranoid diterpenes from the Caribbean gorgonian Pseudoplexaura porosa” Experientia 1993, 49, 179–181.
44. Cantrell, C. L.; Lu, T.; Fronczek, F. R.; Fischer, N. H. “Antimycobacterial Cycloartanes from Borrichia frutescens” J. Nat. Prod. 1996, 59, 1131–1136.
45. 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 withpanels of human tumor cell lines using a microculture tetraxolium assay” Cancer Res. 1988, 48, 589–601.
46. 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.