本研究主要是於2009年4月在東沙環礁採集所得之星形肉質軟珊瑚Sarcophyton stellatum的有機萃取物中分離純化其中天然物成分，並輔以活性測試試驗，檢測所天然物成分是否具有生物活性。
本研究總共分離得到16個雙萜類的天然化合物，其中1–6為新化合物，分別命名為stellatumonone (1)、stellatumolides A–C (2–4) and stellatumonins A–B (5–6)，7–16為已知的化合物。所有化合物的化學構造均由光譜數據的分析(1D與2D NMR、Mass IR、UV、旋光、熔點)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1–16分別進行人類肝癌細胞(HepG2)、人類乳癌細胞(MDA-MB231)和人類肺腺癌細胞(A549)三株癌細胞的細胞毒殺活性測試。活性測試結果顯示只有化合物16對人類肺腺癌細胞(A549)具有細胞毒殺活性。

In order to search for bioactive compounds, we have studied the chemical constituents from the organic extracts of the soft coral Sarcophyton stellatum, collected from Dongsha Atoll. This study had led to the identification of sixteen natural diterpenoids, including six new compounds, stellatumonone (1), stellatumolides A–C (2–4) and stellatumonins A–B (5–6), along with ten known compounds. The structures of compounds 1–16 were established by detailed spectroscopic data (IR, MS, 1D, 2D NMR) and by comparison of the spectral data with those of the related known compounds.
The cytotoxicity of compounds 1–16 against the HepG2 (human liver hepatocellular carninoma cells), MDA-MB231 (human breast cancer cells) and A549 (human respiratory epithelial cells) cancer cell lines were determined. Among them, compound 16 showed cytotoxic activity toward the A549 cancer cells.

中文摘要 I
英文摘要II
化合物1-16化學結構III
第一章、緒論1
第一節、研究動機1
第二節、生物材料型態、分佈與分類地位5
第三節、文獻回顧5
第二章、實驗程序與研究方法52
第一節、Sarcophyton stellatum的採集與分離流程52
第二節、實驗設備儀器及材料56
第三章、化合物之結構解析 58
第一節、 星形肉質軟珊瑚Sarcophyton stellatum 所分離出之化合物的結構解析58
(一)、Stellatumonone (1)的結構解析58
(二)、Stellatumolide A (2)的結構解析67
(三)、Stellatumolide B (3)的結構解析81
(四)、Stellatumolide C (4)的結構解析91
(五)、Stellatumonin A (5)的結構解析99
(六)、Stellatumonin B (6)的結構解析108
(七)、Sarsolilide A (7)的結構解析117
(八)、未命名已知化合物(8)的結構解析121
(九)、Sarcophine (9)的結構解析126
(十)、Laevigatol B (10)的結構解析130
(十一)、Sarcophtonin E (11)的結構解析134
(十二)、Sarcophytonin C (12)的結構解析138
(十三)、Sarcophtonin F (13)的結構解析142
(十四)、7β-Acetoxy-8α-hydroxy-deepoxysarcophine (14)的結構解析146
(十五)、7β,8α-Dihydroxydeepoxy-ent-sarcophine (15)的結構解析150
(十六)、未命名已知化合物(16)的結構解析154
第二節、化合物物理性質 158
第四章、生物活性試驗方法 160
第一節、細胞毒殺檢測原理160
第二節、癌細胞培養160
第三節、細胞毒殺試験160
第四節、細胞毒殺的實験步驟161
第五章、生物活性試驗結果162
第一節、細胞毒殺活性試驗結果 162
第六章、結論 163
第七章、參考文獻164

1.Marty, A. T. “The Complete German Commission E Monographs: Therapeutic Guide to Herbal Medicines” JAMA. 1999,281,1852-1853.
2.Ruggieri, G. D. “Drugs from the Sea” Science 1976, 194, 491–497.
3.Fenical, W. “New Pharmaceuticals from Marine Organisms” Mar. Biotechnol. 1997, 15, 339–341.
4.Kijjoa, A.; Sawangwong, P. “Drugs and cosmetics from the sea” Mar. Durgs 2004, 2, 73–82.
5.Wallace, M. S.; Charapata, S. G.; Fisher, R. “Intrathecal ziconotide in the treatment of chronic non-malignant pain: a randomized double blind placebo-controlled clinical trial.” Neuromodulation 2006, 9, 75–86.
6.Glaser, K.B.; Mayer, A. M. S. “A Renaissance in Marine Pharmacology：From Preclinical Curiosity to Clinical Reality.” Biochem. Pharmacol. 2009, 78, 440–448.
7.Blunt, J. W.; Copp, B. R.; Munro, M. H. G.; Northcote, P. T.; Prinsep, M. R. “Marine Natural Products” Nat. Prod. Rep. 2004, 21, 1–49.
8.Blunt, J. W.; Copp, B. R.; Hu, W.-P.; Munro, M. H. G.; Northcote, P. T.; Prinsep, M. R. “Marine natural products” Nap. Prod. Rep. 2008, 25, 35–94.
9.Bakus, G. J. “Chemical Defense Mechanisms on the Great Barrier Reef, Australia” Science 1981, 211, 497–499.
10.戴昌鳳、洪勝雯，台灣珊瑚圖鑑，貓頭鷹出板社，中華民國98年2月。
11.Ofwegen, L.P. van, S.D. Cairns and J. van der Land “Catalogue of life: 2006 Annual Checklist.” 2006.
12.Bernstein, J.; Shmueli, U.; Zadock, E.; Kashman, Y.; Neeman, I. “Sarcophine, a new epoxy cembranolide from marine origin” Tetrahedron 1974, 30, 2817–2824.
13.Tursch, B. “Some recent developments in the chemistry of Alcyonaceans” Pure＆Appl.Chem. 1976, 48, 1–6.
14.Coll, J. C.; Hawes, G. B.; Liyanage, N.; Oberhansli, W.; Wells, R. J. “Studies of Australian soft corals. I* A new cembranoid diterpene from a Sarcophyton species” Aust. J. Chem. 1977, 30, 1305–1309.
15.Bowden, B. F.; Coll, J. C.; Hicks, W.; Kazlauskas, R.; Mitchell, S. J. “Studies of Australian soft corals. X* The isolation of Epoxyisoneocembrene-A from Sinularia grayi and Isoneocembrene-A from Sarcophyton ehrenbergi” Aust. J. Chem. 1978, 31, 2027–2012.
16.Bowden, B. F.; Coll, J. C.; Mitchell, S. J.; Stokie, G. J. “Studies of Australian soft corals. XI* Two new cembranoid diterpenes from a Sarcophyton species” Aust. J. Chem. 1979, 32, 653–659.
17.Kobayashi, M.; Nakagawa, T.; Mitsuhashi, H. “Marine terpenes and terpenoids. I. Structures of four cembrane-type diterpenes; Sarcophytol-A, Sarcophytol-A acetate Sarcophytol-B, and Sarcophytonin-A, from the soft coral, Sarcophyton glaucum” Chem. Pharm. Bull. 1979, 27, 2382–2387.
18.Bowden, B. F.; Coll, J. C.; Mitchell, S. J. “Studies of Australian soft corals. XVIII* Further cembranoid diterpenes from soft corals of the genus Sarcophyton” Aust. J. Chem. 1980, 33, 879–884.
19.Nakagawa, T.; Kobayashi, M.; Hayashi, K.; Mitsuhashi, H. “Marine terpenes and terpenoids. II. Structures of three cembrane-type diterpenes; Sarcophytol-C, Sarcophytol-D, and Sarcophytol-E, from the soft coral, Sarcophyton glaucum Q. et. G” Chem. Pharm. Bull. 1981, 29, 82–87.
20.Carmely, S.; Groweiss, A.; Kashman, Y. “Decaryiol, a new cembrane diterpene from the marine soft coral Sarcophyton decaryi” J. Org. Chem. 1981, 46, 4279–4284.
21.Blackman, A. J.; Bowden, B. F.; Coll, J. C.; Frick, B.; Mahendran, M.; Mitchell, S. J. “Studies of Australian soft corals. XXIX* Several new cembranoid diterpenes from Nephthea brassica and related diterpenes from a Sarcophyton species” Aust. J. Chem. 1982, 35, 1873–1880.
22.Kazlauskas, R.; Baird-Lambert, J. A.; Murphy, P. T., Wells, R. J. “Two new cembrane from a soft coral (Sarcophyton species)” Aust. J. Chem. 1982, 35, 61–68.
23.Bowden, B. F.; Coll, J. C.; Wells, R. H. “Studies of Australian soft corals. XXVII* Two novel diterpenes from Sarcophyton glaucum” Aust. J. Chem. 1982, 35, 621–627.
24.Uchio, Y.; Nitta. M.; Nakayama, M.; Iwagawa, T.; Hase, T. “Ketoemblide and Sarcophytolide, two new cembranolides with ε-lactone function from the soft coral Sarcophyton elegans” Chem. Lett. 1983, 613–616.
25.Suleimenova, A. M.; Kalinovskii, A. I.; Raldugin, V. A.; Shevtsov, S. A.; Bagryanskaya, I. Yu.; Gatilov, V. Yu; Kuznetsova, T. A; Elyakov, G. B. “New derivatives of (−)-Neocembrene from the soft coral Sarcophyton trocheliophorum” Chem. Nat. Comp. 1988, 24, 453-458.
26.Kuznetsova, T. A.; Popov, A. M.; Agafonova, I. G.; Suleimenova, A. M.; Elyakov, G. B. “Physiological activity of diterpenoids from the soft coral Sarcophyton trocheliophorum” Chem. Nat. Comp.1989, 25, 122-124.
27.Kobayashi, M.; Osabe, K. “Marine terpenes and terpenoids. VII. Minor cembranoid derivatives, structurally related to the potent anti-tumor-promoter Sarcophytol A, from the soft coral Sarcophyton glaucum” Chem. Pharm. Bull. 1989, 37, 631–636.
28.Kobayashi, M.; Osabe, K. “Marine terpenes and terpenoids. VIII. Transannular cyclization of 3,4-Epoxy-1,7,11-cembratriene systems” Chem. Pharm. Bull. 1989, 37, 1192–1196.
29.Kobayashi, M.; Iesaka, T.; Nakano, E. “Marine terpenes and terpenoids. IX. Structures of six new cembranoids, Sarcophytols F, K, P, Q, R and S, from the soft coral Sarcophyton glaucum” Chem. Pharm. Bull. 1989, 37, 2053–2057.
30.Kobayashi, M.; Hirase, T. “Marine terpenes and terpenoids. XI. Structures of new dihydrofuranocembranoids isolated from a Sarcophyton sp. soft coral of Okinawa” Chem. Pharm. Bull. 1990, 38, 2442–2445.
31.Kobayashi, M.; Nakano, E. “Sterochemical course of the transannular cyclization, in chloroform, of epoxycembranoids derived from the geometrical isomers of (14S)-14-Hydroxy-1,3,7,11-cembratetraenes” J. Org. Chem. 1990, 55, 1947–1951.
32.Leone, P. A.; Bowden, B. F.; Carroll, A. R.; Coll, J. C.; Meehan, G. V. “Studies of Australian soft corals. XLIX. A new biscembranoid and its probable biosynthetic precursors from the soft coral Sarcophyton tortuosum” J. Nat. Prod. 1993, 56, 521–526.
33.Greenland, G. J.; Bowden, B. F. “Cembranoid diterpenes related to Sarcophytol A from the soft coral Sarcophyton trocheliophorum (Alcyonacea)” Aust. J. Chem. 1994, 47, 2013–2021.
34.Duh, C.-Y.; Hou, R.-S. “Cytotoxic cembranoids from the soft corals Sinularia gibberosa and Sarcophyton trocheliophorum” J. Nat. Prod. 1996, 59, 595–598.
35.König, G. M.; Wright, A. D. “New cembranoid diterpenes from the soft coral Sarcophyton ehrenbergi” J. Nat. Prod. 1998, 61, 494–496.
36.Badria, F. A.; Guirguis, A. N.; Perovic, S.; Steffen, R.; Müller, W. E. G.; Schröder, H. C. “Sarcophytolide: a new neuroprotective compound from the soft coral Sarcophyton glaucum” Toxicology 1998, 131, 133–143.
37.Iwagawa, T.; Nakashima, R.; Takayama, K.; Okamura, H.; Nakatani, M.; Doe, M.; Shibata, K. “New cembranes from the soft coral Sarcophyton species” J. Nat. Prod. 1999, 62, 1046–1049.
38.Dong, H.; Gou, Y.-L.; Kini, R. M.; Xu, H.-X.; Chen, S.-X.; Teo, L.-M.; But, P.-H. “A new cytotoxic polyhydroxysterol from soft coral Sarcophyton trocheliophorum” Chem. Pharm. Bull. 2000, 48, 1087–1089.
39.Duh, C.-Y.; Wang, S.-K.; Chung, S.-G.; Chou, G.-C.; Dai, C.-F. “Cytotoxic cembrenolides and steroids from the Formosan soft coral Sarcophyton crassocaule” J. Nat. Prod. 2000, 63, 1634–1637.
40.Pham, N. B.; Butler, M. S.; Quinn, R. J. “Naturally occurring cembranes from an Australian Sarcophyton species” J. Nat. Prod. 2002, 65, 1147–1150.
41.Longeon, A.; Bourguet-Kondracki, M.-L.; Guyot, M. “Two new cembrane diterpenoids from a Madagascan soft coral of the genus Sarcophyton” Tetrahedron Lett. 2002, 43, 5937–5939.
42.Gross, H.; Kehraus, S.; Nett, M.; König, G. M.; Beil, W.; Wright, A. D. “New cytotoxic cembrane based diterpenes from the soft corals Sarcophyton cherbonnieri and Nephthea sp.” Org. Biomol. Chem. 2003, 1, 944–949.
43.Kobayashi, M.; Hirase, T. “Marine terpenes and terpenoids. XI. Structures of new dihydrofuranocembranoids isolated from a Sarcophyton sp. soft coral of Okinawa” Chem. Pharm. Bull. 1990, 38, 2442–2445.
44.Sawant, S. S.; Youssef, D. T. A.; Reiland, J.; Ferniz, M.; Marchetti, D,; Sayed, K. A. E. “ Biocatalytic and Antimetastatic Studies of the Marine Cembranoids Sarcophine and 2-epi-16-Deoxysarcophine” J. Nat. Prod. 2006. 69, 1010–1013.
45.Gross, H.; Wright, A. D.; Beil, W.; König, G. M. “Two new bicyclic cembranolides from a new Sarcophyton species and determination of the absolute configuration of Sarcoglaucol-16-one” Org. Biomol. Chem. 2004, 2, 1133–1138.
46.Jia, R.; Guo, Y.-W.; Mollo, E.; Cimino, G. “Sarcophytonolides A-D, four new cembranolides from the Hainan soft coral Sarcophyton sp.” Helv. Chim. Acta. 2005, 88, 1028–1033.
47.Jia, R.; Guo, Y.-W.; Mollo, E.; Gavagnin, M.; Cimino, G. “Sarcophytonolides E-H, cembranolides from the Hainan soft corals Sarcophyton latum” J. Nat. Prod. 2006, 69, 819–822.
48.Zhang, C.; Li, J.; Su, J.; Liang, Y.-J.; Yang, X.-P; Zheng, K.-C; Zeng, L.-M. “Cytotoxic diterpenoids from the soft coral Sarcophyton crassocaule” J. Nat. Prod. 2006, 69, 1476–1480.
49.Huang, H.-C.; Ahmed, A. F.; Su J.-H.; Chao, C.-H.; Wu, Y-C.; Chiang, M.-Y.; Sheu, J.-H. “Crassocolides A-F, cembranoids with a trans-fused lactone from the soft coral Sarcophyton crassocaule” J. Nat. Prod. 2006, 69, 1554–1559.
50.Yan, X.-H.; Li, Z.-Y.; Guo Y.-W. “Further new cembranoid diterpenes from the Hainan soft coral Sarcophyton latum” Helv. Chim. Acta. 2007, 90, 1574–1580.
51.Yan, X.-H.; Gavagnin, M.; Ciminob, G.; Guoa, Y.-W. “Two new biscembranes with unprecedented carbon skeleton and their probable biogenetic precursor from the Hainan soft coral Sarcophyton latum” J. Nat. Prod. 2007, 48, 5313–5316.
52.Jia, R.; Guo, Y.-W.; Chen, P.; Yang, Y.-M.; Mollo, E.; Gavagnin, M.; Cimino, G. “Biscembranoids and their probable biogenetic precursor from the Hainan soft coral Sarcophyton tortuosum” J. Nat. Prod. 2007, 70, 1158–1166.
53.Bishara, A.; Rudi, A.; Benayahu, Y.; Kashman, Y. “Three biscembranoids and their monomeric counterpart cembranoid, a biogenetic diels-alder precursor, from the soft coral Sarcophyton elegans” J. Nat. Prod. 2007, 70, 1951–1954.
54.Cheng, Y.-B.; Shen, Y.-C.; Kuo, Y.-H.; Khalil A.-T. “Cembrane diterpenoids from the Taiwanese soft coral Sarcophyton stolidotum” J. Nat. Prod. 2008, 71, 1141–1145.
55.E. Fridovsky, A. Rudi, Y. Benayahu, Y. Kashman and M. Schleyer, Tetrahedron Lett., 1996, 37, 6909.
56.A. Umeyama, N. Shoji, M. Ozeki and S. Arihara, J. Nat. Prod., 1996, 59, 894.
57.A. S. R. Anjaneyulu, M. V. R. Krishna Murthy and N. S. K. Rao, J. Chem. Res. (S), 1997, 450.
58.A. S. R. Anjaneyulu, M. J. R. V. Venugopal, P. Sarada, G. V. Rao, J. Clardy and E. Lobkovsky, Tetrahedron Lett., 1998, 39, 135.
59.M. Sugano, T. Shindo, A. Sato, Y. Iijima, T. Oshima, H. Kuwano and T. Hata, J. Org. Chem., 1990, 55, 5803.
60.A. S. R. Anjaneyulu, P. M. Gowri and M. V. R. Krishna Murthy, J. Chem. Res. (S), 1999, 140.
61.M. Yasumoto, K. Mada, T. Ooi and T. Kusumi, J. Nat. Prod., 2000, 63, 1534.
62.A. S. R. Anjaneyulu and P. M. Gowri, Indian J. Chem., Sect. B: Org. Chem. Incl. Med. Chem., 2000, 39, 773.
63.V. Anjaneyulu, T. N. Makarieva, S. G. Ilyin, A. S. Dmitrenok, P. Radhika, P. V. Subbarao, V. V. Nesterov, M. Y. Antipin and V. A. Stonik, J. Nat. Prod., 2000, 63, 109.
64.A. S. R. Anjaneyulu, M. V. R. Krishna Murthy and P. M. Gowri, J. Nat. Prod., 2000, 63, 112.
65.T. V. Rezanka and V. M. Dembitsky, Tetrahedron, 2001, 57, 8743.
66.M. Feller, A. Rudi, N. Berer, I. Goldberg, Z. Stein, Y. Benayahu, M. Schleyer and K. Kashman, J. Nat. Prod., 2004, 67, 1303.
67.G.-H. Wang, J.-H. Su, C.-T. Chen, C.-Y. Duh, C.-F. Dai and J.-H. Sheu, J. Chin. Chem. Soc. (Taipei), 2004, 51, 217.
68.L.-M. Zeng,W.-J. Lan, J.-Y. Su, G.-W. Zhang, X.-L. Feng, Y.-J. Liang and X.-P. Yang, J. Nat. Prod., 2004, 67, 1915.
69.V. Sepe, R. D’Orsi, N. Borbone, M. V. D’Auria, G. Bifulco, M. C. Monti, A. Catania and A. Zampella, Tetrahedron, 2006, 62, 833.
70.Y.-C. Shen, Y.-H. Chen, Y.-B. Cheng, Y. H. Kuo and A. T. Khalil, Nat. Prod. Res., 2007, 21, 1171.
71.C.-X. Zhang, S.-J. Yan, G.-W. Zhang, J.-Y. Su and L.-M. Zeng, Chem. J. Chin. Univ., 2007, 28, 686.
72.A. S. R. Anjaneyulu, V. L. Rao, V. G. Sastry and D. V. Rao, J. Asian Nat. Prod. Res., 2008, 10, 597.
73.Yao, L.-G.; Liu, H.-L.; Guo, Y.-W.; Mollo, E. “New Cembranoids from the Hainan Soft Coral Sarcophyton glaucum” Helv. Chim. Acta. 2009, 92, 1085–1091.
74.Zhang, M.; Long, K.; Huang, S.; Shi, K.; Mak, T. C. W. “A Novel Diterpenolide from the Soft Coral Sarcophyton solidun” J. Nat. Prod. 1992, 55, 1672–1675.
75.Kobayashi, M. “Marine Terpenes and Terpenoids. 12. Autoxidation of Dihydrofuranocembranoids” J. Chem. Research (M) 1991, 11, 2840–2852.
76.Quang, T. H.; Ha, T. T.; Minh, C. V.; Kiem, P. V.; Huong, H. T.; Ngan, N. T. T.; Nhiem, N. X.; Tung, N. H.; Tai, B. H.; Thuy, D. T. T.; Song, S. B.; Kang, H.-K.; Kim, Y. H. “Cytotoxic and anti-inflammatory cembranoids from the Vietnamese soft coral Lobophytum laevigatum” Bioorg. Med. Chem. 2011, 19, 2625–2632.
77.陳世斌，台灣海域軟珊瑚Sinularia gibberosa和Sarcophyton sp.所含天然化合物的研究，國立中山大學海洋資源學系博士論文，中華民國96年。
78.Czarkie, D.; Carmely, S.; Groweiss, A.; Kashman, Y. “Attempted acid-catalyzed transannular reactions in the cembranoids” Tetrahedron, 1985 , 41, 1049–1056.
79.Sayed, K. A. E.; Hamann, M. T.; Waddling, C. A.; Jensen, C.; Lee, S. K.; Dunstan, C. A.; Pezzuto, J. M. “Structurally Novel Bioconversion Products of the Marine Natural Product Sarcophine Effectively Inhibit JB6 Cell Transformation”J. Org. Chem., 1998, 63, 7449–7455.
80.林世增，粗厚葉形軟珊瑚Lobophytum crassum二次代謝物之研究，國立中山大學海洋資源學系碩士論文，中華民國98年。
81.Findlay, J. A.; Patil, A. D. “Novel sterols from the finger spong Haliclona oculata” Can. J. Chem., 1985, 63, 2406–2410.
82.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.