本研究分別探討從臺灣海域採集的短指形軟珊瑚 Sinularia humilis 及松球小枝軟珊瑚Cladiella conifera所含之天然化合物成分。從短指形軟珊瑚之中得到1個環八硫化合物 cyclooctasulfur (1) 及3個固醇類化合物5α,8α-epidioxy-24-methylcholesta-6,9(11),-24(28)-trien-3β-ol (2)、5α,8α-epidioxy-24-methylcholesta-6,24(28)-dien-3β-ol (3)、(22R,23R,24R)-5R, 8R-epidioxy-22,23-methylene-24-methyl cholest-6-en-3β-ol (4)。在松球小枝軟珊瑚中總共得到10個化合物，包含了4個新eunicellin類雙萜化合物coniferains A–D (5–8)，1個首次從自然界中獲得的butenolide類化合物 (5R)-3-tetradecyl-5-methyl-2(5H)-furanone (14)，及5個已知eunicellin類化合物 multifloralin (9)、litophynin F (10)、krempfielin J (11)、(1R*,2R*,3R*,6S*,7S*,9R*,10R*,14R*)-3-butanoyloxycladiell-11(17)-en-6,7-diol (12)、litophynin I monoacetate (13)。化合物 1–14 的結構是經由光譜數據分析及比對相關文獻而確定。
本研究將化合物 2–9、12 及 13 行癌細胞毒殺測試以及將化合物 14 進行抗發炎活性測試。化合物2、3 作用下對癌細胞株HT-29 的 IC50 分別為18.9及16.7 μM。化合物 3 作用下對癌細胞株DLD-1的IC50 為33.4 μM。以20 μM的化合物 7、8和 13 作用下對癌細胞株DLD-1具備促細胞生長的活性，分別促進至158.8、145.8、129.5 %。另外以抗發炎活性當中，於10 μM 濃度作用下，化合物 14 抑制發炎相關蛋白COX-2使其表現量下降至 84.5%。

Studies on the chemical constituents of soft corals Sinularia humilis and Cladiella conifera. Cyclooctasulfur (1) and three steroids, 5α,8α-epidioxy-24-methylcholesta-6,9(11),-24(28)-trien-3β-ol (2), 5α,8α-epidioxy-24-methylcholesta-6,24(28)-dien-3β-ol (3), (22R,23R,24R)-5R, 8R-epidioxy-22,23-methylene-24-methylcholest-6-en-3β-ol (4), were isolated from S. humilis. In addition, ten secondary metabolites were purified from the soft coral C. conifera. According to the chemical classifications, there were nine eunicellin-type diterpenoids, including four new compounds, coniferain A–D (5–8), together with five known analogs, multifloralin (9), litophynin F (10), krempfielin J (11), (1R*,2R*,3R*,6S*,7S*,9R*,10R*,14R*)-3-butanoyloxycladiell-11(17) -en-6,7-diol (12) and litophynin I monoactate (13), as well as a new natural butanolide-type compound, (5R)-3-tetradecyl-5-methyl-2(5H)-furanone (14). The structures of compounds 1–14 were established by spectroscopic methods, and comparison of the experimental NMR data and those of literatures.
The cytotoxic activities of 2-9, 12 and 13 were evaluated toward HL-60, HT-29 and DLD-1 cell lines. Compound 14 was evaluated the anti-inflammatory activity. Compounds 2 and 3 have cytotoxic activity against cancer cell line HT-29 with IC50 of 18.9 and 16.7 μM. In addition, compound 3 also have cytotoxic activity against cancer cell line DLD-1 with IC50 of 33.4 μM. However, compounds 7, 8 and 13 against cell lines DLD-1 did promote cell viability in DLD-1 cells at a concentration of 20 μΜ. Moreover, 14 inhibited to 84.5% of COX-2 in LPS stimulated RAW 264.7 macrophage cells at a concentration of 10 μΜ.

中文摘要 I
Abstract III
目錄 VII
圖目錄 IX
表目錄 XIII
第一章、緒論 1
第一節、前言 1
第二節、研究目的 2
第三節、Sinularia 屬帶硫化合物及S. humilis 珊瑚之文獻顧 2
第四節、Cladiella 屬珊瑚所含 eunicellin 類天然物之文獻回顧 8
第二章、生物材料與研究辦法 23
第一節、生物樣品的採集與分類 23
第二節、生物樣品的萃取與分離 25
第三章、實驗器材與有機溶液 29
第一節、儀器 29
第二節、其他器材與有機溶液 31
四章、生物活性試驗 33
第一節、抗發炎活性測試 33
第二節、細胞毒殺活性 34
第五章、結果與討論 37
第一節、短指形珊瑚 Sinularia humilis 所分離出化學成分之結構分析 37
(一)、Cyclooctasulfur (1) 之結構解析 37
(二)、5α,8α-Epidioxy-24-methylcholesta-6,9(11),24(28)-trien-3β-ol (2)之結構解析 41
(三)、5α,8α-Epidioxy-24-methylcholesta-6,24(28)-dien-3β-ol (3)之結構解析 47
(四)、(22R,23R,24R)-5R,8R-epidioxy-22,23-methylene-24-methylcholest-6-en-3β-ol (4)之結構解析 53
第二節、松球小枝軟珊瑚 Cladiella conifera 所分離出化學成分之結構分析 59
(五)、Coniferain A (5) 之結構解析 59
(六)、Coniferain B (6) 之結構解析 69
(七)、Coniferain C (7) 之結構解析 79
(八)、Coniferain D (8) 之結構解析 89
(九)、Multifloralin (9) 之結構解析 99
(十)、Litophynin F (10) 之結構解析 105
(十一)、Krempfielin J (11)之結構解析 111
(十二)、(1R*,2R*,3R*,6S*,7S*,9R*,10R*,14R*)-3-Butanoyloxycladiell-11(17)-en-6,7-diol (12) 之結構析 117
(十三)、Litophynin I monoactate (13) 之結構解析 123
(十四)、(5R)-3Ttetradecyl-5-methyl-2(5H)-furanone (14) 之結構解析 129
第三節、化合物數據實驗整理 137
第三節、化合物活性試驗數據 141
第六章、結論 143
參考文獻 145

1. Solecki, R. S. Shanidar IV, a Neanderthal flower burial in Northern Iraq. Science 1975, 190, 880–881.
2. Stewart, T. D. Shanidar skeletons IV and VI. Sumer. 1963, 19, 8–26.
3. Solomon, P. W. Effects of birch polypore mushroom, Piptoporus betulinus (Agaricomycetes), the "Iceman’s Fungus", on human immune cells. Int. J. Med. Mushrooms 2019, 21, 1135–1147.
4. Atanasov, A. G.; Waltenberger, B.; Pferschy-Wenzig, E. M.; Linder, T.; Wawrosch, C.; Uhrin, P. Discovery and resupply of pharmacologically active plant-derived natural products: A review. Biotechnol. Adv. 2015, 33, 1582–1614.
5. Grene, M.; David, D. The philosophy of biology: an episodic history. C. U. P.: Cambridge, 2004.
6. Unschuld, P. Huang Di Nei Jing Su Wen: Nature, knowledge, imagery in an ancient chinese medical text. U. C. Press: California, 2003.
7. Gilles, B. Marine biodiversity characteristics. C. R. Biol. 2011, 334, 435–440.
8. Roberts, T. R.; Hutson, D. H. Nereistoxin precursors. Metab. Pathw. Argo. 2007, 127–138.
9. Scheuer, P. J.; Lazarovici, P.; Spira, M. E.; Zlotkin.; Eliahu. Eds.; Alaken. Marine metabolites as drug leads-retrospect and prospect. Biochem. Aspect. Mar. Pharmacol. 1996, 1–12.
10. Ruggieri, G. D. Drugs from the sea. Science 1976, 194, 491–497.
11. Nweze, J. A.; Mbaoji, F. N.; Li, Y. M.; Huang, S. S.; Chigor, V. N.; Eze, E. A.; Pan, L. X.; Zhang, T.; Yang, D. F. Potentials of marine natural products against malaria, leishmaniasis, and trypanosomiasis parasites: A review of recent articles. Infect. Dis. Poverty 2021, 10, 9.
12. Chen, B. W.; Chao, C. H.; Su, J. H.; Huang, C. Y.; Dai, C. F.; Wen, Z. H.; Sheu, J. H. A novel symmetric sulfurcontaining biscembranoid from the Formosan soft coral Sinularia flexibilis. Tetrahedron Lett. 2010, 51, 5764–5766.
13. Putra, M. Y.; Ianaro, A.; Panza, E.; Bavestrello, G.; Cerrano, C.; Fattorusso, E.; Taglialarela-Scafati, O. Sinulasulfoxide and sinulasulfone, sulfur-containing alkaloids from the Indonesian soft coral Sinularia sp. Tetrahedron Lett. 2012, 53, 3937–3939.
14. Lei, L. F, et al. Novel cytotoxic nine-membered macrocyclic polysulfur cembranoid lactones from the soft coral Sinularia sp. Tetrahedron 2017, 70, 6851–6858.
15. Sun, P.; Meng, L. Y.; Tang, H.; Liu, B. S.; Li, L.; Yi, Y.; Zhang, W. Sinularosides A and B, bioactive 9, 11-secosteroidal glycosides from the South China Sea soft coral Sinularia humilis Ofwegen. J. Nat. Prod. 2012, 75, 1656–1659.
16. Li, J.; Huan, X. J.; Wu, M. J.; Chen, Z. H.; Chen, B.; Miao, Z. H.; Li, X. W. Chemical constituents from the South China sea soft coral Sinularia humilis. Nat. Prod. Res. 2022, 36, 3324–3330.
17. Sun, L. L.; Li, W. S.; Li, J.; Zhang, H. Y.; Yao, L. G.; Luo, H.; Li, X. W. Uncommon diterpenoids from the south china sea soft coral Sinularia humilis and their stereochemistry. J. Org. Chem. 2021, 86, 3367–3376.
18. Tai, C. J.; Su, J. H.; Huang, M. S.; Wen, Z. H.; Dai, C. F.; Sheu, J. H. Bioactive eunicellin-based diterpenoids from the soft coral Cladiella krempfi. Mar. Drugs 2011, 9, 2036–2045.
19. Tai, C. J.; Su, J. H.; Huang, C. Y.; Huang, M. S.; Wen, Z. H.; Dai, C. F.; Sheu, J. H. Cytotoxic and anti-inflammatory eunicellin-based diterpenoids from the soft coral Cladiella krempfi. Mar. Drugs 2013, 11, 788–799.
20. Lee, Y. N.; Tai, C. J.; Hwang, T. L.; Sheu, J. H. Krempfielins J–M, new eunicellin-based diterpenoids from the soft coral Cladiella krempfi. Mar. Drugs 2013, 11, 2741–2750.
21. Lee, Y. N.; Tai, C. J.; Hwang, T. L.; Sheu, J. H. Krempfielins N–P, new anti-inflammatory eunicellins from a Taiwanese soft coral Cladiella krempfi. Mar. Drugs 2014, 12, 1148–1156.
22. Ahmed, A. F.; Wu, M. H.; Wang, G. H.; Wu, Y. C.; Sheu, J. H. Eunicellin-Based Diterpenoids, Australins A−D, Isolated from the soft Coral Cladiella australis. J. Nat. Prod. 2005, 68, 1051–1055.
23. Chen, B. W.; Chang, S. M.; Huang, C. Y.; Chao, C. H.; Su, J. H.; Wen, Z. H.; Sheu, J. H. Hirsutalins A−H, eunicellin-based diterpenoids from the soft coral Cladiella hirsuta. J. Nat. Prod. 2010, 73, 1785–1791.
24. Chen, B. W.; Wang, S. Y.; Huang, C. Y.; Chen, S. L., Wu, Y. C.; Sheu, J. H. Hirsutalins I–M, eunicellin-based diterpenoids from the soft coral Cladiella hirsuta. Tetrahedron 2013, 69, 2296–2301.
25. Huang, T. Z.; Chen, B. W.; Huang, C. Y.; Hwang, T. L.; Dai, C. F.; Sheu, J. H. Eunicellin-based diterpenoids, hirsutalins N–R, from the Formosan soft coral Cladiella hirsuta. Mar. Drugs 2014, 12, 2446–2457.
26. Huang, T. Z.; Chen, B. W.; Huang, C. Y.; Hwang, T. L.; Uvarani, C.; Dai, C. F.; Sheu, J. H. Eunicellin-Based Diterpenoids, Hirsutalins S–V, from the Formosan Soft Coral Cladiella hirsuta. Mar. Drugs 2015, 13, 2757–2769.
27. Williams, D. E.; Amlani, A.; Dewi, A. S.; Patrick, B. O.; Van Ofwegen, L.; Mui, A. L. F.; Andersen, R. J. Australin E isolated from the soft coral Cladiella sp. collected in pohnpei activates the inositol 5-phosphatase SHIP1. Aust. J. Chem. 2010, 63, 895–900.
28. Chen, Y. H.; Tai, C. Y.; Hwang, T. L.; Weng, C. F.; Li, J. J.; Fang, L. S.; Sung, P. J. Cladielloides A and B: new eunicellin-type diterpenoids from an Indonesian octocoral Cladiella sp. Mar. Drugs 2010, 8, 2936–2945.
29. Tai, C. Y.; Chen, Y. H.; Hwang, T. L.; Fang, L. S.; Wang, W. H.; Liu, M. C.; Sung, P. J. Cladielloides C and D: Novel eunicellin-based diterpenoids from an Indonesian octocoral Cladiella sp. Bull. Chem. Soc. Jpn. 2011, 84, 531–536.
30. Chen, Y. H.; Tai, C. Y.; Kuo, Y. H.; Kao, C. Y.; Li, J. J.; Hwang, T. L.; Sung, P. J. Cladieunicellins A–E, new eunicellins from an Indonesian soft coral Cladiella sp. Chem. Pharm. Bull. 2011, 59, 353–358.
31. Chen, Y. H.; Tai, C. Y.; Su, Y. D.; Chang, Y. C.; Lu, M. C.; Weng, C. F.; Sung, P. J. Discovery of new eunicellins from an Indonesian octocoral Cladiella sp. Mar. Drugs 2011, 9, 934–943.
32. Chen, Y. H.; Hwang, T. L.; Su, Y. D.; Chang, Y. C.; Chen, Y. H.; Hong, P. H.; Sung, P. J. New 6-hydroxyeunicellins from a soft coral Cladiella sp. Chem. Pharm. Bull. 2012, 60, 160–163.
33. Chen, Y. H.; Tai, C. Y.; Hwang, T. L.; Sung, P. J. Cladieunicellin H, a new hemiketal eunicellin-based diterpenoid from the octocoral Cladiella sp. Nat. Prod. Commun. 2012, 7, 481–484.
34. Chen, T. H.; Lu, M. C.; Chang, Y. C.; Su, Y. D.; Chen, Y. H.; Lin, N. C.; Sung, P. J. Discovery of new eunicellin-based diterpenoids from a Formosan soft coral Cladiella sp. Mar. Drugs 2013, 11, 4585–4593.
35. Chen, T. H.; Cheng, C. H.; Chen, Y. H.; Lu, M. C.; Fang, L. S.; Chen, W. F.; Sung, P. J. Cladieunicellin J, a new hydroperoxyeunicellin from Cladiella sp. Nat. Prod. Commun. 2014, 9, 613–614.
36. Shih, F. Y.; Chen, T. H.; Lu, M. C.; Chen, W. F.; Wen, Z. H.; Kuo, Y. H.; Sung, P. J. Cladieunicellins K and L, new eunicellin-based diterpenoids from an octocoral Cladiella sp. Int. J. Mol. Sci. 2013, 14, 21781–21789.
37. Chen, T. H.; Chen, W. F.; Wen, Z. H.; Lu, M. C.; Wang, W. H.; Li, J. J.; Sung, P. J. Cladieunicellins M–Q, new eunicellins from Cladiella sp. Mar. Drugs 2014, 12, 2144–2155.
38. Peng, K. Y.; Chen, N. F.; Chen, Z. C.; Tsui, K. H.; Wen, Z. H.; Su, Y. D.; Sung, P. J. Cladieunicellins R and S, new eunicellins from the Formosan octocoral Cladiella tuberculosa. Tetrahedron Lett. 2016, 57, 4239–4242.
39. Zhang, Z. J.; Peng, B. R.; Hu, C. C.; Lin, N. C.; Yao, J. W.; Lu, M. C.; Sung, P. J. Cladieunicellin T, a new eunicellin-based diterpenoid produced by the octocoral Cladiella sp. Nat. Prod. Commun. 2019, 14, 101–102.
40. Makariewa, T. N.; Stonik, V. A.; Dmitrenok, A. S.; Grebnev, B. B,; Isakov, V. V.; Rebachyk, N. M.; Rashkes, Y. W. Varacin and three new marine antimicrobial polysulfides from the Far-Eastern ascidian Polycitor sp. J. Nat. Prod. 1995, 58, 254–258.
41. Gunatilaka, A. A. L.; Gopichand, Y.; Schmitz, F. J.; Djerassi, C. Minor and trace sterols in marine invertebrates. 26. Isolation and structure elucidation of nine new 5α,8α-epidoxy sterols from four marine organisms. J. Org. Chem. 1981, 46, 3860–3866.
42. Yang, M.; Liang, L. F.; Li, H.; Tang, W.; Guo, Y. W. A new 5α,8α-epidioxysterol with immunosuppressive activity from the south china sea soft coral Sinularia sp. Nat. Prod. Res. 2019, 34, 1–6.
43. Radhika, P. Chemical constituents and biological activities of the soft corals of genus Cladiella: A review. Biochem. Syst. Ecol. 2006, 34, 781–789.
44. Li, G.; Dickschat, J. S.; Guo, Y. W. Diving into the world of marine 2,11-cyclized cembranoids: A summary of new compounds and their biological activities. Nat. Prod. Rep. 2020, 37, 1367–1383.
45. Wang, Z.; Li, P. L.; Luo, X. C.; Wang, Q.; Van Ofwegen, L.; Tang, X. L.; Li, G. Q. Terpenoids from the South China Sea soft coral Sinularia multiflora. Nat. Prod. Res. 2021, 35, 2395–2402.
46. Ochi, M.; Yamada, K.; Futatsugi, K.; Kotsuki, H. Litophynins F, G, and H, Three new diterpenoids from a soft coral Litophyton sp. Heterocycles 1991, 32, 29–32.
47. Rao, C. B.; Rao, D. S.; Satyanarayana, C.; Rao, D. V.; Kassühlke, K. E.; Faulkner, D. J. New cladiellane diterpenes from the soft coral Cladiella australis of the Andaman and Nicobar Islands. J. Nat. Prod. 1994, 57, 574–580.
48. Miyamoto, T.; Yamada, K.; Ikeda, N.; Komori, T.; Higuchi, R. Bioactive terpenoids from Octocorallia, I. Bioactive diterpenoids: litophynols A and B from the mucus of the soft coral Litophyton sp. J. Nat. Prod. 1994, 57, 1212–1219.
49. Gawronski, J. K.; Gawronski, J. K; Van Oeveren, A.; Der Deen, V. H.; Leung, C. W.; Feringa, B. L. Simple circular dichroic method for the determination of absolute configuration of 5-substituted 2(5H)-furanones. J. Chem. Soc. Chem. Commun. 1994, 2371.
50. Zeng, B. B.; Hu, T. S.; Yun, W.; Wu, Y.; Wu, Y. L. Straightforward synthesis of two pairs of enantiomeric butenolides 3-tetradecyl-and 3-hexadecyl-5-methyl-2(5H)-furanone. Enantiomer 2002, 7, 133–137.
51. Ortuño, R. M.; Bigorra, J.; Font, J. Stereocontrolled synthesis of (3S,4S,5S)-3-alkyl-4-hydroxy-5-methyl-2 (3H)-dihydrofubanones and derivatives: Configurational assignment of some Clinostemon mahuba and Plexaura flava metabolites. Tetrahedron 1988, 44, 5139–5144.
52. Pjevac, P.; Kamyshny, A.; Dyksma, S.; Muβmann, M. Microbial consumption of zero-valence sulfur in marine benthic habitats. Environ. Microbiol. 2014, 16, 3416–3430.
53. Tasaka, K.; Mio, M.; Izushi, K.; Akagi, M.; Makino, T. Anti-allergic constituents in the culture medium of Ganoderma lucidum. (II). The inhibitory effect of cyclooctasulfur on histamine release. Agents Actions 1988, 23, 157–160.
54. Hayashi, H.; Tarui, N.; Murao, S. Isolation and identification of cyclooctasulfur, a fruiting-body inducing substance, produced by Streptomyces albulus TO 447. Agric. Biol. Chem. 1985, 49, 101–105.
55. Lognay, G.; Seck, D.; Marlier, M.; Haubruge, E.; Gaspar, C.; Severin, M. Identification of elemental sulfur (S8) in Boscia senegalensis (Pers.) Lam ex poir. Leaves. Bull. Rech. Agron. Gembloux 1993, 28, 501–505.
56. Li, N.; Li, X.; Yang, S. L.; Zhang, P. Isolation and identification of heteroatom-containing and polyols constituents from Comptosorus sibiricus rupr. Chin. J. Med. Chem. 2004, 14, 368–369.
57. Gao, Y. T.; Yang, X. W.; Ai, T. M. Studies on the chemical constituents in herbs of ethanolic extract from herbs of Dicliptera chinensis. China J. Chin. Mater. Med. 2006, 31, 985–987.
58. Faizi, S.; Sumbul, S.; Versiani, M. A.; Saleem, R.; Sana, A.; Siddiqui, H. GC/GCMS analysis of the petroleum ether and dichloromethane extracts of Moringa oleifera roots. Asian Pac. J. Trop. Biomed. 2014, 4,650–654.
59. Holmer, M.; Frederiksen, M. S.; Møllegaard, H. Sulfur accumulation in eelgrass (Zostera marina) and effect of sulfur on eelgrass growth. Aquat. Bot. 2005, 81, 367–379.
60. Kennard, O.; Watson, D. G.; Rivadi Sanseverino, L.; Tursch, B.; Bosmans, R.; Djerassi, C. Eunicellin, A diterpenoid of the Gorgonian Eunicella stricta. X-ray diffraction analysis of eunicellin dibromide. Tetrahedron Lett. 1968, 9, 1879–2884.
61. Sheu, J. H.; Chang, K. C.; Duh, C. Y. A cytotoxic 5α, 8α-epidioxysterol from a Soft Coral Sinularia species. J. Nat. Prod. 2000, 63, 149–151.
62. Chen, B. W.; Chao, C. H.; Su, J. H.; Huang, C. Y.; Dai, C. F.; Wen, Z. H.; Sheu, J. H. A novel symmetric sulfurcontaining biscembranoid from the Formosan soft coral Sinularia flexibilis. Tetrahedron Lett. 2010, 51, 5764–5766.
63. Putra, M. Y.; Ianaro, A.; Panza, E.; Bavestrello, G.; Cerrano, C.; Fattorusso, E.; Taglialatela-Scafati, O. Sinulasulfoxide and sinulasulfone, sulfur-containing alkaloids from the Indonesian soft coral Sinularia sp. Tetrahedron Lett. 2012, 53, 3937–3939.
64. Lei, L. F.; Chen, M. F.; Wang, T. Novel cytotoxic nine-membered macrocyclic polysulfur cembranoid lactones from the soft coral Sinularia sp. Tetrahedron 2017, 70, 6851–6858.
65. Welford, A. J.; Collins, I. The 2, 11-cyclized cembranoids: Cladiellins, asbestinins, and briarellins (period 1998–2010), J. Nat. Prod. 2011, 74, 2318–2328.
66. Chen, W. T., Li, Y.; Guo, Y. W. Terpenoids of Sinularia soft corals: Chemistry and bioactivity. Acta Pharm. Sin. B. 2012, 2, 227–237.