臺灣博碩士論文加值系統

為了尋找具有活性和新穎的二次代謝物，本研究針對台灣台東縣三仙台採集的指形軟珊瑚Sinularia nanolobata進行二次代謝物之研究。在過去的研究發現軟珊瑚S. nanolobata有許多不同的骨架的二次代謝物，大部分屬於雙萜類和倍半萜類化合物，包含amphilectane、xeniaphyllane、norxeniaphyllane、caryophyllane和norcaryophyllane等相關的骨架。本研究從S. nanolobata中分離出9個天然化合物，包含了2個新骨架(1和2)、1個新化合物norditerpenoid (3)、2個新化合物norsesquiterpenes (4和5)以及4個已知化合物，gibberosin A (6)、gibberosin J (7)、nanonorcaryphyllene A (8)與pubinernoids A (9)。
這些天然化合物的結構解析及鑑定是以各種有機光譜的分析，包含了一維和二維光譜解析、紅外線光譜、紫外線光譜、比旋光和質譜，並且和已知化合物的文獻比較資料之後確定結構。同時針對這些分離的化合物進行P-388 (小鼠血癌細胞)、HT-29 (人類直腸癌細胞)和A-549 (人類肺癌細胞)毒殺活性測試，發現只有gibberosin J (7)有較顯著抑制效果(ED50 = 1.0, 1.2和0.5 μg/mL)，然而其他化合物則對於以上癌細胞並無顯著的毒殺活性(ED50 > 50 μg/mL)。

In order to dearch for bo-active and novel secondary metabolites, we have investigated the chemical constituents of the soft coral Sinularia nanolobata collected at San-Shen-Tai Taiwan. Soft coral S. nanolobata was found contain diterpenoids and sesquiterpenoids, such as amphilectane, xeniaphyllane, norxeniaphyllane, caryophyllane, and norcaryophyllane. This investigations resulted in the purification of ten compounds, comprising two novel skeletons (1 and 2), a new norditerpenoid (3), two new norsesquiterpenes (4 and 5), together with four known compounds, designated as gibberosin A (6), gibberosin J (7), nanonorcaryphyllene A (8) and pubinernoids A (9).
The structures of these natural compounds were determined on the basis of their spectroscopic analyses (1H NMR, 13C NMR, 1H–1H COSY, HSQC, HMBC, IR, specific rotation and HRESIMS) and comparison of the physical and spectral data with those of the related known compounds. Meanwhile, the cytotoxicity against of P-388 (murine lymphocytic leukemia), HT-29 (human colon adenocarcinoma) and A-549 (human lung epithelial carcinoma) activities of these compounds were evaluated in vitro. Gibberosin J (7) showed significant activity against P-388, HT-29 and A-549 (ED50 = 1.0, 1.2 and 0.5 μg/mL). However, metabolites 1–6, 8, and 9 were inactive (ED50 >50 μg/mL) toward the above cancer cell lines.

論文審定書 i
謝辭 ii
中文摘要 iii
英文摘要 iv
新化合物1−5結構與已知化合物6−9 v
目錄 vi
表次 viii
圖次 ix
縮寫對照表 xiii


壹、緒論 1
一、前言及研究動機與目的 1
二、指形軟珊瑚Sinularia nanolobata二次代謝物研究回顧 2
1. Amphilectane類 3
2. Caryophyllane類 7
3. Norcaryophyllane類 12
4. Xeniaphyllane類 14
5. Norxeniaphyllane類 20

貳、Sinularia nanolobata樣品採集及分離流程 22
一、指形軟珊瑚Sinularia nanolobata樣品採集時間、地點與鑑定 22
二、指形軟珊瑚Sinularia nanolobata之分類地位 22
三、指形軟珊瑚Sinularia nanolobata之萃取與分離流程 23
四、指形軟珊瑚Sinularia nanolobata之分離純化 25
1. SST09-03-01-3-16 (3)之純化分離 25
2. SST09-12-80%(4)-H#2-H22 (1)、SST09-12-80%(4)-H#6-H10 (2)和
SST09-12-80%(4)-H#2.H#3-wash-H3 (6)之純化分離 26
3. SST09-14-3-R5-H12 (8)之純化分離 28
4. SST09-17-3-M10-H15-H2 (4)之純化分離 29
5. SST09-18-10-H7 (5)、SST09-18-12(1)-R3 (7)和 30
SST09-18-12(1)-R1-6-H3 (9)之純化分離

参、化合物之結構解析 32
一、指形軟珊瑚Sinularia nanolobata之化學成分的結構證明 32
1. SST09-12-80%(4)-H#2-H22化合物1之結構解析 32
2. SST09-12-80%(4)-H#6-H10化合物2之結構解析 49
3. SST09-03-01-3-16化合物3之結構解析 67
4. SST09-17-3-M10-H15-H2化合物4之結構解析 87
5. SST09-18-10-H7化合物5之結構解析 104
6. SST09-12-80%(4)-H#2.H#3-wash-H3已知化合物6之結構鑑定 121
7. SST09-18-12(1)-R3已知化合物7之結構鑑定 127
8. SST09-14-3-R5-H12已知化合物8之結構鑑定 132
9. SST09-18-12(1)-R1-6-H3已知化合物9之結構鑑定 139

肆、生物活性之測試 145
一、細胞毒殺活性測試 145
二、MTT分析原理 145
三、抗 HCMV 病毒的測試 146
四、活性測試結果 146

伍、結論 148
陸、參考文獻 152
柒、附錄 155
一、實驗設備儀器及材料 155
1. 實驗設備儀器 155
2. 實驗材料 156

1. 方力行，”珊瑚學”教育部大學聯合出版委員會，中華民國，1989。
2. 邵廣昭，國際海洋年談台灣的海洋生態保育，科學月刊 1989, 29, 978−984.
3. Collado, I. G.；Hanson, J. R.；Macias-Sanchez, A. J., Recent Advances in the Chemistry of Caryophyllene. Nat. Prod. Res. 1998, 15, 187−204
4. Yamada, K.；Ujiie, T.；Yoshida, K.；Miyamoto, T.；Higuchi, R., Sinulobatins A−D, New Amphilectane-type Diterpenoids from the Japanese Soft Coral Sinularia nanolobata. Tetrahedron 1997, 53, 4569−4578.
5. Wratten, S. J.；Faulkner D. J., Diterpenoid Isocyanides from the Marine Sponge Hymeniacidon amphilecta. Tetrahedron Lett. 1978, 45, 4345−4348.
6. Kazlauskas, R.；Murphy, P. T.；Wells, R. J., New Diterpene Isocyanides from a Sponge. Tetrahedron Lett. 1980, 21, 315−318.
7. Molinski, T. F.；Faulkner, D. J., Three New Diterpene Isonitriles from a Palauan Sponge of the Genus Halichondria. J. Org. Chem. 1987, 52, 3334−3337.
8. Ahmed, A. F.；Su, J. H.；Shiue, R. T.；Pan, X. J.；Dai, C. F.；Kuo, Y. H.；Sheu, J. H., New β-Caryophyllene-Derived Terpenoids from the Soft Coral Sinularia nanolobata. J. Nat. Prod. 2004, 67, 592−597.
9. Ahond, A.；Bowden, B. F.；Coll, J. C.；Fourneron, J. D.；Mitchell, S. J., Studies of Australian Soft Corals. XXV* Several Caryophyllene-Based Diterpenes from a Nephthea Species. Aust. J. Chem. 1981, 34, 2657−2664.
10. Thebtaranonth, C.；Thebtaranonth, Y.；Wanauppathamkul, S.；Yuthavong, Y., Antimalarial Sesquiterpenes from Tubers of Cyperus rotundus: Structure of 10, 12-peroxycalamenene, a Sesquiterpene Endoperoxide. Phytochemistry 1995, 40, 125−128.
11. Chen, S. P.；Ahmed, A. F.；Dai, C. F.；Lu, C. K.；Hu, W. P.；Wang, J. J.；Sheu, J. H., Sinugibberosides A-E, New Terpenoids with Cyclicperoxyhemiketal from the Soft Coral Sinularia gibberosa. Tetrahedron 2006, 62, 6802−6807.
12. Bohlmann, F.；Zdero, C., Neue Furanoeremophilane und Andere Sesquiterpene aus Vertretern der Gattung Euryops. Phytochemistry 1978, 17, 1135−1153.
13. Bohlmann, F.；Ziesche, J., Sesquiterpenes from Three Senecio Species. Phytochemistry 1981, 20, 469−472.
14. Yoshida, T.；Nobuhara, J.；Uchida, O.；Okuda, T., Studies on the Constituents of Buddleja Species. I. Structures of Buddledin A and B, Two New Toxic Sesquiterpenes from Buddleja davidii Franch. Chem. Pharm. Bull. 1978, 26, 2535−2542.
15. Abraham, W. R.；Ernst, L.；Stumpf, B., Biotransformation of Caryophyllene by Diplodia gossypina. Phytochemistry 1990, 29, 115−120.
16. Demirci, B.；Can Baser, K. H.；Demirci, F.；Hamann, M. T., New Caryophyllene Derivatives from Betula litwinowii. J. Nat. Prod. 2000, 63, 902−904.
17. Chen, S. P.；Chao, C. H.；Huang, H. C.；Wu, Y. C.；Lu, C. K.；Dai, C. F.；Sheu, J. H., New β-Caryophyllene-Derived Terpenoids from the Formosan Soft Coral Sinularia gibberosa. Bull. Chem. Soc. Jpn. 2006, 79, 1547−1551.
18. Tseng, Y. J.；Wen, Z. H.；Dai, C. F.；Chiang, M. Y.；Sheu, J. H., Nanolobatolide, a New C18 Metabolite from the Formosan Soft Coral Sinularia nanolobata. Org. Lett. 2009, 11, 5030−5032.
19. Fattorusso, C.；Stendardo, E.；Appendino, E. F.；Luciano, P.；Rpmano, A.；Orazio, T. S., Artarborol, a nor-Caryophyllane Sesquiterpene Alcohol from Artemisia arborescens. Stereostructure Assignment through Concurrence of NMR Data and Computational Analysis. Org. Lett. 2007, 9, 2377−2380.
20. Groweiss, A.；Kashman, Y., Xeniculin, Xeniaphyllenol and Xeniaphyllenol, New Diterpenoids from the Soft Coral Xenia macrospiculata. Tetrahedron Lett. 1978, 25, 2205−2208.
21. Kashman, Y.；Groweiss, A., New Diterpenoids from the Soft Corals Xenia macrospiculata and Xenia obscuronata. J. Org. Chem. 1980, 45, 3814−3824.
22. Groweiss, A.；Kashman, Y., Eight New Xenia Diterpenoids from Three Soft Corals of the Red Sea. Tetrahedron 1983, 39, 3385−3396.
23. Chen, S. P.；Su, J. H.；Ahmed, A. F.；Dai, C. F.；Wu, Y. C.；Sheu, J. H., Xeniaphyllane-Derived Terpenoids from the Formosan Soft Coral Sinularia gibberosa. Chem. Pharm. Bull. 2007, 55, 1471−1475.
24. Chen, S. P.；Su, J. H.；Yeh, H. C.；Ahmed, A. F.；Dai, C. F.；Wu, Y. C.；Sheu, J. H., Novel Norhumulene and Xeniaphyllane-Derived Terpenoids from a Formosan Soft Coral Sinularia gibberosa. Chem. Pharm. Bull. 2009, 57, 162−166.
25. Huang, S. X.；Yang, J.；Xiao, W. L.；Zhu, Y. L.；Li, R. T.；Ni, L. M.；Pu, J. X.；Li, X.；Li, S. H.；Sun, H. D., Three Novel Terpenoids from Schisandra pubescens var. pubinervis. Helv. Chim. Acta 2006, 89, 1169−1175.