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研究生:陳華平
研究生(外文):Hwa-Pyng Chen
論文名稱:葉形軟珊瑚Lobophytum durum二次代謝物之研究
論文名稱(外文):Studies on the Secondary Metabolites from the Soft CoralLobophytum durum
指導教授:杜昌益教授
指導教授(外文):Chang-Yih Duh
學位類別:碩士
校院名稱:國立中山大學
系所名稱:海洋生物科技暨資源學系研究所
學門:自然科學學門
學類:海洋科學學類
論文種類:學術論文
論文出版年:2011
畢業學年度:99
語文別:中文
論文頁數:157
中文關鍵詞:葉型軟珊瑚(Lobophytum durum)抗感染活性海雞頭科(Alcyoniidae)雙萜類(cembrane diterpenes)抗菌活性
外文關鍵詞:Lobophytum durumcembrane diterpenesanti-inflammatory propertiesantibacterial activitiesAlcyoniidae
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軟珊瑚中海雞頭科葉形軟珊瑚屬(Alcyoniidae, Lobophytum)已知富含有許多二次代謝物,由於他們複雜的結構和可能具有值得探討的藥理活性,如:抗癌細胞的毒性、抗菌活性、抗感染及抗病毒活性等,引起了天然物化學家廣泛的興趣。本論文從東沙環礁海域的葉型軟珊瑚(Lobophytum durum)萃取有機層,分離得到十二個雙萜類化合物(cembrane diterpenes),包括六個新的二次代謝物(1−6)。這六個新的二次代謝物的化學結構主要是利用各種光譜進行分析(IR, HRMS, 1D NMR及2D NMR)並運用 1H, 13C, DEPT, HSQC , COSY, HMBC, NOESY, IR及MS等光譜資料而決定。此外這六個新的二次代謝物並進行人類肺癌細胞(A-549)、人類直腸癌細胞(HT-29)、老鼠血癌細胞(P-388)毒殺活性的測試及也進行抗人類巨細胞病毒(HCMV)測試。
Soft corals of the genus Lobophytum (Alcyoniidae) have been well recognized as a rich source of various secondary metabolites that have attracted much interest for the natural products chemists due to their structural complexity and remarkable pharmacological activities such as cytotoxicity, antibacterial activities, anti-inflammatory properties, and antiviral activity. Twelve cembrane diterpenes including six new secondary metabolites 1−6 were isolated organic extracts of soft coral Lobophytum durum collected at Dongsha Atolls. The structures of these six new cembranolides were determined by 1H, 13C, DEPT, COSY, HMBC, HSQC, NOESY, IR and Mass spectra. Furthermore, these six new secondary metabolites 1−6 were evaluated in vitro for the cytotoxicity against A-459 (human lung carcinoma), HT-29 (human colon adenocarcinoma), and P-388 (mouse lymphocytic leukemia) cancer cell lines, and antiviral activity against HCMV (human cytomegalovirus) cells.
目錄
頁次
壹、緒論 1
一、前言 1
二、葉型軟珊瑚屬(Lobophytum) cembrane 型二次
代謝物之研究回顧 3
三、研究動機與目的 16
貳、材料與研究方法 17
一、葉型軟珊瑚Lobophytum durum樣品採集時間
、地點與鑑定 17
二、葉型軟珊瑚Lobophytum durum之分類地位 17
三、葉型軟珊瑚Lobophytum durum之萃取與分離
流程 18
参、化學成分之結構證明 22
一、葉型軟珊瑚Lobophytum durum之化學成分的
結構證明 22
1.化合物1之結構解析 22
2.化合物2之結構解析 41
3.化合物3之結構解析 55
4.化合物4之結構解析 71
5.化合物5之結構解析 88
6.化合物6之結構解析 104
肆、生物活性之測試 125
一、細胞毒殺活性測試方法 125
二、MTT分析原理 126
三、抗HCMV病毒的測試方法 127
四、活性測試結果 128
伍、結論 130
陸、參考文獻 133
柒、附錄 138
一、Mosher醯化反應的原理與製備 138
二、實驗設備儀器及材料 140
1.實驗設備儀器 140
2.實驗材料 141
表目錄
頁次
表一、化合物1之物理性質與圖譜資料(in CDCl3) 26
表二、化合物2之物理性質與圖譜資料(in CDCl3) 43
表三、化合物3之物理性質與圖譜資料(in CDCl3) 59
表四、化合物4之物理性質與圖譜資料(in CDCl3) 74
表五、化合物5之物理性質與圖譜資料(in CDCl3) 91
表六、化合物6之物理性質與圖譜資料(in CDCl3) 108
表七、化合物1−6之癌細胞毒殺活性 128
表八、化合物1、4及6之抗人類細胞病毒之活性 129
圖目錄
頁次
圖A、葉型軟珊瑚Lobophytum durum之萃取與分離
流程 19
圖B、以RP-18 HPLC (250×25mm)分離出化合物2
、3和6之層析圖 20
圖C、以RP-18 HPLC (250×25mm)分離出化合物4
和5之層析圖 21
圖1−1.化合物1之高解析ESI質譜 27
圖1−2.化合物1之低解析ESI質譜 27
圖1−3.化合物1之紅外光譜 28
圖1−4.化合物1之氫譜(in CDCl3, 400 MHz) 29
圖1−5.化合物1之局部放大低場區氫譜(in CDCl3) 30
圖1−6.化合物1之局部放大低場區氫譜 (in CDCl3) 31
圖1−7.化合物1之局部放大高場區氫譜 (in CDCl3) 32
圖1−8.化合物1之碳譜與DEPT圖譜(in CDCl3, 100MHz) 33
圖1−9.化合物1之HSQC關聯圖(in CDCl3) 34
圖1−10.化合物1之局部放大HSQC關聯圖(in CDCl3) 35
圖1−11.化合物1之COSY關聯圖(in CDCl3) 36
圖1−12.化合物1之HMBC關聯圖(in CDCl3) 37
圖1−13.化合物1之局部放大HMBC關聯圖(in CDCl3) 38
圖1−14.化合物1之NOESY關聯圖(in CDCl3) 39
圖1−15.化合物1與已知sinularolide B (73)氫值、碳值與NOE的關聯圖 40
圖2−1.化合物2之高解析ESI質譜 44
圖2−2.化合物2之低解析ESI質譜 44
圖2−3.化合物2之紅外光譜 45
圖2−4.化合物2之氫譜(in CDCl3, 500 MHz) 46
圖2−5.化合物2之局部放大低場區氫譜(in CDCl3) 47
圖2−6.化合物2之局部放大高場區氫譜 (in CDCl3) 48
圖2−7.化合物2之碳譜(in CDCl3, 125MHz) 49
圖2−8.化合物2之HSQC關聯圖(in CDCl3) 50
圖2−9.化合物2之COSY關聯圖(in CDCl3) 51
圖2−10.化合物2之HMBC關聯圖(in CDCl3) 52
圖2−11.化合物2之NOESY關聯圖(in CDCl3) 53
圖2−12.化合物2與已知sinularolide B (73)氫值、碳值與NOE的關聯圖 54
圖3−1.化合物3之高解析ESI質譜 60
圖3−2.化合物3之低解析ESI質譜 60
圖3−3.化合物3之紅外光譜 61
圖3−4.化合物3之氫譜(in CDCl3, 500 MHz) 62
圖3−5.化合物3之局部放大低場區氫譜(in CDCl3) 63
圖3−6.化合物3之局部放大高場區氫譜(in CDCl3) 64
圖3−7.化合物3之碳譜(in CDCl3, 125MHz) 65
圖3−8.化合物3之HSQC關聯圖(in CDCl3) 66
圖3−9.化合物3之COSY關聯圖(in CDCl3) 67
圖3−10.化合物3之HMBC關聯圖(in CDCl3) 68
圖3−11.化合物3與已知13-hydroxylobolide (17)氫值與碳值之比較圖 69
圖3−12.化合物3之NOESY關聯圖(in CDCl3) 70
圖4−1.化合物4之高解析ESI質譜 75
圖4−2.化合物4之低解析ESI質譜 75
圖4−3.化合物4之紅外光譜 76
圖4−4.化合物4之氫譜(in CDCl3, 400 MHz) 77
圖4−5.化合物4之局部放大低場區氫譜(in CDCl3) 78
圖4−6.化合物4之局部放大高場區氫譜(in CDCl3) 79
圖4−7.化合物4之碳譜與DEPT圖譜(in CDCl3, 100MHz) 80
圖4−8.化合物4之HSQC關聯圖(in CDCl3) 81
圖4−9.化合物4之局部放大HSQC關聯圖(in CDCl3) 82
圖4−10.化合物4之COSY關聯圖(in CDCl3) 83
圖4−11.化合物4之HMBC關聯圖(in CDCl3) 84
圖4−12.化合物4之NOESY關聯圖(in CDCl3) 85
圖4−13.化合物4之局部放大NOESY關聯圖(in CDCl3) 86
圖4−14.化合物4與已知sinularolide D (94)氫值、碳值與NOE的關聯圖 87
圖5−1.化合物5之高解析ESI質譜 92
圖5−2.化合物5之低解析ESI質譜 92
圖5−3.化合物5之紅外光譜 93
圖5−4.化合物5之氫譜(in CDCl3, 400 MHz) 94
圖5−5.化合物5之局部放大低場區氫譜(in CDCl3) 95
圖5−6.化合物5之局部放大高場區氫譜(in CDCl3) 96
圖5−7.化合物5之碳譜與DEPT圖譜(in CDCl3, 100MHz) 97
圖5−8.化合物5之HSQC關聯圖(in CDCl3) 98
圖5−9.化合物5之局部放大HSQC關聯圖(in CDCl3) 99
圖5−10.化合物5之COSY關聯圖(in CDCl3) 100
圖5−11.化合物5之HMBC關聯圖(in CDCl3) 101
圖5−12.化合物5之NOESY關聯圖(in CDCl3) 102
圖5−13.化合物5與已知durumolide B (75)氫值、碳值與NOE的關聯圖 103
圖6−1.化合物6之高解析ESI質譜 109
圖6−2.化合物6之低解析ESI質譜 109
圖6−3.化合物6之紅外光譜 110
圖6−4.化合物6之氫譜(in CDCl3, 400 MHz) 111
圖6−5.化合物6之局部放大低場區氫譜(in CDCl3) 112
圖6−6.化合物6之局部放大高場區氫譜(in CDCl3) 113
圖6−7.化合物6之碳譜與DEPT圖譜(in CDCl3, 100MHz) 114
圖6−8.化合物6之HSQC關聯圖(in CDCl3) 115
圖6−9.化合物6之局部放大HSQC關聯圖(in CDCl3) 116
圖6−10.化合物6之COSY關聯圖(in CDCl3) 117
圖6−11.化合物6之HMBC關聯圖(in CDCl3) 118
圖6−12.化合物6之NOESY關聯圖(in CDCl3) 119
圖6−13.化合物6之局部放大NOESY關聯圖(in CDCl3) 120
圖6−14.化合物6之局部放大NOESY關聯圖(in CDCl3) 121
圖6−15.化合物6 (S)-MTPA ester (6a)之氫譜(in D-pyridine, 300MHz) 122
圖6−16.化合物6 (R)-MTPA ester (6b)之氫譜(in D-pyridine, 300MHz) 123
圖6−17.化合物6與已知durumolide J (91)氫值、碳值與NOE的關聯圖 124
圖D、Geranylgeranyl pyrophosphate酵素催化,環化生合成cembrane骨架之立體構型 131
圖 E、以人工合成方式得到在C-15位置上含有methoxymethyl之內 酯(γ-lactone)的混合立體異構物 132
圖F、化合物6之Mosher醯化反應 139

陸、參考文獻

1.M. C. Wani, H. L. Taylor, M. E. Wall, P. Coggon, A. T. McPhail, “ Plant Antitumor Agents. VI. The Isolation and Structure of Taxol, a Novel Antileukemic and Antitumor Agent from Taxus brevifolia.” J. Am. Chem. Soc. 1971, 93, 2325−2327.
2.M. E. Wall, M. C. Wani, C. E. Cook, K. H. Palmer, A. T. McPhail, G. A. Sim, “Plant Antitumor Agents. I. The Isolation and Structure of Camptothecin, a Novel Alkaloidal Leukemia and Tumor Inhibitor from Camptotheca acuminata.” J. Am. Chem. Soc. 1966, 88, 3888−3890.
3.B. Tursch, J. C. Braekman, D. Daloze, M. Herin, R. Karlsson, “Chemical Studies of Marine Invertebrates. X. Lobophytolide, A New Cembranolide Diterpene from the Soft Coral Lobophytum cristagalli (Coelenterata, Octocorallia, Alcyonacea).” Tetrahedron Lett. 1974, 15, 3769−3772.
4.Y. Kashman, A. Groweiss, “Lobolide: A New Epoxy Cembranolide from Marine Origin.” Tetrahedron Lett. 1977, 18, 1159−1160.
5.B. F. Bowden, J. C. Coll, S. J. Mitchell, G. J. Stokie, “Studies of Australian Soft Corals. VΠ. Two New Diterpenes from an Unknown Species of Soft Coral (Genus Lobophytum).” Aust. J. Chem. 1978, 31, 1303−1312.
6.A. Ahond, B. F. Bowden, J. C. Coll, J. D. Fourneron, S. J. Mitchell, “Studies of Australian Soft Corals. ХΠ. Further Cembranolide Diterpenes from Lobophytum crassospiculatum and a Correction of a Previous Stereochemical Assignment.” Aust. J. Chem. 1979, 32, 1273−1280.
7.Y. Yamada, S. Suzuki, K. Iguchi, H. Kikuchi, Y. Tsukitani, H. Horiai, “Studies on Marine Natural Products. Ш. Two New Cembranolides from the Soft Coral Lobophytum pauciflorum (Ehrenberg).” Chem. Pharm. Bull. 1980, 28, 2035−2038.
8.Y. Kashman, S. Carmely, A. Groweiss, “Further Cembranoid Derivatives from the Red Sea Soft Corals Alcyonium flaccidum and Lobophytum crassum.” J. Org. Chem. 1981, 46, 3592−3596.
9.Y. Uchio, J. Toyota, H. Nozaki, M. Nakayama, Y. Nishzono, T. Hase, “Lobohedleolide and (7Z)-lobohedleolide, New Cembranolides from the Soft Coral Lobophytum hedleyi Whitelegge.” Tetrahedron Lett. 1981, 22, 4089−4092.
10.Z. Kinamoni, A. Groweiss, S. Carmely, Y. Kashman, Y. Loya, “Several New Cembranoid Diterpenes from Three Soft Corals of the Red Sea.” Tetrahedron 1983, 39, 1643−1648.
11.B. F. Bowden, J. C. Coll, D. M. Tapiolas, “Studies of Australian Soft Corals. ХХХШ. New Cembranoid Diterpenes from a Lobophytum Species.” Aust. J. Chem. 1983, 36, 2289−2295.
12.B. F. Bowden, J. C. Coll, M. S. L. D. Costa, M. F. Mackay, M. Mahendran, E. D. D. Silva, R. H. Willis, “The Structure Determination of a New Cembranolide Diterpene from the Soft Coral Lobophytum cristigalli (Coelenterata, Octocorallia, Alcyonacea).” Aust. J. Chem. 1984, 37, 542−552.
13.Y. Uchio, S. Eguchi, J. Kuramoto, M. Nakayama, T. Hase, “Denticulatolide, an Ichthyotoxic Peroxide-containing Cembranolide from the Soft Coral Lobophytum denticulatum.” Tetrahedron Lett. 1985, 26, 4487−4490.
14.C. Subrahmanyam, C. V. Rao, V. Anjaneyulu, P. Satyanarayana, P. V. S. Rao, R. S. Ward, A. Pelter, “New Diterpenes from a New Species of Lobophytum Soft Coral of the South Andaman Coast.” Tetrahedron 1992, 48, 3111−3120.
15.S. K. Wang, C. Y. Duh, Y. C. Wu, Y. Wang, M. C. Cheng, K. Soong, L. S. Fang, “Studies on Formosan Soft Corals, П. Cytotoxic Cembranolides from the Soft Coral Lobophytum michaelae.” J. Nat. Prod. 1992, 55, 1430−1435.
16.K. Yamada, K. Ryu, T. Miyamoto, R. Higuchi, “Bioactive Terpenoids from Octocorallia. 4. Three New Cembrane-Type Diterpenoids from the Soft Coral Lobophytum schoedei.” J. Nat. Prod. 1997, 60, 798−801.
17.G. F. Matthée, G. M. König, A. D. Wright, “Three New Diterpenes from the Marine Soft Coral Lobophytum crassum.” J. Nat. Prod. 1998, 61, 237−240.
18.C. Y. Duh, S. K. Wang, B. T. Huang, C. F. Dai, “Cytotoxic Cembrenolide Diterpenes from the Formosan Soft Coral Lobophytum crassum.” J. Nat. Prod. 2000, 63, 884−885.
19.M. A. Rashid, K. R. Gustafson, M. R. Boyd, “HIV-Inhibitory Cembrane Derivatives from a Philippines Collection of the Soft Coral Lobophytum Species.” J. Nat. Prod. 2000, 63, 531−533.
20.S. W. Yin, Y. P. Shi, X. M. Li, B. G. Wang, “A New Cembranoid Diterpene and Other Related Metabolites from the South-China-Sea Soft Coral Lobophytum crassum.” Helv. Chim. Acta. 2006, 89, 567−572.
21.L. T. Wang, S. K. Wang, K. Soong, C. Y. Duh, “New Cytotoxic Cembranolides from the Soft Coral Lobophytum michaelae.” Chem. Pharm. Bull. 2007, 55, 766−770.
22.S. H. Chen, Y. W. Guo, H. Huang, G. Cimino, “Six New Cembranolides from the Hainan Soft Coral Lobophytum sp..” Helv. Chim. Acta. 2008, 91, 873−880.
23.C. H. Chao, Z. H. Wen, Y. C. Wu, H. C. Yeh, J. H. Sheu, “Cytotoxic and Anti-inflammatory Cembranoids from the Soft Coral Lobophytum crassum.” J. Nat. Prod. 2008, 71, 1819−1824.
24.S. H. Chen, H. Huang, Y. W. Guo, “Four New Cembrane Diterpenes from the Hainan Soft Coral Lobophytum sp..” Chin. J. Chem. 2008, 26, 2223−2227.
25.S. Y. Cheng, Z. H. Wen, S. F. Chiou, C. H. Hsu, S. K. Wang, C. F. Dai, M. Y. Chiang, C. Y. Duh, “Durumolides A−E, Anti-inflammatory and Antibacterial Cembranolides from the Soft Coral Lobophytum durum.” Tetrahedron 2008, 64, 9698−9704.
26.W. Zhang, K. Krohn, J. Ding, Z. H. Miao, X. H. Zhou, S. H. Chen, G. Pescitelli, P. Salvadori, T. Kurtan, Y. W. Guo, “Structural and Stereochemical Studies of α-Methylene-γ-lactone-Bearing Cembrane Diterpenoids from a South China Sea Soft Coral Lobophytum crassum.” J. Nat. Prod. 2008, 71, 961−966.
27.S. Y. Cheng, Z. H. Wen, S. K. Wang, S. F. Chiou, C. H. Hsu, C. F. Dai, M. Y. Chiang, C. Y. Duh, “Unprecedented Hemiketal Cembranolides with Anti-inflammatory Activity from the Soft Coral Lobophytum durum.” J. Nat. Prod. 2009, 72, 152−155.
28.S. Y. Cheng, Z. H. Wen, S. K. Wang, S. F. Chiou, C. H. Hsu, C. F. Dai, C. Y. Duh, “Anti-inflammatory Cembranolides from the Soft Coral Lobophytum durum.” Bioorg. Med. Chem. 2009, 17, 3763−3769.
29.S. K. Wang, C. Y. Duh, Y. C. Wu, Y. Wang, M. C. Cheng, K. Soong, L. S. Fang, “Additions and Corrections.” J. Nat. Prod. 2009, 72, 324.
30.G. Li, Y. Zhang, Z. Deng, L. V. Ofwegen, P. Proksch, W. Lin, “Cytotoxic Cembranoid Diterpenes from a Soft Coral Sinularia gibberosa.” J. Nat. Prod. 2005, 68, 649−652.
31.A. D. Rodrfguez, Y. Li, H. Dhasmana, “New Marine Cembrane Diterpenoids Isolated from the Caribbean Gorgonian Eunicea mammosa.” J. Nat. Prod. 1993, 56, 1101−1113.
32.A. D. Rodriguez, I. C. Pina, A. L. Acosta, C. Ramirez, J. J. Soto, “Synthesis of Analogues of Eunicea γ-Cembranolides Containing Cyclic Ethers via Saponification.” J. Org. Chem. 2001, 66, 648−658.
33.I. Ohtani, T. Kusumi, Y. Kashman, H. Kakisawa, “High-Field FT NMR Application of Mosher’ s Method. The Absolute Configurations of Marine Terpenoids.” J. Am. Chem. Soc. 1991, 113, 4092−4096.

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