瓊崖海棠 （Calophyllum inophyllum） 是一種富含天然活性物質的濱海藥用植物。本實驗採集了此樹木的樹皮部份加以萃取分離得到五個 xanthones 類的化合物，分別是 calophinone（29）、caloxanthone I（30）、brasilixanthone B（31）、pyranojacareubin（32）以及 osajaxanthone（33），其中 calophinone（29）為在自然界首次發現的新化合物。另外，為了確定 calophinone（29）的結構也做了乙醯化的反應，得到了 6-acetylcalophinone（34）。而這些化合物及其衍生物的結構鑑定，是依據其 NMR圖譜資料的分析以及其物理性質來做判定。所應用的核磁共振磁譜技術包括 1H、13C、DEPT、COSY、HMQC、HMBC、NOESY 以及其他的分析方法，物理性質的測定則由 UV、IR、EIMS 以及 HREIMS。另外，將這六個化合物做生物毒殺活性的測試（包含 Hepa59T/VGH、KB 以及 Hela），由毒殺活性測試的結果得知這五個化合物對這三種癌細胞均無細胞毒殺的活性。

另一方面，從 Taxus baccata 及 T. mairei紅豆杉中所分離得到的baccatin III（30） 和 13-deacetyl-1-deoxybaccatin VI （31）做一系列衍生物的製備，如去乙醯化反應後再酯化反應，反應後的混合產物經純化分別得到 13-O-camphanyl-7-O-nicotinoylbaccatin III（34）、13-O-camphanyl -1-deoxybaccatin VI （35）、13-O-（4-chlorobenzoyl）-7-O-nicotinoylbaccatin III（36）以及 13-O-benzoyl -7-O -nicotinoylbaccatin III（37）。而這系列衍生物的結構鑑定則由NMR圖譜資料的分析以及其物理性質來做判定，並將這一系列的產物送測活性以探討化合物結構與活性之間的關聯。

在人類的前列腺腫瘤細胞 （PC-3） 之 in vitro 毒殺活性測試結果顯示化合物13-O-camphanyl-7-O-nicotinoylbaccatin III（34）、13-O-camphanyl -1-deoxybaccatin VI（35）具有不錯的細胞毒殺活性，在10 μM的濃度下其細胞的存活率分別為76%以及65%，與標準物 Taxol 的60%相去不遠。而這些化合物的活性來源可能為其具有 nicotinoyl 以及 camphanic acyl 之部份結構的關係。也因此未來在做衍生物的製備時可以將 nicotinoyl chloride 以及 camphanic acyl chloride 這兩個取代基列入考量，也許可以用來改善化合物的生物活性之用。

Calophyllum inophyllum is a medicinal plant that is rich in bioactive natural products. Calophinone（29）、caloxanthone I（30）、brasilixanthone B（31）、pyranojacareubin（32） and osajaxanthone（33）are five compounds which were isolated from the bark in this experiment. It is the first time to isolate calophinone（29） from a matural source. In order to identify calophinone（29）, 6-acetylcalophinone（34） was prepared via acylation. All structures were determined primarily on the basis of 1D, 2D NMR、UV、IR and Mass spectral analyses. Besides, biological studies don’t reveal that Calophinone（29）、caloxanthone I（30）、brasilixanthone B（31）、pyranojacareubin（32）、osajaxanthone（33）and 6-acetylcalophinone（34）, exhibited in vitro cytotoxicity against human liver carcunoma、Human oral epidermoid carcunoma and Human cervical epidermoid carcunoma.

In addition, four new Taxoid derivatives that were 13-O-camphanyl-7-O -nicotinoylbaccatin III（34）、13-O-camphanyl-1-deoxybaccatin VI（35）、13-O-（4-chlorobenzoyl）-7-O-nicotinoylbaccatin III（36）and 13-O-benzoyl-7-O -nicotinoylbaccatin III（37）have been prepared via esterification under sonication starting from 13-deacetyl-7-O-nicotinoylbaccatin III（32） and 13-deacetyl-1-deoxybaccatin VI（31）. All the structures were established primarily on the basis of 1D and 2D NMR techniques including DEPT, COSY, HMBC experiments, as well as comparison with known related compounds. It was deemed quite promising to investigate the structure-activity relationship ( SAR ) for the C - 13 side chain analogues of Taxol with some modification of the baccatin III（30） moiety in order to discover more effective anticancer agents with better pharmacological properties.

Compounds 34 and 35 showed significant cytotoxicity against prostate cancer cell line（PC-3）. Under concentration of 10μM, the cell survival percent was 76% and 65% in case of compounds 34 and 35 compared to 60 % in case of Taxol. According to the structure-activity relationship, nicotinoyl and camphanic acyl group should be the source of activity in compounds 30 and 31. Consequently, it is necessary to introduce nicotinoyl chloride and camphanic acyl chloride groups via chemical reaction to improve the bioactivity.

目 錄
頁次
壹、濱海植物瓊崖海棠之活性天然成分研究

第一部份 緒論

第一章 前言……………………………………………………………11
第二章 研究動機及目標………………………………………………12
第三章 相關化合物之文獻回顧………………………………………13

第二部份 材料與方法

第一章 材料介紹………………………………………...…………….21
第二章 樣品的採集………………………………………...………….23
第三章 分離與純化……………………………………...…………….23
第四章 衍生物的製備…………………………………...…………….27
第五章 生物活性之測試…………………………………...………….27
第六章 結構鑑定及器材……………………………………...……….28

第三部份 結果與討論

第一章 結構解析
Calophinone（29）之結構解析……………………........................30
Caloxanthone I（30）之結構解析……………………....................41
Brasilixanthone B（31）之結構解析…………………....................47
Pyranojacareubin（32）之結構解析………….................................53
6-Deoxyjacareubin（33）之結構解析…………………..................57
Calophinone-monoacetate（34）之結構解析…………...................33

第二章 活性測試結果……………………………….………………...63

第四部份 結論................................................................................................67

第五部份 附錄................................................................................................70

第六部份 文獻................................................................................................73

貳、紅豆杉雙萜類之半合成及抗癌活性研究

第一部份 緒論

第一章 前言............................................................................................77
第二章 研究動機及目標........................................................................77
第三章 相關 Taxoids 化合物之研究....................................................79

第二部份 實驗方法

第一章 衍生物的製備
製備13-deacetyl-7-O-nicotinoylbaccatin III（32）...........................83
製備13-O-camphanyl-7-O-nicotinoylbaccatin III（34）..................84
製備13-O-camphanyl-1-deoxybaccatin VI（35）............................84
製備13-O-（4-chlorobenzoyl）-7-O-nicotinoylbaccatin III（36）....85
製備13-O-benzoyl-7-O-nicotinoylbaccatin III（37）.......................86

第二章 生物活性之測試........................................................................87

第三章 結構鑑定及器材........................................................................87

第三部份 結果與討論

第一章 Taxoids 衍生物之半合成
化合物32、34-37之製備.................................................................90
化合物34-37之結構解析.................................................................93
第二章 活性測試結果..........................................................................106

第四部份 結論..............................................................................................107

第五部份 附錄..............................................................................................109

第六部份 文獻..............................................................................................115

壹、濱海植物瓊崖海棠之活性天然成分研究
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貳、紅豆杉雙萜類之半合成及抗癌活性研究
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13. Surinder Grover; John M. Rimoldi; Anthony A. Molinero; Ashok G. Chaudhary; David G. I. Kingston; Ernest Hamel, Differential Effects of Paclitaxel (Taxol) Analogs Modified at Positions C-2, C-7, and C-3' on Tubulin Polymerization and Polymer Stabilization: Identification of a Hyperactive Paclitaxel Derivative. Biochemistry, 1995, 34, 3927-3934.
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