鷹爪桃與香水樹化學成分及生物活性之研究與固醇類新藥的開發

延續對台灣產番荔枝科植物（Formosan Annonaceous Plants）相關抗血小板凝集、抗腫瘤與抗HIV病毒等之活性成分物質的開發與研究。採集了鷹爪桃 (Artabotrys uncinatus) 之根、莖、葉部及果實部分與香水樹 (Cananga odorata) 之葉及果實部，分別進行化學成分分離及純化與生物活性關係的探討。由鷹爪桃的部分，得到六十四個化合物，包括：三十二個alkaloids、二個triterpenoids、十個steroids、九個flavonoids、四個benzenoids、一個lactam、一個cyclohex-2-en-1-one、一個neolignan、三個lignans。其中uncinine (124)、artabonatine C (84)、-D (85)、-E (87)、 -A (86)、-F (88) 和 -B (97) 有七個新化合物。由香水樹得到二十二個化合物，包括：十三個alkaloids、三個sesquiterpenes、一個benzenoid、二個steroids、一個cyclohexane 及二個amides。其中(+)-ushinsunine--N-oxide (125)、cananodine (128)、cryptomeridiol-11--L-rhamnoside (129) 及 -eudesmol-11--L-rhamnoside (130) 四個為新化合物。經由光譜分析和配合衍生物之製備，加以證明及確認其結構式。因為steroids及triterpenes具有抗癌與抗HIV活性，所以進行一系列衍生物的製備，共計35個化合物，提供活性篩選。從過去文獻得知butenolides相關的化合物，有不錯的抗癌活性。所以我們也合成了新化合物uncinine (124) 相關的中間產物以提供活性的測試。抗血小板凝集活性之研究顯示化合物quercetin (51) 主要對於AA及collagen；7-dehydrocholesteryl succinicc acid (155) 對於collagen所誘發血小板的凝集作用有較佳的抑制效果。化合物atherospermidine (2)、lysicamine (3)、polycarpol (33)、squamolone (101)、6-hydroxystigmasterone (106)、 stigmast-4-en-3,6-dione (107)、stigmast-4,22-dien-3,6-dione (108)、 squamolone (101)、blumenol A (115)、(+)-syringaresinol (117)、(+)-ushinsunine--N-oxide (125)、cananodine (128)、cryptomeridiol 11--L-rhamnoside (129)、-eudesmol (131)、N-trans-feruloytranine (134)、cleistopholine (136) 和 stigmast-5,22-dien-3-one (139) 進行肝癌細胞 (Hep G2 & Hep 2,2,15) 的細胞毒殺測試，顯示部分化合物具有相當不錯的活性。化合物stigmast-4,22-dien-3,6-dione (108) 對於癌細胞HONE-1與NUGC之活性試驗，在50 mΜ濃度下，皆呈現強效的細胞毒殺作用；化合物liriodenine (1)、atherspermidine (2)、luteolin (112)對兩種癌細胞HONE-1與NUGC在50μg/ml濃度下，亦具有毒殺效果。 Steroids的衍生物 -sitosteryl-glycoside (49)、6-hydroxystigmasterone (140)、7-dehydrocholesterol (154) 和6-succinicylstigmasterone (153) 經過篩選，顯示具有抑制HIV活性之效果。利用發酵技術，麵包酵母菌將 (-)-ushinsunine (14) 進行生物轉換，產生兩種具有細胞毒殺的生物鹼類liriodenine (1)及(-)-ushinsunine--N-oxide (63)。雖然這些化合物亦可藉由化合物(-)-ushinsunine (14) 氧化作用的方法來合成以及從香水樹中分離出來，此次我們藉由微生物來證明7-hydroxyaporphine生物鹼的氧化過程。上述化合物的分離與合成，以及其生物活性結果，希望能提供藥理研究者有關方面的探討。

關鍵字

生物鹼；鷹爪桃；香水樹；固醇

並列摘要

As a result of our continuing search for bioactive agents (e.g. antiplatelet aggregation, antitumor and anti-HIV agents) from Formosan Annonaceous plants, we have undertaken the methanolic extracts of the fruits, roots stems and leaves of Artabotrys uncinatus and the methanolic extracts of the fruits and leaves of Cananga odorata for evaluation of their in vitro antitumor activity. Sixty-four compounds, including thirty-two alkaloids, two triterpenoids, ten steroids,nine flavonoids, four benzenoids, one lactam, one cyclohex-2-en-1-one, one neolignan, and three lignans, were isolated from A. uncinatus and their structures were established on the basis of NMR and MS data interpretation. Among them, artabonatine -C (84), -D (85), A (86), -E (87), -F (88), -B (97), and uncinine (124) were new compounds. We have also isolated twenty-two compounds, including thirteen alkaloids, three sesquiterpenes, one guaipyridine sesquiterpene alkaloid, one benzenoid,two steroids, one cyclohexane and two amides from C. odorata. Of these compounds, four new compounds, (+)-ushinsunine--N-oxide (125), cananodine (128), cryptomeridiol-11--L-rhamnoside (129) and -eudesmol-11--L-rhamnoside (130) were characterized by spectral and chemical transformation studies. Since steroids and triterpenes have shown anti-cancer and anti-HIV activity, we synthesized a series of derivatives for bioactivity testing. In previous studies, butenolides revealed anti-cancer activity. As a result, we synthesized several butenolide analogues for screening of antitumor activity. At a concentration of 100 g/mL, quercetin (51) showed significant inhibition of arachidonic acid (AA)-induced and collagen-induced platelet aggregation；7-dehydrocholesteryl succinic acid (155) showed significant inhibition of collagen-induced platelet aggregation. Furthermore, atherospermidine (2), lysicamine (3), polycarpol (33), squamolone (101), 6-hydroxystigmasterone (106), stigmast-4-en-3,6-dione (107), stigmast-4,22-dien-3,6-dione (108), squamolone (101), blumenol A (115), (+)-syringaresinol (117), (+)-ushinsunine--N-oxide (125), cananodine (128), cryptomeridiol 11--L-rhamnoside (129), -eudesmol (131), N-trans-feruloytranine (134), cleistopholine (136) and stigmast-5,22-dien-3-one (139) were found to exhibit significant cytotoxicity against two human hepatocarcinoma cell lines (Hep G2 and 2,2,15). On antitumor bioassay, stigmast-4,22-dien-3,6-dione (108) showed significant cytotoxicities against HONE-1 and NUGC cell lines at (50μg/mL). Liriodenine (1), atherspermidine (2), luteolin (112) showed marginal cytotoxic activity against the HONE-1 and NUGC cell lines (50μg/mL). Derivatives of sterols such as -sitosterylglycoside (49), 6-hydroxystigmasterone (140), 6-succinicylstigmasterone (153), and 7-dehydrocholesterol (154) showed inhibition of HIV. Using a fermentation technique, Saccharomyces cerevisiae produced two cytotoxic alkaloids liriodenine (1) and (-)-ushinsunine--N-oxide (63) by using 14 as precursor. Although compounds 1 and 63 have been synthesized from 14 by chemical oxidation and isolated from C. odorata, we proved the oxidative conversion of alkaloid in the microorganic system. As a result, the isolation and synthesis of compounds and the bioactivities can be provided for further studies.