紅花八角果實和阿里山五味子果實活性成分之研究

Abstract

本論文分成兩大部分：A 部分主要研究臺灣產紅花八角(Illicium arborescens Hayata)果實部位的二氯甲烷和乙酸乙酯萃取物，進行分離並分析鑑定所含的prenylated C6-C3 類化合物和成分，實驗結果共分得六個化合物。其中包括了一個新的prenylated C6-C3 類化合物及一個類似prenylated C6-C3 且具有tricyclic ring system 的新骨架化合物，分別命名為illicaborin D (A1)和illicaborene (A2)。 B 部分主要研究臺灣產阿里山五味子(Schisandra arisanensis Hayata)果實部位的丙酮萃取物以及中國大陸產華中五味子(Schisandra sphenanthera Rehd. et Wils.)果實部位的酒精萃取物，進行分離並分析鑑定所含的木酚素成分，實驗結果共分得十一個木酚素化合物。其中包括了三個新的C18-dibenzocyclooctadiene lignans，分別命名為arisanschinins M 和N (B1 和B2)以及schisphenin A (B3)。此外，化合物B1 在結構上透過一個含氧架橋，將cyclooctadiene 上C-6 的非共軛酯基和biphenyl moiety 上C-14 連接在一起。 以上化合物的結構是利用各種物理數據，包括比旋光度、紅外線、紫外線、CD 光譜、質譜以及配合1D、2D 核磁共振圖譜，並參考相關文獻而建立。 最後，將所分得的十七個化合物進行生物活性檢測。紅花八角果實分離出的六個化合物，在八個月ICR 母鼠的初代骨母細胞其類雌激素功能評估試驗中，根據細胞活性分析方法(MTT assay)，發現化合物A2、已知化合物A4 和A5 分別在濃度為50 nM, 10 nM 和5 nM 的情況下，即具有明顯的促進細胞增生之作用，其促進率分別為146.2 %、152.6 %、163.7 %。另外，在癌細胞毒殺測試中發現化合物A4 和A5 對癌細胞株Daoy 具有些微的細胞毒殺活性，其ED50 值分別為20.87 和12.45 μg/ml。 阿里山五味子和華中五味子果實分離出的十一個化合物，在大鼠肝臟星狀細胞的抗肝纖維化作用評估試驗中，根據螢火蟲冷光 分析方法(luciferase assay)，發現化合物B3、已知化合物B7 和B10，在濃度為50 μg /ml 下，對於NF-kB 的活性壓制效果較為明顯，其抑制率分別為24.4 %、53.8 %和52.5 %。

This thesis can be divided into two parts. Part A focuses on the isolation of prenylated C6-C3 compounds from the dichloromethane and ethyl acetate extracts of the fruits of Illicium arborescens Hayata. Application of a series of chromatographic separation afforded two new compounds, illicaborin D (A1) and illicaborene (A2), along with four known compounds identified as 1-allyl-3,5-dimethoxy-4-(3-methylbut-2-enyloxy)benzene (A3), (–)-illicinone A (A4),(2R, 4R)-(–)-illicinone B (A5) and (–)-illicinone A derivative (A6). It is noted that compound A1 possesses a prenylated C6-C3 skeleton, while compound A2 belongs to an unprecedented skeleton with a tricyclic ring system. Part B is the phytochemical investigation of the acetone extract from the fruits of the Taiwanese medicinal plant Schisandra arisanensis Hayata, and the ethanol extract from the fruits of Schisandra sphenanthera Rehd. et Wils.. The two crude extracts separated by extensive chromatography analysis yielded three new polyoxygenated C18-dibenzocyclooctadiene lignans designated as arisanschinins M and N (B1 and B2) and schisphenin A (B3), together with eight known compounds, (+)-gomisin K3 (B4), interiotherin B (B5), schisantherin D (B6), gomisin S (B7), schisantherin E (B8), gomisin C (B9), arisantetralone A (B10) and pre-gomisin (B11). Among them, in particular, there is an oxygen bridge between C-14 and and C-2' in the structure of compound B1. All the structures of these compounds were elucidated on the basis of spectroscopic analysis such as 2D-NMR techniques, high resolution mass spectroscopy (HR-MS), CD spectrscopy and so on. Additionally, known compounds were determined by comparison with literature values as well. The biological activities of these compounds were also evaluated. The estrogen-like activities of compounds A2, A4 and A5 in primary cell culture of osteoblast cells obtained from eight-month-old female ICR mice were more active than the other compounds isolated from Illicium arborescens Hayata. The highest rates of promotion of cell proliferation for compounds A2, A4 and A5 determined by MTT assay were 146.2 %, 152.6 % and 163.7 % at 50 nM, 10 nM and 5 nM, respectively. Moreover, compounds A4 and A5 exhibited weak cytotoxicity against Daoy tumor cell line with ED50 values of 20.87 and 12.45 μg/ml. As for the compouds isolated from Schisandra arisanensis Hayata and Schisandra sphenanthera Rehd. et Wils., they were elvaluated for anti-fibrotic effects on rat hepatic stellate cells (HSC-T6) by luciferase assay. The preliminary result revealed that compounds B3, B7 and B10 showed suppressive activities in NF-kB activation with rates of inhibition 24.4 %, 53.8 % and 52.5 % at 50μg /ml, respectively.