台灣粗榧樹葉中具生物活性成分之分離與鑑定

Abstract

台灣粗榧是一種常綠樹木，分佈在台灣中海拔山區。台灣粗榧已有三種抗癌成分被分離出來，分別是Taiwanhomoflavone-A，Taiwanhomoflavone-B 和 C-3-epi-wilsonione。研究初步發現由4.6 kg台灣粗榧樹葉的甲醇萃取物可能具有生物活性物質且對HeLa cell具有很強的抑制效果，這與先前研究結果不一致。為了找出此一物質，我們針對粗榧各個不同部位進行一系列甲醇、水、正丁醇的萃取，再以HPLC分離各成份，隨後以MTT assay對分離出17 mg的單一成分進行生物活性確認，並運用各種光譜分析以鑑定結構。根據1H-NMR、13C-NMR、HMQC、HMBC和COSY得到的結構命名為4', 5, 7－trihydroxyflavone，學名是apigenin。Apigenin於先前文獻中已指出此化合物對數種癌細胞有抑制生長之作用。本研究發現apigenin對HeLa，BEAS-2B，AGS，COLO-205的生物活性，其ED50為3.084、無反應、無反應、8.195 μg/ml（11.422，無反應，無反應，30.35μM）。在我們成功的分離並鑑定出此藥物後，在未來將對此化合物的抑癌機制作進一步的探討，並希望能對此化合物作官能基的修飾以增進其抑癌的效果。

CELL ED50 μg/ml μM BEAS-2B x x AGS x x COLO-205 8.195 30.35 HeLa 3.084 11.422

Cephalotaxus wilsoniana Hayata is an evergreen tree distributed over the middle mountains of Taiwan. Several antitumor components, including Taiwanhomoflavone-A, Taiwanhomoflavone-B and C-3-epi-wilsonione, have been reported from C. wilsoniana Hayata recently. A crude methanol extract of C. wilsoniana leaves（4.6 kg） showed potent cytotoxic activity against HeLa cells in vitro. This subdivided cytotoxic activity exhibited different discrepancy than previous research. In order to characterize the chemical constituents in different parts of plant and to find out the bioactivities compounds of C. wilsoniana Hayata, a series of extraction (from H2O to butanol of original methanol extraction layer) was carried out. Moreover, HPLC chromatography was utilized to separate each component. The cytotoxic activity of these extracts against HeLa cells were analyzed by the MTT assay. Structure elucidation, using 1H-NMR, 13C-NMR, HMQC, HMBC, and COSY, revealed the structure as 4', 5, 7－trihydroxyflavone and thatz the scientific name is apigenin（17 mg）. Researches on apigenin has shown inhibitory effect on the growth of various cancer cells. Our study showed that apigenin showed bioactivity ageinst the HeLa, BEAS-2B, AGS, and COLO-205 in concentration-dependent manner with ED50 values of about 3.084, no reaction, no reaction, and 8.195 μg/ml（11.422, no reaction, no reaction, 30.35 μM）. In the future, research will focus on the mode of action of the drug and the functional group modification of the drug for the improvement of anti-cancer efficacy.

CELL ED50 μg/ml μM BEAS-2B x x AGS x x COLO-205 8.195 30.35 HeLa 3.084 11.422