第一部分 臺灣產固有種植物臺灣嗩吶草之多酚類化合物及細胞毒活性之研究;第二部分 臺灣產固有種植物紅莖椒草之內生菌–蘆筍擬莖點黴菌之二次代謝產物與生物活性之研究

Part I Polyphenols and their cytotoxic activity from the endemic plant, Mitella formosana;Part II Secondary metabolites and biological activities of the endophytic fungus– Phomopsis asparagi from the endemic plant of Peperomia sui

彭千柔(Chien-Jou Peng)

指導教授 ： 張訓碩

共同指導教授 ： 鄭銘仁(Ming-Jen Cheng)

高雄醫學大學/藥學院/天然藥物研究所碩士班/碩士(2016年)

摘要

第一部分 臺灣產固有種植物臺灣嗩吶草之多酚類化合物及細胞毒活性之研究
　　臺灣嗩吶草（Mitella formosana (Hay.) Masam.）是虎耳草科（Saxifragaceae）嗩吶草屬（Mitella）的多年生草本植物，為臺灣固有種。本研究發現臺灣嗩吶草全草之甲醇抽出物對於抗藥性卵巢癌細胞 (TOV-21G-RTx)、肝癌細胞 (SK-Hep-1) 及骨肉瘤細胞 (MG-63) 皆具有細胞毒活性，然而在較低濃度時卻僅對於TOV-21G-RTx仍然呈現非常好的抑制效果。利用生物活性導向分離試驗及光譜分析法，由臺灣嗩吶草全草之活性乙酸乙酯層萃取物分離得到30個化合物，包括2個新化合物─mitellaformosin (MF-1)、(S)-(–)-2,3-dihydroxy-1-(4-hydroxy-3-methoxyphenyl)-propan-1-one (MF-2) 與28個已知化合物。其中，活性化合物1,2,3,4,6-penta-O-galloyl-β-D-glucose (MF-23) 與tellimagrandin II (MF-30) 對TOV-21G-RTx的IC50值分別為4.8 μM和6.0 μM。
第二部分 臺灣產固有種植物紅莖椒草之內生菌–蘆筍擬莖點黴菌之二次代謝產物與生物活性之研究
　　蘆筍擬莖點黴菌 (Phomopsis asparagi (Sacc.) Grove) 為臺灣產固有種植物紅莖椒草 (Peperomia sui Lin & Lu) 的植物內生菌，經篩選發現其發酵液具有調節雌性激素受體 (estrogen receptor, ER) 的活性，與乳癌防治、停經後骨質疏鬆及心血管疾病等防治有關。利用生物活性導向分離試驗及圖譜分析法，由蘆筍擬莖點黴菌之乙酸乙酯層萃取物中分離得10個化合物，包含2個新化合物─phomocytochalasin A (PA-1) 和 (R)-(–)-1-(4-hydroxyphenyl)ethane-1,2-diol (PA-2) 及8個已知化合物。然而，目前仍未分離得到具有調節雌激素受體活性之化合物。另一方面，本研究發現phomocytochalasin A (PA-1)與cytochalasin H (MF-6) 對SiHa cells（子宮頸癌細胞）具有卓越地破壞細胞骨架的活性，兩者的IC50值分別為0.34 ± 0.05 μM和0.27 ± 0.02 μM，並透過探討結構與活性之關係發現細胞鬆弛素類的活性作用位為OH-7、6(12)-en-7-ol及乙醯氧基側鏈。

關鍵字

臺灣嗩吶草 ； 虎耳草科 ； 多酚類化合物 ； 抗藥性 ； 細胞毒活性 ； GNMT啟動子活性 ； 蘆筍擬莖點黴菌 ； 紅莖椒草 ； 胡椒科 ； 內生菌 ； 化學成分 ； 雌激素受體調節活性 ； 細胞骨架破壞活性

並列摘要

Part I. Polyphenols and their cytotoxic activity from the endemic plant, Mitella formosana.
　　Mitella formosana (Hay.) Masam. (Saxifragaceae) is a perennial and pubescent herb, endemic to Taiwan and grows at medium to high altitudes. The methanolic extract of the whole plant of this species showed cytotoxic activity against TOV-21G-RTx (paclitaxel-resistant ovarian cancer), SK-Hep-1 (hepatocellular carcinoma), MG-63 (osteosarcoma) cell lines, but only revealed potent cytotoxicity to TOV-21G-RTx at low concentration. Bioassay-guided fractionation of the active ethyl acetate-soluble layer from the whole plant of M. formosana led to the isolation of two new compounds, mitellaformosin (MF-1) and (S)-(–)-2,3-dihydroxy-1-(4-hydroxy-3-methoxyphenyl)propan-1-one (MF-2) as well as 28 known compounds. The structures of these isolates were elucidated by spectral analysis. The predominant compounds were 1,2,3,4,6-penta-O-galloyl-β-D-glucose (MF-23) and tellimagrandin II (MF-30), which showed strong cytotoxic effect on TOV-21G-RTx cancer cell line with IC50 values of 4.8 and 6.0 μM, respectively.
Part II. Secondary metabolites and biological activities of the endophytic fungus– Phomopsis asparagi from the endemic plant of Peperomia sui.
　　Phomopsis asparagi (Sacc.) Grove was the endophytic fungus which isolated from the whole plant of Peperomia sui Lin & Lu. Preliminary data showed that its liquid fermentation had the estrogen receptor modulating activity. Bioassay-guided fractionation of the ethyl acetate-soluble layer from the liquid fermentation of P. asparagi led to the isolation of 10 compounds, including two new compounds, phomocytochalasin A (PA-1), (R)-(–)-1-(4-hydroxyphenyl)ethane-1,2-diol (PA-2) and 8 known compounds. The structures of these isolates were elucidated by spectral analysis. However, the active compounds for estrogen receptor modulating activity have not been isolated yet. On the other hand, phomocytochalasin A (PA-1) and cytochalasin H (PA-6) showed remarkable effect on actin cytoskeleton disruption of SiHa cells with the IC50 values of 0.34 ± 0.05 μM and 0.27 ± 0.02 μM, respectively. The structure-activity relationship of cytochalasins was observed and the OH-7, 6(12)-en-7-ol as well as acetoxy group at C-21 were claimed to be essential to the manifestation of the toxicity.

並列關鍵字

Mitella formosana ； Saxifragaceae ； polyphenols ； drug-resistant ； cytotoxic activity ； GNMT promoter activity ； Phomopsis asparagi ； Peperomia sui ； Piperaceae ； endophytic fungus ； chemical constituents ； estrogen receptor modulating activity ； actin disruption activity

參考文獻

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