海洋真菌Penicillium sp.HD-1-1次级代谢产物研究

目的 研究涠洲岛来源的海洋真菌Penicillium sp. HD-1-1产生的次级代谢产物及其生物活性。方法 利用硅胶柱色谱和半制备液相等技术进行化合物的分离纯化，运用HRESIMS、NMR等方法对化合物的结构进行鉴定，使用CCK-8法评估化合物的细胞毒活性。结果 从涠洲岛海洋真菌Penicilliumsp. HD-1-1次级代谢产物中分离鉴定了1个新的霉酚酸类化合物以及16个已知化合物，分别为6-(5-甲氧羰基-3-甲基戊-2-烯)-7-羟基-3,5-双甲氧基-4-甲基苯酚-1-酮（1）、霉酚酸（2）、甲基麦考酚酸（3）、6-(5-甲氧羰基-3-甲基戊-2-烯)-3,7-二羟基-5-甲氧基-4-甲基苯酚-1-酮（4）、6-(5-羧基-3-甲基戊-2-烯)-3,7-二羟基-5-甲氧基-4-甲基苯酚-1-酮（5）、7-羟基-5-甲氧基-4-甲基-6-(3-甲基-4-氧代戊基)-二氢异苯并呋喃-l-酮（6）、6-(3-羧基丁酯)-7-羟基-5-甲氧基-4-甲基苯酞-1-酮（7）、breynivosamide A（8）、伞形香青酰胺（9）、N-(N''-苯甲酰基-S^*-苯丙氨酰基)-S^*-苯丙氨醇苯甲酸酯（10）、(3S,6R)-6-(对-羟苯基)-1,4-二甲基-3,6-二甲硫基哌嗪-2,5-二酮（11）、(3R,6R)-6-(对-羟苯基)-1,4-二甲基-3,6-二甲硫基哌嗪-2,5-二酮（12）、(+)-新海胆灵A（13）、(3R,8S)-5,7-二羟基-3-(1-羟乙基)苯酚（14）、5,7-二甲氧基-4-甲基异苯并呋喃-1(3H)-酮（15）、saccharonol A（16）、6,8-二羟基-3-羟甲基异香豆素（17）。细胞毒活性结果表明化合物2和3对人胃癌AGS细胞显示出强细胞毒活性，半数抑制浓度（median inhibition concentration，IC₅₀）值分别是0.69、1.12 μmol/L，此外化合物2对人非小细胞肺癌NCI-H1581细胞表现出中等程度的抑制活性，IC₅₀值为17.95 μmol/L，化合物3对人结肠癌HT29细胞也显示出中等抑制活性，IC₅₀值为19.30 μmol/L。结论 化合物1为新的霉酚酸类化合物，命名为甲氧基霉酸酯。化合物2和3具有肿瘤细胞毒活性。

Study on secondary metabolites of marine-derived Penicillium sp. HD-1-1

Objective To study the secondary metabolites and biological activities of marine-derived Penicillium sp. HD-1-1 from Weizhou Island. Methods The compounds were isolated and purified by silica gel column chromatography and semi-preparative liquid chromatography. The structures of the compounds were identified by HRESIMS, NMR and other methods. The cytotoxicity of compounds was evaluated by CCK-8 method. Results A new mycophenolic acid compound and 16 known compounds were isolated and identified from the secondary metabolites of marine fungus Penicillium sp. HD-1-1 and identified as: 6-(5-methoxycarbonyl-3-methylpent-2-enyl)-7-hydroxy-3,5-dimethoxy-4-methylphthalan-1-one (1), mycophenolic acid (2), methyl mycophenolic acid (3), 6-(5-methoxycarbonyl-3-methylpent-2-enyl)-3,7-dihydroxy-5-methoxy-4-methylphthalan-1-one (4), 6-(5- carboxy-3-methylpent-2-enyl)-3,7-dihydroxy-5-methoxy-4-methylphthalan-1-one (5), 7-hydroxy-5-methoxy-4-methyl-6-(3-methyl- 4-oxopentyl)-phthalan-l-one (6), 6-(3-carboxybutyl)-7-hydroxy-5-methoxy-4-methylphthalan-1-one (7), breynivosamide A (8), anabellamide (9), N(N''-benzoyl-S^*-phenylalaninyl)-S^*-phenylalaninol benzoate (10), (3S,6R)-6-(p-hydroxybenzyl)-1,4-dimethyl- 3,6-bis(methylthio) piperazine-2,5-dione (11), (3R,6R)-6-(p-hydroxybenzyl)-1,4-dimethyl-3,6-bis(methylthio) piperazine-2,5-dione (12), (+)-neoechinulin A (13), (3R,8S)-5,7-dihydroxy-3-(1-hydroxyethyl)phthalide (14), 5,7-dimethoxy-4-methylisobenzofuran- 1(3H)-one (15), saccharonol A (16) and 6,8-dihydroxy-3-hydroxymethylisocoumarin (17). The results of cytotoxic activity showed that compounds 2 and 3 showed strong cytotoxic activity against AGS cells, with IC₅₀ values of 0.69 μmol/L and 1.12 μmol/L, respectively. Compound 2 showed moderate inhibitory activity against NCI-H1581 cells with IC₅₀ value of 17.95 μmol/L, and compound 3 also showed moderate inhibitory activity against HT29 cells with IC₅₀ value of 19.30 μmol/L. Conclusion Compound 1 is a new mycophenolic acid compound, named methoxy mycophenolate. Compounds 2 and 3 have tumor cytotoxic activities.