青灰叶下珠内生菌Phomopsis sp.TJ507A的化学成分及生物活性研究

The bioactive secondary metabolites from Phyllanthus glaucus-associated fungus Phomopsis sp. TJ507A

为探究药用植物青灰叶下珠(Phyllanthus glaucus)来源内生真菌拟茎点霉(Phomopsis sp. TJ507A)的化学成分,本实验联合运用硅胶柱色谱、ODS柱色谱、凝胶柱色谱(Sephadex LH-20)、半制备型高效液相色谱(HPLC)等分离技术,从该菌株大米固体发酵产物的乙酸乙酯提取物中分离得到4个倍半萜化合物phomophyllin O(1)、phomophyllin P(2)、7-hydroxy-10-methoxydehydrodihydrobotrydial(3)、plorantinone D(4)和8个甾体化合物fortis-terol(5)、dankasterone B(6)、(14α,22E)-14-hydroxy-ergosta-7,22-diene-3,6-dione(7)、(14α,22E)-14-hydroxyerg-osta-4,7,22-triene-3,6-dione(8)、calvasterol B(9)、isocyathisterol(10)、ergosta-4,6,8(14),22-tetraen-3-one(11)和ganodermaside D(12)。化合物1和2为新化合物,运用现代波谱分析技术、[Rh2(OCOCF3)4]络合诱导ECD法及与文献对比等方法,鉴定了化合物1和2的结构和绝对构型。化合物1～5、7、8和10均为首次从该属真菌中分离得到。通过体外生物活性评价,发现化合物9具较强的NO生成抑制活性(IC508. 7μM)。

Four sesquiterpenoids,named phomophyllin O(1),phomophyllin P(2),7-hydroxy-10-methoxydehydrodihydrobotrydial(3),and plorantinone D(4),and eight steroids,named fortisterol(5),dankasterone B(6),(14α,22E)-14-hydroxy-ergosta-7,22-diene-3,6-dione(7),(14 α,22E)-14-hydroxyergosta-4,7,22-triene-3,6-dione(8),calvasterol B(9),isocyathisterol(10),ergosta-4,6,8(14),22-tetraen-3-one(11),and ganodermaside D(12),were isolated from the ethyl acetate extract of the solid-state fermentation of the endophytic fungus Phomopsis sp.TJ507A,a strain isolated from the medicinal plant Phyllanthus glaucus.These compounds were purified by comprehensive chromatography on silica gel column,Sephadex LH-20,octadecyl silane(ODS),and semi-preparative HPLC.Two new sesquiterpenoids 1 and 2 were identified and their structures were determined by detailed analyses of the NMR spectra,[Rh2(OCOCF3)4]-induced ECD spectra,and comparison with literatures.Compounds 1-5,7,8 and 10 were isolated from the genus Phomopsis for the first time.Evaluation of in vitro anti-inflammatory activity of the isolates showed that compound 9 exhibited significant NO inhibitory activity with the IC50 value at 8.7 μM.