芸薹生链格孢-粒状青霉共培养次级代谢产物及生物活性研究

Studies on the secondary metabolites and their bioactivities from co-culture of Alternaria brassicicola and Penicillium granulatum

目的研究芸薹生链格孢-粒状青霉共培养发酵的次级代谢产物,寻找具有生物活性的先导化合物。方法采用两种真菌共培养方式,通过LC-MS进行监控,综合应用现代色谱分离技术和波谱鉴定手段,获取并鉴定单体化合物;对单体化合物进行抗帕金森病活性评价和抑菌活性评价;运用Discovery Studio对单体化合物进行成药性预测和靶点虚拟筛选。结果与结论从共培养菌株的大米发酵物中分离得到3个四环二萜类化合物,分别为conidiogenone C(1)、conidiogenone E(2)和conidiogenone F(3)。MTT实验结果表明,化合物2能够抑制MPP^(+)对SH-SY5Y细胞的诱导损伤,EC值为4.76μmol·L^(-1);化合物1对表皮葡萄球菌具有一定的抑菌活性,MIC_(50)值为144.63μg·mL^(-1)。Discovery Studio预测结果表明,所有化合物均符合Lipinski成药性五规则;ADEMT预测结果表明,化合物2具有较好的中枢神经系统药物成药性特征;反向虚拟靶点筛选结果表明,化合物2能够选择性地结合在腺苷A_(2A)受体疏水口袋,与氨基酸残基Cys166和Ile66形成较强的氢键作用力。化合物2可开发为腺苷A受体拮抗剂,具有用作抗帕金森病先导化合物的潜在价值。

In this research, three tetracyclic diterpenoids, namely conidiogenone C(1),conidiogenone E(2) and conidiogenone F(3),were discovered from the co-culture fermentation products of Alternaria brassicicola and Penicillium granulatum.The inhibition against MPP-insult SH-SY5 Y cells and the antibacterial activity of all compounds were evaluated.The drug-likeness prediction and the target virtual screening of isolates were performed by Discovery Studio 2020(DS20).The MTT assay results showed that compound 2 had a certain protective effect on MPP-induced SH-SY5 Y cells with an EC_(50)value of 4.76 μmol·L^(-1).The antibacterial assay results exhibited that compound 1 had the potential inhibition against Staphylococcus epidermidis with a MICvalue of 144.63 μg·mL^(-1).The results of computational prediction via DS20 showed that all compounds conformed to Lipinski′s five rules, and the ADEMT prediction showed that compound 2 had the good drug-likeness characteristics of central nervous system.Furthermore, reverse virtual target screening presented that compound 2 could bind to the hydrophobic pocket of adenosine A_(2A)receptor and interact with Cys166 and Ile66 via hydrogen bonds.In conclusion, compound 2 has the potential to be developed as a lead compound against Parkinson′s disease as an adenosine Areceptor antagonist.