网格蛋白抑制剂的虚拟筛选与活性评价

In silico screening and biological evaluation of clathrin inhibitors

目的基于网格蛋白发现内吞抑制剂或具有抗病毒活性的化合物。方法以网格蛋白晶体结构为基础,利用分子对接对自有化合物库进行虚拟筛选,所获得的优选分子采用分子动力学模拟方法对受体-配体相互作用的动态过程进行分析,预测复合物的结合自由能并找出受体蛋白中的关键残基,通过细胞水平抗肠道病毒EV71的活性筛选、毒性测定及转铁蛋白内吞实验,对化合物的抗病毒活性及作用机制进行评价与验证。结果获得了1个结构新颖且对EV71病毒有抗病毒作用的活性分子Cg20(IC50=24.88μmol/L),该化合物的细胞毒性低于阳性对照化合物pitstop2。结论本文报道的虚拟筛选方法与活性预测模型已通过本研究生物学实验获得验证,可继续运用于网格蛋白抑制剂的设计与开发,研究结果为先导化合物Cg20提供了明确的优化方向,为网格蛋白抑制剂的高效开发提供了理论支撑。

ObjectiveTo discover clathrin inhibitors with antiviral ability.MethodsMolecular docking and molecular dynamics simulation were performed on the basis of clathrin crystal structure for the compounds in our self-made real compound library in order to search for potential clathrin inhibitors and explore the modes of their binding interaction with clathrin. The biological assay was conducted for the selected compounds by the EV71 infection,cytotoxicity,and transferrin internalization test in cellular level.ResultsA novel hit compound Cg20 with antiviral activity against EV71 virus(IC50=24.88 μmol/L)was obtained by in silico screening,whose cytotoxicity was lower than that of the positive control compound pitstop2.ConclusionThe in silico screening method and the prediction model for the biological activity,validated by and reported in the present work,might be applied to the future studies on the research and development of clathrin inhibitors. The present results also provide a research direction for further optimization of the hit compound Cg20. With the present method, it will be possible to obtain more effective clathrin inhibitors.