Survivin小分子抑制剂的计算机虚拟筛选及初步活性研究

Virtual screening and preliminary evaluation of small molecule inhibitors of survivin

目的通过计算机虚拟筛选,寻找生存素(survivin)的小分子抑制剂,并对其活性进行初步研究。方法基于sur-vivin与其抑制性配体Smac/Diablo复合物的三维结构,用分子对接的方法对含有149,214个小分子的三维结构数据库进行筛选。通过初步活性测定确定活性较高的化合物,进一步研究其对凋亡,细胞周期及活性氧产生的影响。利用分子图像学方法分析化合物与survivin之间的作用模式。结果通过能量打分并根据结构多样性和类药性原则,最终选择了35个化合物进行初步活性测定,其中有9个化合物显示出明显的抑制活性,3个化合物的半数抑制浓度在30μmol.L-1以下。选择其中活性最高的化合物1-19进行进一步研究,结果显示小剂量的1-19能够促进细胞的早期凋亡,诱导细胞内活性氧产生,导致细胞发生S和G2/M期阻滞。分子图像学分析显示活性最高的1-19能够与survivin配体结合区的71位天门冬氨酸形成两个稳定的氢键。结论通过计算机辅助药物设计、药理活性测试以及分子图像学分析,初步确定了一个针对survivin的全新的先导化合物,为今后survivin小分子抑制剂的研究奠定了基础。

Aim To develop small molecule inhibitors of survivin through structure-based virtual screening.Methods Based on the three dimension structure of survivin in complex with Smac/Diablo,docking procedure against the binding site was performed on a database including 149,214 compounds.Further apoptosis assay,cell cycle analysis and reactive oxygen species（ROS） production assay were performed based on the preliminary activity evaluation.Analysis of molecular graphics was carried out to deduce a probable binding model of the compounds with survivin.Results From the top 1000 compounds with the best DOCK energy score,35 compounds were selected for biological assay based on the further analysis of the structural diversity and drug-like property.Nine of 35 compounds showed notable inhibitory activity and three of them had half maximal inhibition concentration less than 30 μmol·L-1.The compound with the best activity（1-19） showed a dose-dependent induction of apoptosis,S and G2/M cell cycle arrest and increased ROS production.Molecular graphic analysis indicated that two hydrogen bonds may form between compound 1-19 and Asp71 in the binding site of survivin.Conclusions A promising lead compound is discovered to inhibit survivin through virtual screening combined with study of pharmacological activity and analysis of molecular graphics.