筛选人睾丸特异性含溴结构域蛋白(BRDT)的小分子抑制剂  

Discovery of male anti-fertility inhibitors target BRDT through structure-based virtual screening

作　　者：高娜娜[1,2] 侯丽 周越 辛玲 杨朵[1] 李娜[1] 王慧萍 

机构地区：[1]首都医科大学附属北京世纪坛医院,北京100038 [2]国家卫生计生委科学技术研究所,北京100081

出　　处：《生殖医学杂志》2016年第6期550-555,共6页Journal of Reproductive Medicine

基　　金：中央级公益科研院所基本科研业务费专项(2012GJSSJKB01);中国博士后科学基金(2014M550678)

摘　　要：目的睾丸特异性含溴结构域的蛋白(BRDT)在睾丸中特异表达,参与精子生成的染色质重组过程。本文虚拟筛选基于结构的人BRDT小分子抑制剂。方法从PDB(Protein Data Bank)下载BRDT核心结构域与小分子抑制剂JQ1结合结构域的晶体结构(PDB ID:4FLP),利用分子模拟软件Discovery Studio 3.0,构建了其活性位点抑制剂的药效团模型。运用最佳药效团模型对10 000个化合物的数据库进行初筛,随后运用Libdock分子对接方式对初筛得到的潜在目标化合物进行复筛;最后运用BRDT Bromodomain TR-FRET Assay方法,对虚拟筛选得到的化合物进行实物筛选。结果运用虚拟筛选和蛋白水平筛选相结合的方法,从40 000个化合物中最终得到1个对BRDT有较高抑制作用的小分子抑制剂T480。结论利用一种基于结构的集合虚拟筛选与蛋白水平筛选的BRDT抑制剂的高通量筛选方法,筛选到BRDT的一个小分子抑制剂。Objective： To structure-based virtual screen the inhibitors of human testis-specific and bromodomain containing protein（BRDT）.Methods： The crystal structure（PDB ID：4FLP）of core domain of BRDT and binding domain of JQ1 were downloaded from PDB.In this study,the pharmacophor model of BRDT was established based on the three dimensional crystal structure of BRDT and JQ1 cocrystallization by Discovery Studio 3.0.Firstly,the optimal pharmacophore model was used to screen large compound database.Secondly,we used Libdock to screen the foregoing candidate compounds.Thirdly,We re-screened the candidate compounds using screening model at protein level.Finally,virtual screening of compounds was subjected to physical screening by BRDT Bromodomain TR-FRET Assay.Results： Using virtual screening combined with protein level screening,a small molecular inhibitor target human BRDT（T480）was obtained from 40 000 compounds.Conclusions： One positive compound target human BRDT is obtained through structure-based virtual screening combined with protein level screening.

关 键 词：睾丸特异性含溴结构域蛋白 男性抗生育 精子生成 虚拟筛选 

分 类 号：R[医药卫生]