非水溶性药物潜在靶蛋白筛选方法探索  

An exploratory method for screening candidate target proteins of insoluble drugs

作　　者：陶定银[1,2] 夏思敏[1,2] 刘晋湘[1,2] 张丽华[1] 梁振[1] 张玉奎[1] 

机构地区：[1]国家色谱研究分析中心中国科学院大连化学物理研究所分离分析化学重点实验室,大连116023 [2]中国科学院研究生院,北京100039

出　　处：《中国科学：化学》2011年第6期976-981,共6页SCIENTIA SINICA Chimica

基　　金：国家重大科学研究计划(2007CB914100);中国科学院知识创新工程重要方向性项目(KJCX2YW.H09)的资助

摘　　要：针对化学蛋白质组学在筛选非水溶性药物的靶蛋白中存在的问题,建立了以非水溶性药物颗粒为载体的靶蛋白筛选方法.通过避免药物固定化,不仅可以保留全部的药物官能团,而且减少了蛋白质在固载基质上的非特异性吸附,可提高获得数据的可信度.将非溶性药物地塞米松颗粒直接与人小细胞肺癌H446细胞提取蛋白质通过间歇性振荡孵育24h,然后采用缓冲液清洗药物颗粒,最后对药物颗粒特异性结合的蛋白质进行酶解和分离鉴定.结果表明,筛选出41个潜在的药物靶蛋白,参与了与DEX药物作用机理相关的多种蛋白质代谢通路和糖代谢通路,同时还发现部分蛋白质参与了帕金森症疾病过程.To solve the problem in screening the candidate target proteins of poor solubility drugs with limited immobilization function groups by traditional chemical proteomics method,a chemical proteomics method for screening candidate proteins targets,with insoluble drugs particles directly as matrix,was explored.Compared with the traditional methods,the support free protocol could not only avoid the problems caused by matrices and linkers,but also maintain all active sites of drugs without immobilization.The protein extracted from human small cell lung cancer（SCLC）NCI-H446 cells was incubated with dexamethasone（DEX） particles directly for 24 h with shaking in intervals.After incubation,the pellets of DEX were washed with PBS buffer and NaCl to remove other proteins non-specifically adsorbed to particles.Then the proteins adsorbed on DEX particles were processed by thermal denaturation,reduction,alkylation and digestion,followed by μRPLC-ESI MS/MS analysis,41 candidate target proteins were screened which interaction to each other,and several proteins were involved in pathways which related with DEX mechanism,including the one related to Parkinson＇s disease.

关 键 词：地塞米松 化学蛋白质组 药物靶蛋白 非水溶性药物 

分 类 号：R96[医药卫生—药理学]