人源蛋白酪氨酸磷酸酶(PTP1B)抑制剂的高通量筛选  

High-throughput screening of human soluble protein tyrosine phosphatase 1B inhibitors

作　　者：庞晓斌[1,2] 谢欣梅[2] 王守宝[1] 杜冠华[1] 

机构地区：[1]中国医学科学院北京协和医学院药物研究所国家新药筛选中心,北京100050 [2]河南大学药学院,河南开封475004

出　　处：《药学学报》2011年第9期1058-1064,共7页Acta Pharmaceutica Sinica

基　　金：重大新药创制"科技重大专项"(2009ZX09302-003);国际科技合作项目"新药重大创新"(2009DFA32010)

摘　　要：本研究拟建立体外人源蛋白酪氨酸磷酸酶(PTP1B)抑制剂高通量筛选模型,用于PTP1B抑制剂的发现。利用大肠杆菌重组表达PTP1B,以对硝基苯磷酸二钠(PNPP)为特异性的底物,建立了基于酶反应速率的384孔微板为载体的PTP1B抑制剂高通量筛选模型(Z'=0.78)。选择24 240个样品进行筛选,对抑制率大于70%的80个样品作为活性样品进行复筛,最终确定6个具有较强的抑制活性的化合物J5753、J10550、J10551、J10583、J10585和J11101,其IC50值分别为21.58、18.39、15.37、11.92、37.27和36.61μg·mL-1。结果表明,所建立的PTP1B抑制剂高通量筛选模型具有快速、灵敏、稳定、重复性好的特点。To screen potential human soluble protein tyrosine phosphatase 1B（PTP1B） inhibitors,a high-throughput screening（HTS） model in 384-well microplate with total volume of 50 μL was established.Recombinant PTP1B was cloned and expressed in E.coli.with its specific substrate 4-nitrophenyl phosphate disodium salt hexahydrate（PNPP）.The HTS model was based on enzyme reaction rate with enhanced sensitivity and specificity（Z＇ = 0.78）.A total of 24 240 samples were screened,among them 80 samples with inhibition greater than 70% were selected for further rescreening.Finally,six compounds with high inhibitory activity were identified,whose IC 50 values were 21.58,18.39,15.37,11.92,37.27,and 36.61 μg·mL 1,separately.The results indicated that the method was stable,sensitive,reproducible and also suitable for high-throughput screening.

关 键 词：蛋白酪氨酸磷酸酶 抑制剂 高通量筛选 

分 类 号：R965.1[医药卫生—药理学]