仓鼠糜酶2抑制剂高通量筛选模型的建立及其应用  

High-throughput screening for hamster chymase 2 inhibitors

作　　者：王守宝 竺晓鸣 高峰[1] 庞晓斌 杜冠华 

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

出　　处：《药学学报》2012年第2期168-173,共6页Acta Pharmaceutica Sinica

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

摘　　要：本研究拟建立重组仓鼠糜酶2(chymase 2)抑制剂的体外高通量筛选模型,寻找新型的糜酶2抑制剂。首先,利用大肠杆菌表达、制备成熟的重组仓鼠糜酶2蛋白,以384孔微板为媒介,建立基于荧光方法测定酶活性的抑制剂筛选模型,结果表明所建立的糜酶2抑制剂高通量筛选模型具有灵敏、稳定、重复性好的特点(Z'=0.84)。利用建立的模型对40 080种样品(包括28 060种化合物和12 020种天然产物提取物)进行抑制活性筛选,取抑制率大于90%的613种样品进行复筛。最终确定化合物J16647和J16648具有较强的抑制活性,其IC50值分别为0.823和0.690μmol.L?1。To screen potential hamster chymase 2 inhibitors, a high-throughput screening （HTS） model was established. Recombinant hamster chymase 2 with active form was cloned and expressed in E. coll. The HTS model with total volume of 50 μL in 384-well microplate was based on fluorescence analysis and was proved sensitive as well as specific （Z′ = 0.84）. A total of 40 080 samples （including 28 060 compounds and 12 020 natural products） were screened, and 613 samples with inhibition greater than 90% were selected for further rescreening. Finally, compounds J16647 and J16648 were identified with high inhibitory activity on chymase 2, and whose IC50 values were 0.823 and 0.690 μmaol·L^-1, respectively.

关 键 词：仓鼠糜酶2 糜酶抑制剂 高通量筛选 

分 类 号：R965.1[医药卫生—药理学]