Wang Shou-Bao, Guo Jing, Yu Xiao-Ming, Du Guan-Hua
National Center for Pharmaceutical Screening, Institute of Materia Medica, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100050, China.
Yao Xue Xue Bao. 2010 Nov;45(11):1367-72.
To screen potential human soluble epoxide hydrolase (hsEH) inhibitors, a high-throughput screening model in 384-well microplate with total volume of 50 microL was established. Recombinant hsEH was cloned and expressed in E. coli. and its specific substrate PHOME was synthesized. The HTS model was based on fluorescence analysis with enhanced sensitivity and specificity (Z' = 0.65). A total of 47 360 samples (including 25 040 compounds and 22 320 natural products) were screened, of which 950 samples with inhibition greater than 80% were selected for further rescreening. Finally, two compounds with high inhibitory activity were identified, whose IC50 value were 8.56 and 4.31 micromol x L(-1), separately. The results indicated that the method was stable, sensitive, reproducible and also suitable for high-throughput screening.
为筛选潜在的人可溶性环氧化物水解酶（hsEH）抑制剂，建立了一种总反应体积为50微升的384孔微孔板高通量筛选模型。重组hsEH在大肠杆菌中克隆并表达，其特异性底物PHOME被合成。该高通量筛选模型基于荧光分析，具有更高的灵敏度和特异性（Z' = 0.65）。共筛选了47360个样品（包括25040种化合物和22320种天然产物），其中选择了950个抑制率大于80%的样品进行进一步复筛。最终，鉴定出两种具有高抑制活性的化合物，其IC50值分别为8.56和4.31微摩尔/升。结果表明该方法稳定、灵敏、可重复，也适用于高通量筛选。
