Discovery of novel PDEIO inhibitors by a robust homogeneous screening assay  

作　　者：Qun-yi LI Ming-kai XU Gang LIU Claus Tornby CHRISTOFFERSEN Ming-wei WANG 

机构地区：[1]The National Center for Drug Screening and the State Key Laboratory of Drug Research, Shanghai Institute of Materia Medica, Chi nese Academy of Sciences, Shanghai 201203, China [2]Institute of Materia Medica, Chinese Academy of Medical Sciences, Beijing 100050, China [3]H Lundbeck A/S, DK-2500 Copenhagen, Denmark

出　　处：《Acta Pharmacologica Sinica》2013年第8期1116-1120,共5页中国药理学报（英文版）

摘　　要：Aim： To develop a homogeneous assay for high-throughput screening （HTS） of inhibitors of phosphodiesterase 10 （PDEIO）. Methods-Purified human PDEIO enzyme derived from E coil, [3H]-cAMP and yttrium silicate microbeads were used to develop an HTS assay based on the scintillation proximity assay （SPA） technology. This method was applied to a large-scale screening campaign against a diverse compound library and subsequent confirmation studies. Preliminary structure-activity relationship （SAR） studies were initiated through limited structural modifications of the hits. Results： The IC50 value of the control compound （papaverine） assessed with the SPA approach was comparable and consistent with that reported in the literature. Signal to background （S/B） ratio and Z＇ factor of the assay system were evaluated to be 5.24 and 0.71, respectively. In an HTS campaign of 71 360 synthetic and natural compounds, 67 hits displayed reproducible PDEIO inhibition, of which, 8 were chosen as the scaffold for structural modifications and subsequent SAR analysis. Conclusion. The homogeneous PDEIO SPA assay is an efficient and robust tool to screen potential PDEIO inhibitors. Preliminary SAR studies suggest that potent PDEIO inhibitors could be identified and developed through this strategy.Aim： To develop a homogeneous assay for high-throughput screening （HTS） of inhibitors of phosphodiesterase 10 （PDEIO）. Methods-Purified human PDEIO enzyme derived from E coil, [3H]-cAMP and yttrium silicate microbeads were used to develop an HTS assay based on the scintillation proximity assay （SPA） technology. This method was applied to a large-scale screening campaign against a diverse compound library and subsequent confirmation studies. Preliminary structure-activity relationship （SAR） studies were initiated through limited structural modifications of the hits. Results： The IC50 value of the control compound （papaverine） assessed with the SPA approach was comparable and consistent with that reported in the literature. Signal to background （S/B） ratio and Z＇ factor of the assay system were evaluated to be 5.24 and 0.71, respectively. In an HTS campaign of 71 360 synthetic and natural compounds, 67 hits displayed reproducible PDEIO inhibition, of which, 8 were chosen as the scaffold for structural modifications and subsequent SAR analysis. Conclusion. The homogeneous PDEIO SPA assay is an efficient and robust tool to screen potential PDEIO inhibitors. Preliminary SAR studies suggest that potent PDEIO inhibitors could be identified and developed through this strategy.

关 键 词：phosphodiesterase 10 （PDEIO） PAPAVERINE high-throughput screening PSYCHOSIS 

分 类 号：O643.3[理学—物理化学] S511[理学—化学]