High-throughput cell-based screening using scintillation proximity assay for the discovery of inositol phosphatase inhibitors.
Inositol monophosphatase is a potential drug target for developing lithium-mimetic agents for the treatment of bipolar disorder. Enzyme-based assays have been traditionally used in compound screening to identify inositol monophosphatase inhibitors. A cell-based screening assay in which the compound needs to cross the cell membrane before reaching the target enzyme offers a new approach for discovering novel structure leads of the inositol monophosphatase inhibitor. The authors have recently reported a high-throughput measurement of G-protein-coupled receptor activation by determining inositol phosphates in cell extracts using scintillation proximity assay. This cell-based assay has been modified to allow the determination of inositol monophosphatase activity instead of G-protein-coupled receptors. The enzyme is also assayed in its native form and physiological environment. The authors have applied this cell-based assay to the high-throughput screening of a large compound collection and identified several novel inositol monophosphatase inhibitors.
Department of Automated Biotechnology, Merck & Co., North Wales, PA 19454, USA. wei_zheng@merck.com
Wei
W
Zheng
Philip E
PE
Brandish
D Garrett
DG
Kolodin
Edward M
EM
Scolnick
Berta
B
Strulovici
D001285:Atropine / Q000494:pharmacology; D001681:Biological Assay / Q000379:methods; D002217:Carbachol / Q000494:pharmacology; D002478:Cells, Cultured; D002851:Chromatography, High Pressure Liquid; D006801:Humans; D008954:Models, Biological; D010744:Phosphoric Monoester Hydrolases / Q000037:antagonists & inhibitors*; D012588:Scintillation Counting
D001285:Atropine; D002217:Carbachol; D010744:Phosphoric Monoester Hydrolases; C021822:myo-inositol-1 (or 4)-monophosphatase