Identification of small molecules that inhibit GSK-3beta through virtual screening.
Glycogen synthase kinase-3beta (GSK-3beta) is involved in glycogen metabolism, neuronal cell development, osteoblast differentiation. Small molecule inhibitors of GSK-3beta have various therapeutic potential for the treatment of diabetes type II, bipolar disorders, stroke and chronic inflammatory disease. To identify GSK-3beta inhibitors with novel scaffold from chemical library, we primarily screened out putative inhibitors through computer modeling and subsequently evaluated the inhibitory activity of selected compounds against GSK-3beta by in vitro Z'-LYTEtrade mark assay. A series of compound KRMs strongly inhibited phosphorylation of its substrate with IC(50) value of approximately 0.5microM. Also, we demonstrated that KRM-189 and KRM-191 competed with ATP for GSK-3beta, leading to decreased Vmax and constant Km with increasing concentrations of ATP as determined from Lineweaver-Berk equation. Moreover, they showed the selectivity for GSK-3beta over other kinases with IC(50) values of 2 to 10microM or more Incubation of cells with KRM-191 with highly selective and potent inhibitory activity caused accumulation of beta-catenin, downstream of GSK-3beta signaling pathway, indicating that small molecule can prevent degradation of beta-catenin via GSK-3beta inhibition. Our results suggest that modeling in combination with in vitro assays can be used for the identification of selective and potent inhibitors.
Kang
Nam Sook
NS
Center for Drug Discovery Technologies, Korea Research Institute of Chemical Technology, PO Box 107, Yuseong-gu, Daejeon 305-600, Republic of Korea.
Lee
Gil Nam
GN
Kim
Chi Hyun
CH
Bae
Myung Ae
MA
Kim
Ikyon
I
Cho
Young Sik
YS
D001667:Binding, Competitive; D003198:Computer Simulation; D004791:Enzyme Inhibitors / Q000378:metabolism / Q000494:pharmacology*; D038362:Glycogen Synthase Kinase 3 / Q000037:antagonists & inhibitors* / Q000378:metabolism; D000071679:Glycogen Synthase Kinase 3 beta; D010766:Phosphorylation; D015398:Signal Transduction
D004791:Enzyme Inhibitors; D000071679:Glycogen Synthase Kinase 3 beta; D038362:Glycogen Synthase Kinase 3