title: Small molecule inhibitors of phospholipase C from a novel high-throughput screen.
abstract: Phospholipase C (PLC) isozymes are important signaling molecules, but few small molecule modulators are available to pharmacologically regulate their function. With the goal of developing a general approach for identification of novel PLC inhibitors, we developed a high-throughput assay based on the fluorogenic substrate reporter WH-15. The assay is highly sensitive and reproducible: screening a chemical library of 6280 compounds identified three novel PLC inhibitors that exhibited potent activities in two separate assay formats with purified PLC isozymes in vitro. Two of the three inhibitors also inhibited G protein-coupled receptor-stimulated PLC activity in intact cell systems. These results demonstrate the power of the high-throughput assay for screening large collections of small molecules to identify novel PLC modulators. Potent and selective modulators of PLCs will ultimately be useful for dissecting the roles of PLCs in cellular processes, as well as provide lead compounds for the development of drugs to treat diseases arising from aberrant phospholipase activity.
authors.affiliation: Division of Chemical Biology and Medicinal Chemistry, University of North Carolina, Chapel Hill, North Carolina 27599, USA.
authors.forename: Weigang
authors.initials: W
authors.lastname: Huang
authors.forename: Matthew
authors.initials: M
authors.lastname: Barrett
authors.forename: Nicole
authors.initials: N
authors.lastname: Hajicek
authors.forename: Stephanie
authors.initials: S
authors.lastname: Hicks
authors.forename: T Kendall
authors.initials: TK
authors.lastname: Harden
authors.forename: John
authors.initials: J
authors.lastname: Sondek
authors.forename: Qisheng
authors.initials: Q
authors.lastname: Zhang
mesh_terms: D001681:Biological Assay / Q000379:methods; D002626:Chemistry, Pharmaceutical / Q000379:methods; D000242:Cyclic AMP / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D004353:Drug Evaluation, Preclinical / Q000379:methods; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D057809:HEK293 Cells; D006801:Humans; D007527:Isoenzymes; D008954:Models, Biological; D008956:Models, Chemical; D010740:Phospholipases / Q000737:chemistry; D043562:Receptors, G-Protein-Coupled / Q000378:metabolism; D015398:Signal Transduction; D054852:Small Molecule Libraries; D013379:Substrate Specificity; D010738:Type C Phospholipases / Q000037:antagonists & inhibitors* / Q000378:metabolism
chemical_list: D004791:Enzyme Inhibitors; D007527:Isoenzymes; D043562:Receptors, G-Protein-Coupled; D054852:Small Molecule Libraries; D000242:Cyclic AMP; D010740:Phospholipases; D010738:Type C Phospholipases