title: In vivo activity of a phospholipase C inhibitor, 1-(6-((17beta-3-methoxyestra-1,3,5(10)-trien-17-yl)amino)hexyl)-1H-pyrrole-2,5-dione (U73122), in acute and chronic inflammatory reactions.
abstract: To investigate the role of phospholipase C (PLC) in inflammatory processes, we tested 1-(6-((17beta-3-methoxyestra-1,3,5(10)-trien-17-yl)amino)hexyl)-1H-pyrrole-2,5-dione (U73122), a widely used PLC inhibitor, in several in vitro and in vivo assays. We first examined the effects of U73122 on human phospholipase C-beta (PLC-beta) isozymes and found that U73122 significantly inhibited recombinant human PLC-beta2, with an IC(50) of approximately 6 microM. U73122 had little effect on PLC-beta1, PLC-beta3, or PLC-beta4. Consistent with its ability to inhibit PLC-beta2 enzymatic activity, U73122 reduced interleukin-8 and leukotriene B(4)-induced Ca(2+) flux and chemotaxis in human neutrophils in a concentration-dependent manner. In vivo, U73122 blocked carrageenan-induced hind paw edema in rats, carrageenan-induced macrophage and lymphocyte accumulation into subcutaneous chambers in dogs, lipopolysaccharide-induced macrophage, lymphocyte infiltration and prostaglandin E(2) production in a mouse peritonitis model, and 12-O-tetradecanoylphorbol-13-acetate-induced ear edema in mice. These results implicate PLC-dependent signaling pathways in the development of acute and chronic inflammatory responses in vivo.
authors.affiliation: Department of Drug Discovery, Johnson and Johnson Pharmaceutical Research and Development, L.L.C., Raritan, NJ 08869, USA. chou@prdus.jnj.com
authors.forename: Cuifen
authors.initials: C
authors.lastname: Hou
authors.forename: Thomas
authors.initials: T
authors.lastname: Kirchner
authors.forename: Monica
authors.initials: M
authors.lastname: Singer
authors.forename: Michele
authors.initials: M
authors.lastname: Matheis
authors.forename: Dennis
authors.initials: D
authors.lastname: Argentieri
authors.forename: Druie
authors.initials: D
authors.lastname: Cavender
mesh_terms: D000700:Analgesics / Q000494:pharmacology; D000818:Animals; D001665:Binding Sites; D002118:Calcium / Q000378:metabolism; D002351:Carrageenan; D002465:Cell Movement / Q000187:drug effects; D051546:Cyclooxygenase 2; D015232:Dinoprostone / Q000378:metabolism; D004285:Dogs; D004347:Drug Interactions; D004487:Edema / Q000139:chemically induced / Q000188:drug therapy; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D004963:Estrenes / Q000494:pharmacology*; D006801:Humans; D016209:Interleukin-8 / Q000494:pharmacology; D007527:Isoenzymes / Q000037:antagonists & inhibitors* / Q000378:metabolism; D007975:Leukotriene B4 / Q000494:pharmacology; D008070:Lipopolysaccharides / Q000494:pharmacology; D017737:Macrophages, Peritoneal / Q000187:drug effects; D008565:Membrane Proteins; D051379:Mice; D009504:Neutrophils / Q000166:cytology / Q000187:drug effects*; D010533:Peritoneal Lavage; D054799:Phospholipase C beta; D010739:Phospholipase D / Q000378:metabolism; D011451:Prostaglandin-Endoperoxide Synthases / Q000378:metabolism; D011760:Pyrrolidinones / Q000494:pharmacology*; D051381:Rats; D011965:Receptors, Glucocorticoid / Q000378:metabolism; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D010738:Type C Phospholipases / Q000037:antagonists & inhibitors*
chemical_list: D000700:Analgesics; D004791:Enzyme Inhibitors; D004963:Estrenes; D016209:Interleukin-8; D007527:Isoenzymes; D008070:Lipopolysaccharides; D008565:Membrane Proteins; D011760:Pyrrolidinones; D011965:Receptors, Glucocorticoid; C060229:1-(6-((3-methoxyestra-1,3,5(10)-trien-17-yl)amino)hexyl)-1H-pyrrole-2,5-dione; D007975:Leukotriene B4; D002351:Carrageenan; D051546:Cyclooxygenase 2; C496540:PTGS2 protein, human; D011451:Prostaglandin-Endoperoxide Synthases; D010738:Type C Phospholipases; C518033:PLCB1 protein, human; C518036:PLCB2 protein, human; C518039:PLCB3 protein, human; C518042:PLCB4 protein, human; D054799:Phospholipase C beta; C518034:Plcb1 protein, mouse; C518035:Plcb1 protein, rat; C518037:Plcb2 protein, mouse; C518038:Plcb2 protein, rat; C518040:Plcb3 protein, mouse; C518041:Plcb3 protein, rat; C518043:Plcb4 protein, mouse; C518044:Plcb4 protein, rat; D010739:Phospholipase D; D015232:Dinoprostone; D013755:Tetradecanoylphorbol Acetate; D002118:Calcium