TY - JOUR T1 - KLYP956 Is a Non-Imidazole-Based Orally Active Inhibitor of Nitric-Oxide Synthase Dimerization JF - Molecular Pharmacology JO - Mol Pharmacol SP - 153 LP - 162 DO - 10.1124/mol.109.055434 VL - 76 IS - 1 AU - Kent T. Symons AU - Mark E. Massari AU - Phan M. Nguyen AU - Tom T. Lee AU - Jeffrey Roppe AU - CĂ©line Bonnefous AU - Joseph E. Payne AU - Nicholas D. Smith AU - Stewart A. Noble AU - Marciano Sablad AU - Natasha Rozenkrants AU - Yan Zhang AU - Tadimeti S. Rao AU - Andrew K. Shiau AU - Christian A. Hassig Y1 - 2009/07/01 UR - http://molpharm.aspetjournals.org/content/76/1/153.abstract N2 - Nitric-oxide synthases (NOS) generate nitric oxide (NO) through the oxidation of l-arginine. Inappropriate or excessive production of NO by NOS is associated with the pathophysiology of various disease states. Efforts to treat these disorders by developing arginine mimetic, substrate-competitive NOS inhibitors as drugs have met with little success. Small-molecule-mediated inhibition of NOS dimerization represents an intriguing alternative to substrate-competitive inhibition. An ultra-high-throughput cell-based screen of 880,000 small molecules identified a novel quinolinone with inducible NOS (iNOS) inhibitory activity. Exploratory chemistry based on this initial screening hit resulted in the synthesis of KLYP956, which inhibits iNOS at low nanomolar concentrations. The iNOS inhibitory potency of KLYP956 is insensitive to changes in concentrations of the substrate arginine, or the cofactor tetrahydrobiopterin. Mechanistic analysis suggests that KLYP956 binds the oxygenase domain in the vicinity of the active site heme and inhibits iNOS and neuronal NOS (nNOS) by preventing the formation of enzymatically active dimers. Oral administration of KLYP956 [N-(3-chlorophenyl)-N-((8-fluoro-2-oxo-1,2-dihydroquinolin-4-yl)methyl)-4-methylthiazole-5-carboxamide] inhibits iNOS activity in a murine model of endotoxemia and blocks pain behaviors in a formalin model of nociception. KLYP956 thus represents the first nonimidazole-based inhibitor of iNOS and nNOS dimerization and provides a novel pharmaceutical alternative to previously described substrate competitive inhibitors. ER -