Development of LC/MS/MS, high-throughput enzymatic and cellular assays for the characterization of compounds that inhibit kynurenine monooxygenase (KMO).
Kynurenine monooxygenase (KMO) catalyzes the conversion of kynurenine to 3-hydroxykynurenine. Modulation of KMO activity has been implicated in several neurodegenerative diseases, including Huntington disease. Our goal is to develop potent and selective small-molecule KMO inhibitors with suitable pharmacokinetic characteristics for in vivo proof-of-concept studies and subsequent clinical development. We developed a comprehensive panel of biochemical and cell-based assays that use liquid chromatography/tandem mass spectrometry to quantify unlabeled kynurenine and 3-hydroxykynurenine. We describe assays to measure KMO inhibition in cell and tissue extracts, as well as cellular assays including heterologous cell lines and primary rat microglia and human peripheral blood mononuclear cells.
['Winkler|Dirk|D|', 'Beconi|Maria|M|', 'Toledo-Sherman|Leticia M|LM|', 'Prime|Michael|M|', 'Ebneth|Andreas|A|', 'Dominguez|Celia|C|', 'Muñoz-Sanjuan|Ignacio|I|']
D000818:Animals D016466:CHO Cells D002460:Cell Line D002853:Chromatography, Liquid D003412:Cricetulus D004285:Dogs D055808:Drug Discovery D057075:Enzyme Assays D057809:HEK293 Cells D006801:Humans D006816:Huntington Disease D007737:Kynurenine D050601:Kynurenine 3-Monooxygenase D007963:Leukocytes, Mononuclear D008252:Macaca fascicularis D051379:Mice D008810:Mice, Inbred C57BL D051381:Rats D017208:Rats, Wistar D053719:Tandem Mass Spectrometry
LC/MS/MS kynurenine 3-hydroxykynurenine microglia human PBMCs kynurenine 3-monooxygenase