title: High-throughput screening for the discovery of inhibitors of fatty acid amide hydrolase using a microsome-based fluorescent assay.
abstract: Fatty acid amide hydrolase (FAAH) is a membrane-associated enzyme that catalyzes the hydrolysis of several endogenous bioactive lipids, including anandamide (AEA), N-palmitoylethanolamine (PEA), oleamide, and N-oleoylethanolamine (OEA). These fatty acid amides participate in many physiological activities such as analgesia, anxiety, sleep modulation, anti inflammatory responses, and appetite suppression. Because FAAH plays an essential role in controlling the tone and activity of these endogenous bioactive lipids, this enzyme has been implicated to be a drug target for the therapeutic management of pain, anxiety, and other disorders. In an effort to discover FAAH inhibitors, the authors have previously reported the development of a novel fluorescent assay using purified FAAH microsomes as an enzyme source and a fluorogenic substrate, arachidonyl 7-amino, 4-methyl coumarin amide (AAMCA). Herein, the authors have adapted this assay to a high-throughput format and have screened a large library of small organic compounds, identifying a number of novel FAAH inhibitors. These data further verify that this fluorescent assay is sufficiently robust, efficient, and low-cost for the identification of FAAH inhibitory molecules and open this class of enzymes for therapeutic exploration.
authors.affiliation: Neuroscience Discovery Research and Chemical and Screening Sciences, Wyeth Research Princeton, NJ 08543-8000, USA. wangy4@wyeth.com
authors.forename: Yuren
authors.initials: Y
authors.lastname: Wang
authors.forename: Fernando
authors.initials: F
authors.lastname: Ramirez
authors.forename: Girija
authors.initials: G
authors.lastname: Krishnamurthy
authors.forename: Adam
authors.initials: A
authors.lastname: Gilbert
authors.forename: Nina
authors.initials: N
authors.lastname: Kadakia
authors.forename: Jun
authors.initials: J
authors.lastname: Xu
authors.forename: Gary
authors.initials: G
authors.lastname: Kalgaonkar
authors.forename: Manjunath K
authors.initials: MK
authors.lastname: Ramarao
authors.forename: Wade
authors.initials: W
authors.lastname: Edris
authors.forename: Kathryn E
authors.initials: KE
authors.lastname: Rogers
authors.forename: Philip G
authors.initials: PG
authors.lastname: Jones
mesh_terms: D000581:Amidohydrolases / Q000037:antagonists & inhibitors*; D000818:Animals; D001331:Automation / Q000379:methods; D016466:CHO Cells; D006224:Cricetinae; D004353:Drug Evaluation, Preclinical / Q000379:methods*; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D005456:Fluorescent Dyes / Q000032:analysis*; D006801:Humans; D020128:Inhibitory Concentration 50; D008861:Microsomes / Q000201:enzymology*; D008954:Models, Biological; D014162:Transfection; D000099252:Fatty Acid Amide Hydrolases
chemical_list: D000581:Amidohydrolases; D004791:Enzyme Inhibitors; D005456:Fluorescent Dyes; D000099252:Fatty Acid Amide Hydrolases