Wang Yuren, Ramirez Fernando, Krishnamurthy Girija, Gilbert Adam, Kadakia Nina, Xu Jun, Kalgaonkar Gary, Ramarao Manjunath K, Edris Wade, Rogers Kathryn E, Jones Philip G
Neuroscience Discovery Research and Chemical and Screening Sciences, Wyeth Research Princeton, NJ 08543-8000, USA.
J Biomol Screen. 2006 Aug;11(5):519-27. doi: 10.1177/1087057106288188. Epub 2006 Jun 7.
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.
脂肪酸酰胺水解酶（FAAH）是一种与膜相关的酶，可催化多种内源性生物活性脂质的水解，包括花生四烯酸乙醇胺（AEA）、N-棕榈酰乙醇胺（PEA）、油酰胺和N-油酰乙醇胺（OEA）。这些脂肪酸酰胺参与许多生理活动，如镇痛、焦虑调节、睡眠调节、抗炎反应和食欲抑制。由于FAAH在控制这些内源性生物活性脂质的水平和活性方面起着至关重要的作用，该酶已被认为是治疗疼痛、焦虑和其他疾病的药物靶点。为了发现FAAH抑制剂，作者此前报道了一种新型荧光测定法的开发，该方法使用纯化的FAAH微粒体作为酶源和荧光底物花生四烯酰7-氨基、4-甲基香豆素酰胺（AAMCA）。在此，作者将该测定法调整为高通量形式，并筛选了一个大型有机小分子化合物库，鉴定出了一些新型FAAH抑制剂。这些数据进一步证实，这种荧光测定法对于鉴定FAAH抑制分子具有足够的稳健性、高效性和低成本性，并为治疗探索打开了这类酶的大门。
