The endocannabinoid anandamide is removed from the synaptic space by a selective transport system, expressed in neurons and astrocytes, that remains molecularly uncharacterized. Here we describe a partly cytosolic variant of the intracellular anandamide-degrading enzyme fatty acid amide hydrolase-1 (FAAH-1), termed FAAH-like anandamide transporter (FLAT), that lacked amidase activity but bound anandamide with low micromolar affinity and facilitated its translocation into cells. Known anandamide transport inhibitors, such as AM404 and OMDM-1, blocked these effects. We also identified a competitive antagonist of the interaction of anandamide with FLAT, the phthalazine derivative ARN272, that prevented anandamide internalization in vitro, interrupted anandamide deactivation in vivo and exerted profound analgesic effects in rodent models of nociceptive and inflammatory pain, which were mediated by CB1 cannabinoid receptors. The results identify FLAT as a critical molecular component of anandamide transport in neural cells and a potential target for therapeutic drugs.
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We thank G. Colombano, J. Kim, S. Mandrup-Bertozzi, D. Thongkham, G. Moreno-Sanz and R. Scarpelli for help with experiments and discussions about the project. This research was supported by grants from the US National Institute on Drug Abuse and the US National Institute on Alcohol Abuse and Alcoholism (DA-012413 to D.P.) and the US National Institute on General Medicine (1R01GM093937-01 to W.R.). The support provided by the Agilent/University of California Irvine Analytical Discovery Facility is gratefully acknowledged.
A patent was filed by the University of California, Irvine, and the Italian Institute of Technology on behalf of D.P., J.F., G.B. and A.C.
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