hu-243 and anandamide

Topics: Amides; Analgesics; Animals; Arachidonic Acids; Benzoxazines; Calcium Channel Blockers; Cannabinoids; Endocannabinoids; Fatty Acids, Unsaturated; Humans; Ligands; Morpholines; Naphthalenes; Polyunsaturated Alkamides; Receptors, Cannabinoid; Receptors, Drug

A putative endogenous cannabinoid ligand, arachidonylethanolamide (termed "anandamide"), was isolated recently from porcine brain. Here we demonstrate that this compound is a specific cannabinoid agonist and exerts its action directly via the cannabinoid receptors. Anandamide specifically binds to membranes from cells transiently (COS) or stably (Chinese hamster ovary) transfected with an expression plasmid carrying the cannabinoid receptor DNA but not to membranes from control nontransfected cells. Moreover, anandamide inhibited the forskolin-stimulated adenylate cyclase in the transfected cells and in cells that naturally express cannabinoid receptors (N18TG2 neuroblastoma) but not in control nontransfected cells. As with exogenous cannabinoids, the inhibition by anandamide of the forskolin-stimulated adenylate cyclase was blocked by treatment with pertussis toxin. These data indicate that anandamide is an endogenous agonist that may serve as a genuine neurotransmitter for the cannabinoid receptor.

Topics: Adenylyl Cyclase Inhibitors; Amides; Animals; Arachidonic Acids; Brain; Cannabinoids; CHO Cells; Cricetinae; Endocannabinoids; Fatty Acids, Unsaturated; Polyunsaturated Alkamides; Receptors, Cannabinoid; Receptors, Drug; Transfection

Arachidonylethanolamide, an arachidonic acid derivative in porcine brain, was identified in a screen for endogenous ligands for the cannabinoid receptor. The structure of this compound, which has been named "anandamide," was determined by mass spectrometry and nuclear magnetic resonance spectroscopy and was confirmed by synthesis. Anandamide inhibited the specific binding of a radiolabeled cannabinoid probe to synaptosomal membranes in a manner typical of competitive ligands and produced a concentration-dependent inhibition of the electrically evoked twitch response to the mouse vas deferens, a characteristic effect of psychotropic cannabinoids. These properties suggest that anandamide may function as a natural ligand for the cannabinoid receptor.

Topics: Amides; Animals; Arachidonic Acids; Binding, Competitive; Brain; Brain Chemistry; Cannabinoids; Chromatography, Thin Layer; Endocannabinoids; Fatty Acids, Unsaturated; Gas Chromatography-Mass Spectrometry; Kinetics; Polyunsaturated Alkamides; Receptors, Cannabinoid; Receptors, Drug; Swine