title: Inhibition of anandamide hydrolysis in rat brain tissue by (E)-6-(bromomethylene) tetrahydro-3-(1-naphthalenyl)-2H-pyran-2-one.
abstract: Anandamide, an endogenous canabinoid substance, is hydrolyzed by an amidohydrolase activity present in rat brain and liver. We report that the bromoenol lactone, (E)-6-(bromomethylene) tetrahydro-3-(1-naphthalenyl)-2H-pyran-2-one (BTNP), is a potent inhibitor of this enzyme activity. BTNP prevented anandamide hydrolysis in rat brain microsomes with an IC50 of 0.8 +/- 0.3 microM. Kinetic and dialysis experiments indicated that this effect was non-competitive and irreversible. After chromatographic fractionation of the enzyme activity, BTNP was still effective, suggesting that it interacts directly with the enzyme. Anandamide hydrolysis was 12-fold greater in rat cortical neurons (1.94 +/- 0.1 pmol/min/mg protein) than in cortical astrocytes (0.16 +/- 0.01 pmol/min/mg protein) and, in either cell type, it was inhibited by BTNP (IC50 = 0.1 microM in neurons). These results suggest that BTNP may provide a useful lead for the development of novel inhibitors of anandamide hydrolysis.
authors.affiliation: Neurosciences Institute, San Diego, CA 92121, USA.
authors.forename: M
authors.initials: M
authors.lastname: Beltramo
authors.forename: E
authors.initials: E
authors.lastname: di Tomaso
authors.forename: D
authors.initials: D
authors.lastname: Piomelli
mesh_terms: D000581:Amidohydrolases / Q000037:antagonists & inhibitors*; D000818:Animals; D001095:Arachidonic Acids / Q000378:metabolism*; D001253:Astrocytes / Q000201:enzymology; D001921:Brain / Q000201:enzymology*; D002540:Cerebral Cortex / Q000166:cytology / Q000201:enzymology; D063388:Endocannabinoids; D004791:Enzyme Inhibitors / Q000494:pharmacology; D006868:Hydrolysis / Q000187:drug effects; D007476:Ionophores / Q000494:pharmacology; D008862:Microsomes, Liver / Q000201:enzymology; D009281:Naphthalenes / Q000494:pharmacology*; D009474:Neurons / Q000201:enzymology; D010713:Phosphatidylcholines / Q000378:metabolism; D010714:Phosphatidylethanolamines / Q000378:metabolism; D053284:Polyunsaturated Alkamides; D011753:Pyrones / Q000494:pharmacology*; D051381:Rats; D017208:Rats, Wistar; D000099252:Fatty Acid Amide Hydrolases
chemical_list: D000581:Amidohydrolases; D001095:Arachidonic Acids; D063388:Endocannabinoids; D004791:Enzyme Inhibitors; D007476:Ionophores; D009281:Naphthalenes; D010713:Phosphatidylcholines; D010714:Phosphatidylethanolamines; D053284:Polyunsaturated Alkamides; D011753:Pyrones; D000099252:Fatty Acid Amide Hydrolases; C068314:6-(bromomethylene)tetrahydro-3-(1-naphthaleneyl)-2H-pyran-2-one; C078814:anandamide