title: Lack of selectivity of URB602 for 2-oleoylglycerol compared to anandamide hydrolysis in vitro.
abstract: OBJECTIVE
  Two compounds, URB602 and URB754, have been reported in the literature to be selective inhibitors of monoacylglycerol lipase, although a recent study has questioned their ability to prevent 2-arachidonoyl hydrolysis by brain homogenates and cerebellar membranes. In the present study, the ability of these compounds to inhibit monoacylglycerol lipase and fatty acid amide hydrolase has been reinvestigated.
  
  
  METHODS
  Homogenates and cell lines were incubated with test compounds and, thereafter, with either [(3)H]-2-oleoylglycerol or [(3)H]-anandamide. Labelled reaction products were separated from substrate using chloroform: methanol extraction.
  
  
  RESULTS
  In cytosolic fractions from rat brain, URB602 and URB754 inhibited the hydrolysis of 2-oleoylglycerol with IC(50) values of 25 and 48 microM, respectively. Anandamide hydrolysis by brain membranes was not sensitive to URB754, but was inhibited by URB602 (IC(50) value 17 microM). Hydrolysis of 2-oleoylglycerol by human recombinant monoacylglycerol lipase was sensitive to URB602, but not URB754. The lack of selectivity of URB602 for 2-oleoylglycerol compared to anandamide hydrolysis was also observed for intact RBL2H3 basophilic leukaemia cells. C6 glioma expressed mRNA for monoacylglycerol lipase, and hydrolyzed 2-oleoylglycerol in a manner sensitive to inhibition by methyl arachidonoyl fluorophosphonate but not URB754 or URB597. MC3T3-E1 mouse osteoblastic cells, which did not express mRNA for monoacylglycerol lipase, hydrolyzed 2-oleoylglycerol in the presence of URB597, but the hydrolysis was less sensitive to methyl arachidonoyl fluorophosphonate than for C6 cells.
  
  
  CONCLUSIONS
  The data demonstrate that the compounds URB602 and URB754 do not behave as selective and/or potent inhibitors of monoacylglycerol lipase.
authors.affiliation: Department of Pharmacology and Clinical Neuroscience, Umeå University, Umeå, Sweden.
authors.forename: S
authors.initials: S
authors.lastname: Vandevoorde
authors.forename: K-O
authors.initials: KO
authors.lastname: Jonsson
authors.forename: G
authors.initials: G
authors.lastname: Labar
authors.forename: E
authors.initials: E
authors.lastname: Persson
authors.forename: D M
authors.initials: DM
authors.lastname: Lambert
authors.forename: C J
authors.initials: CJ
authors.lastname: Fowler
mesh_terms: D000581:Amidohydrolases / Q000037:antagonists & inhibitors*; D000814:Aniline Compounds / Q000494:pharmacology*; D000818:Animals; D001095:Arachidonic Acids / Q000378:metabolism*; D048588:Benzoxazines / Q000494:pharmacology*; D001713:Biphenyl Compounds / Q000494:pharmacology*; D045744:Cell Line, Tumor; D002478:Cells, Cultured; D002531:Cerebellum / Q000378:metabolism; D003600:Cytosol / Q000378:metabolism; D063388:Endocannabinoids; D005989:Glycerides / Q000378:metabolism*; D006801:Humans; D006868:Hydrolysis; D066298:In Vitro Techniques; D008297:Male; D008566:Membranes / Q000378:metabolism; D051379:Mice; D008994:Monoacylglycerol Lipases / Q000037:antagonists & inhibitors*; D053284:Polyunsaturated Alkamides / Q000378:metabolism*; D051381:Rats; D017208:Rats, Wistar; D011994:Recombinant Proteins / Q000037:antagonists & inhibitors; D000099252:Fatty Acid Amide Hydrolases
chemical_list: D000581:Amidohydrolases; D000814:Aniline Compounds; D001095:Arachidonic Acids; D048588:Benzoxazines; D001713:Biphenyl Compounds; D063388:Endocannabinoids; D005989:Glycerides; D008994:Monoacylglycerol Lipases; D053284:Polyunsaturated Alkamides; D011994:Recombinant Proteins; D000099252:Fatty Acid Amide Hydrolases; C514879:URB 754; C501835:URB602; C505247:2-oleoylglycerol; C078814:anandamide