title: PET imaging of fatty acid amide hydrolase in the brain: synthesis and biological evaluation of an 11C-labelled URB597 analogue.
abstract: BACKGROUND
  Fatty acid amide hydrolase (FAAH) is part of the endocannabinoid system (ECS) and has been linked to the aetiology of several neurological and neuropsychiatric disorders. So far no useful PET or SPECT tracer for in vivo visualisation of FAAH has been reported. We synthesized and evaluated a carbon-11-labeled URB597 analogue, biphenyl-3-yl [(11)C]-4-methoxyphenylcarbamate or [(11)C]-1, as potential FAAH imaging agent.
  
  
  METHODS
  The inhibitory activity of 1 was determined in vitro using recombinant FAAH. Radiosynthesis of [(11)C]-1 was performed by methylation using [(11)C]-CH(3)I, followed by HPLC purification. Biological evaluation was done by biodistribution studies in wild-type and FAAH knock-out mice, and by ex vivo and in vivo metabolite analysis. The influence of URB597 pretreatment on the metabolisation profile was assessed.
  
  
  RESULTS
  [(11)C]-1 was obtained in good yields and high radiochemical purity. Biodistribution studies revealed high brain uptake in wild-type and FAAH knock-out mice, but no retention of radioactivity could be demonstrated. Metabolite analysis and URB597 pretreatment confirmed the non-FAAH-mediated metabolisation of [(11)C]-1. The inhibition mechanism was determined to be reversible. In addition, the inhibition of URB597 appeared slowly reversible.
  
  
  CONCLUSIONS
  Although [(11)C]-1 inhibits FAAH in vitro and displays high brain uptake, the inhibition mechanism seems to deviate from the proposed carbamylation mechanism. Consequently, it does not covalently bind to FAAH and will not be useful for mapping the enzyme in vivo. However, it represents a potential starting point for the development of in vivo FAAH imaging tools.
authors.affiliation: Department of Radiopharmacy, Ghent University, Harelbekestraat 72, 9000 Ghent, Belgium.
authors.forename: Leonie
authors.initials: L
authors.lastname: Wyffels
authors.forename: Giulio G
authors.initials: GG
authors.lastname: Muccioli
authors.forename: Coco N
authors.initials: CN
authors.lastname: Kapanda
authors.forename: Geoffray
authors.initials: G
authors.lastname: Labar
authors.forename: Sylvie
authors.initials: S
authors.lastname: De Bruyne
authors.forename: Filip
authors.initials: F
authors.lastname: De Vos
authors.forename: Didier M
authors.initials: DM
authors.lastname: Lambert
mesh_terms: D000581:Amidohydrolases / Q000037:antagonists & inhibitors / Q000172:deficiency / Q000235:genetics / Q000378:metabolism*; D000818:Animals; D001549:Benzamides / Q000138:chemical synthesis* / Q000378:metabolism / Q000494:pharmacology; D001921:Brain / Q000000981:diagnostic imaging* / Q000378:metabolism*; D002219:Carbamates / Q000138:chemical synthesis* / Q000378:metabolism / Q000494:pharmacology; D002250:Carbon Radioisotopes; D055786:Gene Knockout Techniques; D006801:Humans; D008297:Male; D051379:Mice; D049268:Positron-Emission Tomography / Q000379:methods*; D011786:Quality Control; D000099252:Fatty Acid Amide Hydrolases
chemical_list: D000581:Amidohydrolases; D001549:Benzamides; D002219:Carbamates; D002250:Carbon Radioisotopes; D000099252:Fatty Acid Amide Hydrolases; C500528:cyclohexyl carbamic acid 3'-carbamoylbiphenyl-3-yl ester