PET imaging of fatty acid amide hydrolase in the brain: synthesis and biological evaluation of an 11C-labelled URB597 analogue.
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.
Wyffels
Leonie
L
Department of Radiopharmacy, Ghent University, Harelbekestraat 72, 9000 Ghent, Belgium.
Muccioli
Giulio G
GG
Kapanda
Coco N
CN
Labar
Geoffray
G
De Bruyne
Sylvie
S
De Vos
Filip
F
Lambert
Didier M
DM
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
D000581:Amidohydrolases; D001549:Benzamides; D002219:Carbamates; D002250:Carbon Radioisotopes; D000099252:Fatty Acid Amide Hydrolases; C500528:cyclohexyl carbamic acid 3'-carbamoylbiphenyl-3-yl ester