Development of an In Vivo Method to Estimate Effective Drug Doses and Quantify Fatty Acid Amide Hydrolase in Rodent Brain using Positron Emission Tomography Tracer [C]DFMC.

Fatty acid amide hydrolase (FAAH) is a key enzyme in the endocannabinoid system. -(3,4-Dimethylisoxazol-5-yl)piperazine-4-[4-(2-fluoro-4-[C]methylphenyl)thiazol-2-yl]-1-carboxamide ([C]DFMC) was developed as an irreversible-type positron emission tomography (PET) tracer for FAAH. Here, we attempted to noninvasively estimate rate constant k (rate of transfer to the specifically-bound compartment) as a direct index for FAAH in the rat brain. First, the two-tissue compartment model analysis including three parameters [K-k, two-tissue compartment model for the irreversible-type radiotracer (2TCMi)] in PET study with [C]DFMC was conducted, which provided 0.21 ± 0.04 ml·cm·min of the net uptake value (K), an indirect index for FAAH, in the FAAH-richest region (the cingulate cortex). Subsequently, to noninvasively estimate K value, the reference model analysis (Patlak graphical analysis reference model) was tried using a time-activity curve of the spinal cord. In that result, the noninvasive K value (K) was concisely estimated with high correlation ( > 0.95) to K values based on 2TCMi. Using estimated K value, we tried to obtain calculated-k based on previously defined equations. The calculated k was successfully estimated with high correlation ( = 0.95) to direct k in 2TCMi. Finally, the dose relationship study using calculated k demonstrated that in vivo ED value of [3-(3-carbamoylphenyl)phenyl] -cyclohexylcarbamate, a major inhibitor of FAAH, was 66.4 µg/kg in rat brain. In conclusion, we proposed the calculated k as an alternative index corresponding to regional FAAH concentrations and suggested that PET with [C]DFMC enables occupancy study for new pharmaceuticals targeting FAAH. SIGNIFICANCE STATEMENT: In the present study, we proposed calculated k as an alternative index corresponding with fatty acid amide hydrolase concentration. By using calculated k, in vivo ED of [3-(3-carbamoylphenyl)phenyl] -cyclohexylcarbamate was successfully estimated to be 66.4 µg/kg for rats. Thus, we demonstrated the pharmacological utility of positron emission tomography with -(3,4-dimethylisoxazol-5-yl)piperazine-4-[4-(2-fluoro-4-[C]methylphenyl)thiazol-2-yl]-1-carboxamide.