Abstract
The prevalence of cannabis use and the concentrations of the psychoactive cannabinoid in cannabis, (-)-∆9-tetrahydrocannabinol (THC), are rising. Physiologically based pharmacokinetic modeling and simulations (PBPK M&S) can mechanistically predict exposure of THC and its major and active metabolite, 11-hydroxy-THC (11-OH-THC). To build a THC/11-OH-THC PBPK model, mechanistic information about the disposition of these compounds is necessary, including the drug-metabolizing enzymes (DMEs) involved and the fraction metabolized (fm) and metabolic kinetic parameters (intrinsic clearance, maximal formation rate, and Km) via the identified enzymes. We previously identified and quantified the fm of DMEs involved in hepatic depletion of THC and 11-OH-THC. In this study, we extend this work to characterize the enzyme kinetics of THC and 11-OH-THC by monitoring their depletion and formation of some of their metabolites in pooled human liver microsomes. A P450 and UDP-glucuronosyltransferase (UGT) kinetic model was fitted to the concentration-time depletion/formation profiles to establish the contribution and kinetics of the individual DME pathways. CYP2C9 pathway was the major pathway for depletion of THC (fm = 0.91, Km,u = 3 nM) and formation of 11-OH-THC. The remaining THC depletion pathway was attributed to CYP2D6. 11-OH-THC was depleted by UGTs (fm = 0.67 and Km,u = 39 nM), CYP3A4 (fm = 0.18, Km,u = 824 nM), and CYP2C9 (fm = 0.15, Km,u = 33 nM). These mechanistic in vitro data can be used to predict the exposure of THC and 11-OH-THC in healthy and special populations, including in the presence of drug-drug interactions, via PBPK M&S.