Identification of CYP3A4 and CYP2C8 as the major cytochrome P450 s responsible for morphine N -demethylation in human liver microsomes

Abstract

1. The aim was to identify the cytochrome P450 (CYP) enzymes responsible for the N -demethylation of morphine in vitro. 2. In human liver microsomes, normorphine formation followed Michaelis-Menten kinetics with mean K m and V max of 12.4 ± 2.2 mM and 1546 ± 121 pmol min − 1 mg − 1, respectively. In microsomes from a panel of 14 human livers phenotyped for 10 CYP enzymes, morphine N -demethylation correlated with testosterone 6 β -hydroxylation (r = 0.91, p <0.001) and paclitaxel 6- α hydroxylation (r = 0.72, p <0.001), two specific markers of CYP3A4 and CYP2C8, respectively. Normorphine formation decreased when incubated in the presence of troleandomycin or quercetin (by 46 and 33-36%, respectively), which further corroborates the contribution of CYP3A4 and CYP2C8. 3. Among eight recombinant human CYP enzymes tested, CYP3A4 and CYP2C8 exhibited the highest intrinsic clearance. More than 90% of morphine N -demethylation could be accounted for via the action of both CYP3A4 and CYP2C8. 4. The in vitro findings suggest that hepatic CYP3A4, and to a lesser extent CYP2C8, play an important role in the metabolism of morphine into normorphine.