In vivo study on the roles of cytochrome P450 enzymes for metabolism of 3,4-methylenedioxymethamphetamine (Ecstasy) in rats

Some major metabolic pathways of 3,4-methylenedioxymethamphetamine (MDMA) have been shown to be dependent on cytochrome P450 (CYP) isozymes by in vitro studies. The aim of this study was to clarify the roles of these CYP enzymes for in vivo metabolism of MDMA with respect to two pathways using rats: N-demethylation of MDMA to 3,4-methylenedioxyamphetamine (MDA) and O-demethylenation of MDMA to 3,4-dihydroxymethamphetamine (HHMA)followed by O-methylation to 4-hydroxy-3-methoxymethamphetamine (HMMA). Rats were pretreated with phenobarbital (PB, 80 mg/kg i.p.) or β-naphthoflavone (BNF, 80 mg/kg i.p.) once a day for 3 days before administration of MDMA (10 mg/kg i.p.). Metabolic changes were monitored by measuring the urinary excretion of MDMA and its metabolites. Twenty-four hours after MDMA administration, MDA in rat urine was significantly decreased by 43% and 70%, and HMMA was significantly increased by 33% and 64% in urine samples from PB-pretreated and BNF-pretreated rats, respectively, as compared with the control values. Testosterone 6β-hydroxylase (CYP3A), pentoxyresorufin O-dealkylase (CYP2B1), ethoxyresorufin O-deethylase (CYP1A1), and methoxyresorufin O-demethylase (CYP1A2) activities were increased 2–6 fold in both PB-pretreated and BNF-pretreated rat liver microsomes sampled at 24 h after MDMA administration as compared with the control values. These results suggest that PB-induced and BNF-induced CYP enzymes have inhibitory effects on N-demethylation of MDMA to MDA in vivo in rats. If HHMA is the precursor of HMMA in rats, there is a possibility that the O-demethylenation of MDMA to HHMA is increased by the induced CYP enzymes. The decreased urinary concentration of MDMA and very low percent recoveries of MDA, HMMA, and (4-hydroxy-3-methoxyphenyl)acetone (HMPA) in the inducer-pretreated groups suggest that other metabolic pathways of MDMA exist and are activated under the present experimental conditions.