New heterocyclic modifiers of oxidative drug metabolism--II. Steric factors in the interaction of isomeric 2-(naphthyl)methylbenzimidazoles with rat hepatic microsomal cytochrome P-450 and monooxygenase activities.

The inhibitory potency of the two isomeric 2-(naphthyl)methylbenzimidazoles towards three monooxygenase activities (aminopyrine N-demethylase, 7-ethoxycoumarin O-deethylase and aniline p-hydroxylase) was assessed in hepatic microsomal fractions from untreated, phenobarbitone-induced and beta-naphthoflavone-induced rats. The isomers were essentially equipotent with each other as inhibitors of the phenobarbitone-induced monooxygenases (the ratio of the I50s of the isomers was about 1.0 in each case) but differences between the isomers were noted in the inhibition potencies against three monooxygenase activities from beta-naphthoflavone-induced liver. The isomer 2-(1'-naphthyl)methylbenzimidazole was approximately twice as potent as the 2'-naphthyl isomer against 7-ethoxyresorufin O-deethylase activity, whereas the opposite was observed with respect to 7-ethoxycoumarin O-deethylase inhibition; aniline p-hydroxylase was poorly inhibited by both isomers. The binding affinity and extent of binding, assessed from double-reciprocal plots of spectral binding studies, of the 1'-isomer was much greater than that of the 2'-isomer in beta-naphthoflavone-induced microsomes. Inhibition data in untreated hepatic microsomes were more complex and the finding of principal interest was that the 1'-isomer was poorly inhibitory towards aniline p-hydroxylase activity whereas the 2'-isomer enhanced this activity. These studies suggest that the steric conformations of the isomeric naphthylmethylbenzimidazoles at the cytochrome P-450 active centre determines the extent to which the inhibitors modulate a specific monooxygenase activity, and that multiple binding sites with the capacity to interact to different extents with benzimidazole derivatives are present in P-450 in beta-naphthoflavone-induced hepatic microsomes. The apparent importance of steric conformation as a determinant of inhibition and enhancement of aniline p-hydroxylase in untreated microsomal fractions may well reflect specific interactions with multiple binding sites.