Selective inhibition of rat hepatic microsomal cytochrome P-450. II. Effect of the in vitro administration of cimetidine.

The purpose of this study was to determine whether the differential inhibition of rat hepatic microsomal cytochrome P-450 in adult male rats by in vivo cimetidine administration is observed when the drug is administered in vitro. Cimetidine, at concentrations of up to 10 mM, did not affect the catalytic function of cytochrome P-450IIA1 as measured by testosterone 7 alpha-hydroxylase activity. In contrast, it did inhibit activities that are specific for cytochromes P-450IIC11 (testosterone 2 alpha-hydroxylase activity), P-450IIB1/2 (testosterone 16 beta-hydroxylase activity) and P-450IIA1/2 (testosterone 2 beta- and 6 beta-hydroxylase activities), with IC50 values in the range of 1.0 to 7.4 mM. To further investigate the inhibition of cytochrome P-450 enzymes by in vitro cimetidine, preincubation experiments were performed. Hepatic microsomes were preincubated with a low concentration (0.05 mM) of cimetidine and 1 mM NADPH for 15 min before the initiation of substrate (testosterone) oxidation. Under these conditions, cimetidine resulted in the inhibition of the enzyme activities specific for cytochrome P-450IIC11, but it had no effect on those specific for cytochromes P-450IIA1, P-450IIB1/2 and P-450IIIA1/2. This differential inhibition by in vitro cimetidine required the presence of NADPH in the preincubation medium, suggesting that a catalysis-dependent process is involved. Therefore, preincubation of hepatic microsomes with NADPH and a relatively low concentration (0.05 mM) of cimetidine in vitro results in a pattern of inhibition of cytochrome P-450 enzymes similar to that which occurs after the in vivo administration of cimetidine reported in the previous study (Chang et al., 1992).