Metabolism of lidocaine by liver microsomes from streptozotocin-diabetic rats.
The aim of the present study was to determine optimum conditions for the study of thioridazine metabolism in rat liver microsomes and to investigate the influence of specific cytochrome P-450 inhibitors on 2- and 5-sulfoxidation, and N-demethylation of thioridazine. Basing on the developed method, the thioridazine metabolism in liver microsomes was studied at linear dependence of the product formation on time, and protein and substrate concentrations (incubation time was 15 min, concentration of microsomal protein was 0.5 mg/ml, substrate concentrations were 25, 50 and 75 nmol/ml). Dixon analysis of tioridazine metabolism carried out in the control liver microsomes, in the absence and presence of specific cytochrome P-450 inhibitors, showed that quinine (CYP2D1 inhibitor), metyrapone (CYP2B1/B2 inhibitor) and alpha-naphthoflavone (CYP1A2 inhibitor) affected while erythromycin (CYP3A inhibitor) and sulfaphenazole (CYP2C9 inhibitor) did not affect the neuroleptic biotransformation. Thus, quinine and metyrapone inhibited competitively thioridazine N-demethylation and mono-2-sulfoxidation. As reflected by Ki values, N-demethylation was inhibited to a higher degree (Ki = 16.5 and 43 microM, respectively) than mono-2-sulfoxidation (Ki = 25 and 137 microM, respectively). On the other hand, alpha-naphthoflavone inhibited competitively not only N-demethylation and mono-2-sulfoxidation, but also 5-sulfoxidation of thioridazine. The calculated Ki values showed that the highest potency of alpha-naphthoflavone to inhibit thioridazine metabolism was observed for N-demethylation and it descended in the following order: N-demethylation (Ki = 13.8 microM) > mono-2-sulfoxidation (Ki = 34 microM) > 5-sulfoxidation (Ki = 70.4 microM). In conclusion, it can be assumed that N-demethylation and mono-2-sulfoxidation are catalyzed by the isoenzymes 2D1, 2B and 1A2 while 5-sulfoxidation only by 1A2; isoenzymes belonging to the subfamilies 2C and 3A seem not to be involved in the metabolism of thioridazine. The obtained results are discussed in the view of species and structure differences in the enzymatic catalysis of phenothiazines' metabolism as well as in relation to their pharmacological and clinical significance.