The effect of antioxidants on electrocatalytic activity of cytochrome P450 3A4

The electrochemical analysis of cytochrome P450 3A4 catalytic activity has shown that vitamins C, A and E influence reduction of cytochrome P450 3A4. These data suggest a possibility of cross effects and interference of vitamins-antioxidants with drugs metabolised by cytochrome P450 3A4, during complex therapy of patients. These vitamins demonstrate antioxidant properties that lead to the increase of the cathodic current corresponding to heme reduction of this functionally significant hemoprotein. Ascorbic acid (0.028–0.56 mM) stimulated the cathodic peak (an electrochemical signal) of cytochrome P450 3A4. In the presence of diclofenac (Voltaren), a typical substrate of cytochrome P450 3A4, the increase in the catalytic current suggesting electrocatalysis and stimulating action of ascorbic acid was observed. In the presence of vitamins A and E the dose-dependent increase in the catalytic current of cytochrome P450 3A4 was observed in the range of vitamin concentrations from 10 to 100 μM. The maximal increase of 229 ± 20 and 162 ± 10% was observed at 100 μM vitamin A and vitamin E, respectively. In contrast to vitamin A, vitamin E in the presence of the cytochrome P450 inhibitor itraconazole did not increase the catalytic current. The latter implies existence of some substrate properties in vitamin E. The electrochemical approach for the analysis of catalytic activity of cytochrome P450 3A4 and studies of the effect of biologically active compounds on electrocatalysis is the sensitive and effective sensor approach, allowing to use low concentration of protein on an electrode (up to 10–15 mol/electrode), to carry out the analysis without involvement of protein redox partners, and to reveal drug-drug or drug-vitamins interaction in pre-clinical experiments.