In vitro oxidation of oxicam NSAIDs by a human liver cytochrome P450

Abstract

The nature of the enzyme(s) catalyzing the major metabolic pathway (5′-hydroxylation) of oxicam NSAIDs was investigated in subcellular preparations of human liver tissue. Microsomal, but not cytosolic, fractions catalyzed the 5′-hydroxylation of tenoxicam. This reaction required NADPH and was inhibited by various nonselective P450 inhibitors (CO, SKF-525A, ketoconazole), but not by the peroxidase inhibitor NaN 3. Tenoxicam 5′-hydroxylation exhibited simple Michaelis-Menten kinetics compatible with catalysis by a single enzyme, but it strongly inhibited its own oxidation at concentrations higher than 100–150 μM. Piroxicam competitively inhibited tenoxicam 5′-hydroxylation and, conversely, tenoxicam competitively inhibited piroxicam 5′-hydroxylation. Tenoxicam 5′-hydroxylation kinetics were similar in microsomes from one poor and five extensive metabolizers of debrisoquin (CYP2D6). Dextromethorphan (CYP2D6 prototype substrate) and midazolam (CYP3A prototype substrate) had no influence on tenoxicam 5′-hydroxylation, whereas mephenytoin, tolbutamide and sulfaphenazole (K i ≈ 0.1 μM) inhibited it. This indicates that the 5′-hydroxylation of both piroxicam and tenoxicam is predominantly catalyzed by at least one cytochrome P450 isozyme of the CYP2C subfamily.