Activation of mianserin and its metabolites by human liver microsomes

Human liver microsomes metabolise mianserin to the stable 8-hydroxymianserin, desmethylmianserin and mianserin-2-oxide and in addition to one or more chemically reactive metabolites which bind, irreversibly, to microsomal protein. The stable metabolites were isolated by HPLC and characterized by mass spectrometry. The generation of each of these metabolites showed substantial inter-individual variation between eight sets of human liver microsomes studied. Inhibition of irreversible binding was observed with SKF-525A together with concomitant decrease in the formation of 8-hydroxymianserin and desmethylmianserin but not mianserin-2-oxide. Methimazole inhibited binding and the formation of each of the metabolites at a low concentration. Quinidine did not significantly inhibit irreversible binding but did inhibit the formation of 8-hydroxymianserin. Sulphaphenazole had no effect on irreversible binding or metabolism. The irreversible binding of mianserin was inhibited by ascorbic acid, glutathione and N-acetyl cysteine, whereas N-acetyl lysine and trichloropropane oxide had no effect. The irreversible binding of mianserin, 8-hydroxymianserin and desmethylmianserin was of the same order of magnitude however significantly greater binding was observed with the desmethyl metabolite. Incubations with [10- 3H/ 14C]mianserin showed no change in the 3H/ 14C ratio when irreversible binding occurred. Inhibition of irreversible binding was demonstrated with sodium cyanide at concentrations which did not inhibit total metabolism, which suggest that metabolic activation by the cytochrome P-450 enzyme system may lead to the formation of a reactive iminium intermediate that can bind to nucleophilic groups on proteins.