A comparative study of the formation of chemically reactive drug metabolites by human liver microsomes

1. The metabolism of amodiaquine (A), ethinyloestradiol (E), mianserin (M), phenytoin (Ph), sulphanilamide (S) and paracetamol (Pa) to both stable and chemically reactive, i.e. irreversibly protein bound, metabolites was investigated using microsomes prepared from histologically normal human liver obtained from eight kidney donors. 2. All drugs, except amodiaquine, were metabolized by NADPH-dependent microsomal enzymes to chemically reactive metabolites. The degree of NADPH-dependent binding varied between drugs (E, 11.5 +/- 5.8% incubated drug; M, 3.0 +/- 1.9%; Ph, 0.10 +/- 0.09%; S, 0.57 +/- 0.38%; Pa, 1.2 +/- 1.2%; mean of eight livers +/- s.d.). 3. Inclusion of glutathione (1 mM) or ascorbic acid (1 mM) in the incubation reduced the NADPH-dependent binding for all substrates, indicating the involvement of electrophilic oxidation products. 4. Binding of M and Pa correlated with each other (Spearman's r = 0.86) and with total cytochrome P-450 content (r = 0.76 and 0.78 respectively). E binding also correlated with the binding of M (r = 0.79) and Pa (r = 0.81) but not with cytochrome P-450. Binding of Ph and S did not correlate with any of the other measured metabolic parameters.