Mechanism-based inhibition of CYP activities in rat liver by fluoxetine and structurally similar alkylamines.
1. The inhibition of cytochrome P450 (CYP)-mediated substrate oxidations by alkylamine-based drugs was investigated in rat hepatic microsomes. The effects of pre-incubation of the drugs with NADPH-fortified microsomes on inhibition potency was evaluated in relation to the formation of metabolite intermediate (MI) complexes with CYP in vitro. 2. The selective serotonin-reuptake inhibitor fluoxetine (FLU) emerged as a potent and preferential inhibitor of CYP2C11 in rat liver microsomes. After FLU biotransformation in NADPH-supplemented microsomes, IC50 values of 2 and 1 microM were determined against CYP2C11-dependent testosterone 2alpha- and 16alpha-hydroxylation; in the absence of pre-incubation, the corresponding IC50 values were 47 and 39 microM. MI complexation of CYP appeared to contribute significantly to inhibition by FLU, as evidenced by the 21% decrease in apparent microsomal CYP content produced by 50 microM FLU in the presence of NADPH. 3. The secondary amines nisoxetine (NIS), and especially, desipramine (DES) and nortriptyline (NOR), also inhibited CYP2C11 and generated MI complexes with microsomal CYP. In contrast, with the exception of SKF-525-A, tertiary alkylamines (10 compounds) inhibited specific CYP activities but did not form MI complexes. Pre-incubation of these agents with NADPH-supplemented microsomes did not enhance inhibition of CYP activities, thus suggesting that formation of inhibitory metabolites was minimal for these compounds. 4. These findings implicate drug-mediated MI complexation of CYPs in the inhibition of hepatic biotransformation processes by secondary alkylamines. In contrast, tertiary amines did not generate significant quantities of CYP-MI complexes under the test conditions. Despite their diffusion from the CYP active site, inhibition produced by tertiary amines and stable metabolites of other drugs may be significant. However, such inhibition would be of shorter duration than that from MI complexation, which involves quasi-covalent binding to the haem and prevention of oxygen activation.
Murray
M
M
Department of Physiology and Pharmacology, University of New South Wales, Australia. michaelm@pharm.usyd.edu.au
Murray
K
K
D018759:Adrenergic Uptake Inhibitors / Q000494:pharmacology; D000588:Amines / Q000494:pharmacology*; D000818:Animals; D001189:Aryl Hydrocarbon Hydroxylases / Q000037:antagonists & inhibitors*; D001665:Binding Sites; D000072467:Cytochrome P450 Family 2; D003891:Desipramine / Q000494:pharmacology; D004058:Diffusion; D004305:Dose-Response Relationship, Drug; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005473:Fluoxetine / Q000031:analogs & derivatives* / Q000494:pharmacology*; D006418:Heme / Q000737:chemistry; D006900:Hydroxylation; D020128:Inhibitory Concentration 50; D007700:Kinetics; D008099:Liver / Q000187:drug effects / Q000201:enzymology*; D008297:Male; D008862:Microsomes, Liver / Q000187:drug effects / Q000201:enzymology*; D008956:Models, Chemical; D009249:NADP / Q000378:metabolism; D009661:Nortriptyline / Q000494:pharmacology; D010100:Oxygen / Q000378:metabolism; D011485:Protein Binding; D051381:Rats; D017208:Rats, Wistar; D017367:Selective Serotonin Reuptake Inhibitors / Q000494:pharmacology; D039181:Steroid 16-alpha-Hydroxylase / Q000037:antagonists & inhibitors*; D013256:Steroids / Q000378:metabolism; D013997:Time Factors
D018759:Adrenergic Uptake Inhibitors; D000588:Amines; D001189:Aryl Hydrocarbon Hydroxylases; D000072467:Cytochrome P450 Family 2; D003891:Desipramine; D004791:Enzyme Inhibitors; D005473:Fluoxetine; D006418:Heme; D009249:NADP; D009661:Nortriptyline; D010100:Oxygen; D017367:Selective Serotonin Reuptake Inhibitors; D039181:Steroid 16-alpha-Hydroxylase; D013256:Steroids; C011386:nisoxetine; C450279:CYP2C11 protein, rat