Different Effects of Desipramine on Bufuralol 1″-Hydroxylation by Rat and Human CYP2D Enzymes

• Published in: Biological & Pharmaceutical Bulletin 2005, Vol.28(4), pp.634-640
• Main Authors: Isobe, Takashi, Hichiya, Hiroyuki, Hanioka, Nobumitsu, Yamamoto, Shigeo, Shinoda, Sumio, Funae, Yoshihiko, Satoh, Tetsuo, Yamano, Shigeru, Narimatsu, Shizuo
• Format: Article
• Language: English
• Subjects: Adult; Animals; Antidepressive Agents, Tricyclic - pharmacology; Aryl Hydrocarbon Hydroxylases - antagonists & inhibitors; Aryl Hydrocarbon Hydroxylases - metabolism; binding; CYP2D2; CYP2D6; desipramine; Desipramine - pharmacology; Ethanolamines - metabolism; Humans; Hydroxylation; Imipramine - pharmacology; In Vitro Techniques; inactivation; Kinetics; Male; Microsomes, Liver - drug effects; Microsomes, Liver - metabolism; Middle Aged; Rats; Rats, Wistar; reactive metabolite; Species Specificity
• ISSN: 0918-6158; 1347-5215
• DOI: 10.1248/bpb.28.634

Inhibitory effects of desipramine (DMI) on rat and human CYP2D enzymes were studied using bufuralol (BF) 1″-hydroxylation as an index. Inhibition was examined under the following two conditions: 1) DMI was co-incubated with BF and NADPH in the reaction mixture containing rat or human liver microsomes or yeast cell microsomes expressing rat CYP2D1, CYP2D2 or human CYP2D6 (co-incubation); 2) DMI was preincubated with NADPH and the same enzyme sources prior to adding the substrate (preincubation). When either rat liver microsomes or recombinant CYP2D2 was employed, the preincubation with DMI (0.3 μM) caused a greater inhibition of BF 1″-hydroxylation than the co-incubation did, whereas BF 1″-hydroxylation by rat CYP2D1 was not markedly affected under the same conditions. The inhibitory effect of DMI on BF 1″-hydroxylation by human liver microsomal fractions or recombinant CYP2D6 was much lower than that on the hydroxylation by rat liver microsomes or CYP2D2. Kinetic studies demonstrated that the inhibition-type changed from competitive for the co-incubation to noncompetitive for the preincubation in the case of CYP2D2, whereas the inhibition-type was competitive for both the co-incubation and the preincubation in the case of CYP2D6. Furthermore, the loss of activity of rat CYP2D2 under the preincubation conditions followed pseudo-first-order kinetics. Binding experiments employing the recombinant enzymes and [3H]-DMI revealed that CYP2D2 and CYP2D6 were the only prominent proteins to which considerable radioactive DMI metabolite(s) bound. These results indicate that rat CYP2D2 biotransforms DMI into reactive metabolite(s), which covalently bind to CYP2D2, resulting in inactivation of the enzyme. In contrast, human CYP2D6 may also biotransform DMI into some metabolite(s) that covalently bind to CYP2D6, but that do not inactivate the enzyme.