Abstract
The metabolism of chlorpromazine by expressed recombinant cytochromes P450 4F4 and 4F5 cloned from rat brain was analyzed to characterize the individual activities of the isoforms. Both isoforms metabolized chlorpromazine to both the N-demethylated and the S-oxide products. When isoforms were incubated with chloropromazine in the presence of increasing concentrations of imipramine, imipramine significantly inhibited both N-demethylation and S-oxidation of chlorpromazine. A dilution of the serum fraction of anti-4F antibody was also found to significantly inhibit both S-oxidation and N-demethylation of chlorpromazine by both 4F4 and 4F5.
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Bennett, B. M., McDonald, B. J., Nigam, R. and Simon, W. C. (1994). Biotransformation of organic nitrates and vascular smooth muscle cell function.Trends Pharmacol Sci,15, 245–9.
Boehme, C. L. and Strobel, H. W. (1998) High-performance liquid chromatographic methods for the analysis of haloperidol and chlorpromazine metabolism in vitro by purified cytochrome P450 isoforms.J Chromatogr B Biomed Sci Appl,718, 259–66.
Bowlby, M. R. (1993) Pregnenolone sulfate potentiation of N-methyl-D-aspartate receptor channels in hippocampal neurons.Mol Pharmacol,43, 813–9.
Chen, L. and Hardwick, J. P. (1993) Identification of a new P450 subfamily, CYP4F1, expressed in rat hepatic tumors.Arch Biochem Biophys,300, 18–23.
Corpechot, C, Synguelakis, M., Talha, S., Axelson, M., Sjo-vall, J., Vihko, R., Baulieu, E. E. and Robel, P. (1983) Pregnenolone and its sulfate ester in the rat brain.Brain Res,270, 119–25.
Delaforge, M., Piffeteau, A., Boucher, J. L. and Viger, A. (1995) Nitric oxide formation during the cytochrome P-450-dependent reductive metabolism of 18-nitro-oxy-androstenedione.J Pharmacol Exp Ther,274, 634–40.
Flood, J. F., Morley, J.E. and Roberts, E. (1992) Memory-enhancing effects in male mice of pregnenolone and steroids metabolically derived from it.Proc Natl Acad Sci USA,89, 1567–71.
Fonne-Pfister, R., Bargetzi, M. J. and Meyer, U. A. (1987) MPTP, the neurotoxin inducing Parkinson’s disease, is a potent competitive inhibitor of human and rat cytochrome P450 isozymes (P450bufl, P450dbl) catalyzing debrisoquine 4-hydroxylation.Biochem Biophys Res Commun,148, 1144–50.
Hu, Z. Y., Bourreau, E., Jung-Testas, I., Robel, P. and Baulieu, E. E. (1987) Neurosteroids: oligodendrocyte mitochondria convert cholesterol to pregnenolone.Proc Natl Acad Sci USA,84, 8215–9.
Kawashima, H., Kusunose, E., Kubota, I., Maekawa, M. and Kusunose, M. (1992) Purification and NH2-terminal amino acid sequences of human and rat kidney fatty acid omega-hydroxylases.Biochim Biophys Acta,1123, 156–62.
Kawashima, H., Kusunose, E., Thompson, C. M. and Strobel, H. W. (1997) Protein expression, characterization and regulation of CYP4F4 and CYP4F5 cloned from rat brain.Arch Biochem Biophys,347, 148–54.
Kawashima, H., Sequeira, D. J., Nelson, D. R. and Strobel, H. W. (1996) Genomic cloning and protein expression of a novel rat brain cytochrome P-450 CYP2D18 catalyzing imipramine N-demethylation.J Biol Chem,271, 28176–80.
Kawashima, H. and Strobel, H. W. (1995) cDNA cloning of three new forms of rat brain cytochrome P450 belonging to the CYP4F subfamily.Biochem Biophys Res Commun,217, 1137–44.
Kikuta, Y., Kusunose, E., Endo, K., Yamamoto, S., Sogawa, K., Fujii-Kuriyama, Y. and Kusunose, M. (1993) A novel form of cytochrome P-450 family 4 in human polymorphonuclear leukocytes. cDNA cloning and expression of leukotriene B4 omega-hydroxylase.J Biol Chem,268, 9376–80.
Kusunose, E., Sawamura, A., Kawashima, H., Kubota, I. and Kusunose, M. (1989) Isolation of a new form of cytochrome P-450 with prostaglandin A and fatty acid omega-hydroxylase activities from rabbit kidney cortex microsomes.J Biochem (Tokyo),106, 194–6.
Langston, J. W., Ballard, P., Tetrud, J. W. and Irwin, I. (1983) Chronic Parkinsonism in humans due to a product of meperidine-analog synthesis.Science,219, 979–80.
Matsubara, S., Yamamoto, S., Sogawa, K., Yokotani, N., Fujii-Kuriyama, Y., Haniu, M., Shively, J.E., Gotoh, O., Kusunose, E. and Kusunose, M. (1987) cDNA cloning and inducible expression during pregnancy of the mRNA for rabbit pulmonary prostaglandin omega-hydroxylase (cytochrome P-450p-2).J Biol Chem,262, 13366–71.
Murray, M. (1992) P450 enzymes. Inhibition mechanisms, genetic regulation and effects of liver disease.Clin Phar-macokinet,23, 132–6.
Pollock, B. G., Mulsant, B. H., Sweet, R. A., Rosen, J., Altieri, L. P. and Perel, J. M. (1995) Prospective cytochrome P450 phenotyping for neuroleptic treatment in dementia.Psychopharmacol Bull,31, 327–31.
Renaud, J.P., Boucher, J.L., Vadon, S., Delaforge, M. and Mansuy, D. (1993) Particular ability of liver P450s3A to catalyze the oxidation of N omega-hydroxyarginine to citrulline and nitrogen oxides and occurrence in no synthases of a sequence very similar to the heme-binding sequence in P450s.Biochem Biophys Res Commun,192, 53–60.
Sawamura, A., Kusunose, E., Satouchi, K. and Kusunose, M. (1993) Catalytic properties of rabbit kidney fatty acid omega-hydroxylase cytochrome P-450ka2 (CYP4A7).Biochim Biophys Acta,1168, 30–6.
Sequeira, D. J. and Strobel, H. W. (1995) High-performance liquid chromatographic method for the analysis of imipramine metabolism in vitro by liver and brain microsomes.J Chromatogr B Biomed Appl,673, 251–8.
Stromstedt, M., Hayashi, S., Zaphiropoulos, P. G. and Gus-tafsson, J. A. (1990) Cloning and characterization of a novel member of the cytochrome P450 subfamily IVA in rat prostate.DNA Cell Biol,9, 569–77.
Thompson, C. M., Kawashima, H. and Strobel, H. W. (1998) Isolation of partially purified P450 2D 18 and characterization of activity toward the tricyclic antidepressants imipramine and desipramine.Arch Biochem Biophys,359, 115–21.
Yokotani, N., Kusunose, E., Sogawa, K., Kawashima, H., Kinosaki, M., Kusunose, M. and Fujii-Kuriyama, Y. (1991) cDNA cloning and expression of the mRNA for cytochrome P-450kd which shows a fatty acid omega-hydroxylating activity.Eur J Biochem,196, 531–6.
Youngster, S. K., Gluck, M. R., Heikkila, R. E. and Nicklas, W. J. (1990) 4’-alkylated analogs of l-methyl-4-phe-nylpyridinium ion are potent inhibitors of mitochondrial respiration.Biochem Biophys Res Commun,169, 758–64.
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The data presented here form part of the dissertation of Christopher L. Boehme submitted to the faculty of The University of Texas Houston Graduate School of Biomedical Sciences in partial fulfillment of the requirements for the Doctor of Philosophy.
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Boehme, C.L., Strobel, H.W. In vitro metabolism of chlorpromazine by cytochromes P450 4F4 and 4F5 and the inhibitory effect of imipramine. neurotox res 3, 329–337 (2001). https://doi.org/10.1007/BF03033194
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DOI: https://doi.org/10.1007/BF03033194