Comparison of catalytical activity and stereoselectivity between the recombinant human cytochrome P450 2D6.1 and 2D6.10

Polymorphisms of the cytochrome P450 2D6 (CYP2D6) gene play a major role in pharmacokinetic variability in human, while CYP2D6*10 is an important subtype in Asian people. In this study, the co-expression enzyme of human recombinant CYPOR, CYPb5 and CYP2D6.1 or CYP2D6.10 with the Bac-to-Bac (TM) system in baculovirus-infected insect cells was used to study the catalytical activity to imipramine metabolism and stereoselective metabolism of propranolol. The metabolites of imipramine were identified of hydroxyl imipramine and desipramine by LC-MS/MS. There are some differences between CYP2D6.1 and CYP2D6.10 activity. The kinetics parameters K-m, V-max, and CLint are 11.77 +/- 0.91 mu mol/L, 0.4235 +/- 0.05 nmol/nmol CYP2D6.1/min and 3.60 x 10(-5) ml/min/nmol CYP2D6.1 (n = 3) for CYP2D6.1, respectively, and 9.05 +/- 0.87 mu mol/L, 0.42 +/- 0.03 nmol/nmol CYP2D6.10/min, and 4.60 x 10(-5) ml/min/nmol CYP2D6.10 (n = 3) for CYP2D6.10. For propranolol, two metabolites were identified to be hydroxyl and N-desisopropylation propranolol by LC-MS/MS. When the substrate concentration was 0.20 mu mol/L, CYP2D6.1 and CYP2D6.10 exhibited significant stereoseletivity. Furthermore, enan-tioselective formation has been detected. Both of CYP2D6.1 and CYP2D6.10 produced more hydroxyl propranolol from the R-(+)-isomer than from the S-(-)-isomer while there was no obvious difference for N-desisopropylation propranolol production between R-(+)- and S-(-)- isomer. In summary, there is a some-what different catalytical activity and stereoselectivity between the human recombinant CYP2D6.1 and CYP2D6.10. The data we got will be helpful in preclinical research and clinical use of CYP2D6 substrates.