PREDICTION OF DRUG DISPOSITION BY HUMAN DRUG METABOLIZING ENZYMES

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Drug oxidation activities of twelve recombinant human cytochrome P450 (P450) coexpressed with human NADPH-P450 reductase (NPR) in bacterial membranes (P450/NPR membranes) were determined and compared with those of other recombinant systems and of human liver microsomes. Addition of exogenous membrane-bound NPR to the P450/NPR membranes enhanced the catalytic activities of CYP2C8, CYP2C9, CYP2C19, CYP3A4, and CYP3A5 ; however, enhancement of activities of CYP1A1, CYP1A2, CYP1B1, CYP2A6, CYP2B6, CYP2D6, and CYP2E1 in membranes was not observed after the addition of NPR in 4-molar excess to each P450. Exogenous purified human cytochrome b_5 (b_5) further enhanced catalytic activities of CYP2A6, CYP2B6, CYP2C8, CYP2E1, CYP3A4, and CYP3A5/NPR membranes. Catalytic activities of CYP2C9 and CYP2C19 were enhanced by addition of b_5 in reconstitution systems but not in the P450/NPR membranes. Apo b_5 (devoid of heme) enhanced catalytic activities when added to the both systems, except for CYP2E1/NPR membranes and the reconstituted systems containing purified CYP2C8 or CYP2E1 in comparison with b_5. Catalytic activities in P450/NPR membranes plus b_5 systems were roughly similar to those measured with microsomes of insect cells coexpressing P450 with NPR (and b_5) and/or of human liver microsomes, based on equivalent P450 contents. These results suggest that interactions of P450 and NPR coexpressed in membranes and reconstituted systems appear to be different in some human CYP2 family enzymes, possibly due to a conformational role of b_5. P450/NPR membrane systems containing b_5 are useful models for prediction of the rates for liver microsomal P450-dependent drug oxidations.