Metabolism of A-ring diastereomers of 1α,25-dihydroxyvitamin D3 by CYP24A1

The metabolism of 1alpha,25(OH)(2)D-3 (1alpha,3beta) and its A-ring diastereomers, 1beta,25(OH)(2)D-3 (1beta,3beta), 1alpha,25(OH)(2)-3-epi-D-3 (1alpha,3alpha), and 1beta,25(OH)(2)-3-epi-D-3 (1beta,3alpha), was examined to compare the substrate specificity and reaction specificity of CYP24A1 between humans and rats. The ratio between C-23 and C-24 oxidation pathways in human CYP24A1 -dependent metabolism of (1alpha,30() and (1beta,3) was 1:1, although the ratio for (1alpha,3beta) and (1beta,3beta) was 1:4. These results indicate that the orientation of the hydroxyl group at the C-3 position determines the ratio between C-23 and C-24 oxidation pathways. A remarkable increase of metabolites in the C-23 oxidation pathway was also observed in rat CYP24A1 -dependent metabolism. The binding affinity of human CYP24A1 for A-ring diastereomers was (1alpha,3beta) > (1alpha,3alpha) > (1beta,3beta) > (1beta,3alpha), indicating that both hydroxyl groups at C-1 and C-3 positions significantly affect substrate-binding. The information obtained in this study is quite useful for understanding substrate recognition of CYP24A1 and designing new vitamin D analogs. (C) 2004 Elsevier Inc. All rights reserved.