Stereoselective Conjugation of Oxazepam by Human UDP-Glucuronosyltransferases (UGTs): S-Oxazepam Is Glucuronidated by UGT2B15, While R-Oxazepam Is Glucuronidated by UGT2B7 and UGT1A9

doi:10.1124/dmd.30.11.1257

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Vol. 30, Issue 11, 1257-1265, November 2002

Stereoselective Conjugation of Oxazepam by Human UDP-Glucuronosyltransferases (UGTs): S-Oxazepam Is Glucuronidated by UGT2B15, While R-Oxazepam Is Glucuronidated by UGT2B7 and UGT1A9

Michael H. Court, Su X. Duan, Chantal Guillemette, Kim Journault, Soundararajan Krishnaswamy, Lisa L. von Moltke, and David J. Greenblatt

Comparative and Molecular Pharmacogenetics Laboratory (M.H.C., S.X.D., S.K.) and Clinical Pharmacology Laboratory (L.L.v.M., D.J.G.), Department of Pharmacology and Experimental Therapeutics, Tufts University School of Medicine, Boston, Massachusetts; and Pharmacogenetics Laboratory, Centre Hospitalier de l'Université Laval Research Center, University of Laval, Faculty of Pharmacy, Quebec, Canada (C.G., K.J.)

(R,S)-Oxazepam is a 1,4-benzodiazepine anxiolytic drug that is metabolized primarily by hepatic glucuronidation. In previousstudies, S-oxazepam (but not R-oxazepam) was shown to be polymorphicallyglucuronidated in humans. The aim of the present study was toidentify UDP-glucuronosyltransferase (UGT) isoforms mediatingR- and S-oxazepam glucuronidation in human liver, with the longterm objective of elucidating the molecular genetic basis forthis drug metabolism polymorphism. All available recombinant UGTisoforms were screened for R- and S-oxazepam glucuronidation activities.Enzyme kinetic parameters were then determined in representativehuman liver microsomes (HLMs) and in UGTs that showed significantactivity. Of 12 different UGTs evaluated, only UGT2B15 showedsignificant S-oxazepam glucuronidation. Furthermore, the apparentK_m for UGT2B15 (29-35 µM) was similar to values determined forHLMs (43-60 µM). In contrast, R-oxazepam was glucuronidated byUGT1A9 and UGT2B7. Although apparent K_m values for HLMs (256-303µM) were most similar to UGT2B7 (333 µM) rather than UGT1A9 (12µM), intrinsic clearance values for UGT1A9 were 10 times higherthan for UGT2B7. A common genetic variation results in aspartate(UGT2B15*1) or tyrosine (UGT2B15*2) at position 85 of the UGT2B15protein. Microsomes from human embryonic kidney (HEK)-293 cellsoverexpressing UGT2B15*1 showed 5 times higher S-oxazepam glucuronidationactivity than did UGT2B15*2 microsomes. Similar results were obtainedfor other substrates, including eugenol, naringenin, 4-methylumbelliferone,and androstane-3 $alpha$ -diol. In conclusion, S-oxazepam is stereoselectivelyglucuronidated by UGT2B15, whereas R-oxazepam is glucuronidatedby multiple UGT isoforms. Allelic variation associated with theUGT2B15 gene may explain polymorphic S-oxazepam glucuronidationinhumans.

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