Vol. 29, Issue 3, 289-295, March 2001

Characterization of 2-[[4-[[2-(1H-Tetrazol-5-ylmethyl)phenyl]methoxy]phenoxy]methyl] Quinoline N-Glucuronidation by in Vitro and in Vivo Approaches

Jeffrey C. Stevens,¹ Jessica L. Fayer,² and Kenneth C. Cassidy²

Department of Drug Metabolism and Pharmacokinetics, Aventis Pharma, Collegeville, Pennsylvania

RG 12525 is a new chemical entity recently evaluated for the treatment of type II diabetes. Clinical studies have previously identified the tetrazole N2-glucuronide conjugate of RG 12525 as the predominant metabolite in plasma following oral administration of RG 12525. Species differences in RG 12525 glucuronidation were first investigated with incubations of RG 12525 with rat, monkey, and human hepatocytes. The results showed the N2-glucuronide to be the major metabolite in human and monkey samples, with only low levels observed for the rat. The formation of this glucuronide by human liver microsomes was subsequently characterized. RG 12525 N2-glucuronidation was found to have a pH optimum of 7.0 to 7.5 and demonstrated a high affinity with a K_m range of 16.6 to 21.1 µM RG 12525 (n = 3). The rate of N2-glucuronide formation ranged from 2.5 to 15.4 nmol of RG 12525 N2-glucuronide formed/min/mg of protein (~6-fold) in the 21 samples assayed. The reaction was inhibited by known substrates for glucuronidation, with imipramine (62%), naringenin (44%), and scopoletin (38%) producing the largest degree of inhibition at equimolar concentrations of substrate and inhibitor. Of the eight expressed UDP-glucuronosyltransferase (UGT) forms assayed, UGT1A1 and 1A3 displayed the highest rate of RG 12525 N2-glucuronidation (0.109 and 0.125 nmol/min/mg, respectively). Finally, low levels of N2-glucuronidation of RG 12525 by human jejunum microsomes were demonstrated, suggesting that presystemic clearance via glucuronidation may constitute a barrier to bioavailability.