In Vitro and In Vivo Correlation of Hepatic Fraction of Metabolism by P450 in Dogs

• Published in: Journal of pharmaceutical sciences 2019-02, Vol.108 (2), p.1017-1026
• Main Authors: Zhou, Xin, Hui, Yu-Hua, Hudson, Loyd, Katyayan, Kishore K., Mohutsky, Michael A., Hao, Junliang, Schober, Douglas A., Hembre, Erik J.
• Format: Article
• Language: English
• Subjects: Animals; Canagliflozin - blood; Canagliflozin - metabolism; Cytochrome P-450 Enzyme Inhibitors - blood; Cytochrome P-450 Enzyme Inhibitors - metabolism; Cytochrome P-450 Enzyme System - metabolism; cytochrome P450 (P450); Dogs - blood; Dogs - metabolism; drug-drug interaction(s); Glucuronosyltransferase - metabolism; Hepatocytes - metabolism; Liver - cytology; Liver - metabolism; Male; Oxidation-Reduction; phase I metabolism; phase II metabolism; Triazoles - blood; Triazoles - metabolism; UDP-glucuronosyltransferase (UGT)
• ISSN: 0022-3549; 1520-6017
• DOI: 10.1016/j.xphs.2018.09.012

1-Aminobenzotriazole (ABT) has been widely used as a nonspecific mechanism-based inhibitor of cytochrome P450 (P450) enzymes. It is extensively used in preclinical studies to determine the relative contribution of oxidative metabolism mediated by P450 in vitro and in vivo. The aim of present study was to understand the translation of fraction metabolized by P450 in dog hepatocytes to in vivo using ABT, for canagliflozin, known to be cleared by P450-mediated oxidation and UDP-glucuronosyltransferases–mediated glucuronidation, and 3 drug discovery project compounds mainly cleared by hepatic metabolism. In a dog hepatocyte, intrinsic clearance assay with and without preincubation of ABT, 3 Lilly compounds exhibited a wide range of fraction metabolized by P450. Subsequent metabolite profiling in dog hepatocytes demonstrated a combination of metabolism by P450 and UDP-glucuronosyltransferases. In vivo, dogs were pretreated with 50 mg/kg ABT or vehicle at 2 h before intravenous administration of canagliflozin and Lilly compounds. The areas under the concentration-time curve (AUC) were compared for the ABT-pretreated and vehicle-pretreated groups. The measured AUCABT/AUCveh ratios were correlated to fraction of metabolism by P450 in dog hepatocytes, suggesting that in vitro ABT inhibition in hepatocytes is useful to rank order compounds for in vivo fraction of metabolism assessment.