Prediction of fraction metabolized via CYP3A in humans utilizing cryopreserved human hepatocytes from a set of 12 single donors

Abstract

1. It has previously been demonstrated that metabolism of drugs via a single enzymatic pathway, particularly CYP3A4, is associated with increased risk for drug-drug interactions (DDI). Quantitative experimental systems as well as integrated prediction models to assess such risk during the preclinical phase are highly warranted. 2. The present study was designed to systematically investigate the performance of human cryopreserved hepatocytes in suspension to predict fraction metabolized via CYP3A (fmCYP3A) by assessing the ketoconazole sensitive intrinsic clearance (CLint) for five prototypical CYP3A substrates with varying degree of CYP3A dependent CLint in twelve individual hepatocyte batches. 3. We demonstrate that in contrast to well predicted mean hepatic metabolic clearance (CLH) and mean fmCYP3A data, the variability in CYP3A contribution for compounds having multiple metabolic pathways cannot be predicted from inhibition experiments using ketoconazole as inhibitor. Instead, data in the present paper indicate that the variability is larger after inhibition of CYP3A for compounds having multiple metabolic pathways. 4. It is therefore recommended to estimate the average CLint and fmCYP3A for a given test compound in a series (n = 10) of individual human hepatocyte batches.

Translational Biomedical Informatics and Pharmacometrics Approaches in the Drug Interactions Research

“…Physiologically, cryopreserved human hepatocyte is closer to the human hepatic metabolism than the other in vitro system does. Desbans et al . used cryopreserved human hepatocytes from 12 donors to estimate fm of CYP3A for 5 prototypical CYP3A substrates.…”

“…Physiologically, cryopreserved human hepatocyte is closer to the human hepatic metabolism than the other in vitro system does. Desbans et al 48 used cryopreserved human hepatocytes from 12 donors to estimate fm of CYP3A for 5 prototypical CYP3A substrates. After hepatocytes were incubated with test compounds and/or the inhibitor, the intrinsic clearance was estimated from the parent compound depletion profile.…”

Drug biokinetic and toxicity assessments in rat and human primary hepatocytes and HepaRG cells within the EU-funded Predict-IV project

“…The HepaRG cell line was chosen as one of the most advanced hepatocyte cell culture models that unifies ease of handling, availability and physiological drug metabolism (Aninat et al, 2006). In addition, cryopreserved human hepatocytes that are well characterized for drug metabolizing enzymes (DME) are also useful to explore if donor-donor variability in DME expression is the cause of donordonor variability in the sensitivity to drug-drug interactions and toxicity (Desbans et al, 2014). It is of note that these hepatic models may reflect liver-specific toxicity but predominantly reflect general toxicity induced/reduced by metabolic activation, be it a toxic metabolite or the increased metabolic activity that can affect viability of hepatocytes.…”

The Cancer Drug Fraction of Metabolism Database

“…Physiologically, the cryopreserved human hepatocyte is closer to human hepatic Fm 3A4 = 1 − AUC control AUC inhibited 513 www.psp-journal.com The Cancer Drug Fm Database Hua et almetabolism than the other in vitro system. Desbanset al 36 used cryopreserved human hepatocytes from 12 donors to estimate Fm 3A for five prototypical CYP3A substrates with varying degrees of CYP3A-dependent in vivo CL using intrinsic metabolic stability measurements in the presence and absence of the CYP3A probe inhibitor ketoconazole. After hepatocytes were incubated with test compounds and/or the inhibitor, the intrinsic CL was estimated from the parent compound depletion profile.…”