Comparison of Cryopreserved HepaRG Cells with Cryopreserved Human Hepatocytes for Prediction of Clearance for 26 Drugs

Abstract

ABSTRACT:Prediction of clearance in drug discovery currently relies on human primary hepatocytes, which can vary widely in drug-metabolizing enzyme activity. Potential alternative in vitro models include the HepaRG cell (from immortalized hepatoma cells), which in culture can express drug-metabolizing enzymes to an extent comparable to that of primary hepatocytes. Utility of the HepaRG cell will depend on robust performance, relative to that of primary hepatocytes, in routine high-throughput analysis. In this study, we compared intrinsic clearance (CL int ) in the recently developed cryopreserved HepaRG cell system with CL int in human cryopreserved pooled hepatocytes and with CL int in vivo for 26 cytochrome P450 substrate drugs. There was quantitative agreement between CL int in HepaRG cells and human hepatocytes, which was linear throughout the range of CL int (1-2000 ml ⅐ min ؊1 ⅐ kg ؊1 ) and not dependent on particular cytochrome P450 involvement. Prediction of CL int in HepaRG cells was on average within 2-fold of in vivo CL int (using the well stirred liver model), but average fold error was clearance-dependent with greater underprediction (up to at least 5-fold) for the more highly cleared drugs. Recent reporting of this phenomenon in human hepatocytes was therefore confirmed with the hepatocytes used in this study, and hence the HepaRG cell system appears to share an apparently general tendency of clearance-limited CL int in cell models. This study shows the cryopreserved HepaRG cell system to be quantitatively comparable to human hepatocytes for prediction of clearance of drug cytochrome P450 substrates and to represent a promising alternative in vitro tool.

Metabolic Profiling of Human Long-Term Liver Models and Hepatic Clearance Predictions from In Vitro Data Using Nonlinear Mixed-Effects Modeling

“…The underprediction for CYP2D6 metabolized compounds in HepaRG™ has already been reported due to a CYP2D6-deficient donor (11,30,31). However, the fold of underprediction seems to vary between the applied enzyme markers due to potential metabolism by multiple CYP enzymes (11,(30)(31)(32). For example, the intrinsic clearances in HepaRG™ as compared to suspension cultures were decreased more for dextromethorphan and metoprolol than for propranolol (11,30).…”

Comparison of Drug Metabolism and Its Related Hepatotoxic Effects in HepaRG, Cryopreserved Human Hepatocytes, and HepG2 Cell Cultures

“…[12][13][14][15] Furthermore, HepaRG cells have been shown to be quantitatively comparable to human hepatocytes for predicting the clearance of CYP drug substrates and as alternative in vitro tools. 16) Toxicological studies have reports the usefulness of HepaRG cells as in vitro hepatic models. 17,18) Several cytotoxicity and toxicogenomic studies have shown that HepaRG cells could be a more accurate human hepatocyte-like model than other hepatic cell lines and would be useful for toxicological studies.…”

Non-animal approaches for toxicokinetics in risk evaluations of food chemicals

“…Nonetheless, these developments do not have to restrain the quantitative use of Caco-2 results for chemicals that fall into the domain for which adequate in vivo predictive value is already obtained. Moreover, physiologically based kinetic computer modelling to integrate different types Zanelli et al, 2011). In addition, also the HepaRG cell line provides an adequate predictive value of in vivo metabolic clearance rates (r 2 = 0.53) (Zanelli et al, 2011), with a predictivity equal to that of cryopreserved primary human hepatocytes in the same study.…”

“…Moreover, physiologically based kinetic computer modelling to integrate different types Zanelli et al, 2011). In addition, also the HepaRG cell line provides an adequate predictive value of in vivo metabolic clearance rates (r 2 = 0.53) (Zanelli et al, 2011), with a predictivity equal to that of cryopreserved primary human hepatocytes in the same study. In comparison to in vitro methods for absorption or luminal digestion, there is little use of in vitro metabolism data within the risk evaluations of food chemicals (11-17% of the evaluations contained such data) (Fig.…”

“…Work on the human hepatoma cell line HepaRG is particularly promising. HepaRG cells express various cytochrome P450 and phase II enzymes when maintained in a differentiated state (Harwood et al, 2013;Zanelli et al, 2011).…”