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 (CLint) in the recently developed cryopreserved HepaRG cell system with CLint in human cryopreserved pooled hepatocytes and with CLint in vivo for 26 cytochrome P450 substrate drugs. There was quantitative agreement between CLint in HepaRG cells and human hepatocytes, which was linear throughout the range of CLint (1–2000 ml · min−1 · kg−1) and not dependent on particular cytochrome P450 involvement. Prediction of CLint in HepaRG cells was on average within 2-fold of in vivo CLint (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 CLint 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.
Footnotes
This work was supported by funding from the European Union Sixth Framework Programme for Optimisation of Liver and Intestine In Vitro Models for Pharmacokinetics [Liintop/STREP 037499]; and by a consortium of pharmaceutical companies (GlaxoSmithKline, Lilly, Pfizer, and Servier) within the Centre for Applied Pharmacokinetic Research at the University of Manchester.
Article, publication date, and citation information can be found at http://dmd.aspetjournals.org.
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ABBREVIATIONS:
- CLint
- intrinsic clearance
- DMSO
- dimethyl sulfoxide
- WME
- Williams' medium E
- AFE
- average fold error
- RMSE
- root mean square error.
- Received August 16, 2011.
- Accepted October 13, 2011.
- Copyright © 2012 by The American Society for Pharmacology and Experimental Therapeutics
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