Humanized-liver TK-NOG mice lacking plasma carboxylesterase 1c improve the prediction of human carboxylesterase 1-mediated drug metabolism

Chimeric NOD/Shi-Scid IL2 receptor gamma-null mice expressing the HSVtk mutant 30 with humanized livers (humanized-liver mice) are valuable models for predicting drug metabolism in humans. However, carboxylesterase 1c (Ces1c) secreted from residual mouse hepatocytes occasionally prevents the precise evaluation of human drug metabolism. To address this issue, we established a Ces1c knockout (KO) humanized-liver mouse model, characterized its plasma enzymatic activity, and investigated its usefulness in predicting drug metabolism in humans. Ces1c KO mice were successfully generated by deleting exon 5 of Ces1c, which encodes the amino acid residue Ser221 in the catalytic triad. The hydrolytic activities of irinotecan, oseltamivir, and temocapril in plasma were significantly lower in Ces1c KO mice than in normal mice, both untransplanted and transplanted with human hepatocytes, similar to those observed in humans. Drug-metabolizing activities in plasma and liver microsomes were comparable between Ces1c KO humanized-liver mice and humans. Following intravenous administration of irinotecan, the plasma concentration of the hydrolytic metabolite 7-ethyl-10-hydroxycamptothecin was significantly reduced in Ces1c KO humanized-liver mice compared to that in normal humanized-liver mice. Furthermore, the plasma concentrations of cytochrome P450 3A-mediated oxidative metabolites 7-ethyl-10-[4-N-(5-aminopentanoic acid)-1-piperidino] carbonyloxycamptothecin and 7-ethyl-10-(4-amino-1-piperidino)carbonyloxycamptothecin were higher in normal and Ces1c KO humanized-liver mice than in wild-type and Ces1c KO mice. The area under the curve values of irinotecan-related compounds after intravenous administration showed the strongest correlation between humans and Ces1c KO humanized-liver mice. These results demonstrate that Ces1c KO humanized-liver mice are a highly suitable model for the accurate prediction of CES1-mediated drug metabolism in humans. SIGNIFICANT STATEMENT: Carboxylesterase 1c (Ces1c) knockout humanized-liver mice that lacked plasma Ces1c were generated. This model reproduced plasma hydrolytic enzyme activity of irinotecan, oseltamivir, and temocapril observed in humans. Ces1c knockout humanized-liver mice can improve the accuracy of predicting human drug metabolism.