CYP2C9-Catalyzed Metabolism of S-Warfarin to 7-Hydroxywarfarin in Vivo and in Vitro in Chimeric Mice with Humanized Liver
CYP2C9-Catalyzed Metabolism of S-Warfarin to 7-Hydroxywarfarin in Vivo and in Vitro in Chimeric Mice with Humanized Liver
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DOI：
10.1124/dmd.108.022830
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发表时间：
2008-12-01
影响因子：
3.9
通讯作者：
Ohta, Shigeru
中科院分区：
文献类型：
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作者：
Inoue, Tae;Nitta, Kayoko;Ohta, Shigeru
通过移植人肝细胞构建具有人源化肝脏的嵌合小鼠。在本研究中，我们通过检查S-华法林的羟基化，研究了这些小鼠是否具有与人类相似的药物代谢能力，S-华法林主要代谢为S-7-羟基华法林，由CYP 2C 9催化，在人类而非小鼠中。嵌合小鼠肝微粒体对S-华法林的7-羟化活性约为对照（尿激酶型纤溶酶原激活物转基因严重联合免疫缺陷）小鼠的10倍。嵌合小鼠肝微粒体的7-羟化酶活性被磺胺苯吡唑显着抑制，是人肝微粒体，而对照小鼠的活动不受影响。嵌合小鼠肝脏中的CYP 2C亚型也通过免疫印迹分析证实为人亚型CYP 2C 9。在本体内研究中，嵌合小鼠血浆中S-7-羟基华法林的水平约为对照小鼠的7倍，与体外数据一致。因此，嵌合小鼠中的CYP 2C同种型在体内和体外作为人同种型CYP 2C 9发挥功能。这些结果表明，具有人源化肝脏的嵌合小鼠可用于预测人类中的药物代谢，至少关于CYP 2C 9依赖性代谢。
Chimeric mice having humanized livers were constructed by transplantation of human hepatocytes. In this study, we investigated whether these mice have a capacity for drug metabolism similar to that of humans by examining hydroxylation of S-warfarin, which is predominantly metabolized to S-7-hydroxywarfarin, catalyzed by CYP2C9, in humans but not mice. The 7-hydroxylating activity of chimeric mouse liver microsomes toward S-warfarin was approximately 10-fold higher than that of control (urokinase-type plasminogen activator-transgenic severe combined immunodeficient) mice. The 7-hydroxylase activity of chimeric mouse liver microsomes was markedly inhibited by sulfaphenazole, as was that of human liver microsomes, whereas the activity of control mice was unaffected. The CYP2C isoform in chimeric mouse liver was also confirmed to be the human isoform, CYP2C9, by immunoblot analysis. In the present in vivo study, the level of S-7-hydroxywarfarin in plasma of chimeric mice was approximately 7-fold higher than that in control mice, in agreement with the in vitro data. Thus, the CYP2C isoform in chimeric mice functions in vivo and in vitro as a human isoform, CYP2C9. These results suggest that chimeric mice with humanized liver could be useful for predicting drug metabolism in humans, at least regarding CYP2C9-dependent metabolism.