Uehara Shotaro, Uno Yasuhiro, Yuki Yukako, Inoue Takashi, Sasaki Erika, Yamazaki Hiroshi
Laboratory of Drug Metabolism and Pharmacokinetics, Showa Pharmaceutical University, Machida, Tokyo, Japan (S.U., Y.Y., H.Y.); Pharmacokinetics and Bioanalysis Center, Shin Nippon Biomedical Laboratories, Ltd., Kainan, Wakayama, Japan (Y.U.); Department of Applied Developmental Biology (T.I.) and Center of Applied Developmental Biology (E.S.), Central Institute for Experimental Animals, Kawasaki, Japan; and Keio Advanced Research Center, Keio University, Minato-ku, Tokyo, Japan (E.S.).
Laboratory of Drug Metabolism and Pharmacokinetics, Showa Pharmaceutical University, Machida, Tokyo, Japan (S.U., Y.Y., H.Y.); Pharmacokinetics and Bioanalysis Center, Shin Nippon Biomedical Laboratories, Ltd., Kainan, Wakayama, Japan (Y.U.); Department of Applied Developmental Biology (T.I.) and Center of Applied Developmental Biology (E.S.), Central Institute for Experimental Animals, Kawasaki, Japan; and Keio Advanced Research Center, Keio University, Minato-ku, Tokyo, Japan (E.S.)
Drug Metab Dispos. 2016 Jun;44(6):833-41. doi: 10.1124/dmd.116.070367. Epub 2016 Apr 4.
Common marmosets (Callithrix jacchus) are attracting attention as animal models in preclinical studies for drug development. However, cytochrome P450s (P450s), major drug-metabolizing enzymes, have not been fully identified and characterized in marmosets. In this study, based on the four novel P450 4F genes found on the marmoset genome, we successfully isolated P450 4F2, 4F3B, 4F11, and 4F12 cDNAs in marmoset livers. Deduced amino acid sequences of the four marmoset P450 4F forms exhibited high sequence identities (87%-93%) to the human and cynomolgus monkey P450 4F homologs. Marmoset P450 4F3B and 4F11 mRNAs were predominantly expressed in livers, whereas marmoset P450 4F2 and 4F12 mRNAs were highly expressed in small intestines and livers. Four marmoset P450 4F proteins heterologously expressed in Escherichia coli catalyzed the ω-hydroxylation of leukotriene B4 In addition, marmoset P450 4F12 effectively catalyzed the hydroxylation of antiallergy drug ebastine, a human P450 2J/4F probe substrate. Ebastine hydroxylation activities by small intestine and liver microsomes from marmosets and cynomolgus monkeys showed greatly higher values than those of humans. Ebastine hydroxylation activities by marmoset and cynomolgus monkey small intestine microsomes were inhibited (approximately 60%) by anti-P450 4F antibodies, unlike human small intestine microsomes, suggesting that contribution of P450 4F enzymes for ebastine hydroxylation in the small intestine might be different between marmosets/cynomolgus monkeys and humans. These results indicated that marmoset P450 4F2, 4F3B, 4F11, and 4F12 were expressed in livers and/or small intestines and were functional in the metabolism of endogenous and exogenous compounds, similar to those of cynomolgus monkeys and humans.
普通狨猴（Callithrix jacchus）作为药物开发临床前研究的动物模型正受到关注。然而，主要的药物代谢酶细胞色素P450（P450s）在狨猴中尚未得到充分鉴定和表征。在本研究中，基于在狨猴基因组上发现的四个新的P450 4F基因，我们成功地从狨猴肝脏中分离出P450 4F2、4F3B、4F11和4F12的cDNA。四种狨猴P450 4F形式的推导氨基酸序列与人类和食蟹猴的P450 4F同源物具有高度的序列同一性（87%-93%）。狨猴P450 4F3B和4F11 mRNA主要在肝脏中表达，而狨猴P450 4F2和4F12 mRNA在小肠和肝脏中高表达。在大肠杆菌中异源表达的四种狨猴P450 4F蛋白催化白三烯B4的ω-羟基化。此外，狨猴P450 4F12有效地催化了抗过敏药物依巴斯汀（一种人类P450 2J/4F探针底物）的羟基化。来自狨猴和食蟹猴的小肠和肝脏微粒体的依巴斯汀羟基化活性显示出比人类高得多的值。与人类小肠微粒体不同，狨猴和食蟹猴小肠微粒体的依巴斯汀羟基化活性被抗P450 4F抗体抑制（约60%），这表明P450 4F酶对小肠中依巴斯汀羟基化的贡献在狨猴/食蟹猴和人类之间可能不同。这些结果表明，狨猴P450 4F2、4F3B、4F11和4F12在肝脏和/或小肠中表达，并且在内源性和外源性化合物的代谢中具有功能，类似于食蟹猴和人类。
