Uehara Shotaro, Uno Yasuhiro, Okamoto Eriko, Inoue Takashi, Sasaki Erika, Yamazaki Hiroshi
Laboratory of Drug Metabolism and Pharmacokinetics, Showa Pharmaceutical University, Machida, Tokyo, Japan (S.U., E.O., 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, Tokyo, Japan (E.S.).
Laboratory of Drug Metabolism and Pharmacokinetics, Showa Pharmaceutical University, Machida, Tokyo, Japan (S.U., E.O., 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, Tokyo, Japan (E.S.)
Drug Metab Dispos. 2017 Aug;45(8):883-886. doi: 10.1124/dmd.117.076042. Epub 2017 May 9.
Common marmosets (), New World monkeys, are a promising primate model for preclinical drug metabolism studies because of the similarities of cytochrome P450 (P450) enzyme function to those of humans. Despite an increasing number of drug candidates catalyzed by non-P450 enzymes, drug metabolizing enzymes other than P450s have been hardly identified or characterized in marmosets. In this study, we identified () gene by marmoset genome analysis. AOX1 cDNA was cloned from marmoset livers by reverse transcription-polymerase chain reaction. Deduced amino acid sequences of AOX1 cDNA showed high sequence identities (92-93%) with primate AOX1s. Phylogenetic analysis showed that marmoset AOX1 was closely clustered with primate AOX1s, unlike nonprimate animal model AOX1s of pig, rabbit, rat, and mouse used in drug metabolism. The tissue expression analyses by real-time reverse-transcription polymerase chain reaction and immunoblotting showed that marmoset AOX1 mRNA and protein were abundantly expressed in livers, similar to cynomolgus monkeys and humans. Marmoset AOX1 heterologously expressed in catalyzed the oxidation of carbazeran and phthalazine, typical AOX1 substrates, similar to cynomolgus monkey and human AOX1s. Human and marmoset AOX1 effectively catalyzed phthalazine oxidation when assessed with Michaelis-Menten kinetics, but cynomolgus monkey AOX1 catalyzed this reaction with cooperative kinetics with high capacity. These results indicated that tissue distribution and enzymatic function of AOX1 enzyme is similar between marmosets and humans, suggesting that marmosets are a suitable primate model for AOX-dependent drug metabolism in preclinical studies.
普通狨猴，作为新世界猴，因其细胞色素P450（P450）酶功能与人类相似，是临床前药物代谢研究中一种很有前景的灵长类动物模型。尽管由非P450酶催化的候选药物数量不断增加，但除P450酶外的药物代谢酶在狨猴中几乎未被鉴定或表征。在本研究中，我们通过狨猴基因组分析鉴定了（）基因。通过逆转录-聚合酶链反应从狨猴肝脏中克隆了AOX1 cDNA。AOX1 cDNA推导的氨基酸序列与灵长类动物的AOX1具有高度的序列同一性（92-93%）。系统发育分析表明，与用于药物代谢的猪、兔、大鼠和小鼠等非灵长类动物模型的AOX1不同，狨猴AOX1与灵长类动物的AOX1紧密聚类。通过实时逆转录聚合酶链反应和免疫印迹进行的组织表达分析表明，狨猴AOX1 mRNA和蛋白在肝脏中大量表达，类似于食蟹猴和人类。在（）中异源表达的狨猴AOX1催化了典型的AOX1底物卡巴泽兰和酞嗪的氧化，类似于食蟹猴和人类的AOX1。当用米氏动力学评估时，人和狨猴的AOX1有效地催化了酞嗪氧化，但食蟹猴AOX1以高容量的协同动力学催化了该反应。这些结果表明，狨猴和人类之间AOX1酶的组织分布和酶功能相似，这表明狨猴是临床前研究中依赖AOX的药物代谢的合适灵长类动物模型。
