Joint Faculty of Veterinary Medicine, Kagoshima University, Kagoshima, Japan (Y.U., M.Ij., O.Y., M.S., A.A.); Showa Pharmaceutical University, Machida, Japan (N.M., H.Y.); School of Veterinary Medicine, Kitasato University, Towadashi, Japan (H.K.); School of Veterinary Medicine, Rakuno Gakuen University, Ebetsu, Japan (H.T.); Graduate School of Veterinary Medicine, Hokkaido University, Sapporo, Japan (H.M., S.M.M.N., Y.I., M.Is.); and Graduate School of Agriculture, Ehime University, Matsuyama, Japan (H.M.)
Joint Faculty of Veterinary Medicine, Kagoshima University, Kagoshima, Japan (Y.U., M.Ij., O.Y., M.S., A.A.); Showa Pharmaceutical University, Machida, Japan (N.M., H.Y.); School of Veterinary Medicine, Kitasato University, Towadashi, Japan (H.K.); School of Veterinary Medicine, Rakuno Gakuen University, Ebetsu, Japan (H.T.); Graduate School of Veterinary Medicine, Hokkaido University, Sapporo, Japan (H.M., S.M.M.N., Y.I., M.Is.); and Graduate School of Agriculture, Ehime University, Matsuyama, Japan (H.M.).
Drug Metab Dispos. 2022 Nov;50(11):1434-1441. doi: 10.1124/dmd.122.000930. Epub 2022 Jun 14.
Cytochrome P450s (P450s) have been identified and analyzed in dogs and pigs, species that are often used in preclinical drug studies. Moreover, P450s are clinically important for drug therapy not only in humans, but also in species under veterinary care, including dogs and cats. In the present study, seven P450s homologous to human CYP2J2, namely, dog CYP2J2; cat CYP2J2; and pig CYP2J33, CYP2J35, CYP2J91, and CYP2J93, were newly identified and characterized, along with pig CYP2J34 previously identified. The cDNAs of these CYP2Js contain open reading frames of 502 amino acids, except for CYP2J35 (498 amino acids), and share high sequence identity (77%-80%) with human CYP2J2. Phylogenetic analysis revealed that dog and cat CYP2J2 were closely related, whereas pig CYP2Js formed a cluster. All seven genes contain nine coding exons and are located in corresponding genomic regions, with the pig genes forming a gene cluster. These CYP2J2 mRNAs were predominantly expressed in the small intestine with additional expression in the kidney and brain for dog CYP2J2 and pig CYP2J91 mRNAs, respectively. All seven CYP2Js metabolized human CYP2J2 substrates terfenadine, ebastine, and astemizole, indicating that they are functional enzymes. Dog CYP2J2 and pig CYP2J34 and CYP2J35 efficiently catalyzed ebastine primary hydroxylation and secondary carebastine formation at low substrate concentrations, just as human CYP2J2 does. Velocityversus-substate plots exhibited sigmoidal relationships for dog CYP2J2, cat CYP2J2, and pig CYP2J33, indicating allosteric interactions. These results suggest that dog, cat, and pig CYP2Js have similar functional characteristics to human CYP2J2, with slight differences in ebastine and astemizole oxidations. SIGNIFICANCE STATEMENT: Dog CYP2J2; cat CYP2J2; and pig CYP2J33, CYP2J34, CYP2J35, CYP2J91, and CYP2J93, homologous to human CYP2J2, were identified and characterized by sequence, phylogenetic, and genomic structure analyses. Intestinal expression patterns of CYP2J mRNAs were characteristic in dogs, cats, and pigs. Dog, cat, and pig CYP2Js likely play roles as drug-metabolizing enzymes in the small intestine, similar to human CYP2J2.
细胞色素 P450s（P450s）已在狗和猪中被鉴定和分析，这两种动物通常用于临床前药物研究。此外，P450s 不仅在人类，而且在兽医护理下的物种（包括狗和猫）中，对药物治疗具有重要的临床意义。在本研究中，新鉴定并表征了七种与人 CYP2J2 同源的 P450，即狗 CYP2J2；猫 CYP2J2；和猪 CYP2J33、CYP2J35、CYP2J91 和 CYP2J93，以及先前鉴定的猪 CYP2J34。这些 CYP2Js 的 cDNA 包含 502 个氨基酸的开放阅读框，除了 CYP2J35（498 个氨基酸）外，与人类 CYP2J2 具有高度的序列同一性（77%-80%）。系统进化分析表明，狗和猫 CYP2J2 密切相关，而猪 CYP2Js 则形成一个聚类。所有七个基因均包含九个编码外显子，位于相应的基因组区域，猪基因形成基因簇。这些 CYP2J2 mRNA 主要在小肠中表达，狗 CYP2J2 和猪 CYP2J91 mRNA 分别在肾脏和大脑中有额外表达。所有七种 CYP2J 均代谢人 CYP2J2 底物特非那定、依巴斯汀和阿司咪唑，表明它们是功能性酶。狗 CYP2J2 和猪 CYP2J34 和 CYP2J35 在低底物浓度下有效催化依巴斯汀的初级羟化和次级卡巴斯汀形成，就像人 CYP2J2 一样。速度-底物图显示狗 CYP2J2、猫 CYP2J2 和猪 CYP2J33 呈 S 型关系，表明存在变构相互作用。这些结果表明，狗、猫和猪 CYP2Js 具有与人类 CYP2J2 相似的功能特征，在依巴斯汀和阿司咪唑氧化方面略有差异。意义说明：通过序列、系统发育和基因组结构分析，鉴定并表征了与人类 CYP2J2 同源的狗 CYP2J2；猫 CYP2J2；和猪 CYP2J33、CYP2J34、CYP2J35、CYP2J91 和 CYP2J93。狗、猫和猪 CYP2J mRNA 的肠道表达模式具有特征性。狗、猫和猪 CYP2Js 可能在小肠中作为药物代谢酶发挥作用，类似于人类 CYP2J2。
