Interspecies Variation in NCMN-O-Demethylation in Liver Microsomes from Various Species
Interspecies Variation in NCMN-O-Demethylation in Liver Microsomes from Various Species
复制标题
不同物种肝微粒体中 NCMN-O-去甲基化的种间变异
DOI：
10.3390/molecules24152765
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发表时间：
2019
期刊：
影响因子：
4.6
通讯作者：
Yang Ling
中科院分区：
文献类型：
--
作者：
Dai Ziru;Sun Guibo;Yang Jiada;Hou Jie;Zhou Ping;Xie Weijie;Ge Guangbo;Sun Xiaobo;Yang Ling
NCMN（N-（3-羧基丙基）-4-甲氧基-1，8-萘二酰亚胺）是一种新开发的人细胞色素P450 1A（CYP 1A）比率双光子荧光探针，具有实时检测酶活性的最佳特异性和反应性组合复杂生物系统中的CYP 1A。本研究旨在研究来自人、小鼠、大鼠、比格犬、小型猪和食蟹猴的市售肝微粒体中NCMN-O-去甲基化的种间差异。代谢物谱分析表明，NCMN可以在所有种属的肝微粒体中进行O-脱甲基化，但反应速率差异很大。基于化学抑制试验，CYP 1A是所有检查的肝微粒体中参与NCMN-O-去甲基化的主要亚型。呋喃茶碱，哺乳动物CYP 1A的特异性抑制剂，在所有测试种属中显示对NCMN-O-去甲基化的不同抑制作用。动力学分析表明，大鼠、小型猪和食蟹猴肝微粒体中的NCMN-O-去甲基化遵循双相动力学，而人、小鼠和比格犬肝微粒体中的NCMN-O-去甲基化遵循Michaelis-Menten动力学，不同种属的动力学参数差异较大，而MLM中的NCMN-O-去甲基化与HLM中的NCMN-O-去甲基化在特异性、动力学行为和内在清除率方面具有最高的相似性。这些发现将有助于合理使用NCMN作为一种实用工具来解读哺乳动物CYP 1A的功能或研究CYP 1A相关的药物相互作用。
NCMN (N-(3-carboxy propyl)-4-methoxy-1,8-naphthalimide), a newly developed ratiometric two-photon fluorescent probe for human Cytochrome P450 1A (CYP1A), shows the best combination of specificity and reactivity for real-time detection of the enzymatic activities of CYP1A in complex biological systems. This study aimed to investigate the interspecies variation in NCMN-O-demethylation in commercially available liver microsomes from human, mouse, rat, beagle dog, minipig and cynomolgus monkey. Metabolite profiling demonstrated that NCMN could be O-demethylated in liver microsomes from all species but the reaction rate varied considerably. CYP1A was the major isoform involved in NCMN-O-demethylation in all examined liver microsomes based on the chemical inhibition assays. Furafylline, a specific inhibitor of mammalian CYP1A, displayed differential inhibitory effects on NCMN-O-demethylation in all tested species. Kinetic analyses demonstrated that NCMN-O-demethylation in liver microsomes form rat, minipig and cynomolgus monkey followed biphasic kinetics, while in liver microsomes form human, mouse and beagle dog obeyed Michaelis-Menten kinetics, the kinetic parameters from various species are much varied, while NCMN-O-demethylation in MLM exhibited the highest similarity of specificity, kinetic behavior and intrinsic clearance as that in HLM. These findings will be very helpful for the rational use of NCMN as a practical tool to decipher the functions of mammalian CYP1A or to study CYP1A associated drug-drug interactions in vivo.