Deciphering the species differences in CES1A-mediated hydrolytic metabolism by using a bioluminescence substrate
Deciphering the species differences in CES1A-mediated hydrolytic metabolism by using a bioluminescence substrate
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DOI：
10.1016/j.cbi.2022.110197
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发表时间：
2022-10-07
影响因子：
5.1
通讯作者：
Ge, Guang-Bo
中科院分区：
文献类型：
--
作者：
Jin, Qiang;Li, Zan;Ge, Guang-Bo
羧酸酯酶1A（Carboxylesterases 1A，CES1A）是一种重要的酶，负责水解代谢大量含酯键或酰胺键的内源性和外源性底物。本研究旨在通过使用新开发的生物发光CES1A传感器（称为NLMe）作为探针底物，而来自六种不同哺乳动物物种（人、食蟹猴、狗、小型猪、大鼠和小鼠）的肝微粒体作为酶源，来破译CES1A介导的水解代谢的物种差异。代谢物谱表明，所有供试的不同物种的肝微粒体都能催化NLMe水解，但水解速率存在显着差异。不同种属肝微粒体中NLMe水解的动力学图显示，人肝微粒体（HLM）、食蟹猴肝微粒体（CyLM）和猪肝微粒体（PLM）中NLMe的固有清除率（克林特）相当，而犬肝微粒体（DLM）、小鼠肝微粒体（MLM）和大鼠肝微粒体（RLM）中NLMe的克林特值相对较小。此外，化学抑制试验表明，在所有测试的肝微粒体中的NLMe水解可以被BNPP（一种有效的广谱CES抑制剂）完全抑制，但是CUA（一种选择性的人CES1A抑制剂）仅抑制人肝微粒体和狗肝微粒体中的NLMe水解。总之，从代谢产物谱、水解动力学和抑制剂响应的相似性的角度仔细研究了CES1A催化NLMe水解的物种差异。这些发现为研究CES1A介导的水解代谢的种属差异提供了新的视角，并提示药理学家有必要选择合适的动物模型来代替人类来评价CES1A抑制剂的体内效应。
Carboxylesterases 1A (CES1A) is a key enzyme responsible for the hydrolytic metabolism of a great deal of endogenous and exogenous substrates bearing ester-or amide-bond(s). This study aimed to decipher the species difference in CES1A-mediated hydrolytic metabolism by using a newly developed bioluminescence CES1A sensor (termed NLMe) as the probe substrate, while the liver microsomes from six different mammalian species (human, cynomolgus monkey, dog, minipig, rat and mouse) were used as the enzyme sources. Metabolite profiling demonstrated that all tested liver microsomes from various species could catalyze NLMe hydrolysis, but signif-icant difference in hydrolytic rate was observed. Kinetic plots of NLMe hydrolysis in liver microsomes from different species showed that the inherent clearance rates (Clint) of NLMe in human liver microsomes (HLM), cynomolgus monkey liver microsomes (CyLM), and pig liver microsome (PLM) were comparable, while the Clint values of NLMe in dog liver microsomes (DLM), mouse liver microsomes (MLM), and rat liver microsomes (RLM) were relatively small. Moreover, chemical inhibition assays showed that NLMe hydrolysis in all tested liver microsomes could be competently inhibited by BNPP (a potent broad-spectrum inhibitor of CES), but CUA (a selective inhibitor of human CES1A) only inhibited NLMe hydrolysis in human liver microsomes and dog liver microsomes. In summary, the species differences in CES1A-catalyzed NLMe hydrolysis were carefully investi-gated from the views of the similarities in metabolite profile, hydrolytic kinetics and inhibitor response. All these findings provide new insights into the species differences in CES1A-mediated hydrolytic metabolism and suggest that it is necessary for the pharmacologists to choose appropriate animal models to replace humans for evalu-ating the in vivo effects of CES1A inhibitors.