Screening of Specific Inhibitors for Human Carboxylesterases or Arylacetamide Deacetylase
Screening of Specific Inhibitors for Human Carboxylesterases or Arylacetamide Deacetylase
复制标题
DOI：
10.1124/dmd.114.056994
复制
发表时间：
2014-07-01
影响因子：
3.9
通讯作者：
Yokoi, Tsuyoshi
中科院分区：
文献类型：
--
作者：
Shimizu, Mai;Fukami, Tatsuki;Yokoi, Tsuyoshi
酯酶催化在其结构中含有酯或酰胺的治疗药物的水解。人羧酸酯酶（CES）和芳基乙酰胺脱乙酰酶（AADAC）是肝脏中催化药物水解的主要酶。表征负责药物代谢的酶是药物开发和实现最佳药物治疗所必需的。由于多种酶可能对给定化合物显示出代谢效力，因此使用化学抑制剂的抑制研究是确定每种酶在人体组织制备物中的贡献的有用工具。本研究的目的是寻找人CES 1、CES 2和AADAC的特异性抑制剂。我们筛选了542种化学物质对对硝基苯基乙酸酯水解酶活性的抑制效力，通过重组CES 1，CES 2和AADAC。我们发现毛地黄皂苷和替米沙坦分别特异性抑制CES 1和CES 2酶活性。长春碱可有效抑制AADAC和CES 2，但未发现AADAC的特异性抑制剂，通过监测各酶的探针化合物（CES 1：利多卡因，CES 2：CPT-11，AADAC：非那西丁）对人肝微粒体水解酶活性的影响，评价了这些化合物的抑制效力和特异性。替米沙坦和长春碱强烈抑制CPT-11和/或非那西丁的水解，但毛地黄皂苷没有强烈抑制利多卡因的水解，表明重组CES 1和肝微粒体中毛地黄皂苷的抑制效力不同。虽然我们没有找到AADAC的特异性抑制剂，但联合使用替米沙坦和长春碱可以预测人AADAC对药物水解的反应性。
Esterases catalyze the hydrolysis of therapeutic drugs containing esters or amides in their structures. Human carboxylesterase (CES) and arylacetamide deacetylase (AADAC) are the major enzymes that catalyze the hydrolysis of drugs in the liver. Characterization of the enzyme(s) responsible for drug metabolism is required in drug development and to realize optimal drug therapy. Because multiple enzymes may show a metabolic potency for a given compound, inhibition studies using chemical inhibitors are useful tools to determine the contribution of each enzyme in human tissue preparations. The purpose of this study was to find specific inhibitors for human CES1, CES2, and AADAC. We screened 542 chemicals for the inhibition potency toward hydrolase activities of p-nitrophenyl acetate by recombinant CES1, CES2, and AADAC. We found that digitonin and telmisartan specifically inhibited CES1 and CES2 enzyme activity, respectively. Vinblastine potently inhibited both AADAC and CES2, but no specific inhibitor of AADAC was found. The inhibitory potency and specificity of these compounds were also evaluated by monitoring the effects on hydrolase activity of probe compounds of each enzyme (CES1: lidocaine, CES2: CPT-11, AADAC: phenacetin) in human liver microsomes. Telmisartan and vinblastine strongly inhibited the hydrolysis of CPT-11 and/or phenacetin, but digitonin did not strongly inhibit the hydrolysis of lidocaine, indicating that the inhibitory potency of digitonin was different between recombinant CES1 and liver microsomes. Although we could not find a specific inhibitor of AADAC, the combined use of telmisartan and vinblastine could predict the responsibility of human AADAC to drug hydrolysis.