Development of a high-throughput screen for soluble epoxide hydrolase inhibition
Development of a high-throughput screen for soluble epoxide hydrolase inhibition
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DOI：
10.1016/j.ab.2006.04.045
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发表时间：
2006-08-01
影响因子：
2.9
通讯作者：
Hammock, Bruce D.
中科院分区：
文献类型：
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作者：
Wolf, Nicola M.;Morisseau, Christophe;Hammock, Bruce D.
哺乳动物可溶性环氧化物水解酶（sEH）是一个非常有前途的药物开发新靶点。在动物模型中，这种酶的化学抑制剂被证明可以治疗高血压和血管炎症以及相关综合征。现有的sEH抑制剂是相对有效和特异的。然而，所描述的sEH抑制剂的低溶解度和相对快速的代谢使它们在治疗上不那么有效，说明需要新的抑制剂结构。因此，一系列的α-氰基酯和α-氰基碳酸酯环氧化物被评估为潜在的人类sEH（HsEH）的高通量筛选（HTS）的化合物库的基板。选择表现出最高水稳定性和溶解度的（3-苯基-环氧乙烷基）-乙酸氰基-（6-甲氧基-萘-2-基）-甲酯（PHOME）用于开发在室温下长孵育时间的HTS测定。优化HsEH和PHOME的浓度以确保测定灵敏度、可靠性和再现性。采用这些优化浓度的测定验证产生了良好的准确度（60-100%）和高精密度（< 7%相对标准偏差）。此外，0.7的总Z'值证明了系统的鲁棒性和HTS的潜力。开发的测定系统将是一个有价值的工具，发现新的结构的治疗性抑制sEH治疗各种心血管疾病。(c)2006年爱思唯尔公司All rights reserved.
Mammalian soluble epoxide hydrolase (sEH) represents a highly promising new target for drug development. Chemical inhibition of this enzyme in animal models was shown to treat hypertension and vascular inflammation as well as related syndromes. Existing sEH inhibitors are relatively potent and specific. However, the low solubility and relatively fast metabolism of described sEH inhibitors make them less than therapeutically efficient, stating the need for novel inhibitor structures. Therefore, a series of alpha-cyanoester and alpha-cyanocarbonate epoxides were evaluated as potential human sEH (HsEH) substrates for the high-throughput screen (HTS) of compound libraries. (3-Phenyl-oxiranyl)-acetic acid cyano-(6-methoxy-naphthalen-2-yl)-methyl ester (PHOME), which displayed the highest aqueous stability and solubility, was selected for the development of an HTS assay with long incubation times at room temperature. Concentrations of HsEH and PHOME were optimized to ensure assay sensitivity, reliability, and reproducibility. Assay validation, which employed these optimized concentrations, resulted in good accuracy (60-100%) and high precision (< 7% relative standard deviation). In addition, an overall Z' value of 0.7 proved the system's robustness and potential for HTS. The developed assay system will be a valuable tool to discover new structures for the therapeutic inhibition of sEH to treat various cardiovascular diseases. (c) 2006 Elsevier Inc. All rights reserved.