Cyanohydrin based fluorescent substrates of sEH (RMI)

描述（由申请人提供）：最近的研究表明，增加体内花生四烯酸环氧化物（EEl’s）的水平似乎是治疗高血压和血管炎症的一种新的和优秀的手段。我们已经证明，在动物模型中，可溶性环氧化物水解酶（sEH）的体内抑制导致EETs水平升高，并降低血压和炎症。由于第一代sEH抑制剂的有效性和作用时间都不是最佳的，因此我们当前工作的总体目标是开发具有更好体内效力的sEH抑制剂或前药。为此，我们将首先建立一种基于环氧化物水解释放氰醇的sEH荧光检测方法。该分析将朝着建立96孔和384孔格式的高通量筛选进行优化。该分析将针对我们的sEH抑制剂文库和更大的商业组合文库进行验证。最终，该试验将用于开发新的有效的sEH抑制剂。这将通过筛选来自“NIH路线图”的组合库，从商业供应商处签约的库，以及内部生成的更明确的库来完成。

DESCRIPTION (provided by applicant): Recent findings indicate that increasing the in vivo levels of arachidonic acid epoxides (EEl's) appears to be a new and excellent means to treat both hypertension and vascular inflammation. We have demonstrated that in vivo inhibition of soluble epoxide hydrolase (sEH) resulted in higher levels of EETs and in the reduction of blood pressure and inflammation in animal models. Because the effectiveness and length of action were less than optimal for the first generation of sEH inhibitors, the overall objective of our current work is to develop sEH inhibitors or prodrugs with improved r in vivo potency. Towards this end, we will first produce a fluorescent assay for sEH based on the liberation of cyanohydrin upon epoxide hydrolysis. The assay will be optimized toward establishing a high throughput screen in 96- and 384-well formats. The assay will be validated against our library of sEH inhibitors and a larger commercial combinatorial library. Ultimately, this assay will be used to develop new potent inhibitors of sEH. This will be done by screening the combinatorial libraries from the "NIH Roadmap", libraries contracted from commercial vendors, and more defined libraries generated in house.