Cyanohydrin based fluorescent substrates of sEH (RMI)

描述（由申请人提供）：最近的发现表明，增加花生四烯酸环氧化物（EE1）的体内水平似乎是治疗高血压和血管炎症的新的和极好的手段。 我们已经证明，在动物模型中，可溶性环氧化物水解酶（sEH）的体内抑制导致更高水平的雌二醇，并降低血压和炎症。 由于第一代sEH抑制剂的有效性和作用长度低于最佳，因此我们目前工作的总体目标是开发具有改善的体内效力的sEH抑制剂或前药。 为此，我们将首先基于环氧化物水解后氰醇的释放产生sEH的荧光测定。将对测定进行优化，以建立96孔和384孔格式的高通量筛选。 将针对我们的sEH抑制剂文库和更大的商业组合文库验证该测定。 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“NIH路线图”中的组合文库、与商业供应商签订合同的文库以及内部生成的更明确的文库来完成。

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.