The in vivo oxidation of arachidonic acid produces a multitude of compounds which have been found to act as modulators of vascular physiology. Arachidonic acid epoxides (EETs), endogenous regulators of blood pressure and vascular inflammation, are hydrolyzed by the soluble epoxide hydrolase (sEH). Preliminary evidence suggests that sEH inhibition by acyclic urea compounds drastically attenuates urinary albumin excretion (UAE) in Zucker rat models. These inhibitors however are difficult to formulate due to their poor solubility. In addition, the current absorption based in vitro assays cannot distinguish between those inhibitors with the highest potency. Therefore, it is hypothesized that 1) fluorescent substrates will increase the sensitivity of the current enzymatic inhibitor assay and 2) heterocyclic compounds are potent inhibitors of sEH with improved physical properties. These hypotheses will be tested using the following specific aims:
Specific Aim I : Test the hypothesis that a fluorescent based kinetic assay for sEH activity provides a highly sensitive platform for the rapid, high-throughput evaluation of inhibitors.
Specific Aim II : Test the hypothesis that inhibitors, which have reduced rotational freedom within the primary pharmacophore, are potent inhibitors of sEH with increase water solubility.
