Screening And Anti-inflammatory Effect Evaluation Of Novel Soluble Epoxide Hydrolase Inhibitors

Epoxy-fatty acids are biologically active lipid signaling molecules,and can effectively inhibit the progression of various diseases.However,epoxy-fatty acids can be rapidly hydrolyzed to the corresponding diol fatty acids under the catalysis of soluble epoxide hydrolase（sEH）,and lose their biological activity.The small molecule sEH inhibitors have attracted widespread attention because they can keep or increase the biological activity of the epoxy-fatty acids.This subject belongs to the basic research of new drug development.The purity,enzyme inhibitory activity,physicochemical properties,and anti-inflammatory activity in vitro and in vivo of the compound were used as indicators to screen new and potential sEH inhibitors.In this study,a diode array detector and gradient elution program were used to establish a general method for determining the purity of compounds.The inhibitory activity of compounds on sEH was tested by the sEH inhibitor screening kit.The test method was determined by optimizing the test conditions,including sample loading method,incubation time,and detection concentration.Physicochemical properties of the compound such as equilibrium solubility and oil-water partition coefficient were investigated.A model of infammation was established using lipopolysaccharide（LPS）to induce BV2 microglia activation,and the effect of sEH inhabitors was evaluated in vitro.Using the degree of swelling of the foot as an index,the effect of sEH inhabitors in carrageenan-induced inflammation in vivo was evaluated.Through the above tests,a new potential sEH inhibitor was successfully screened.The results showed that the compound ZH-A-030 had an enzyme inhibition rate of60.50%and an IC₅₀value of 5.87 nmol·L^-1.Its equilibrium solubility in water was 2370.73μg·mL^-1,its oil-water partition coefficient was 0.74,and its stability was good.Meanwhile,it could inhibit microglia producing inflammation and reduce the swelling degree of the foot,exhibiting good anti-inflammatory activity both in vitro and in vivo.All these results confirmed that ZH-A-030 was a new and potential sEH inhabitor.