Discovery of benzamide derivatives containing urea moiety as soluble epoxide hydrolase inhibitors

• Benzamide derivatives containing urea moiety were designed and synthesized, and the structure–activity relationship was fully explored. • Compound A34 exhibited outstanding inhibitory activity against human sEH, with an IC 50 value of 0.04 ± 0.01 nM. • Compound A34 showed moderate systemic drug exposure and oral bioavailability in vivo metabolism studies. • In carrageenan-induced inflammatory pain rat model, compound A34 exhibited a better therapeutic effect compared to t -AUCB and Celecoxib. The elevation of epoxy-fatty acids through inhibition of soluble epoxide hydrolase (sEH) is efficient for the treatment of inflammatory and pain-related diseases. Herein, we reported the discovery of a series of benzamide derivatives containing urea moiety as sEH inhibitors. Intensive structural modifications led to the identification of compound A34 as a potent sEH inhibitor with good physicochemical properties. Molecular docking revealed an additional hydrogen-bonding interaction between the unique amide scaffold and Phe497, contributing to sEH inhibition potency enhancement. Compound A34 exhibited outstanding inhibitory activity against human sEH, with an IC 50 value of 0.04 ± 0.01 nM and a K i value of 0.2 ± 0.1 nM. It also showed moderate systemic drug exposure and oral bioavailability in vivo metabolism studies. In carrageenan-induced inflammatory pain rat model, compound A34 exhibited a better therapeutic effect compared to t -AUCB and Celecoxib. Metabolism studies in vivo together with an inflammatory pain evaluation suggest that A34 may be a viable lead compound for the development of highly potent sEH inhibitors.