The treatment of inflammatory bowel disease (IBD) may rely on the comprehensive regulation of immune inflammatory factors to reach a favorable therapeutic effect. In the current study, we discovered a series of sEH inhibitors containing a squaryl sulfonamide scaffold that potently inhibited the primary proinflammatory signaling pathways of NF-kappaB. Especially, lead compounds A1 and A9 showed potent inhibitory activities against sEH (A1 and A9; hsEH IC(50) = 0.1 nM, msEH IC(50) = 0.1 nM). The compound A1 had medium pharmacokinetic characteristics in rats. In vivo, A1 showed a strong anti-inflammatory activity in acute enteritis models and decreased the release of various proinflammatory factors, including IL-6 and TNF-alpha. More importantly, A1 maintained the integrity of the intestinal barrier. Besides, A1 possessed good in vivo tolerability in subacute safety evaluation. Collectively, this study provided valuable lead compounds for the treatment of IBD that were worthy of further development.