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Objectives To determine the protective role of a soluble epoxide hydrolase(sEH) inhibitor, trans‐4‐{4‐[3‐(4‐trifluoromethoxyphenyl)‐ureido] cyclohexyloxy} benzoic acid (t‐TUCB), in isoproterenol (ISO)‐induced myocardial ischaemic injury in vivo. Methods Cardioprotective activity of t‐TUCB was studied against ISO‐induced myocardial ischaemic injury in male Wistar rats. Cardioprotection was assessed by measuring elecrocardiographic (EKG), serum lactate dehydrogenase (LDH) and creatine kinase (CK‐MB) levels, cardiac calcium and antioxidant levels, and also by measuring infarct size in the cardiac tissue. Key findings  Pretreatment with t‐TUCB at 3, 10 and 30 mg/kg orally for a period of 14 days significantly prevented the changes in EKG parameters (QTc interval prolongation, ST height depression, pathological Q waves formation and T‐wave inversion), serum cardiac biomarkers (CK‐MB and LDH), relative heart weight, myocardial calcium levels, infarct size and the oxidative status in the cardiac tissue (lipid peroxidation, catalase and superoxide dismutase levels) when compared with the untreated control animals (P < 0.05). Conclusion The sEH inhibitor t‐TUCB significantly prevents ISO‐induced myocardial ischaemic injury in rats. This study provides a preliminary confirmation of the efficacy of t‐TUCB by oral administration in rats.

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