Many cardiac interventional procedures, such as coronary angioplasty, stenting, and thrombolysis, attempt to reintroduce blood-flow (reperfusion) to an ischemic region of myocardium. However, the reperfusion is accompanied by a complex cascade of cellular and molecular events resulting in oxidative damage, termed myocardial ischemia-reperfusion (I-R) injury. In this study, we evaluated the ability of HO-4038, an N-hydroxypiperidine derivative of verapamil, on the modulation of myocardial tissue oxygenation (pO₂), I-R injury, and key signaling molecules involved in cardioprotection in an in vivo rat model of acute myocardial infarction (MI). MI was created in rats by ligating the left anterior descending coronary artery (LAD) for 30 min followed by 24-h reperfusion. Verapamil or HO-4038 was infused through jugular vein 10 min before the induction of ischemia. Myocardial pO₂ and free-radical scavenging ability of HO-4038 were measured using EPR spectroscopy. HO-4038 showed a significantly better scavenging ability of reactive-oxygen radicals when compared to verapamil. The cardiac contractile functions in the I-R hearts were significantly higher recovery in HO-4038 group compared to verapamil group. A significant decrease in the plasma levels of creatine kinase (CK) and lactate dehydrogenase (LDH) was observed in HO-4038 group compared to verapamil or untreated I-R groups. The left-ventricular infarct size was significantly less in HO-4038 group (23±2%) compared to untreated I-R group (36±4%). HO-4038 significantly attenuated the hyperoxygenation (36±1 mmHg) during reperfusion compared to untreated I-R group (44±2 mmHg). HO-4038-treated group also markedly attenuated superoxide production, increased nitric oxide generation, and enhanced Akt and Bcl-2 levels in the reperfused myocardium. Overall, the results demonstrated that HO-4038 significantly protected hearts against I-R-induced cardiac dysfunction and damage through combined beneficial actions of calcium-channel blocking, antioxidant, and pro-survival signaling activities.