Pharmacological inhibition of matrix metalloproteinase-2 (MMP-2) is a promising target for acute cardioprotection against ischemia/reperfusion injury. Therefore, here we investigated if the MMP inhibitor ilomastat administered either before ischemia or before reperfusion is able to reduce infarct size via inhibition of MMP-2, the most abundant MMP in the rat heart. Infarct-size limiting effect of ilomastat (0.3–6.0 μmol/kg) was tested in an in vivo rat model of myocardial infarction induced by 30 min coronary occlusion/120 min reperfusion. Ilomastat at 0.75 and 1.5 μmol/kg decreased infarct size significantly as compared to the vehicle-treated (dimethyl sulfoxide) group (from 66.1 ± 4.6% to 45.3 ± 7.0% and 46.7 ± 5.5% of area at risk, p