Several casein kinase 2 inhibitors have been prepared using some protocols which require dangerous reagents and specific conditions. The aim of this study was to synthesize a new chloroacetamide-steroid derivative using some chemical tools. In addition, the effect exerted by chloroacetamide-steroid derivative on casein kinase 2 (CK2) was determinate in an ischemia-reperfusion injury model using quinalizarin and N-tertbuthyl-2-chloroacetamide as controls. The results showed that 1) the chloroacetamide-steroid derivative significantly decrease the ischemia-reperfusion injury translated as infarct area compared with quinalizarin and N-tertbuthyl-2-chloroacetamide. In conclusion, in this study, is reported a facile synthesis of a new chloroacetamide-steroid derivative with biological activity against CK2.