Background: Cancer is the leading cause of death around the world, and the number of cancer deaths is expected to significantly rise in the coming years mainly due to aging of world population. The cornerstone of cancer treatment since a long time ago was anthracyclines, mainly doxorubicin. Despite doxorubicin importance in cancer therapy, there are several side effects induced by doxorubicin. Methodology: The toxicity of doxorubicin, calpeptin and combination was studied in different cancer cell lines HepG2, EA.HY629, MDA and A549 using 3-(4,5-dimethylthazolk-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay. Scratch and colony assay was studied in HepG2 and EA.HY629. Histological examinations were studied in Balb/c mice injected with doxorubicin and the combination of doxorubicin and calpeptin. Result: The combination between the two drugs didn't affect the cytotoxicity of DOX. The IC50 of combination treated cells in HepG2, EA.HY629, MDA and A549 was 56.91 µM, 1.692 µM, 2.173 µM and 2.09 µM respectively. In migration assay, the DOX treated cells had scratch closure in HepG2 and EA.HY629 with 109.02 + 9.13 % and 131.89 + 2.91%, respectively. Calpeptin treated cells had scratch closure in HepG2 and EH.HY629 with 105.42 + 1.71 % and 105.42 + 6.38% , respectively. DOX and calpeptin reduced colony formation, HepG2 and EA.HY629 had zero colonization after 14 days of treatment. In mice treated with combination therapy, there was a mild lymphocyte infiltration and necrosis in comparison to mic injected with DOX alone. Conclusion: Combination of DOX and calpeptin didn't affect the cytotoxicity of DOX, also calpeptin showed to prevent migration and colony formation. This work revealed that combination of both drugs could have the potential to reduce heart toxicity in patients receiving DOX therapy.