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Objective:To evaluate the effect of doxorubicin and its pegylated liposomal formulation (Doxil, Caelyx) on in vitro susceptibility of promastigote and amastigote stages of Leishmania major.Methods:Throughout in vitro assays the IC50 was calculated in the promastigotes and amastigotes forms in J774 macrophage cell line. Also as cytotoxicity in J774 cell line macrophages.Results: Doxorubicin and Doxil showed the same activity against promastigote form with IC50 values of 10.49 μg/mL and 9.63 μg/mL, respectively. Similarly, the amastigote stage was susceptible at concentration of at least 1 μg/mL when compared to positive control (P<0.0001). Also, cytotoxicity assay against macrophage revealed no toxicity on the host cells at IC50concentrations.Conclusions: Our findings demonstrated the efficacy of both doxorubicin and its pegylated liposomal formulation on Leishmania major at low concentrations. Further researches are needed for evaluating the safety of drugs in animal model particularly as topical formulation.