Leishmania parasites are the causative agent of leishmaniasis. Many studies are inspecting chemical drugs, including the use of miltefosine and amphotericin B, but curative values may be limited for these drugs with side effects due to the chemical origin, therefore, investigating less toxic therapies is essential. The aim of this study was to investigate the effectiveness of artemisinin on Iraqi strain of Leishmania tropica, by experimental macrophage ex vivo infection of amastigotes into mouse macrophage cell-line RAW264.7. Different concentrations (100, 200, 300, 400, 500)μM of artemisinin (ART) were screened to examine the susceptibility of L. tropica amastigotes to invade macrophage cell line along three times of follow up (24, 48 and 72) hours. Results showed that artemisinin had a cytotoxic effect on the parasite in which a significance difference (P < 0.05) in cell viability was observed and IC50 was calculated as 182.6 μM after 48 hours treatment. In addition, percentage of infectivity of intracellular amastigotes was significantly decreased. These findings revealed the potential efficacy of artemisinin against the infectious amastigotes and can be further studied to screen its effectiveness in vivo for exploring a safer herbal compound to treat cutaneous leishmaniasis.