Natural products are a very important source of bio-molecules for drug to treat infectious diseases. New antileishmanial agents are timely needed, due to leishmania toxicity, drug-resistant strains and high costs treatments. In this study, we report the chemical characterization and antileishmanial activity of oil extracts from three medicinal plants belonging to genus Artemisia (Asteracea), that grow in central Tunisia. The chemical composition of volatile compounds had been elucidated by gas chromatography-mass spectroscopy analysis. Chemical analysis revealed that essential oils were composed of 34 compounds: α-thujone (29.3%), chamazulene (39.2%) and β-pinene (32%), were the main constituents for the essential oils of Artemesia herba alba, Artemesia absinthium and Artemesia campestris, respectively. The hydrodistilled essential oils from these three species of Artemisia showed significant anti-leishmanial activities against Leishmania major. Oils from A. herba alba, A. absinthium and A. campestris exhibited IC₅₀ values of 1.20 ± 0.043 μg/mL, 1.49 ± 0.05 μg/mL and 2.20 ± 0.11 μg/mL respectively against promastigotes of Leishmania major. Among three oils tested, A. campestris exerted a remarkable antileishmanial activity and it has the lowest cytotoxicity effect compared to amphotericin B. Overall, it was proved that our investigated essential oils possess potential antileishmanial properties and could be used as a promising alternative treatment for leishmaniasis disease in the future.