Leishmaniasis is a tropical disease caused by different species of Leishmania. The available treatments have limitations linked to high toxicity and low efficacy. Thus, the search for new antileishmanial compounds is needed. Since Matricaria chamomilla L. have several activities described in literature, the aim of this search was to obtain crude extract and fractions of M. chamomilla and to evaluate the activity against the evolutionary forms of Leishmania amazonensis. Floral chapters of M. chamomilla were preserved in 95% ethanol for 12 days and lyophilized to obtain the crude extract (CE). Then, with the aid of a separating funnel and organic solvents, hexane (FHE), ethyl acetate (FEA) and hydromethanolic (FHM) fractions were obtained. The CE and the fractions were tested in vitro against the promastigote and intracellular amastigotes forms and the macrophage cell line J774A.1. The results were plotted on graphs and the IC50 was determined in relation to the control. In view of the evolutionary forms of L. amazonensis, FHE presented the best results, with the IC50 for promastigotes of 14.2 µg/mL, with a selectivity index (IS) of 8.15; Against amastigotes, FHE showed an IC50 of 53.84 and IS of 2.13. Further studies should be conducted to elucidate FHE's antileishmanial actions.