The objective of this study was to carry out phytochemical analysis of the bark extract of Casearia javitensis and to evaluate its antipromastigote activity against Leishmania amazonensis. The ethanol extract (EE) was fractionated in open-column chromatography to obtain the hexane (FrHex), dichloromethane (FrDcm), ethyl acetate (FrAcOET) and methanol (FrMeOH) fractions. The EE and its fractions were analyzed by thin layer chromatography (TLC). FrDcm was analyzed in high performance liquid chromatography with diode array detection (HPLC-DAD). The antipromastigote assay of Leishmania amazonensis and the cytotoxicity test against the acute monocytic leukemia cell line (THP-1) were performed by the MTT colorimetric assay. The phytochemical profile in TLC detected terpenes in all samples. However, in the ultraviolet spectrum (HPLC-DAD) for FrDcm suggested the presence of a phenolic compound. FrHex and FrDcm showed antipromastigote activity (IC50 = 116.6 ± 0.9 and 59.4 ± 1.1 μg mL-1, respectively) and low cytotoxicity (CC50 = 333.4 ± 3.2 and 241.2 ± 1.9 μg mL-1, respectively). The selectivity index for FrDcm was 4.1. We conclude that the FrDcm of C. javitensis has potantial as a leishmanicide, and this activity may be related to the presence of phenolic compound.