The cells were differentiated into macrophages by adding 40 ng/mL of phorbol-myristate 13acetate (PMA, Sigma, Saint Louis, MI, USA) for 24 hours, followed by replacement with fresh medium for another 24 hours. Subsequently, the cells were incubated with 100 µL of compounds, ranging from 100 to 12.5 µM, diluted in RPMI complete medium. Each condition was tested in triplicate. After 72 hours of incubation at 37 • C with 5% CO 2 , the medium was removed and 200 µL of 0.5 mg/mL MTT solution diluted in RPMI was added. The plates were incubated for an additional 4 hours, after which 160 µL of the media was removed and replaced with 160 µL of 2-propanol. Absorbance was read at 570 nm using a Synergy 2 Activity against L. infantum intracellular amastigotes. The activity against Leishmania infantum intracellular amastigotes was evaluated following the method described with modifications . THP-1 cells were differentiated into macrophages with PMA as described above. The cells were then infected with L. infantum axenic amastigotes expressing episomal luciferase at a macrophage-to-amastigote ratio of 1:10 for 4 hours at 37 • C and 5% CO 2 .