The anti-Trypanosoma cruzi activity of 5-nitro-2-furfuriliden derivatives as well as the cytotoxicity of these compounds on J774 macrophages cell line and FN1 human fibroblast cells were investigated in this study. The most active compounds of series I and II were 4-butyl-[N'-(5-nitrofuran-2-yl) methylene] benzidrazide (3g; IC50=1.05μM±0.07) and 3-acetyl-5-(4-butylphenyl)-2-(5-nitrofuran-2-yl)-2,3-dihydro,1,3,4-oxadiazole (4g; IC50=8.27μM±0.42), respectively. Also, compound 3g was more active than the standard drugs, benznidazole (IC50=22.69μM±1.96) and nifurtimox (IC50=3.78μM±0.10). Regarding the cytotoxicity assay, the 3g compound presented IC50 value of 28.05μM (SI=26.71) against J774 cells. For the FN1 fibroblast assay, 3g showed IC50 value of 98μM (SI=93.33). On the other hand, compound 4g presented a cytotoxicity value on J774 cells higher than 400μM (SI >48), and for the FN1 cells its IC50 value was 186μM (SI=22.49). Moreover, an exploratory data analysis, which comprises hierarchical cluster (HCA) and principal component analysis (PCA), was carried out and the findings were complementary. The molecular properties that most influenced the compounds' grouping were ClogP and total dipole moment, pointing out the need of a lipophilic/hydrophilic balance in the designing of novel potential anti-T. cruzi molecules.