Chagas disease is derived from the infection by the protozoanTrypanosoma cruzi. In many countries, benznidazole is the only drug approved for clinical use despite several side effects and the emergence of resistant parasite strains. In this context, our group has previously pointed out that two novel aminopyridine derivatives complexed with Cu2+, namely,cis-aquadichloro(N-[4-(hydroxyphenyl)methyl]-2-pyridinemethamino)copper (3a) and its glycosylated ligandcis-dichloro (N-{[4-(2,3,4,6-tetra-O-acetyl-β-D-glucopyranosyloxy)pheny]lmethyl}-2-pyridinemethamino)copper (3b), are effective againstT. cruzitrypomastigote forms. With this result in mind, the present work aimed to investigate the effects of both compounds on trypomastigotes physiology and on the interaction process with host cells. Apart from loss of plasma membrane integrity, an increased generation of reactive oxygen species (ROS) and decreased mitochondrial metabolism were observed. Pretreatment of trypomastigotes with these metallodrugs inhibited the association index with LLC-MK2cells in a typical dose-dependent manner. Both compounds showed low toxicity on mammalian cells (CC50> 100 µM), and the IC50values calculated for intracellular amastigotes were determined as 14.4 µM for3aand 27.1 µM for3b. This set of results demonstrates the potential of these aminopyridines complexed with Cu2+as promising candidates for further antitrypanosomal drug development.