CONICET | Buscador de Institutos y Recursos Humanos

Chagas disease is transmitted by the Trypanosoma cruzi kinetoplastid parasite, which possess a complex life cycle alternating between an invertebrate and a vertebrate hosts. Looking to identify new chemical entities, N,N?-substituted diamines have been proposed as an interesting scaffold for antiparasite drug development. Previously, we have prepared a collection of derivatives with that scaffold. The incorporation of 4-benzyloxy group onto the benzyl group resulted in a noticeable enhancement of the growth inhibitory activity on CL Brener T. cruzi epimastigotes. The compounds showed negligible cytotoxicity towards Vero cells. In this work we compared the activity on Dm28c strain with the seven most active analogues previously reported. The best candidate was N1,N6-bis(4-(benzyloxy)benzyl)hexane-1,6-diamine (AR-Tc-3) with an IC50 of 0,99 µM. Looking to elucidate the action mechanism, metabolomic study was performed tracing secreted metabolites variations on parasites under 1 µM AR-Tc-3 treatment. Parasites (0.5 x 106 epimastigotes/mL) were incubated over 96 h in DMEM-high glucose supplemented with 5 µM hemin and 2 % fetal bovine serum (FBS) with or without the compound. The conditioned media were separated from epimastigotes by centrifugation and the supernatants were filtered through 0.22 µm acetate membranes. These samples were analysed by 1H NMR adding deutered DMSO as internal standard and deutered water to lock the sample. The metabolites were unequivocally identified by spiking the culture medium with the pure metabolites to properly determine its chemical shift in these experimental conditions. We observed a three-time fold decrease in the amount of acetate and alanine released to the medium by AR-Tc-3 treated epimastigotes. No changes in ethanol and lactate excretions were observed. Conclusion: N.N?-substituted diamines, are promising candidates to develop new anti-T. cruzi chemotherapies and AR-Tc3 affects the energy metabolism.