Antiprotozoal Activity of Benzoylthiourea Derivatives against Trypanosoma cruzi: Insights into Mechanism of Action Lopes Pereira, Patrícia Morais Carreira de Paula, Jessica Autophagy-dependent pathway Cell death Chagas disease Epimastigote forms Mechanism of action Ultrastructural changes For decades, only two nitroheterocyclic drugs have been used as therapeutic agents for Chagas disease. However, these drugs present limited effectiveness during the chronic phase, possess unfavorable pharmacokinetic properties, and induce severe adverse effects, resulting in low treatment adherence. A previous study reported that N-(cyclohexylcarbamothioyl) benzamide (BTU-1), N-(tertbutylcarbamothioyl) benzamide (BTU-2), and (4-bromo-N-(3-nitrophenyl) carbamothioyl benzamide (BTU-3) present selective antiprotozoal activity against all developmental forms of Trypanosoma cruzi Y strain. In this study, we investigated the mechanism of action of these compounds through microscopy and biochemical analyses. Transmission electron microscopy analysis showed nuclear disorganization, changes in the plasma membrane with the appearance of blebs and extracellular arrangements, intense vacuolization, mitochondrial swelling, and formation of myelin-like structures. Biochemical results showed changes in the mitochondrial membrane potential, reactive oxygen species content, lipid peroxidation, and plasma membrane fluidity. In addition, the formation of autophagic vacuoles was observed. These findings indicate that BTU-1, BTU-2, and BTU-3 induced profound morphological, ultrastructural, and biochemical alterations in epimastigote forms, triggering an autophagic-dependent cell death pathway 2023-10-17T08:40:18Z 2023-10-17T08:40:18Z 2023-08-03 journal article Pereira, P.M.L.; Fernandes, B.T.; dos Santos, V.R.; Cabral,W.R.C.; Lovo-Martins, M.I.; Alonso, L.; Lancheros, C.A.C.; de Paula, J.C.; Camargo, P.G.; Suzukawa, H.T.; et al. Antiprotozoal Activity of Benzoylthiourea Derivatives against Trypanosoma cruzi: Insights into Mechanism of Action. Pathogens 2023, 12, 1012. [https://doi.org/10.3390/ pathogens12081012] https://hdl.handle.net/10481/85043 10.3390/ pathogens12081012 eng http://creativecommons.org/licenses/by/4.0/ open access Atribución 4.0 Internacional MDPI