Chagas disease, caused by Trypanosoma cruzi parasite, was described thousands of years ago. Currently, it affects millions of people, mostly in Latin America, and there are not suitable drugs for treating it. As an attempt to find appropriate drugs to deal with this problem, we report here on the design, synthesis, and characterization of 82 new compounds. Trypanosomicidal behavior in vitro showed more than 20 outstanding derivatives with anti-Trypanosoma cruzi activity. Furthermore, we studied the nonspecific toxicity against mammalian cells determining their selectivity and also performed mutagenicity studies. Proof of concept, in vivo studies, was conducted with two of the most promising derivatives (77 and 80). They were identified as candidates because they have (i) very simple and cost-effective syntheses; (ii) activity against different stages and strains of the parasite showing excellent in vivo behavior during the acute phase of Chagas disease; and (iii) neither nonspecific toxicity nor mutagenic activity.Iberoamerican Program for Science and Technology (CYTED)network RIDIMEDCHAGViceministerio de Ciencia y Tecnologia (El Salvador)Agenda Uruguaya de Cooperacion Internacional (Uruguay)ANII (Uruguay)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Programa de Apoio ao Desenvolvimento Cientifico (PADC-FCF-UNESP)Univ Republica, Grp Quim Med, Lab Quim Organ, Fac Ciencias,Fac Quim, Montevideo 11400, UruguayCtr Nacl Invest Cient El Salvador, San Salvador, El SalvadorUniv Nacl La Plata, Fac Ciencias Exactas, Dept Fis, RA-1900 La Plata, Buenos Aires, ArgentinaConsejo Nacl Invest Cient & Tecn, Inst IFLP, CCT La Plata, RA-1900 La Plata, Buenos Aires, ArgentinaUNESP, Sch Pharmaceut Sci, Dept Drugs & Pharmaceut, Araraquara, BrazilUniv Ind Santander, Dept Ciencias Basicas, Ctr Invest Enfermedades Tropicales, Bucaramanga, ColombiaUniv Nacl Asuncion, Inst Ciencias Salud, Dept Med Trop, Asuncion, ParaguayUNESP, Sch Pharmaceut Sci, Dept Drugs & Pharmaceut, Araraquara, Brazil