 Celso Vataru Nakamura cvnakamura@uem.br 1 Programa de Pós-graduação em Ciências Biológicas, Laboratório de Inovação Tecnológica no Desenvolvimento de Fármacos e Cosméticos, Universidade Estadual de Maringá, Bloco B-08, Av. Colombo 5790, Maringá, PR CEP 87020-900, Brazil 2 LaBioMMi, Departamento de Química, Universidade Federal de São Carlos, CP 676, São Carlos, SP 13.565-905, Brazil 3 CERMAV, CNRS, Université Grenoble Alpes, 38000 Grenoble, France © Springer Science +Business Media New York 2016 A3K2A3-induced apoptotic cell death of occurs through caspase- and ATP-dependent mitochondrial dysfunction Francielle Pelegrin Garcia 1 · Jean Henrique da Silva Rodrigues 1 · Zia Ud Din 2 · Edson Rodrigues-Filho 2 · Tânia Ueda-Nakamura 1 · Rachel Auzély-Velty 3 · Celso Vataru Nakamura 1 Apoptosis DOI 10.1007/s10495-016-1308-4 triphosphate ratio, phosphatidylserine exposure, a decrea in cell volume, caspase production, and DNA fragmenta - tion. Altogether, these fndings indicate that apoptosis can indeed be triggered by chemotherapeutic agents. Keywords  · Dibenzylideneacetone · Mitochondria · Cell death · Apoptosis Introduction Leishmaniasis remains endemic in several parts of the world and is a serious health problem in numerous devel - oping countries. It occurs as a complex and clinicall diverse illness that is caused by protozoan species that are transmitted through the bite of sandfies. Despite recent advances, the treatment of leishmaniasis still problematic because of the high toxicity and adverse side effects of therapeutic drugs. The need to identify new molecular targets to improve therapy is clearly justifed. Target-identifcation and mechanism-of-action studies play important roles in drug discovery. The putative targ should be either absent in the host or markedly different from the host homolog so that it can be exploited as a dru target [ 1]. Interestingly, programmed cell death in protists appea to share some morphological features with apoptosis in multicellular organisms, including cell shrinkage, the loss of mitochondrial membrane potential, and the externali - tion of phosphatidylserine [ 2, 3]. A better understanding of the mechanistic machinery of apoptosis-like programmed cell death in protists would thus prove immensely benefcial in the design of rational chemotherapeutic interventions i target-dependent manner [ 4]. Abstract Leishmaniasis is a neglected tropical dis - ease that affects millions of people worldwide. Current therapies mainly rely on antimonial drugs that are inad - equate because of their high toxicity and increased drug resistance. An urgent need exists to discover new, more effective, more affordable, and more target-specifc drugs. Pathways that are associated with apoptosis-like cell death have been identifed in unicellular eukaryotes, includ- ing protozoan parasites. In the present study, we studied the mechanism of cell death that is induced by A3K2A3 against  . A3K2A3 is a dibenzylideneac - etone that has an acyclic dienone that is attached to aryl groups in both β-positions, which is similar to curcumi- noids and chalcone structures. This compound was pre - viously shown to be safe with regard to cytotoxicity and active against the parasite. Biochemical and morphologi - cal approaches were used in the present study. The results suggested that A3K2A3 caused mitochondrial dysfunction in  promastigotes, leading to mechanisms of cell death that share some common phenotypic features with metazoan apoptosis, such as an increase in reactive oxygen species production, a decrease in the adenosine 1 3