BACKGROUND: The challenge in anti-malarial chemotherapy is based on the 
emergence of resistance to drugs and the search for medicines against all stages 
of the life cycle of Plasmodium spp. as a therapeutic target. Nowadays, many 
molecules with anti-malarial activity are reported. However, few studies about 
the cellular and molecular mechanisms to understand their mode of action have 
been explored. Recently, new primaquine-based hybrids as new molecules with 
potential multi-acting anti-malarial activity were reported and two hybrids of 
primaquine linked to quinoxaline 1,4-di-N-oxide (PQ-QdNO) were identified as the 
most active against erythrocytic, exoerythrocytic and sporogonic stages.
METHODS: To further understand the anti-malarial mode of action (MA) of these 
hybrids, hepg2-CD81 were infected with Plasmodium yoelii 17XNL and treated with 
PQ-QdNO hybrids during 48 h. After were evaluated the production of ROS, the 
mitochondrial depolarization, the total glutathione content, the DNA damage and 
proteins related to oxidative stress and death cell.
RESULTS: In a preliminary analysis as tissue schizonticidals, these hybrids 
showed a mode of action dependent on peroxides production, but independent of 
the activation of transcription factor p53, mitochondrial depolarization and 
arrest cell cycle.
CONCLUSIONS: Primaquine-quinoxaline 1,4-di-N-oxide hybrids exert their 
antiplasmodial activity in the exoerythrocytic phase by generating high levels 
of oxidative stress which promotes the increase of total glutathione levels, 
through oxidation stress sensor protein DJ-1. In addition, the role of HIF1a in 
the mode of action of quinoxaline 1,4-di-N-oxide is independent of biological 
activity.