The spectrum and efficacy of available antileishmanial drugs is limited. In the present work we evaluated in vitro the antiproliferative activity of 11 compounds based on tetradentate polyamines compounds against three species (, and ) and the possible mechanism of action. We identified six compounds (, , , , and ) effective against all three spp both on extracellular and intracellular forms. These six most active leishmanicidal compounds also prevent the infection of host cells. Nevertheless, only compound is targeted against the SOD. Meanwhile, on the glucose metabolism the tested compounds have a species-specific effect on spp.: was affected mainly by and , by , and by and . Finally, the cellular ultrastructure was mainly damaged by in the three spp. studied. These identified antileishmania candidates constitute a good alternative treatment and will be further studied.