It was recently demonstrated that 5-(5-nitrofuran-2-yl)-1, 3, 4-thiadiazoles with piperazinyl-linked benzamidine substituents are effective in vitro against Leishmania major. Following on this evidence, we used colorimetric assay of acid phosphatase activity in the promastigotes as an indicator for cell viability. Also we studied the effect of these compounds on induction of nitric oxide (NO) in macrophage and production of reactive oxygen species (ROS) in lymphocyte that have important role in activation of immune response against Leishmania and elimination of parasite. Results showed that these compounds decrease the viability of the parasite and increase ROS and NO production in lymphocyte and macrophage respectively. These compounds can induce parasite killing, directly by decreasing the parasite viability and indirectly by exhibiting a significant increase on immune system.