4-Amino Bis-Pyridinium Derivatives as Novel Antileishmanial Agents

Abstract

ABSTRACTThe antileishmanial activity of a series of bis-pyridinium derivatives that are analogues of pentamidine have been investigated, and all compounds assayed were found to display activity against promastigotes and intracellular amastigotes ofLeishmania donovaniandLeishmania major, with 50% effective concentrations (EC50s) lower than 1 μM in most cases. The majority of compounds showed similar behavior in bothLeishmaniaspecies, being slightly more active againstL. majoramastigotes. However, compound VGP-106 {1,1′-(biphenyl-4,4′-diylmethylene)bis[4-(4-bromo-N-methylanilino)pyridinium] dibromide} exhibited significantly higher activity againstL. donovaniamastigotes (EC50, 0.86 ± 0.46 μM) with a lower toxicity in THP-1 cells (EC50, 206.54 ± 9.89 μM). As such, VGP-106 was chosen as a representative compound to further elucidate the mode of action of this family of inhibitors in promastigote forms ofL. donovani. We have determined that uptake of VGP-106 inLeishmaniais a temperature-independent process, suggesting that the compound crosses the parasite membrane by diffusion. Transmission electron microscopy analysis showed a severe mitochondrial swelling in parasites treated with compound VGP-106, which induces hyperpolarization of the mitochondrial membrane potential and a significant decrease of intracellular free ATP levels due to the inhibition of ATP synthesis. Additionally, we have confirmed that VGP-106 induces mitochondrial ROS production and an increase in intracellular Ca2+levels. All these molecular events can activate the apoptotic process inLeishmania; however, propidium iodide assays gave no indication of DNA fragmentation. These results underline the potency of compound VGP-106, which may represent a new avenue for the development of novel antileishmanial compounds.