CRYSTAL VIOLET CHEMICAL ANALOGUES INHIBIT THE Trypanosoma cruzi PROLINE TRANSPORTER TcAAAP069 AND EXERT TRYPANOCIDAL EFFECT.

MARTÍNEZ SAYÉ M; GAUNA L; REIGADA C; MIRANDA MR; PEREIRA CA

Congreso; Reunión Conjunta de Sociedades de Biociencias (XXIX Reunión de la Sociedad Argentina de Protozoología); 2017

Sociedad Argentina de Protozoología

Trypanosoma cruzi, the etiological agent of Chagas disease, has a metabolism largely based on the consumption of glucose and proline. Proline is also involved in differentiation processes, cellular invasion and resistanceto oxidative, metabolic and osmotic stress. We have previously identified the proline permease TcAAAP069 in T. cruzi and we showed the importance of this transporter on parasite survival. Crystal violet (CV) was used for several years as a blood additive for prevention of transfusion transmitted Chagas disease. Among the trypanocidal mechanisms proposed for CV, it was reported the protein synthesis inhibition related to inhibition of methionine and proline transport. The aim of this work was to validate CV as a proline transport inhibitor in order to use it as reference in a virtual screening protocol. CV entered at least in part through the proline permease TcAAAP069 and it also inhibited proline transport. Transgenic parasites overexpressing proline transporter(Tc069 parasites) were more sensitive to CV trypanocidal action (12.7μM for control parasites and 0.27μM for Tc069 parasites). The protective effect of proline in oxidative stress situations led to a 3-fold increase resistancefor Tc069 parasites when proline was added along with CV. In order to obtain other drugs that might have similar effects to CV, a similarity-based virtual screening was performed, using the CV as query and different drug databases of compounds approved for use in humans. So far, three of the obtained compounds proved to be effective as proline transport inhibitors that also had trypanocidal effect. Taken together, these results show that it is possible to identify potential trypanocidal compounds by virtual screening using CV as a reference. Drug repositioning is a recommended strategy by the World HealthOrganization to fight neglected diseases, like Chagas disease, since costs and development time are significantly reduced for its application in therapy.