Crystal violet chemical analogues inhibit the Trypanosoma cruzi proline transporter TcAAAP069 and exert trypanocidal effect.

SAYE, MELISA; GAUNA, LUCRECIA; REIGADA, CHANTAL; MIRANDA, MARIANA; PEREIRA, CA

CABA

Congreso; XXIX Reunión Anual de la Sociedad Argentina de Protozoología (SAP).; 2017

Sociedad Argentina de Protozoología (SAP)

Trypanosoma cruzi, the etiological agent of Chagas disease,has a metabolism largely based on the consumptionof glucose and proline. Proline is also involved in differentiationprocesses, cellular invasion and resistanceto oxidative, metabolic and osmotic stress. We havepreviously identified the proline permease TcAAAP069in T. cruzi and we showed the importance of this transporteron parasite survival. Crystal violet (CV) was usedfor several years as a blood additive for prevention oftransfusion transmitted Chagas disease. Among the trypanocidalmechanisms proposed for CV, it was reportedthe protein synthesis inhibition related to inhibition ofmethionine and proline transport. The aim of this workwas to validate CV as a proline transport inhibitor in orderto use it as reference in a virtual screening protocol.CV entered at least in part through the proline permeaseTcAAAP069 and it also inhibited proline transport.Transgenic parasites overexpressing proline transporter(Tc069 parasites) were more sensitive to CV trypanocidalaction (12.7μM for control parasites and 0.27μMfor Tc069 parasites). The protective effect of proline inoxidative stress situations led to a 3-fold increase resistancefor Tc069 parasites when proline was added alongwith CV. In order to obtain other drugs that might havesimilar effects to CV, a similarity-based virtual screeningwas performed, using the CV as query and different drugdatabases of compounds approved for use in humans.So far, three of the obtained compounds proved to beeffective as proline transport inhibitors that also had trypanocidaleffect. Taken together, these results show thatit is possible to identify potential trypanocidal compoundsby virtual screening using CV as a reference. Drug repositioningis a recommended strategy by the World HealthOrganization to fight neglected diseases, like Chagasdisease, since costs and development time are significantlyreduced for its application in therapy.