Ponente
Descripción
Chagas disease or American Trypanosomosis is caused by the protozoan parasite Trypanosoma cruzi.The current chemotherapeutics is restricted to only two drugs, benznidazole and nifurtimox, both sharing low efficacy and serious side effects. The major leucyl aminopeptidase of Trypanosoma cruzi (LAPTc) is an attractive target. Herein we identify a LAPTc´s inhibitor with selectivity toward human aminopeptidase (LAP3). Besides, the LAPTc inhibitor identified in this work showed in vitro inhibition of the intracellular amastigote form using Vero cell as host. Firstly a LAPTc recombinant variant (rLAPTc) was obtained in the bacterium Escherichia coli BL21(DE3) pLysS. The enzyme was purified from the bacterial soluble extract by immobilized metallic ion affinity and gel filtration chromatography, until 92 % purity. Secondly, a high-throughput screening for rLAPTc´s inhibition was performed using RapidFire-Mass Spectrometry enzymatic assay. The high throughput screening revealed a competitive rLAPTc inhibitor, with a Ki value of 0.27 M. The competitive inhibition and selective enzymatic activity was explained by bioinformatics methods.These properties allow considering compound 4 as hit structure against Chagas disease.
