Ensaios celulares baseados em replicon de Chikungunya para identificação de antivirais

Abstract

Chikungunya virus (CHIKV), which causes Chikungunya fever, is a arbovirus transmitted by the Aedes aegypti mosquito, which periodically causes epidemics around the world and can become a serious public health problem. However, there are no approved vaccines or drugs against CHIKV. So it's research to identify antivirals is fundamental, which can be carried out in a quickly and efficiently by high-throughput screening (HTS) assays, such as replicon-based assays. These are subgenomic systems in which genes encoding structural proteins are replaced by a reporter gene, allowing the identification of inhibitors of viral replication by measuring the activity of the reporter protein, in an easier and safer than traditional methods. In this study, the replicon strain was used. CHIKV reporter that contains the Gaussia luciferase (GLuc) gene as a reporter, called BHK-21-Rep-T7-GLuc-nSP-CHIKV-99650, for the screening of three MMV/DNDi small molecule libraries. Through tests based on replicon it was possible to identify 4 compounds, Itraconazole, GSK-983, Rubitecan and MMV676270, as inhibitors of CHIKV replication with low to moderate toxicity. Two of these molecules, the antifungal Itraconazole and the antiviral Rubitecan, were the most promising with selectivity indices >312.5 and >714.3, respectively, values considerably higher than those observed for the too much. In addition, the inhibition of replication was confirmed by the decrease in relative expression of the replicon RNA in cells treated with the concentrations effects of the 4 compounds. To better characterize the activity of these molecules against CHIKV, viral infection assays will be carried out in collaboration with Prof. Ana Carolina Jardim from the Federal University of Uberlândia (UFU).