Evaluation of broad-spectrum antiviral compounds against chikungunya infection using a phenotypic screening strategy

Abstract

Chikungunya fever is an emerging disease and a significant public health problem in tropical countries. Recently reported outbreaks in Brazil in 2015 drew attention to the need to develop prevention and treatment options, as no antiviral chemotherapy or vaccines are currently available for this disease. Two strategies have been proved to accelerate the discovery of new anti-infectives: phenotypic screening and drug repurposing. Phenotypic screening can support the fast interrogation of compounds without the need for a pre-validated drug target, which is not available for the chikungunya virus (CHIKV) and has the additional advantage of facilitating the discovery of antiviral with novel mechanism of action. Drug repurposing can save time and resources in drug development by enabling secondary uses for drugs that are already approved for human treatment, thus precluding the need for several of the mandatory preclinical and clinical studies necessary for drug approval. A phenotypic screening assay was developed by infecting the human hepatoma Huh-7 cells with CHIKV 181/25 and quantifying infection through indirect immunofluorescence. The compound 6-azauridine was used as a positive control drug. The screening assay was validated by testing a commercial library of 1,280 compounds, including FDA-approved drugs, and used to screen a panel of broad-spectrum antiviral compounds for anti-CHIKV activity.  A high content assay was set up in Huh-7 cells-infected with CHIKV. The maximum rate of infection peaked at 48 hours post-infection, after which the host cell number was greatly reduced due to a strong cytopathic effect. Assay robustness was confirmed with Z’-factor values >0.8 and high correlation coefficient between independent runs, demonstrating that the assay is reliable, consistent and reproducible. Among tested compounds, sofosbuvir, an anti-hepatitis C virus drug, exhibited good selectivity against CHIKV with an EC50of 11 µM, suggesting it is a promising candidate for repurposing.