Identification and evaluation of compounds with antiviral proprieties against chikungunya infection using a phenotypic screening strategy

Author(s)

Rafaela Milan Bonotto

• Document type: Doctoral Thesis
• Institution: Universidade Estadual de Campinas (UNICAMP). Instituto de Biologia
• Defense date: 2019-04-29

Examining board members

Laura Helena Vega Gonzales Gil; José Luiz Proença Módena; Edison Luiz Durigon; Juliana Velasco de Castro Oliveira

• Advisor: Holanda Gondim de Freitas Junior

Abstract

Chikungunya fever is an emerging disease caused by Chikungunya virus, which is associated with significant public health problem in tropical countries. Recently reported outbreaks in Brazil in 2016 drew attention to the need to develop prevention and treatment options, as no antiviral chemotherapy or vaccines are currently available for this disease. Thereby, to address drug discovery needs, the present work proposes the development a phenotypic screening strategy for CHIKV by screening a collection of compounds pharmacologically activity compounds, including FDA-approval drugs, and a panel of broad-spectrum antiviral compounds. The assay was based a high content screening methodology (HCS), which was developed by infecting the human hepatoma Huh-7 cells with CHIKV 181/25 and quantifying infection through indirect immunofluorescence. Assay robustness was confirmed with Z¿-factor values >0.8 , coefficient of variation < 10% and high correlation coefficient between independent runs with r of 0.9 , demonstrating that the assay is reliable, consistent and reproducible. In total were selected 4 compounds with promising activity were selected to conduct complementary in vitro assay. The compounds pentamidine isethionate, brequinar sodium, dequalinium chloride and sofosbuvir showed activity dependent on concentration in different CHIKV strains, with high selective indexes in Huh 7 cell line. Moreover, all compound treatment significantly reduced CHIKV infectivity titer in vitro showing a promising antiviral candidates. Thus, further evolution of pentamidine isethionate and dequalinium chloride point they might act an early CHIKV stage viral infection cycle, such as binding, internalization and initial replication phase. Here we presented a robust and reliable HCS phenotypic method to screening compounds against CHIKV which provide to identified and characterized promising compounds candidate as starting point therapeutic development against CHIKV (AU)

• FAPESP's process: 16/03780-5 - A cell-based high throughput screening strategy for Chikungunya drug discovery
• Grantee: Rafaela Milan Bonotto
• Support Opportunities: Scholarships in Brazil - Doctorate (Direct)