Test-system in vitro for screening of therapeutic drugs with IL-17A inhibitory activity

Abstract

To achieve greater clinical relevance of the newly discovered compounds, modern drug discovery requires disease-targeted assays based on human cells. The specific aim of this study was to design and develop a new cell-based assay for screening of compounds with IL-17A inhibitory activity. Human foreskin fibroblasts (HFF) were treated with IL-17A alone (experimental conditions I) or a mixture of IL-17A inhibitor netakimab and IL-17A (experimental conditions II). IL-17A - dependent production of inflammatory mediators IL-6, IL-8, MCP-1 was evaluated by ELISA (enzyme-linked immunosorbent assay). The study demonstrated the ability of HFF subcultured in vitro for a long time (>20 passages) to respond to IL-17A treatment by increased production of inflammatory cytokines IL-6, IL-8, MCP-1. Neutralization of IL-17A by netakimab (IL-17A inhibitor) resulted in a dose-dependent decrease of inflammatory cytokines production into cell growth medium. Thus, a new cell-based assay to evaluate the biological activity of Il-17A inhibitors has been developed and tested. The assay is based on the analysis of IL-17A-dependent production of inflammatory cytokines synthesized by human dermal fibroblasts. Netakimab has been shown to be a highly potent inhibitor of IL-17A.