A high throughput screening assay for inhibitors of SARS-CoV-2 pseudotyped particle entry

• DOI: 10.1016/j.slasd.2021.12.005
• Publication Date: 2022-01-25T05:28:29Z
• Authors: Miao Xu; Manisha Pradhan; Kirill Gorshkov; Jennifer D. Petersen; Min Shen; Hui Guo; Wei Zhu; Carleen Klumpp-Th...; Sam Michael; Misha Itkin; Zina Itkin; Marco R. Straus; Joshua Zimmerberg; Wei Zheng; Gary R. Whittaker; Catherine Z. Chen
• Keywords: High-Throughput Screening; HEK 293 cells

Effective small molecule therapies to combat the SARS-CoV-2 infection are still lacking as COVID-19 pandemic continues globally. High throughput screening assays needed for lead discovery and optimization of inhibitors. In this work, we have applied viral pseudotyping establish a cell-based entry assay. Here, pseudotyped particles (PP) contain spike in membrane enveloping both murine leukemia virus (MLV) gag-pol polyprotein luciferase reporter RNA. Upon addition PP HEK293-ACE2 cells, protein binds ACE2 receptor on cell surface, resulting priming by host proteases trigger endocytosis these particles, fusion between particle envelope membrane. The internalized gene is then expressed luminescent readout surrogate spike-mediated into cells. This assay can be executed biosafety level 2 containment lab high screening. From collection 5,158 approved drugs drug candidates, our efforts identified 7 active compounds that inhibited SARS-CoV-2-S entry. Of seven, six were against live replicating cytopathic effect Our results demonstrated utility development inhibitors well mechanistic study anti-SARS-CoV-2 compounds. Additionally, with proteins from B.1.1.7 B.1.351 variants prepared used evaluate therapeutic effects