학술논문
Identification of novel antiviral drug candidates using an optimized SARS-CoV-2 phenotypic screening platform
Document Type
article
Author
Denisa Bojkova; Philipp Reus; Leona Panosch; Marco Bechtel; Tamara Rothenburger; Joshua D. Kandler; Annika Pfeiffer; Julian U.G. Wagner; Mariana Shumliakivska; Stefanie Dimmeler; Ruth Olmer; Ulrich Martin; Florian W.R. Vondran; Tuna Toptan; Florian Rothweiler; Richard Zehner; Holger F. Rabenau; Karen L. Osman; Steven T. Pullan; Miles W. Carroll; Richard Stack; Sandra Ciesek; Mark N. Wass; Martin Michaelis; Jindrich Cinatl, Jr.
Source
iScience, Vol 26, Iss 2, Pp 105944- (2023)
Subject
Language
English
ISSN
2589-0042
Abstract
Summary: Reliable, easy-to-handle phenotypic screening platforms are needed for the identification of anti-SARS-CoV-2 compounds. Here, we present caspase 3/7 activity as a readout for monitoring the replication of SARS-CoV-2 isolates from different variants, including a remdesivir-resistant strain, and of other coronaviruses in numerous cell culture models, independently of cytopathogenic effect formation. Compared to other models, the Caco-2 subline Caco-2-F03 displayed superior performance. It possesses a stable SARS-CoV-2 susceptibility phenotype and does not produce false-positive hits due to drug-induced phospholipidosis. A proof-of-concept screen of 1,796 kinase inhibitors identified known and novel antiviral drug candidates including inhibitors of phosphoglycerate dehydrogenase (PHGDH), CDC like kinase 1 (CLK-1), and colony stimulating factor 1 receptor (CSF1R). The activity of the PHGDH inhibitor NCT-503 was further increased in combination with the hexokinase II (HK2) inhibitor 2-deoxy-D-glucose, which is in clinical development for COVID-19. In conclusion, caspase 3/7 activity detection in SARS-CoV-2-infected Caco-2-F03 cells provides a simple phenotypic high-throughput screening platform for SARS-CoV-2 drug candidates that reduces false-positive hits.