tetano
Editor, Senior Moderator
iScience
. 2023 Jan 7;105944.
doi: 10.1016/j.isci.2023.105944. Online ahead of print.
Identification of novel antiviral drug candidates using an optimized SARS-CoV-2 phenotypic screening platform
Denisa Bojkova[SUP] 1 [/SUP], Philipp Reus[SUP] 1 2 [/SUP], Leona Panosch[SUP] 1 [/SUP], Marco Bechtel[SUP] 1 [/SUP], Tamara Rothenburger[SUP] 1 [/SUP], Joshua D Kandler[SUP] 1 [/SUP], Annika Pfeiffer[SUP] 1 [/SUP], Julian U G Wagner[SUP] 3 4 5 [/SUP], Mariana Shumliakivska[SUP] 3 [/SUP], Stefanie Dimmeler[SUP] 3 4 5 [/SUP], Ruth Olmer[SUP] 6 [/SUP], Ulrich Martin[SUP] 6 [/SUP], Florian W R Vondran[SUP] 7 8 [/SUP], Tuna Toptan[SUP] 1 [/SUP], Florian Rothweiler[SUP] 1 9 [/SUP], Richard Zehner[SUP] 10 [/SUP], Holger F Rabenau[SUP] 1 [/SUP], Karen L Osman[SUP] 11 [/SUP], Steven T Pullan[SUP] 11 [/SUP], Miles W Carroll[SUP] 12 13 [/SUP], Richard Stack[SUP] 14 [/SUP], Sandra Ciesek[SUP] 1 2 8 [/SUP], Mark N Wass[SUP] 14 [/SUP], Martin Michaelis[SUP] 14 [/SUP], Jindrich Cinatl Jr[SUP] 1 9 [/SUP]
Affiliations
Abstract
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 read-out 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 1796 kinase inhibitors identified known and novel antiviral drug candidates including inhibitors of PHGDH, CLK-1, and CSF1R. The activity of the PHGDH inhibitor NCT-503 was further increased in combination with the 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.
Keywords: COVID-19; Caco-2-F03; SARS-CoV-2; antiviral therapy; drug discovery.
. 2023 Jan 7;105944.
doi: 10.1016/j.isci.2023.105944. Online ahead of print.
Identification of novel antiviral drug candidates using an optimized SARS-CoV-2 phenotypic screening platform
Denisa Bojkova[SUP] 1 [/SUP], Philipp Reus[SUP] 1 2 [/SUP], Leona Panosch[SUP] 1 [/SUP], Marco Bechtel[SUP] 1 [/SUP], Tamara Rothenburger[SUP] 1 [/SUP], Joshua D Kandler[SUP] 1 [/SUP], Annika Pfeiffer[SUP] 1 [/SUP], Julian U G Wagner[SUP] 3 4 5 [/SUP], Mariana Shumliakivska[SUP] 3 [/SUP], Stefanie Dimmeler[SUP] 3 4 5 [/SUP], Ruth Olmer[SUP] 6 [/SUP], Ulrich Martin[SUP] 6 [/SUP], Florian W R Vondran[SUP] 7 8 [/SUP], Tuna Toptan[SUP] 1 [/SUP], Florian Rothweiler[SUP] 1 9 [/SUP], Richard Zehner[SUP] 10 [/SUP], Holger F Rabenau[SUP] 1 [/SUP], Karen L Osman[SUP] 11 [/SUP], Steven T Pullan[SUP] 11 [/SUP], Miles W Carroll[SUP] 12 13 [/SUP], Richard Stack[SUP] 14 [/SUP], Sandra Ciesek[SUP] 1 2 8 [/SUP], Mark N Wass[SUP] 14 [/SUP], Martin Michaelis[SUP] 14 [/SUP], Jindrich Cinatl Jr[SUP] 1 9 [/SUP]
Affiliations
- PMID: 36644320
- PMCID: PMC9822553
- DOI: 10.1016/j.isci.2023.105944
Abstract
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 read-out 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 1796 kinase inhibitors identified known and novel antiviral drug candidates including inhibitors of PHGDH, CLK-1, and CSF1R. The activity of the PHGDH inhibitor NCT-503 was further increased in combination with the 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.
Keywords: COVID-19; Caco-2-F03; SARS-CoV-2; antiviral therapy; drug discovery.