In vitro evaluation of the activity of teriflunomide against SARS-CoV-2 and the human coronaviruses 229E and OC43

Biochem Biophys Rep. 2023 Mar:33:101395. doi: 10.1016/j.bbrep.2022.101395. Epub 2022 Nov 23.

Abstract

Previous data have suggested an antiviral effect of teriflunomide, including against severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2), the agent underlying the ongoing COVID-19 pandemic. We undertook an in vitro investigation to evaluate the inhibitory activity of teriflunomide against SARS-CoV-2 in a cell-based assay. Teriflunomide was added to Vero (kidney epithelial) cells that had been infected with SARS-CoV-2. A nucleocapsid immunofluorescence assay was performed to examine viral inhibition with teriflunomide and any potential cytotoxic effect. The 50% effective concentration (EC50) for teriflunomide against SARS-CoV-2 was 15.22 μM. No cytotoxicity was evident for teriflunomide in the Vero cells (i.e., the 50% cytotoxic concentration [CC50] was greater than the highest test concentration of 100 μM). The data were supported by additional experiments using other coronaviruses and human cell lines. In the SARS-CoV-2-infected Vero cells, the prodrug leflunomide had an EC50 of 16.49 μM and a CC50 of 54.80 μM. Our finding of teriflunomide-mediated inhibition of SARS-CoV-2 infection at double-digit micromolar potency adds to a growing body of evidence for a broad-ranging antiviral effect of teriflunomide.

Keywords: Antiviral; CC50, 50% cytotoxic concentration; COVID-19; COVID-19, coronavirus disease 2019; CPE, cytopathic effect; Coronavirus; EC50, 50% effective concentration; HCoV, human coronavirus; Human cell line; MS, multiple sclerosis; SARS-CoV-2; SARS-CoV-2, severe acute respiratory syndrome coronavirus-2; TCID50, 50% tissue culture infective dose; Teriflunomide; qPCR, quantitative real-time polymerase chain reaction.