In vitro evaluation of antiviral activity of single and combined repurposable drugs against SARS-CoV-2

Andrés Pizzorno, Blandine Padey, Julia Dubois, Thomas Julien, Aurélien Traversier, Victoria Dulière, Pauline Brun, Bruno Lina, Manuel Rosa-Calatrava, Olivier Terrier, Andrés Pizzorno, Blandine Padey, Julia Dubois, Thomas Julien, Aurélien Traversier, Victoria Dulière, Pauline Brun, Bruno Lina, Manuel Rosa-Calatrava, Olivier Terrier

Abstract

In response to the current pandemic caused by the novel SARS-CoV-2, identifying and validating effective therapeutic strategies is more than ever necessary. We evaluated the in vitro antiviral activities of a shortlist of compounds, known for their cellular broad-spectrum activities, together with drugs that are currently under evaluation in clinical trials for COVID-19 patients. We report the antiviral effect of remdesivir, lopinavir, chloroquine, umifenovir, berberine and cyclosporine A in Vero E6 cells model of SARS-CoV-2 infection, with estimated 50% inhibitory concentrations of 0.99, 5.2, 1.38, 3.5, 10.6 and 3 μM, respectively. Virus-directed plus host-directed drug combinations were also investigated. We report a strong antagonism between remdesivir and berberine, in contrast with remdesivir/diltiazem, for which we describe high levels of synergy, with mean Loewe synergy scores of 12 and peak values above 50. Combination of host-directed drugs with direct acting antivirals underscore further validation in more physiological models, yet they open up interesting avenues for the treatment of COVID-19.

Keywords: Antivirals; Berberine; COVID-19; Diltiazem; Drug combination; Remdesivir.

Copyright © 2020 The Author(s). Published by Elsevier B.V. All rights reserved.

Figures

Fig. 1
Fig. 1
Evaluation of antiviral activity of six drug candidates against SARS-CoV-2 in vitro. Vero E6 cells were infected by SARS-CoV-2 at a MOI of 0.01 and treated 1 h post infection (hpi) with serial dilutions of remdesivir, lopinavir, chloroquine, berberine, cyclosporine A and arbidol (umifenovir). Viral production was measured 48 hpi and expressed in relative values compared to the vehicle-treated control. The impact of treatment on cell viability was assessed by MTS assay (CellTiter 96® AQueous One Solution Cell Proliferation Assay, Promega), and expressed in relative values compared to control. Cell viability measurements were performed to ensure that molecular viral quantification was performed within a non-cytotoxic concentration range that could bias the measurement.
Fig. 2
Fig. 2
Evaluation of remdesivir-berberine combined treatment against SARS-CoV-2 in vitro. A. Vero E6 cells were infected by SARS-CoV-2 at a MOI of 0.01 and treated 1 hpi with serial dilutions of remdesivir in the presence of different fixed concentrations of berberine (A, left panel) or, alternatively, serial dilutions of berberine in the presence of different fixed concentrations of remdesivir (A, right panel). Viral production was measured 48 hpi and expressed in relative values compared to the vehicle-treated control. B. Dose-response percent inhibition matrix of single and combined remdesivir-berberine treatments C. Interaction landscape between remdesivir and berberine as calculated using the Loewe additive model. Areas with synergy scores of −10 or lower (green) represent zones of drug antagonism.
Fig. 3
Fig. 3
Synergistic effect of remdesivir-diltiazem combined treatment against SARS-CoV-2 in vitro. A. Vero E6 cells were infected by SARS-CoV-2 at a MOI of 0.01 and treated 1 hpi with serial dilutions of remdesivir in the presence of different fixed concentrations of diltiazem (A, left panel) or, alternatively, serial dilutions of diltiazem in the presence of different fixed concentrations of remdesivir (A, right panel). Viral production was measured 48 hpi and expressed in relative values compared to the vehicle-treated control. B. Dose-response percent inhibition matrix of single and combined remdesivir-diltiazem treatments C. Interaction landscape between remdesivir and diltiazem as calculated using the Loewe additive model. Areas with synergy scores of 10 or higher (red) represent zones of drug synergy.

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