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. 2020 Jul 7;32(3):107940. doi: 10.1016/j.celrep.2020.107940

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© 2020 The Authors

Since January 2020 Elsevier has created a COVID-19 resource centre with free information in English and Mandarin on the novel coronavirus COVID-19. The COVID-19 resource centre is hosted on Elsevier Connect, the company's public news and information website. Elsevier hereby grants permission to make all its COVID-19-related research that is available on the COVID-19 resource centre - including this research content - immediately available in PubMed Central and other publicly funded repositories, such as the WHO COVID database with rights for unrestricted research re-use and analyses in any form or by any means with acknowledgement of the original source. These permissions are granted for free by Elsevier for as long as the COVID-19 resource centre remains active.

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The Prodrug Remdesivir (RDV) and Parent Nucleoside GS-441524 (’524) Potently Inhibit SARS-CoV-2 Replication

(A) Vero E6, Vero CCL-81 (Vero), Huh7, and Calu3 2B4 cells were infected with an MOI of 0.01 and/or 0.1 PFU/cell SARS-CoV-2 (2019-nCoV/USA-WA1/2020), and infectious viral titers were determined by plaque assay 0.5, 24, 48, and 72 h post-infection (hpi). Viral yields were calculated by subtracting the average 0.5 h (post-adsorption, pre-incubation) titer from each subsequent time point. Data represent the average of three replicates from one experiment. Error bars indicate SD. B.D., below detection. Calu3 2B4 cells were infected with 0.1 PFU/cell SARS-CoV-2, and Vero cells were infected with 0.01 PFU/cell SARS-CoV-2 and treated with RDV, GS-441524 (’524), or DMSO only (control) in cell culture medium. Supernatants were collected 48 hpi (Vero E6 cells) or 72 hpi (Calu3 2B4 cells).

(B and C) Reduction of SARS-CoV-2 replication by RDV in Calu3 2B4 cells, as determined by infectious viral titer (B) and qRT-PCR (C).

(D) Percent inhibition of SARS-CoV-2 replication by RDV and GS-441524 in Calu3 2B4 cells, as determined by infectious viral titer (RDV: EC₅₀ = 0.28 μM, EC₉₀ = 2.48 μM; GS-441524: EC₅₀ = 0.62 μM, EC₉₀ = 1.34 μM). No significant cytotoxicity of either compound was detected in Calu3 cells.

(E) Percent inhibition of SARS-CoV-2 replication by RDV and GS-441524 in Calu3 2B4 cells, as determined by qRT-qPCR (RDV: EC₅₀ = 0.60 μM, EC₉₀ = 1.28 μM; GS-441524: EC₅₀ = 1.09 μM, EC₉₀ = 1.37 μM).

(F and G) Reduction of SARS-CoV-2 replication by RDV in Vero E6 cells, as determined by infectious viral titer (F) and qRT-PCR (G).

(H) Percent inhibition of SARS-CoV-2 replication by RDV and GS-441524 in Vero E6 cells, as determined by infectious viral titer (RDV: EC₅₀ = 1.65 μM, EC₉₀ = 2.40 μM; GS-441524: EC₅₀ = 0.47 μM, EC₉₀ = 0.71 μM). No significant cytotoxicity of either compound was detected in Vero E6 cells.

(I) Percent inhibition of SARS-CoV-2 replication by RDV and GS-441524 in Vero E6 cells, as determined by qRT-PCR (RDV: EC₅₀ = 1.49 μM, EC₉₀ = 3.03 μM; GS-441524: EC₅₀ = 0.47 μM, EC₉₀ = 0.80 μM).

Data represent means of 2–4 independent experiments with 2–3 replicates each. Error bars represent SEM. See also Figure S5.