An orally bioavailable broad-spectrum antiviral inhibits SARS-CoV-2 and multiple endemic, epidemic and bat coronavirus

  1. Timothy P. Sheahan
  2. Amy C. Sims
  3. Shuntai Zhou
  4. Rachel L. Graham
  5. Collin S. Hill
  6. Sarah R. Leist
  7. Alexandra Schäfer
  8. Kenneth H. Dinnon
  9. Stephanie A. Montgomery
  10. Maria L. Agostini
  11. Andrea J. Pruijssers
  12. James D. Chapell
  13. Ariane J. Brown
  14. Gregory R. Bluemling
  15. Michael G. Natchus
  16. Manohar Saindane
  17. Alexander A. Kolykhalov
  18. George Painter
  19. Jennifer Harcourt
  20. Azaibi Tamin
  21. Natalie J. Thornburg
  22. Ronald Swanstrom
  23. Mark R. Denison
  24. Ralph S. Baric

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Abstract

Coronaviruses (CoVs) traffic frequently between species resulting in novel disease outbreaks, most recently exemplified by the newly emerged SARS-CoV-2. Herein, we show that the ribonucleoside analog β-D-N 4 -hydroxycytidine (NHC, EIDD-1931) has broad spectrum antiviral activity against SARS-CoV 2, MERS-CoV, SARS-CoV, and related zoonotic group 2b or 2c Bat-CoVs, as well as increased potency against a coronavirus bearing resistance mutations to another nucleoside analog inhibitor. In mice infected with SARS-CoV or MERS-CoV, both prophylactic and therapeutic administration of EIDD-2801, an orally bioavailable NHC-prodrug (b-D-N 4 -hydroxycytidine-5’-isopropyl ester), improved pulmonary function, and reduced virus titer and body weight loss. Decreased MERS-CoV yields in vitro and in vivo were associated with increased transition mutation frequency in viral but not host cell RNA, supporting a mechanism of lethal mutagenesis. The potency of NHC/EIDD-2801 against multiple coronaviruses, its therapeutic efficacy, and oral bioavailability in vivo , all highlight its potential utility as an effective antiviral against SARS-CoV-2 and other future zoonotic coronaviruses.

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  1. ScreenIT

    SciScore for 10.1101/2020.03.19.997890: (What is this?)

    Please note, not all rigor criteria are appropriate for all manuscripts.

    Table 1: Rigor

    NIH rigor criteria are not applicable to paper type.

    Table 2: Resources

    Experimental Models: Cell Lines
    SentencesResources
    To measure cell viability to determine if there was any NHC induced cytotoxicity, Calu3 2B4 cells were plated and treated with NHC only as described above.
    Calu3 2B4
    suggested: RRID:CVCL_YZ47)
    Briefly, 500,000 Vero E6 cells/well were seeded in 6-well plates.
    Vero E6
    suggested: None
    Third, to titer lungs by plaque assay, Vero CCL81 cells were used and plaques were enumerated 3 days post infection.
    Vero CCL81
    suggested: None
    Experimental Models: Organisms/Strains
    SentencesResources
    For SARS-CoV, in cohorts of equivalent numbers of male and female 20-29 week old SPF C57BL/6J (Stock 000664 Jackson Labs) mice (n = 10/dose group), we administered vehicle (10% PEG, 2.5% Cremophor RH40 in water) or 50, 150 or 500mg/kg EIDD-2801 by oral gavage 2hr prior to intranasal infection with 1E+04 PFU mouse-adapted SARS-CoV strain MA15 in 50μl.
    C57BL/6J
    suggested: None
    We performed all in vivo studies with EIDD-2801 in equivalent numbers of 10-14 week old female and male C57BL/6J hDPP4 mice.
    C57BL/6J hDPP4
    suggested: None
    Software and Algorithms
    SentencesResources
    Data was analyzed using GraphPad Prism 8.0 (La Jolla, CA).
    GraphPad
    suggested: (GraphPad Prism, RRID:SCR_002798)
    The RUBY package viral_seq version 1.0.6 (https://rubygems.org/gems/viral_seq) was used to calculate the mutation rate at each position.
    viral_seq
    suggested: (viral_seq, RRID:SCR_018515)
    Amino acid conservation scores of coronavirus RdRp were generated using ConSurf Server (https://consurf.tau.ac.il/) using the protein alignment described above and visualized on the SARS-CoV RdRp structure (PDB: 6NUR) in PyMol (version 1.8.6.0)20,52.
    PyMol
    suggested: (PyMOL, RRID:SCR_000305)
    Statistical analysis: All statistical data analyses were performed in Graphpad Prism 8.
    Graphpad Prism
    suggested: (GraphPad Prism, RRID:SCR_002798)

    Results from OddPub: We did not detect open data. We also did not detect open code. Researchers are encouraged to share open data when possible (see Nature blog).

    Results from LimitationRecognizer: An explicit section about the limitations of the techniques employed in this study was not found. We encourage authors to address study limitations.

    Results from TrialIdentifier: No clinical trial numbers were referenced.

    Results from Barzooka: We did not find any issues relating to the usage of bar graphs.

    Results from JetFighter: We did not find any issues relating to colormaps.

    Results from rtransparent:
    • No conflict of interest statement was detected. If there are no conflicts, we encourage authors to explicit state so.
    • Thank you for including a funding statement. Authors are encouraged to include this statement when submitting to a journal.
    • No protocol registration statement was detected.

    About SciScore

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  2. Version published to 10.1101/2020.03.19.997890v1 on bioRxiv