Emetine as an antiviral agent suppresses SARS-CoV-2 replication by inhibitinginteraction of viral mRNAwith eIF4E: An in vitro study

  1. Ram Kumar
  2. Nitin Khandelwal
  3. Yogesh Chander
  4. Thachamvally Riyesh
  5. Baldev R. Gulati
  6. Yash Pal
  7. Bhupendra N. Tripathi
  8. Sanjay Barua
  9. Naveen Kumar

Reviewed by

Read the full article See related articles

Discuss this preprint

Start a discussion What are Sciety discussions?

Listed in

Log in to save this article

Abstract

Emetine is a FDA-approved drug for the treatment of amebiasis. In the recent times we had also demonstrated the antiviral efficacy of emetine against some RNA and DNA viruses. Following emergence of the COVID-19, we further evaluated the in vitro antiviral activity of emetine against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). The therapeutic index of emetine was determined to be 10910.4, at a cytotoxic concentration 50 (CC 50 ) of 1603.8 nM and effective concentration 50 (EC 50 ) of 0.147 nM.Besides, we also demonstrated the protective efficacy of emetine against lethal challenge with infectious bronchitis virus (IBV; a chicken coronavirus) in the embryonated chicken egg infection model. Emetine treatment was shown to decrease viral RNA and protein synthesis without affecting other steps of viral life cycle such as attachment, entry and budding.In a chromatin immunoprecipitation (CHIP) assay, emetine was shown to disrupt the binding of SARS-CoV-2 RNA with eIF4E (eukaryotic translation initiation factor 4E, a cellular cap-binding protein required for initiation ofprotein translation). Further, SARS-CoV-2 was shown to exploit ERK/MNK1/eIF4E signalling pathwayfor its effective replication in the target cells. To conclude, emetine targets SARS-CoV-2 protein synthesis which is mediated via inhibiting the interaction of SARS-CoV-2 RNA with eIF4E. This is a novel mechanistic insight on the antiviral efficacy of emetine. In vitro antiviral efficacy against SARS-CoV-2 and its ability to protect chicken embryos against IBV suggests that emetine could be repurposed to treat COVID-19.

Article activity feed

  1. ScreenIT

    SciScore for 10.1101/2020.11.29.401984: (What is this?)

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

    Table 1: Rigor

    Institutional Review Board StatementIRB: The work was approved from the Institute Biosafety Committee and Review Committee on Genetic Manipulation (RCGM), Department of Biotechnology, Government of India (No. BT/BS/17/436/2011-PID).
    Consent: Human serumfrom a COVID-19 confirmed patientwascollected at 2 weeks following recovery and obtained after due consent from the patient. 2.4.
    Randomizationnot detected.
    Blindingnot detected.
    Power Analysisnot detected.
    Sex as a biological variablenot detected.
    Cell Line Authenticationnot detected.

    Table 2: Resources

    Antibodies
    SentencesResources
    eIF4E monoclonal antibody (5D11) and Phospho-eIF4E (Ser209) polyclonal antibodies were received from Invitrogen (South San Francisco, CA, USA)
    Ser209
    suggested: None
    Glyceraldehyde 3-phosphate dehydrogenase, house-keeping control protein) primary antibody, Anti-Mouse IgG (whole molecule)−Alkaline Phosphatase antibody (produced in goat) and Anti-Rabbit IgG (whole molecule)–Peroxidase antibody (produced in goat) were received from Sigma-Aldrich (St. Louis, USA)
    Glyceraldehyde 3-phosphate dehydrogenase , house-keeping control protein
    suggested: None
    Anti-Mouse IgG
    suggested: None
    Anti-Rabbit IgG ( whole molecule)–Peroxidase
    suggested: None
    The clarified cell lysateswere mixed with 10 units of RiboLock RNase Inhibitor (Thermo Scientific, USA) and then incubated with the phospho-eIF4E antibody (reactive antibody),phospho ERK antibody (nonreactive antibody) or equivalent volume of IP buffer (beads control) for 45 minutes at room temperature.
    phospho-eIF4E
    suggested: None
    Experimental Models: Cell Lines
    SentencesResources
    Briefly, Vero cells, in triplicates were infected with SARS-CoV-2 at MOI of 5.
    Vero
    suggested: CLS Cat# 605372/p622_VERO, RRID:CVCL_0059)

    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 found bar graphs of continuous data. We recommend replacing bar graphs with more informative graphics, as many different datasets can lead to the same bar graph. The actual data may suggest different conclusions from the summary statistics. For more information, please see Weissgerber et al (2015).

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

    Results from rtransparent:
    • Thank you for including a conflict of interest statement. Authors are encouraged to include this statement when submitting to a journal.
    • 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

    SciScore is an automated tool that is designed to assist expert reviewers by finding and presenting formulaic information scattered throughout a paper in a standard, easy to digest format. SciScore checks for the presence and correctness of RRIDs (research resource identifiers), and for rigor criteria such as sex and investigator blinding. For details on the theoretical underpinning of rigor criteria and the tools shown here, including references cited, please follow this link.

  2. Version published to 10.1101/2020.11.29.401984 on bioRxiv