NUDT18 catalyzes the hydrolysis of active metabolites of the antivirals Remdesivir, Ribavirin and Molnupiravir

  1. Ann-Sofie Jemth
  2. Emma Rose Scaletti
  3. Evert Homan
  4. Pål Stenmark
  5. Thomas Helleday
  6. Maurice Michel

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Abstract

Remdesivir and Molnupiravir have gained considerable interest due to their activity against SARS-CoV-2. Cellular hydrolysis of their active triphosphate forms, Remdesivir-TP and Molnupiravir-TP, would decrease drug efficiency. We therefore tested Remdesivir-TP as a substrate against a panel of human hydrolases and found that NUDT18 catalyzes the hydrolysis of Remdesivir-TP. The k cat value of NUDT18 for Remdesivir-TP was determined to 2.6 s -1 and the K m value was 156 μM, suggesting that NUDT18 catalyzed hydrolysis of Remdesivir-TP occurs in cells. We demonstrate that the triphosphates of the antivirals Ribavirin and Molnupiravir are hydrolyzed by NUDT18, albeit with a lower efficiency compared to Remdesivir-TP. NUDT18 also hydrolyses the triphosphates of Sofosbuvir and Aciclovir although with significantly lower activity. These results suggest that NUDT18 can act as a cellular sanitizer of modified nucleotides and may influence the antiviral efficacy of Remdesivir, Molnupiravir and Ribavirin. NUDT18 is expressed in respiratory epithelial cells and may limit the antiviral efficacy of Remdesivir and Molnupiravir against SARS-CoV2 replication by decreasing the intracellular concentration of their active metabolites at their intended site of action.

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  1. Version published to 10.1101/2021.11.16.468802v2 on bioRxiv
  2. ScreenIT

    SciScore for 10.1101/2021.11.16.468802: (What is this?)

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

    Table 1: Rigor

    Ethicsnot detected.
    Sex as a biological variablenot detected.
    Randomizationnot detected.
    Blindingnot detected.
    Power Analysisnot detected.

    Table 2: Resources

    Software and Algorithms
    SentencesResources
    The Michaelis-Menten equation was fitted to the data points and kinetic parameters of NUDT18 were determined using GraphPad Prism 8.0.
    GraphPad Prism
    suggested: (GraphPad Prism, RRID:SCR_002798)
    The panel included the triphosphate of Sofosbuvir used for the treatment of hepatitis C (Sofosbuvir-TP, (2R)-(2’-deoxy-2’-fluoro-2-methyl-uridine-5’-TP, Sierra bioresearch), the triphosphate of Ganciclovir used to treat cytomegalovirus infection treatment (Ganciclovir-TP, Sierra bioresearch) and the triphosphate of Aciclovir used for Herpes simplex virus infection therapy, (Aciclovir-TP, Sierra bioresearch).
    Sierra
    suggested: (Sierra, RRID:SCR_011831)
    Hydrogen bonding networks were optimized automatically by optimization of hydroxyls, Asn, Gln, and His residue states using ProtAssign.
    ProtAssign
    suggested: None
    Binding site analysis: SiteMap was used to probe the prepared NUDT18 structure for possible ligand binding sites.
    SiteMap
    suggested: (Biositemaps, RRID:SCR_001976)
    Ligand structure preparation: The structure of Remdesivir-TP was copied as isomeric smiles from PubChem (CID: 56832906) and pasted as 3D structure into Maestro.
    PubChem
    suggested: (PubChem, RRID:SCR_004284)
    Maestro
    suggested: (Maestro, RRID:SCR_016748)

    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:
    • 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.

    Results from scite Reference Check: We found no unreliable references.

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  3. Version published to 10.1101/2021.11.16.468802v1 on bioRxiv