In Vitro Activity of Itraconazole Against SARS-CoV-2

  1. Ellen Van Damme
  2. Sandra De Meyer
  3. Denisa Bojkova
  4. Sandra Ciesek
  5. Jindrich Cinatl
  6. Steven De Jonghe
  7. Dirk Jochmans
  8. Pieter Leyssen
  9. Christophe Buyck
  10. Johan Neyts
  11. Marnix Van Loock

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Abstract

Background

As long as there is no vaccine available, having access to inhibitors of SARS-CoV-2 will be of utmost importance. Antivirals against coronaviruses do not exist, hence global drug re-purposing efforts have been carried out to identify agents that may provide clinical benefit to patients with COVID-19. Itraconazole, an antifungal agent, has been reported to have potential activity against animal coronaviruses.

Methods

Using cell-based phenotypic assays, the in vitro antiviral activity of itraconazole and 17-OH itraconazole was assessed against clinical isolates from a German and Belgian patient infected with SARS-CoV-2.

Results

Itraconazole demonstrated antiviral activity in human Caco-2 cells (EC 50 = 2.3 μM; MTT assay). Similarly, its primary metabolite, 17-OH itraconazole, showed inhibition of SARS-CoV-2 activity (EC 50 = 3.6 μM). Remdesivir inhibited viral replication with an EC 50 = 0.4 μM. Itraconazole and 17-OH itraconazole resulted in a viral yield reduction in vitro of approximately 2-log 10 and approximately 1-log 10 , as measured in both Caco-2 cells and VeroE6-eGFP cells, respectively. The viral yield reduction brought about by remdesivir or GS-441524 (parent nucleoside of the antiviral prodrug remdesivir; positive control) was more pronounced, with an approximately 3 log 10 drop and >4 log 10 drop in Caco-2 cells and VeroE6-eGFP cells, respectively.

Discussion

Itraconazole and 17-OH itraconazole exert in vitro low micromolar activity against SARS-CoV-2. Despite the in vitro antiviral activity, itraconazole did not result in a beneficial effect in hospitalized COVID-19 patients in a clinical study (EudraCT Number: 2020-001243-15).

Highlights

  • Itraconazole exerted in vitro low micromolar activity against SARS-CoV-2 (EC 50 = 2.3 μM)

  • Remdesivir demonstrated potent antiviral activity, confirming validity of the assay

  • Itraconazole has since shown no efficacy in a clinical study in hospitalized COVID-19 patients

Article activity feed

  1. Version published to 10.1101/2020.11.13.381194v2 on bioRxiv
  2. ScreenIT

    SciScore for 10.1101/2020.11.13.381194: (What is this?)

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

    Table 1: Rigor

    Institutional Review Board Statementnot detected.
    Randomizationnot detected.
    Blindingnot detected.
    Power Analysisnot detected.
    Sex as a biological variablenot detected.
    Cell Line Authenticationnot detected.

    Table 2: Resources

    Experimental Models: Cell Lines
    SentencesResources
    Assessment of Cell Viability: Cell viability in Caco-2 cells was measured following administration of each of the compounds or metabolites under investigation over the range of concentrations in the absence of virus using the Rotitest Vital (Roth, Karlsruhe, Germany) test according to manufacturer’s instructions, as previously described.
    Caco-2
    suggested: None
    Antiviral activity was also assessed by reduction of viral yield in VeroE6-eGFP cells.
    VeroE6-eGFP
    suggested: None
    Experimental Models: Organisms/Strains
    SentencesResources
    VeroE6-eGFP (African green monkey kidney cell line; provided by Dr. K. Andries J&JPRD; Beerse, Belgium) were cultured in Dulbecco’s Modified Eagle Medium (DMEM) supplemented with 10% FBS, 0.075% sodium bicarbonate, penicillin and streptomycin (100 μg/mL) at 37°C in a 5% CO2 atmosphere.
    VeroE6-eGFP
    suggested: None
    Software and Algorithms
    SentencesResources
    Data were analyzed by four-parameter curve-fitting from a dose-response curve using GraphPad Prism (version 7.00) to calculate the CC50 (cytotoxic concentration of the compound that reduced cell viability to 50%).
    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:
    • 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.

  3. Version published to 10.1101/2020.11.13.381194v1 on bioRxiv