Nanodroplet-Benzalkonium Chloride Formulation Demonstrates In Vitro and Ex-Vivo Broad-Spectrum Antiviral Activity Against SARS-CoV-2 and other Enveloped Viruses

  1. Jessie Pannu
  2. Susan Ciotti
  3. Shyamala Ganesan
  4. George Arida
  5. Chad Costley
  6. Ali Fattom

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Abstract

The Covid-19 pandemic has highlighted the importance of aerosolized droplets inhaled into the nose in the transmission of respiratory viral disease. Inactivating pathogenic viruses at the nasal portal of entry may reduce viral loads, thereby reducing transmission and contagion. We have developed an oil-in-water nanoemulsion ( nanodroplet ) formulation containing the potent antiseptic 0.13% Benzalkonium Chloride (NE-BZK) which demonstrates safe and broad anti-viral activity. While The Centers for Disease Control and Prevention (CDC) have reported that BZK may have less reliable activity than ethyl alcohol against certain viruses, including coronaviruses, we have demonstrated that NE-BZK exhibits broad-spectrum, long-lasting antiviral activity with >99.99% in vitro killing of enveloped viruses including SARS-CoV-2, human coronavirus, RSV and influenza B. Furthermore, in vitro studies demonstrated that NE-BZK continues to kill >99.99% of human coronavirus even when diluted 20-fold, while 0.13% aqueous BZK solution (AQ-BZK) did not. Ex vivo studies of NE-BZK on human cadaver skin demonstrated persistent >99.99% killing of human coronavirus for at least 8 hours after application. AQ-BZK failed to demonstrate durable antimicrobial activity on skin over time. The repeated application of NE-BZK, twice daily for 2 weeks on to rabbit nostrils indicated safety with no irritation. These findings demonstrate that formulating BZK on the surface of proprietary nanodroplets offers a safe and effective antiviral as a significant addition to strategies to combat the spread of respiratory viral infectious diseases.

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  1. ScreenIT

    SciScore for 10.1101/2020.11.12.377598: (What is this?)

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

    Table 1: Rigor

    Institutional Review Board StatementIACUC: The exploratory study was performed at IITRI in accordance with protocols approved by the animal care and use committee.
    RandomizationThe animals were randomly assigned to either naïve control or NE-BZK treatment groups with 2/sex in control group and 4/sex in NE-BZK group.
    Blindingnot detected.
    Power Analysisnot detected.
    Sex as a biological variablenot detected.
    Cell Line Authenticationnot detected.

    Table 2: Resources

    Experimental Models: Cell Lines
    SentencesResources
    Briefly, serially diluted samples were plated onto 80-90% confluent Vero E6 cells for SARS-CoV-2, MRC-5 cells for HCoV229E, MDCK for Influenza B and Vero cells for RSV.
    Vero E6
    suggested: None
    MRC-5
    suggested: None
    MDCK
    suggested: CLS Cat# 602280/p823_MDCK_(NBL-2, RRID:CVCL_0422)
    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: We detected the following sentences addressing limitations in the study:
    However, this approach has major limitations including patient experience of using povidone iodine in the nose, allergic potential, and thyroid disease concerns. Povidone Iodine can become ineffective when diluting from available commercial 5% or 10% solutions and can be effective only up to 4 hours post application25. NE-BZK has an added advantage that safety has been previously demonstrated in humans through two different phase 1 clinical trials using the same nanoemulsion technology with a similar cationic surfactant.26, 27 BZK at concentrations similar to those in NE-BZK has been used in human skin products since the 1940’s and is currently marketed in numerous nasal products.(reference) Hirose et al has shown the survival of SARS-CoV-2 on skin surfaces for 9 hours28 which may require a more durable antiseptic activity. Our ex-vivo data has shown that antimicrobial effect of NE-BZK was seen even 8-12 hours post application, and even when diluting 20-fold, suggesting that application every 8 hours may be useful in protecting against disease. Our in vivo safety study in rabbits also demonstrated no safety issues with twice daily application of NE-BZK for two weeks, and BZK was not detected in the serum samples two-hours post application, indicating no absorption into the systemic circulation. We believe that NE-BZK antiseptic might fulfill a role in reducing viral loads in SARS-Cov-2 infected persons, potentially reducing the risk of disease progression and increasing survi...

    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.
    • No funding statement was detected.
    • 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 10.1101/2020.11.12.377598v1 on bioRxiv