AUG-3387, a Human-Derived Monoclonal Antibody Neutralizes SARS-CoV-2 Variants and Reduces Viral Load from Therapeutic Treatment of Hamsters In Vivo

  1. Christopher J. Emig
  2. Marco A. Mena
  3. Steven J. Henry
  4. Adela Vitug
  5. Christian John Ventura
  6. Douglas Fox
  7. Xammy Huu Nguyenla
  8. Haiyue Xu
  9. Chaeho Moon
  10. Sawittree Sahakijjpijarn
  11. Philip J. Kuehl
  12. David Revelli
  13. Zengrong Cui
  14. Robert O. Williams
  15. Dale J. Christensen

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Abstract

Infections from the SARS-CoV-2 virus have killed over 4.6 million people since it began spreading through human populations in late 2019. In order to develop a therapeutic or prophylactic antibody to help mitigate the effects of the pandemic, a human monoclonal antibody (mAb) that binds to the SARS-CoV-2 spike protein was isolated from a convalescent patient following recovery from COVID-19 disease. This mAb, designated AUG-3387, demonstrates a high affinity for the spike protein of the original viral strains and all variants tested to date. In vitro pseudovirus neutralization and SARS-CoV-2 neutralization activity has been demonstrated in vitro. In addition, a dry powder formulation has been prepared using a Thin-Film Freezing (TFF) process that exhibited a fine particle fraction (FPF) of 50.95 ± 7.69% and a mass median aerodynamic diameter (MMAD) and geometric standard deviation (GSD) of 3.74 ± 0.73 µm and 2.73 ± 0.20, respectively. The dry powder is suitable for delivery directly to the lungs of infected patients using a dry powder inhaler device. Importantly, AUG-3387, administered as a liquid by intraperitoneal injection or the dry powder formulation delivered intratracheally into Syrian hamsters 24 hours after intranasal SARS-CoV-2 infection, demonstrated a dose-dependent reduction in the lung viral load of the virus. These data suggest that AUG-3387 formulated as a dry powder demonstrates potential to treat COVID-19.

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    SciScore for 10.1101/2021.10.12.464150: (What is this?)

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

    Table 1: Rigor

    EthicsConsent: Informed consent was obtained from all patients, and all patient samples were collected after a full recovery from illness and under IRB approval.
    IACUC: Animal work was performed at Lovelace Biomedical Research Institute (LBRI), with approval from the Institutional Animal Care and Use Committee (IACUC) and within Animal Biosafety Level 3 (ABSL3) containment.
    Euthanasia Agents: All animals were euthanized with an euthanasia solution consisting of 390 mg of sodium pentobarbital and 50 mg of phenytoin per mL.
    Sex as a biological variableThe FPF of delivered dose was calculated as the total amount of sugar or sugar alcohol (e.g., trehaose, mannitol) collected with an aerodynamic diameter below 5 µm as a percentage of the total amount of sugar or sugar alcohol deposited on the adapter, the induction port, stages 1–7 and Micro-Orifice Collector. 2.8. Efficacy of AUG-3387 in and in vivo model of SARS-CoV-2 infected Syrian Hamsters: An in vivo efficacy study was performed with male Syrian Hamsters (Mesocricetus auratus) approximately 9 weeks of age with a weight range of 110-134 g, at time of randomization, were sourced from Charles River Laboratory.
    RandomizationThe FPF of delivered dose was calculated as the total amount of sugar or sugar alcohol (e.g., trehaose, mannitol) collected with an aerodynamic diameter below 5 µm as a percentage of the total amount of sugar or sugar alcohol deposited on the adapter, the induction port, stages 1–7 and Micro-Orifice Collector. 2.8. Efficacy of AUG-3387 in and in vivo model of SARS-CoV-2 infected Syrian Hamsters: An in vivo efficacy study was performed with male Syrian Hamsters (Mesocricetus auratus) approximately 9 weeks of age with a weight range of 110-134 g, at time of randomization, were sourced from Charles River Laboratory.
    Blindingnot detected.
    Power Analysisnot detected.
    Cell Line Authenticationnot detected.

    Table 2: Resources

    Antibodies
    SentencesResources
    A fluorescently labelled anti-human IgG/IgA/IgM secondary antibody was also added to each well.
    anti-human IgG/IgA/IgM
    suggested: (Sigma-Aldrich Cat# S4893, RRID:AB_10625701)
    For identification of surface bound antibodies binding to SARS-CoV-2, SARS-CoV-2 antigens conjugated to Alexa Fluor 488 stained SARS-CoV-2 antigen was used as a staining agent for single cell sorting of antigen reactive memory B cells into 96 well plates on a Sony SH-800 Cell Sorter.
    SARS-CoV-2
    suggested: (Thermo Fisher Scientific Cat# 53-6490-82, RRID:AB_2884046)
    AUG-3705 was attached to the LSA flow cell via interaction with its V5 tag and a surface bound anti-V5 antibody.
    anti-V5
    suggested: None
    The cells were enriched 4 times until 97% of the cells showed signal above the negative control as read out by staining with anti-ACE-2 and secondary antibodies.
    anti-ACE-2
    suggested: None
    Experimental Models: Cell Lines
    SentencesResources
    Assay: 2.4.1 ACE-2 Expressing HEK293T Cell Line Construction: An ACE2 expressing HEK293T cell line (“LentiX ACE2.S4”) was constructed by packaging pCMV-AC-GFP (Origene) into lentivirus and transducing HEK293T’s (ATCC).
    ACE-2
    suggested: None
    HEK293T
    suggested: None
    Infected cell culture supernatant was diluted with 950 µL D10 media, and then serial diluted before 50 µL of each dilution was added to 8 wells of Vero E6 cells.
    Vero E6
    suggested: RRID:CVCL_XD71)
    After 96 hours, 100 µL CellTiterGlo reagent was added to each well of the infected Calu-3 cells to assay for live cells.
    Calu-3
    suggested: BCRJ Cat# 0264, RRID:CVCL_0609)
    Recombinant DNA
    SentencesResources
    Assay: 2.4.1 ACE-2 Expressing HEK293T Cell Line Construction: An ACE2 expressing HEK293T cell line (“LentiX ACE2.S4”) was constructed by packaging pCMV-AC-GFP (Origene) into lentivirus and transducing HEK293T’s (ATCC).
    pCMV-AC-GFP
    suggested: None

    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: We found the following clinical trial numbers in your paper:

    IdentifierStatusTitle
    NCT04872231CompletedSingle Ascending Dose and Multiple Ascending Dose Study of V…
    NCT04576325RecruitingPharmacokinetic Profile of Voriconazole Inhalation Powder in…

    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.

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

    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 10.1101/2021.10.12.464150v1 on bioRxiv