Identifying SARS-CoV-2 antiviral compounds by screening for small molecule inhibitors of nsp13 helicase
This article has been Reviewed by the following groups
Listed in
- Evaluated articles (ScreenIT)
Abstract
The coronavirus disease 2019 (COVID-19) pandemic, which is caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), is a global public health challenge. While the efficacy of vaccines against emerging and future virus variants remains unclear, there is a need for therapeutics. Repurposing existing drugs represents a promising and potentially rapid opportunity to find novel antivirals against SARS-CoV-2. The virus encodes at least nine enzymatic activities that are potential drug targets. Here, we have expressed, purified and developed enzymatic assays for SARS-CoV-2 nsp13 helicase, a viral replication protein that is essential for the coronavirus life cycle. We screened a custom chemical library of over 5000 previously characterized pharmaceuticals for nsp13 inhibitors using a fluorescence resonance energy transfer-based high-throughput screening approach. From this, we have identified FPA-124 and several suramin-related compounds as novel inhibitors of nsp13 helicase activity in vitro. We describe the efficacy of these drugs using assays we developed to monitor SARS-CoV-2 growth in Vero E6 cells.
Article activity feed
-
-
SciScore for 10.1101/2021.04.07.438808: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Institutional Review Board Statement not detected. Randomization not detected. Blinding not detected. Power Analysis not detected. Sex as a biological variable not detected. Cell Line Authentication not detected. Table 2: Resources
Antibodies Sentences Resources Anti-FLAG M2 resin was added to the FH-nsp13 supernatant and Glutathione Sepharose 4 Fast Flow resin was added to the GST-nsp13 supernatant, and gently rotated for 2 h at 4°C. Anti-FLAGsuggested: NoneFinally, the cells were infected by adding 20µl of SARS-CoV-2 with a final MOI of 0.5 PFU/cell. 22h post infection, cells were fixed, permeabilised, and stained for SARS-CoV-2 N protein using Alexa488-labelled-CR3009 antibody produced in-house (see section for … SciScore for 10.1101/2021.04.07.438808: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Institutional Review Board Statement not detected. Randomization not detected. Blinding not detected. Power Analysis not detected. Sex as a biological variable not detected. Cell Line Authentication not detected. Table 2: Resources
Antibodies Sentences Resources Anti-FLAG M2 resin was added to the FH-nsp13 supernatant and Glutathione Sepharose 4 Fast Flow resin was added to the GST-nsp13 supernatant, and gently rotated for 2 h at 4°C. Anti-FLAGsuggested: NoneFinally, the cells were infected by adding 20µl of SARS-CoV-2 with a final MOI of 0.5 PFU/cell. 22h post infection, cells were fixed, permeabilised, and stained for SARS-CoV-2 N protein using Alexa488-labelled-CR3009 antibody produced in-house (see section for Recombinant mAb production) and cellular DNA using DRAQ7 (Abcam). DRAQ7suggested: NoneExperimental Models: Cell Lines Sentences Resources Viral inhibition assay: 1.5*10^3 Vero E6 cells (NIBC, UK) resuspended in DMEM containing 10% FBS were seeded into each well of 96-well imaging plates (Greiner 655090) and cultured overnight at 37C and 5% CO2. Vero E6suggested: NoneSoftware and Algorithms Sentences Resources MATLAB was used to process data. MATLABsuggested: (MATLAB, RRID:SCR_001622)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.
-