Identification of potent small molecule inhibitors of SARS-CoV-2 entry

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1. Version published to 10.1016/j.slasd.2021.10.012

   Jan 1, 2022
2. 

   ScreenIT

   Aug 10, 2021

   SciScore for 10.1101/2021.08.05.455262: (What is this?)

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

   Table 1: Rigor

   Ethics	not detected.
   Sex as a biological variable	not detected.
   Randomization	Data reporting: The number of samples tested was completely unbiased and all compounds were formatted in a randomized manner using an acoustic dispenser for analysis in the primary HTS campaigns.
   Blinding	All other downstream experiments were not randomized nor blinded as structures were known and hits were chosen using statistical parameters described below.
   Power Analysis	not detected.
   Cell Line Authentication	Authentication: For the ReFRAME library, all structural information was withheld until primary hits had been validated.

   Table 2: Resources

   Antibodies
   Sentences	Resources
   ACE2 expression was confirmed by immunofluorescence …

   SciScore for 10.1101/2021.08.05.455262: (What is this?)

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

   Table 1: Rigor

   Ethics	not detected.
   Sex as a biological variable	not detected.
   Randomization	Data reporting: The number of samples tested was completely unbiased and all compounds were formatted in a randomized manner using an acoustic dispenser for analysis in the primary HTS campaigns.
   Blinding	All other downstream experiments were not randomized nor blinded as structures were known and hits were chosen using statistical parameters described below.
   Power Analysis	not detected.
   Cell Line Authentication	Authentication: For the ReFRAME library, all structural information was withheld until primary hits had been validated.

   Table 2: Resources

   Antibodies
   Sentences	Resources
   ACE2 expression was confirmed by immunofluorescence staining using mouse monoclonal antibody against c-Myc antibody 9E10 (Thermo Fisher) and Goat-anti-mouse FITC (Jackson ImmunoResearch Laboratories, Inc).	c-Myc suggested: None Goat-anti-mouse FITC suggested: None
   Vero E6 cells enriched in ACE2 were selected by FACS with antibody against human ACE2.	human ACE2 suggested: None
   Vero E6 cells were stained with Goat anti-Human Phycoerythrin-conjugated ACE2 Polyclonal Antibody (R&D Systems) for 30 min at 4 °C in dark.	anti-Human Phycoerythrin-conjugated ACE2 suggested: None
   Experimental Models: Cell Lines
   Sentences	Resources
   The HEK293T cell line expressing human ACE2 was created by transduction with VSV-G protein-pseudotyped MLV containing pQCXIP-myc-ACE2-c9, as previously described18.	HEK293T suggested: None
   HEK293T-ACE2-TMPRSS2 cells were kept under selection with 1.3 µg/mL of puromycin (Sigma).	HEK293T-ACE2-TMPRSS2 suggested: None
   Vero E6 cells enriched in ACE2 were selected by FACS with antibody against human ACE2.	Vero E6 suggested: RRID:CVCL_XD71)
   The HEK293T-ACE2 cells were seeded at 2,000 cells/well, using an Aurora FRD (Aurora Discovery), at 2.5 µL/well.	HEK293T-ACE2 suggested: None
   Time-of-drug addition assay: Time-of-drug addition assay was performed in Vero CCL81 cells.	Vero CCL81 suggested: None
   Recombinant DNA
   Sentences	Resources
   The HEK293T cell line expressing human ACE2 was created by transduction with VSV-G protein-pseudotyped MLV containing pQCXIP-myc-ACE2-c9, as previously described18.	pQCXIP-myc-ACE2-c9 suggested: None
   Briefly, HEK293T cells were co-transfected with three plasmids, pMLV-gag-pol, pCAGGS-VSV-G and pQCXIP-myc-ACE2-c9 and the medium was replaced the next day.	pMLV-gag-pol suggested: None pCAGGS-VSV-G suggested: None
   Briefly, HEK293T cells were co-transfected with three plasmids, pMLV-gag-pol, pQC-Fluc and pCAGGS-SARS2-SFmut-cflag or pCAGGS-VSV-G using PEI 40K (Polysciences, Inc.), and the medium was refreshed 6 hours later.	pQC-Fluc suggested: None pCAGGS-SARS2-SFmut-cflag suggested: None
   The 3CLpro assay followed the exact same procedure albeit utilized the 3CLpro vector.	3CLpro suggested: None
   Software and Algorithms
   Sentences	Resources
   Statistical analysis was performed using GraphPad Prism software (San Diego, CA, USA).	GraphPad Prism suggested: (GraphPad Prism, RRID:SCR_002798)

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   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).

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   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.

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   Results from TrialIdentifier: No clinical trial numbers were referenced.

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   Results from Barzooka: We did not find any issues relating to the usage of bar graphs.

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   Results from JetFighter: We did not find any issues relating to colormaps.

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   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.

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

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3. 

   Version published to 10.1101/2021.08.05.455262v1 on bioRxiv

   Aug 5, 2021