Glycopeptide Antibiotic Teicoplanin Inhibits Cell Entry of SARS-CoV-2 by Suppressing the Proteolytic Activity of Cathepsin L

Abstract

Since the outbreak of the coronavirus disease 2019 (COVID-19) caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), public health worldwide has been greatly threatened. The development of an effective treatment for this infection is crucial and urgent but is hampered by the incomplete understanding of the viral infection mechanisms and the lack of specific antiviral agents. We previously reported that teicoplanin, a glycopeptide antibiotic that has been commonly used in the clinic to treat bacterial infection, significantly restrained the cell entry of Ebola virus, SARS-CoV, and MERS-CoV by specifically inhibiting the activity of cathepsin L (CTSL). Here, we found that the cleavage sites of CTSL on the spike proteins of SARS-CoV-2 were highly conserved among all the variants. The treatment with teicoplanin suppressed the proteolytic activity of CTSL on spike and prevented the cellular infection of different pseudotyped SARS-CoV-2 viruses. Teicoplanin potently prevented the entry of SARS-CoV-2 into the cellular cytoplasm with an IC ₅₀ of 2.038 μM for the Wuhan-Hu-1 reference strain and an IC ₅₀ of 2.116 μM for the SARS-CoV-2 (D614G) variant. The pre-treatment of teicoplanin also prevented SARS-CoV-2 infection in hACE2 mice. In summary, our data reveal that CTSL is required for both SARS-CoV-2 and SARS-CoV infection and demonstrate the therapeutic potential of teicoplanin for universal anti-CoVs intervention.

SciScore for 10.1101/2020.02.05.935387: (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

Experimental Models: Cell Lines
Sentences	Resources
HIV-1/2019-nCoV-S/pseudoviruses were produced by the co-transfection of pHIV-luciferase, psPAX2, and plasmids expressing different envelope or S proteins into HEK293T cells as previously described 27.	HEK293T suggested: None
HEK293T, A549 and Huh7 cell lines were maintained in Dulbecco’s modified Eagle’s medium (Gibco) with 10% fetal calf serum (Gibco), 100 units/ml penicillin, and 100 μg/ml streptomycin (Gibco) at 37 °C and 5% CO2.	Huh7 su…

SciScore for 10.1101/2020.02.05.935387: (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

Experimental Models: Cell Lines
Sentences	Resources
HIV-1/2019-nCoV-S/pseudoviruses were produced by the co-transfection of pHIV-luciferase, psPAX2, and plasmids expressing different envelope or S proteins into HEK293T cells as previously described 27.	HEK293T suggested: None
HEK293T, A549 and Huh7 cell lines were maintained in Dulbecco’s modified Eagle’s medium (Gibco) with 10% fetal calf serum (Gibco), 100 units/ml penicillin, and 100 μg/ml streptomycin (Gibco) at 37 °C and 5% CO2.	Huh7 suggested: None
A549 cells were seeded in 105 cells per ml in each well of 12-well-microtiter plates and transiently transfected with siRNA (10–20 pmol/well) using lipofectinRNAimax reagent according to the manufacturer’s instructions in serum-free medium with suitable scrambled siRNA control.	A549 suggested: None
Software and Algorithms
Sentences	Resources
The IC50 curve was determined by using software from GraphPad (San Diego). siRNA transfection, RNA isolation, and RT-PCR: Sequences of siRNA against cathepsin Lor TMPRSS2 were predesigned by Ribobio Company (Guangzhou, China).	GraphPad suggested: (GraphPad Prism, RRID:SCR_002798)
The sequence datasets were aligned using the ClustalW program implemented in MEGA X software28.	ClustalW suggested: (ClustalW, RRID:SCR_017277) MEGA X suggested: None
Consensus sequences were created using the BioEdit software (http://www.mbio.ncsu.edu/bioedit/bioedit.html) based on the multiple alignment of SARS-CoV and 2019-nCoV, respectively.	BioEdit suggested: (BioEdit, RRID:SCR_007361)
Statistics: Statistical analysis was performed with GraphPad Prism 7.	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 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:

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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.

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Glycopeptide Antibiotic Teicoplanin Inhibits Cell Entry of SARS-CoV-2 by Suppressing the Proteolytic Activity of Cathepsin L

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