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Unfractionated heparin inhibits live wild type SARS-CoV-2 cell infectivity at therapeutically relevant concentrations

Unfractionated heparin inhibits live wild type SARS-CoV-2 cell infectivity at therapeutically relevant concentrations
Unfractionated heparin inhibits live wild type SARS-CoV-2 cell infectivity at therapeutically relevant concentrations

BACKGROUND AND PURPOSE: Currently, there are no licensed vaccines and limited antivirals for the treatment of COVID-19. Heparin (delivered systemically) is currently used to treat anticoagulant anomalies in COVID-19 patients. Additionally, in the United Kingdom, Brazil and Australia, nebulised unfractionated heparin (UFH) is being trialled in COVID-19 patients as a potential treatment. A systematic comparison of the potential antiviral effect of various heparin preparations on live wild type SARS-CoV-2, in vitro, is needed.

EXPERIMENTAL APPROACH: Seven different heparin preparations including UFH and low MW heparins (LMWH) of porcine or bovine origin were screened for antiviral activity against live SARS-CoV-2 (Australia/VIC01/2020) using a plaque inhibition assay with Vero E6 cells. Interaction of heparin with spike protein RBD was studied using differential scanning fluorimetry and the inhibition of RBD binding to human ACE2 protein using elisa assays was examined.

KEY RESULTS: All the UFH preparations had potent antiviral effects, with IC50 values ranging between 25 and 41 μg·ml-1 , whereas LMWHs were less inhibitory by ~150-fold (IC50 range 3.4-7.8 mg·ml-1 ). Mechanistically, we observed that heparin binds and destabilizes the RBD protein and furthermore, we show heparin directly inhibits the binding of RBD to the human ACE2 protein receptor.

CONCLUSION AND IMPLICATIONS: This comparison of clinically relevant heparins shows that UFH has significantly stronger SARS-CoV-2 antiviral activity compared to LMWHs. UFH acts to directly inhibit binding of spike protein to the human ACE2 protein receptor. Overall, the data strongly support further clinical investigation of UFH as a potential treatment for patients with COVID-19.

Angiotensin-Converting Enzyme 2/metabolism, Animals, Antiviral Agents/pharmacology, COVID-19/drug therapy, Chlorocebus aethiops, Heparin, Low-Molecular-Weight/pharmacology, Heparin/metabolism, Protein Binding/drug effects, SARS-CoV-2/growth & development, Spike Glycoprotein, Coronavirus/metabolism, Viral Plaque Assay
0007-1188
626-635
Tree, Julia A.
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Turnbull, Jeremy E.
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Buttigieg, Karen R.
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Elmore, Michael J.
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Coombes, Naomi
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Hogwood, John
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Mycroft-West, Courtney J.
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Lima, Marcelo A.
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Skidmore, Mark A.
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Karlsson, Richard
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Chen, Yen-Hsi
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Zhang, Yang
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Spalluto, Cosma Mirella
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Staples, Karl J.
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Yates, Edwin A.
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Gray, Elaine
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Singh, Dave
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Wilkinson, Tom
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Page, Clive P.
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Carroll, Miles W.
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Tree, Julia A.
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Turnbull, Jeremy E.
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Buttigieg, Karen R.
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Elmore, Michael J.
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Coombes, Naomi
699ffcfe-0e4d-4098-9ddc-308dce0ce685
Hogwood, John
c88b210e-280d-453c-ac2c-93f5d0658b39
Mycroft-West, Courtney J.
2acb0b1f-146c-48a8-86f0-5a850b64fdbf
Lima, Marcelo A.
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Skidmore, Mark A.
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Karlsson, Richard
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Chen, Yen-Hsi
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Zhang, Yang
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Spalluto, Cosma Mirella
6802ad50-bc38-404f-9a19-40916425183b
Staples, Karl J.
e0e9d80f-0aed-435f-bd75-0c8818491fee
Yates, Edwin A.
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Gray, Elaine
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Singh, Dave
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Wilkinson, Tom
8c55ebbb-e547-445c-95a1-c8bed02dd652
Page, Clive P.
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Carroll, Miles W.
73c6ebde-75fe-4ddb-bf67-1baaa68b775b

Tree, Julia A., Turnbull, Jeremy E., Buttigieg, Karen R., Elmore, Michael J., Coombes, Naomi, Hogwood, John, Mycroft-West, Courtney J., Lima, Marcelo A., Skidmore, Mark A., Karlsson, Richard, Chen, Yen-Hsi, Zhang, Yang, Spalluto, Cosma Mirella, Staples, Karl J., Yates, Edwin A., Gray, Elaine, Singh, Dave, Wilkinson, Tom, Page, Clive P. and Carroll, Miles W. (2021) Unfractionated heparin inhibits live wild type SARS-CoV-2 cell infectivity at therapeutically relevant concentrations. British Journal of Pharmacology, 178 (3), 626-635. (doi:10.1111/bph.15304).

Record type: Article

Abstract

BACKGROUND AND PURPOSE: Currently, there are no licensed vaccines and limited antivirals for the treatment of COVID-19. Heparin (delivered systemically) is currently used to treat anticoagulant anomalies in COVID-19 patients. Additionally, in the United Kingdom, Brazil and Australia, nebulised unfractionated heparin (UFH) is being trialled in COVID-19 patients as a potential treatment. A systematic comparison of the potential antiviral effect of various heparin preparations on live wild type SARS-CoV-2, in vitro, is needed.

EXPERIMENTAL APPROACH: Seven different heparin preparations including UFH and low MW heparins (LMWH) of porcine or bovine origin were screened for antiviral activity against live SARS-CoV-2 (Australia/VIC01/2020) using a plaque inhibition assay with Vero E6 cells. Interaction of heparin with spike protein RBD was studied using differential scanning fluorimetry and the inhibition of RBD binding to human ACE2 protein using elisa assays was examined.

KEY RESULTS: All the UFH preparations had potent antiviral effects, with IC50 values ranging between 25 and 41 μg·ml-1 , whereas LMWHs were less inhibitory by ~150-fold (IC50 range 3.4-7.8 mg·ml-1 ). Mechanistically, we observed that heparin binds and destabilizes the RBD protein and furthermore, we show heparin directly inhibits the binding of RBD to the human ACE2 protein receptor.

CONCLUSION AND IMPLICATIONS: This comparison of clinically relevant heparins shows that UFH has significantly stronger SARS-CoV-2 antiviral activity compared to LMWHs. UFH acts to directly inhibit binding of spike protein to the human ACE2 protein receptor. Overall, the data strongly support further clinical investigation of UFH as a potential treatment for patients with COVID-19.

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Tree et al 2020 BJP - Accepted Manuscript
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Accepted/In Press date: 18 October 2020
e-pub ahead of print date: 30 October 2020
Published date: February 2021
Keywords: Angiotensin-Converting Enzyme 2/metabolism, Animals, Antiviral Agents/pharmacology, COVID-19/drug therapy, Chlorocebus aethiops, Heparin, Low-Molecular-Weight/pharmacology, Heparin/metabolism, Protein Binding/drug effects, SARS-CoV-2/growth & development, Spike Glycoprotein, Coronavirus/metabolism, Viral Plaque Assay

Identifiers

Local EPrints ID: 444485
URI: http://eprints.soton.ac.uk/id/eprint/444485
ISSN: 0007-1188
PURE UUID: 7cdc326e-1418-4f7f-a048-b56cb3db52f5
ORCID for Karl J. Staples: ORCID iD orcid.org/0000-0003-3844-6457

Catalogue record

Date deposited: 21 Oct 2020 16:31
Last modified: 18 Oct 2021 04:01

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Contributors

Author: Julia A. Tree
Author: Jeremy E. Turnbull
Author: Karen R. Buttigieg
Author: Michael J. Elmore
Author: Naomi Coombes
Author: John Hogwood
Author: Courtney J. Mycroft-West
Author: Marcelo A. Lima
Author: Mark A. Skidmore
Author: Richard Karlsson
Author: Yen-Hsi Chen
Author: Yang Zhang
Author: Cosma Mirella Spalluto
Author: Karl J. Staples ORCID iD
Author: Edwin A. Yates
Author: Elaine Gray
Author: Dave Singh
Author: Tom Wilkinson
Author: Clive P. Page
Author: Miles W. Carroll

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