Pentosan Polysulfate Inhibits Attachment and Infection by SARS-CoV-2 In Vitro: Insights into Structural Requirements for Binding

Sabrina Bertini; Anna Alekseeva; Stefano Elli; Isabel Pagani; Serena Zanzoni; Giorgio Eisele; Ravi Krishnan; Klaus P. Maag; Christian Reiter; Dominik Lenhart; Rudolf Gruber; Edwin A Yates; Elisa Vicenzi; Annamaria Naggi; Antonella Bisio; Marco Guerrini

doi:10.1055/a-1807-0168

Thrombosis and Haemostasis, Table of Contents

CC BY-NC-ND 4.0 · Thromb Haemost 2022; 122(06): 984-997
DOI: 10.1055/a-1807-0168

Blood Cells, Inflammation and Infection

Pentosan Polysulfate Inhibits Attachment and Infection by SARS-CoV-2 In Vitro: Insights into Structural Requirements for Binding

Sabrina Bertini^*

¹Istituto di Ricerche Chimiche e Biochimiche G. Ronzoni, Milan, Italy

,

Anna Alekseeva^*

²Centro Alta Tecnologia “Istituto di Ricerche Chimiche e Biochimiche G. Ronzoni” Srl, Milan, Italy

,

Stefano Elli^*

¹Istituto di Ricerche Chimiche e Biochimiche G. Ronzoni, Milan, Italy

,

Isabel Pagani^*

³Viral Pathogenesis and Biosafety Unit, IRCCS San Raffaele Scientific Institute, Milan, Italy

,

Serena Zanzoni^*

⁴Centro Piattaforme Tecnologiche, University of Verona, Verona, Italy

,

Giorgio Eisele

²Centro Alta Tecnologia “Istituto di Ricerche Chimiche e Biochimiche G. Ronzoni” Srl, Milan, Italy

,

Ravi Krishnan

⁵Paradigm Biopharmaceuticals Ltd., Melbourne, Victoria, Australia

,

Klaus P. Maag

⁶Bene PharmaChem GmbH & Co.KG, Geretsried, Germany

,

Christian Reiter

⁵Paradigm Biopharmaceuticals Ltd., Melbourne, Victoria, Australia

,

Dominik Lenhart

⁶Bene PharmaChem GmbH & Co.KG, Geretsried, Germany

,

Rudolf Gruber

⁶Bene PharmaChem GmbH & Co.KG, Geretsried, Germany

,

Edwin A Yates^*

⁷Department of Biochemistry and Systems Biology, ISMIB, University of Liverpool, Liverpool, United Kingdom

,

Elisa Vicenzi^*

³Viral Pathogenesis and Biosafety Unit, IRCCS San Raffaele Scientific Institute, Milan, Italy

,

Annamaria Naggi^**

¹Istituto di Ricerche Chimiche e Biochimiche G. Ronzoni, Milan, Italy

,

Antonella Bisio^**

¹Istituto di Ricerche Chimiche e Biochimiche G. Ronzoni, Milan, Italy

,

Marco Guerrini^**

¹Istituto di Ricerche Chimiche e Biochimiche G. Ronzoni, Milan, Italy

› Author Affiliations

Abstract

Two years since the outbreak of the novel coronavirus SARS-CoV-2 (severe acute respiratory syndrome coronavirus 2) pandemic, there remain few clinically effective drugs to complement vaccines. One is the anticoagulant, heparin, which in 2004 was found able to inhibit invasion of SARS-CoV (CoV-1) and which has been employed during the current pandemic to prevent thromboembolic complications and moderate potentially damaging inflammation. Heparin has also been shown experimentally to inhibit SARS-CoV-2 attachment and infection in susceptible cells. At high therapeutic doses however, heparin increases the risk of bleeding and prolonged use can cause heparin-induced thrombocytopenia, a serious side effect. One alternative, with structural similarities to heparin, is the plant-derived, semi-synthetic polysaccharide, pentosan polysulfate (PPS). PPS is an established drug for the oral treatment of interstitial cystitis, is well-tolerated, and exhibits weaker anticoagulant effects than heparin. In an established Vero cell model, PPS and its fractions of varying molecular weights inhibited invasion by SARS-CoV-2. Intact PPS and its size-defined fractions were characterized by molecular weight distribution and chemical structure using nuclear magnetic resonance spectroscopy and liquid chromatography–mass spectrometry, then employed to explore the structural basis of interactions with SARS-CoV-2 spike protein receptor-binding domain (S1 RBD) and the inhibition of Vero cell invasion. PPS was as effective as unfractionated heparin, but more effective in inhibiting cell infection than low-molecular-weight heparin (on a weight/volume basis). Isothermal titration calorimetry and viral plaque-forming assays demonstrated size-dependent binding to S1 RBD and inhibition of Vero cell invasion, suggesting the potential application of PPS as a novel inhibitor of SARS-CoV-2 infection.

Keywords

pentosan polysulfate - SARS-CoV-2 - COVID-19 - antiviral agent

Full Text

References

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Supplementary Material

Supplementary Material