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CC BY-NC-ND 4.0 · Thromb Haemost
DOI: 10.1055/a-2483-0107
Coagulation and Fibrinolysis

Dabigatran Attenuates the Binding of Thrombin to Platelets—A Novel Mechanism of Action

Tomas L. Lindahl   *
1   Division of Clinical Chemistry and Pharmacology, Department of Biomedical and Clinical Sciences, Linköping University, Linköping, Sweden
,
Aishwarya Prasanna Kumar   *
1   Division of Clinical Chemistry and Pharmacology, Department of Biomedical and Clinical Sciences, Linköping University, Linköping, Sweden
,
Teresia Hallström
2   NanoTemper Technologies, Munich, Germany
,
Ahmed Al-Hashimi
1   Division of Clinical Chemistry and Pharmacology, Department of Biomedical and Clinical Sciences, Linköping University, Linköping, Sweden
,
Anna du Rietz
3   Division of Molecular Surface Physics and Nanoscience, Department of Physics, Chemistry and Biology, Linköping University, Linköping, Sweden
,
Elena Arlaman
1   Division of Clinical Chemistry and Pharmacology, Department of Biomedical and Clinical Sciences, Linköping University, Linköping, Sweden
,
Kajsa Uvdal
3   Division of Molecular Surface Physics and Nanoscience, Department of Physics, Chemistry and Biology, Linköping University, Linköping, Sweden
,
Ankit S. Macwan
1   Division of Clinical Chemistry and Pharmacology, Department of Biomedical and Clinical Sciences, Linköping University, Linköping, Sweden
› Author Affiliations Funding This study was supported by grants with TL as main and AM as co-applicant from the Swedish Research Council no. 2020–01002 and the Swedish Heart-Lung foundation no. 2019037022 and 20220205 and Swedish Research Council Grant no. 2019–02409 to KU as main applicant, TL was one of the co-applicants.
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Abstract

Background Thrombin is a multifunctional regulatory enzyme of the haemostasis and has both pro- and anticoagulant roles. It has, therefore, been a main target for drug discovery over many decades. Thrombin is a serine protease and possesses two positively charged regions called exosites, through which it is known to bind to many substrates. Dabigatran is a thrombin inhibitor and is widely used as an oral anticoagulant for the antithrombotic treatment of atrial fibrillation and venous thromboembolism. The mechanism by which dabigatran inhibits thrombin is the blockage of the active site, however, its effect on thrombin binding to its substrates has not been studied thoroughly and is thus poorly understood.

Material and Methods The effect of dabigatran on thrombin binding to platelets was evaluated by flow cytometry using fluorescently labelled thrombin and washed platelets. Further, to confirm the results we utilized modern techniques for biomolecular binding studies, microscale thermophoresis (MST) and surface plasmon resonance (SPR), which validated the results.

Results Dabigatran inhibited thrombin binding to platelets as analysed by flow cytometry. The inhibition was dose dependent with IC50 of 118 nM which was slightly lower than for inhibition of platelet activation and is close to the clinically relevant plasma concentration of dabigatran. MST and SPR also confirmed inhibitory effect of dabigatran on thrombin binding to platelets.

Conclusion Apart from blocking the active site, dabigatran also inhibits thrombin binding to platelets. Since thrombin has numerous functions beyond the cardiovascular system, this finding may have important implications.

Keywords

thrombin - dabigatran - microscale thermophoresis - platelets

Authors' Contribution

A.M. and T.L. conceived the study and designed the flow cytometry and MST experiments. A.M., T.H., A.P.K., and A.A.H. performed MST experiments. A.P.K. and E.A. performed flow cytometry experiments. A.R. and K.U. designed the SPR experiments, analyzed the data, and prepared the figures. A.R. performed the SPR experiments. A.M. and T.L. wrote the manuscript. All authors have read, commented, and approved the final version of the manuscript.


* These authors contributed equally.


Supplementary Material



Publication History

Received: 11 May 2024

Accepted: 09 October 2024

Accepted Manuscript online:
25 November 2024

Article published online:
19 December 2024

© 2024. The Author(s). This is an open access article published by Thieme under the terms of the Creative Commons Attribution-NonDerivative-NonCommercial License, permitting copying and reproduction so long as the original work is given appropriate credit. Contents may not be used for commercial purposes, or adapted, remixed, transformed or built upon. (https://creativecommons.org/licenses/by-nc-nd/4.0/)

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