BACKGROUND AND PURPOSE: Although the amphiphilic nature of the widely used antithrombotic drug Ticagrelor is well known, it was never considered as a membranotropic agent capable of interacting with the lipid bilayer in a receptor-independent way. In this study, we investigated the influence of Ticagrelor on plasma membrane lipid order in platelets and if this modulates the potency of Ticagrelor at the P2Y12 receptor. EXPERIMENTAL APPROACH: We combined fluorescent in situ, in vitro and in silico approaches to probe the interactions between the plasma membrane of platelets and Ticagrelor. The influence of Ticagrelor on the lipid order of the platelet plasma membrane and large unilamellar vesicles was studied using the advanced fluorescent probe NR12S. Furthermore, the properties of model lipid bilayers in the presence of Ticagrelor were characterized by molecular dynamics simulations. Finally, the influence of an increased lipid order on the dose-response of platelets to Ticagrelor was studied. KEY RESULTS: Ticagrelor incorporates spontaneously into lipid bilayers and affects the lipid order of the membranes of model vesicles and isolated platelets, in a nontrivial composition and concentration-dependent manner. We showed that higher plasma membrane lipid order in platelets leads to a lower IC50 value for Ticagrelor. It is shown that membrane incorporation of Ticagrelor increases its potency at the P2Y12 receptor, by increasing the order of the platelet plasma membrane. CONCLUSION AND IMPLICATIONS: A novel dual mechanism of Ticagrelor action is suggested that combines direct binding to P2Y12 receptor with simultaneous modulation of receptor-lipid microenvironment.

CNRS Chrono environnement Université de Franche Comté Besançon France

Czech Academy of Sciences Institute of Organic Chemistry and Biochemistry Prague Czech Republic

Department of Integrative Biology and Pharmacology The University of Texas Health Science Center at Houston Houston Texas USA

Department of Neurochemistry Palladin Institute of Biochemistry of the NAS of Ukraine Kyiv Ukraine

Department of Physical Chemistry Faculty of Science Palacký University Olomouc Olomouc Czech Republic

Department of Physics of Biological Systems Institute of Physics of the National Academy of Sciences of Ukraine Kyiv Ukraine

Receptor AI Inc London UK

SINERGIES Université de Franche Comté Besançon France

Université de Franche Comté CHU Besançon SINERGIES Besançon France

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