NADPH oxidase 2 inhibitor GSK2795039 exerts antiplatelet and antithrombotic activity

Abstract

Abstract Reactive oxygen species (ROS) production by NADPH oxidase (NOX) is a key promoter of platelet activation, making NOX inhibition an attractive antiplatelet strategy. This study evaluated the effects of the novel NOX2 inhibitor, GSK2795039, on human platelet functional responses and ROS-mediated signaling pathways. GSK2795039 effectively suppressed both extracellular and intracellular ROS production in collagen-stimulated platelets. Consequently, GSK2795039 treatment significantly inhibited collagen-induced platelet aggregation, dense-granule ATP release, and α-granule P-selectin exposure. Furthermore, GSK2795039 attenuated tyrosine phosphorylation-mediated activation of spleen tyrosine kinase (Syk), linker for the activation of T cells (LAT), vav guanine nucleotide exchange factor 1 (Vav1), and Bruton tyrosine kinase (Btk) within the collagen receptor signaling pathway, leading to decreased phospholipase Cγ2 (PLCγ2) activation and calcium mobilization. GSK2795039 also inhibited collagen-induced integrin αIIbβ3 activation, associated with increased cyclic guanosine monophosphate (cGMP) levels and vasodilator-stimulated phosphoprotein (VASP) phosphorylation. Additionally, the inhibitor reduced collagen-induced p38 mitogen-activated protein kinase (MAPK) activation, which led to lower cytosolic phospholipase A2 (cPLA2) phosphorylation and thromboxane production. GSK2795039 further decreased extracellular signal regulated kinase 5 (ERK5) activation, thereby limiting procoagulant phosphatidylserine exposure. Notably, thrombus formation induced by platelet adhesion to collagen was abolished in the presence of GSK2795039. In vivo, GSK2795039 administration markedly inhibited arterial thrombosis. This is the first study to demonstrate that GSK2795039 suppresses collagen-induced ROS production, platelet activation, and thrombus formation, highlighting its potential as a therapeutic agent for thrombotic and cardiovascular diseases.