Jang Joara, Yu Hyunseong, Oh Eun Bee, Park Ji Won, Kim Solee, Kim Taeryeong, Sohn Jisue, Jin Bo-Ram, Chang Tong-Shin
College of Pharmacy, Seoul National University, Seoul, Republic of Korea.
Division of Hematology, Department of Medicine, Washington University School of Medicine, St. Louis, Missouri, USA.
Antioxid Redox Signal. 2025 Aug;43(4-6):288-307. doi: 10.1089/ars.2024.0695. Epub 2025 Apr 4.
NADPH oxidase (NOX)-derived reactive oxygen species (ROS) are critical for platelet activation and thrombus formation. We hypothesized that inhibiting NOX-mediated ROS production with a pan-NOX inhibitor, APX-115, could effectively suppress platelet activation and thrombus formation, potentially serving as a novel antiplatelet therapeutic. This study aimed to explore the effects of APX-115 on human platelet functional responses and ROS-mediated signaling pathways. APX-115 inhibited intracellular and extracellular ROS production in collagen-stimulated platelets, suppressing aggregation, P-selectin exposure, and ATP release. By preserving protein tyrosine phosphatase activity, APX-115 reduced tyrosine phosphorylation-dependent pathways inhibition, including spleen tyrosine kinase, LAT, Vav1, Bruton's tyrosine kinase, and phospholipase Cγ2, leading to decreased PKC activation and calcium mobilization. APX-115 also suppressed collagen-induced integrin αIIbβ3 activation, accompanied by elevated cGMP and vasodilator-stimulated phosphoprotein phosphorylation levels. In addition, APX-115 reduced p38 MAPK and ERK5 activation, leading to diminished phospholipase A2 phosphorylation, thromboxane production, and the exposure of procoagulant phosphatidylserine. These inhibitory effects extended to thrombus development caused by platelet adherence under shear and arterial thrombosis without prolonging bleeding time in murine models. This study is the first to demonstrate that APX-115 inhibits NOX-mediated ROS production, platelet activation, and thrombus formation. By uncovering its effects on collagen receptor glycoprotein VI-mediated pathways, the work highlights the promise of APX-115 as an antiplatelet and antithrombotic agent. Our findings highlight the therapeutic potential of APX-115 in treating thrombotic and cardiovascular disorders by targeting NOX-mediated ROS production to mitigate platelet hyperreactivity and thrombus formation. 43, 288-307.
烟酰胺腺嘌呤二核苷酸磷酸氧化酶（NOX）衍生的活性氧（ROS）对于血小板活化和血栓形成至关重要。我们推测，用泛NOX抑制剂APX-115抑制NOX介导的ROS产生可有效抑制血小板活化和血栓形成，有可能成为一种新型抗血小板治疗方法。本研究旨在探讨APX-115对人血小板功能反应和ROS介导的信号通路的影响。APX-115抑制胶原刺激的血小板中细胞内和细胞外ROS的产生，抑制聚集、P-选择素暴露和ATP释放。通过保留蛋白酪氨酸磷酸酶活性，APX-115减少了酪氨酸磷酸化依赖性途径的抑制，包括脾酪氨酸激酶、LAT、Vav1、布鲁顿酪氨酸激酶和磷脂酶Cγ2，导致蛋白激酶C活化和钙动员减少。APX-115还抑制胶原诱导的整合素αIIbβ3活化，同时伴有cGMP升高和血管舒张刺激磷蛋白磷酸化水平升高。此外，APX-115降低p38丝裂原活化蛋白激酶和ERK5活化，导致磷脂酶A2磷酸化、血栓素产生减少以及促凝磷脂酰丝氨酸暴露减少。这些抑制作用扩展到小鼠模型中由剪切力下血小板黏附引起的血栓形成和动脉血栓形成，而不会延长出血时间。本研究首次证明APX-115抑制NOX介导的ROS产生、血小板活化和血栓形成。通过揭示其对胶原受体糖蛋白VI介导途径的影响，这项工作突出了APX-115作为抗血小板和抗血栓药物的前景。我们的研究结果突出了APX-115通过靶向NOX介导的ROS产生以减轻血小板高反应性和血栓形成来治疗血栓性和心血管疾病的治疗潜力。43, 288 - 307。
