Identification of an inhibitor of PDI-GPIbalpha signaling as a novel antithromboinflammatory agent

鉴定 PDI-GPIbalpha 信号传导抑制剂作为新型抗血栓炎症剂

• 批准号: 10242945
• 负责人: Jaehyung Cho
• 金额: $ 55.13万
• 依托单位: WASHINGTON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-04 至 2023-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10242945
• 关键词: Affect; Animal Model; Animals; Anti-Inflammatory Agents; Antibodies; Asia; Back; Binding; Bleeding time procedure; Blocking Antibodies; Blood Platelets; Blood Vessels; Blood flow; Canis familiaris; Cardiovascular Diseases; Cessation of life; Chemicals; Computer Assisted; Data; Development; Disease; Dose; Drug Design; Drug Kinetics; Endothelium; Europe; Exhibits; Extracellular Protein; Future; Glycoproteins; Goals; Hemorrhage; Hemostatic function; Immune response; Impairment; In Vitro; Inflammation; Injections; Integrins; Investigational New Drug Application; Knockout Mice; Lead; Leukocytes; Ligand Binding; Ligands; Macrophage-1 Antigen; Mediating; Megakaryocytes; Molecular Conformation; Mus; Myocardial Infarction; Neutrophil Infiltration; Pathologic; Pathology; Patients; Peripheral; Pharmaceutical Preparations; Platelet Adhesiveness; Platelet aggregation; Protein Disulfide Isomerase; Protein Inhibition; Risk; Series; Signal Pathway; Signal Transduction; Signaling Molecule; Specificity; Stroke; Surface; Tail; Techniques; Technology; Tertiary Protein Structure; Testing; Therapeutic; Thrombosis; Thrombus; Tissues; Toxic effect; Vasculitis; atherothrombosis; conditional knockout; drug metabolism; efficacy study; high throughput screening; improved; in vivo; inhibitor/antagonist; mouse model; neutrophil; nonhuman primate; novel; novel therapeutics; platelet function; preclinical study; prevent; receptor; scaffold; small molecule; small molecule inhibitor; targeted treatment; therapeutically effective; thromboinflammation; vascular inflammation

Project Summary Thromboinflammatory diseases, including atherothrombosis, stroke and peripheral vasculitis, result in >30% of all deaths globally. Underlying the pathology of thromboinflammation is increased adhesiveness of platelets and leukocytes. Although many antiplatelet and anti-inflammatory therapies have been used for disease treatment, these drugs increase the risk of major bleeding or impair immune responses. Using protein disulfide isomerase (PDI) conditional knockout (CKO) mice and inhibitors, we and others have shown that extracellular PDI is crucial for platelet thrombus formation in arterial thrombosis and neutrophil recruitment to inflamed endothelium in vascular inflammation. However, inhibition of extracellular PDI with a function-blocking antibody prolongs tail bleeding times in mice, raising a concern that specific inhibition of PDI may perturb hemostatic function. Our recent studies have demonstrated that platelet-released PDI promotes the ligand-binding function of glycoprotein Ibα (GPIbα) and enhances GPIbα-mediated platelet adhesiveness, platelet-neutrophil aggregation and vascular occlusion under thromboinflammatory conditions. These results suggest that inhibitors blocking the PDI-GPIbα signaling axis may be a novel antithromboinflammatory drug. Using high throughput screening, we have identified one compound that specifically inhibits PDI-GPIbα binding and GPIbα-mediated platelet aggregation. We have found that iv injection of the compound into mice abolishes platelet-neutrophil interactions and improves blood flow rates in microvessels under thromboinflammatory conditions. Unlike a conventional inhibitor of PDI or GPIbα, treatment with our compound does not prolong tail bleeding times in mice. These results have provided evidence for the feasibility of identifying small-molecule inhibitors targeting a specific PDI signaling pathway. In Aim 1, using the computer-aided drug design technique and a series of in vitro studies, we will identify small- molecule compounds that specifically block PDI-GPIbα signaling and GPIbα-mediated platelet functions. In Aim 2, we will synthesize derivatives of hits, test them in animal studies and examine DMPK profiles of the selected compounds. The proposed study will prove that compared to conventional inhibition of PDI or GPIbα, specific inhibition of the PDI-GPIbα signaling axis might be a safer and effective therapeutic strategy for treating thromboinflammatory disease.

项目概要 血栓炎性疾病，包括动脉粥样硬化血栓形成、中风和外周血管炎，导致>30% 全球所有死亡。血栓炎症病理学的基础是血小板和血小板粘附力的增加。 白细胞。尽管许多抗血小板和抗炎疗法已被用于疾病治疗， 这些药物会增加大出血的风险或损害免疫反应。使用蛋白质二硫键异构酶 (PDI) 条件敲除 (CKO) 小鼠和抑制剂，我们和其他人已经证明细胞外 PDI 至关重要 用于动脉血栓形成中的血小板血栓形成和中性粒细胞募集至发炎的内皮细胞 血管炎症。然而，用功能阻断抗体抑制细胞外 PDI 会延长尾部 小鼠的出血时间，引起人们对 PDI 的特异性抑制可能会干扰止血功能的担忧。我们的 最近的研究表明，血小板释放的PDI可促进糖蛋白的配体结合功能 Ibα (GPIbα) 并增强 GPIbα 介导的血小板粘附性、血小板中性粒细胞聚集和血管 血栓炎症条件下的闭塞。这些结果表明抑制剂阻断 PDI-GPIbα 信号轴可能是一种新型的抗血栓炎症药物。通过高通量筛选，我们有 鉴定出一种特异性抑制 PDI-GPIbα 结合和 GPIbα 介导的血小板聚集的化合物。 我们发现，向小鼠静脉注射该化合物可以消除血小板与中性粒细胞的相互作用，并改善 血栓炎症条件下微血管的血流量。与传统的 PDI 抑制剂或 GPIbα，用我们的化合物治疗不会延长小鼠尾部出血时间。这些结果提供了 鉴定针对特定 PDI 信号通路的小分子抑制剂的可行性证据。在 目标1，利用计算机辅助药物设计技术和一系列体外研究，我们将识别小 特异性阻断 PDI-GPIbα 信号传导和 GPIbα 介导的血小板功能的分子化合物。瞄准 2、我们将合成命中的衍生物，在动物研究中对其进行测试，并检查所选药物的 DMPK 配置文件 化合物。拟议的研究将证明，与 PDI 或 GPIbα 的常规抑制相比，特异性 抑制 PDI-GPIbα 信号轴可能是一种更安全有效的治疗策略 血栓炎性疾病。