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.

项目摘要 血栓炎性疾病，包括动脉粥样硬化，中风和周围血管炎，导致30％> 30％ 全球所有死亡。血栓炎的基础病理学是血小板的粘附性提高 尽管许多抗血小板和抗炎疗法已用于疾病治疗，但 这些药物增加了严重出血或损害免疫反应的风险。使用蛋白质二硫异构酶 （PDI）条件敲除（CKO）小鼠和抑制剂，我们和其他人表明细胞外PDI至关重要 用于动脉血栓形成和中性粒细胞募集的血小板血栓形成，以发炎 血管炎症。但是，用功能阻滞抗体抑制细胞外PDI延长尾巴 小鼠的出血时间引起了人们的关注，即对PDI的特定抑制可能会扰动止血功能。我们的 最近的研究表明，血小板发行的PDI促进了糖蛋白的配体结合功能 IBα（GPIBα）并增强了GPIBα介导的血小板粘附性，血小板 - 中性嗜性和血管 血栓炎条件下的阻塞。这些结果表明，阻断PDI-GPIBα的抑制剂 信号轴可能是一种新型的抗滴注药物。使用高吞吐量筛选，我们有 确定了一种特异性抑制PDI-GPIBα结合和GPIBα介导的血小板聚集的化合物。 我们发现，将化合物注射到小鼠中，消除了血小板中性嗜性相互作用并改善 在血栓炎条件下微血管中的血流量。与PDI的常规抑制剂或 GPIBα，用我们的化合物的治疗不会延长小鼠的尾巴出血时间。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的证据。在 AIM 1，使用计算机辅助药物设计技术和一系列体外研究，我们将确定小型 分子化合物专门阻断PDI-GPIBα信号传导和GPIBα介导的血小板功能。目标 2，我们将合成命中的衍生物，在动物研究中测试它们并检查所选的DMPK曲线 化合物。拟议的研究将证明，与常规抑制PDI或GPIBα相比 抑制PDI-GPIBα信号传导轴可能是治疗的更安全，有效的治疗策略 血栓炎性疾病。