A NOVEL APPROACH TO TARGET INFLAMMATION AND THROMBOSIS

针对炎症和血栓形成的新方法

• 批准号: 6952334
• 负责人: HARSHAL H NANDURKAR
• 金额: $ 21.6万
• 依托单位: ST. VINCENT'S HOSPITAL, MELBOURNE LTD
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-09-24 至 2008-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6952334
• 关键词: CHO cells; adenosinetriphosphatase; affinity chromatography; antiinflammatory agents; atherosclerosis; biotechnology; cardiovascular disorder chemotherapy; cardiovascular injury; chimeric proteins; drug design /synthesis /production; enzyme activity; genetically modified animals; human subject; inflammation; intermolecular interaction; laboratory mouse; ligands; nonhuman therapy evaluation; platelet aggregation; recombinant proteins; selectins; thromboembolism; thrombosis; venous thrombosis

DESCRIPTION (provided by applicant): Cardiovascular disease is currently the leading cause of morbidity and mortality in developed countries. Chronic lesions of atherosclerosis form the base for the development of acute thrombi, which lead to heart attack and stroke. Currently available therapeutic maneuvers have their limitations. It is now known that atherosclerosis has an important inflammatory component. Interaction between Pselectin expressed on endothelial cells and platelets, and P-selectin glycoprotein ligand-1 (counter receptor) displayed on leukocytes, forms an important mechanism to recruit inflammatory cells to the vessel wall. Inhibition of this interaction helps to maintain vascular patency. CD39/NTPase-1 is a key enzyme expressed on endothelial cells and platelets, which inhibits platelet reactivity and maintains endothelial integrity and CD39-null mice display thrombotic sequelae. Transgenic mice over expressing human CD39 display prolonged bleeding times and protection from collagen-induced vein thrombosis. This proposal will characterize the usefulness of a novel therapeutic agent that will target the platelet inhibitory activity of soluble CD39 to the activated endothelial-platelet microenvironment by incorporation of a PSGL-1 tag (rSolPSGL-Cd39). It is designed to reduce inflammation and thrombus formation in the proximity of injured vessel wall. The novel therapeutic agent with antithrombotic and anti-inflammatory activities will be produced as a recombinant protein and its function will be characterized by in vitro tests of platelet aggregation, binding to activated platelet and endothelial cells, and in vivo testing such as tail bleeding time, and monitoring of pharmacokinetics. The potency of rSolPSGL-CD39 to maintain vascular patency, will be assessed in animal models of mechanical arterial injury and also deep vein thrombosis. The ability of CD39 expression to modify atherosclerosis, will be analyzed in the CD39 transgenic mice and CD39-null mice bred on a background of apolipoprotein E-null and low density lipoprotein receptor-null mice.

描述（由申请人提供）： 心血管疾病目前是发达国家发病和死亡的主要原因。动脉粥样硬化的慢性病变形成急性血栓形成的基础，从而导致心脏病发作和中风。目前可用的治疗方法有其局限性。 现在已知动脉粥样硬化具有重要的炎症成分。内皮细胞和血小板上表达的 P 选择素与白细胞上显示的 P-选择素糖蛋白配体 1（反受体）之间的相互作用，形成了将炎症细胞募集到血管壁的重要机制。 抑制这种相互作用有助于维持血管通畅。 CD39/NTPase-1 是在内皮细胞和血小板上表达的关键酶，可抑制血小板反应性并维持内皮完整性，CD39 缺失小鼠会出现血栓后遗症。过度表达人 CD39 的转基因小鼠表现出延长的出血时间和对胶原诱导的静脉血栓形成的保护。 该提案将描述一种新型治疗剂的有用性，该治疗剂通过掺入 PSGL-1 标签 (rSolPSGL-Cd39)，将可溶性 CD39 的血小板抑制活性靶向激活的内皮-血小板微环境。它旨在减少受伤血管壁附近的炎症和血栓形成。具有抗血栓和抗炎活性的新型治疗剂将以重组蛋白的形式生产，其功能将通过血小板聚集、与活化血小板和内皮细胞结合的体外测试以及尾部出血时间等体内测试来表征。和药代动力学监测。 rSolPSGL-CD39 维持血管通畅的效力将在机械动脉损伤和深静脉血栓形成的动物模型中进行评估。 CD39表达改变动脉粥样硬化的能力将在CD39转基因小鼠和在载脂蛋白E缺失和低密度脂蛋白受体缺失小鼠背景下饲养的CD39缺失小鼠中进行分析。