BC-mediated delivery of thromboprophylaxis

BC 介导的血栓预防

• 批准号: 10652489
• 负责人: Vladimir R Muzykantov
• 金额: $ 66.66万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-01 至 2025-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10652489
• 关键词: Abnormal Red Blood Cell; Acute; Address; Adhesions; Adhesiveness; Animal Model; Animals; Antiinflammatory Effect; Benign; Binding; Biodistribution; Biological Products; Biomechanics; Blood; Blood Cells; Blood Platelets; Blood Vessels; Brain Injuries; COVID-19; Cells; Circulation; Clinic; Clinical; Clinical Research; Coagulation Process; Complement Activation; Complex; Coupled; Coupling; Cytolysis; Data; Development; Disease; Dose; Drug Delivery Systems; Drug Kinetics; Embolism; Endothelium; Endotoxemia; Epitopes; Erythrocytes; Fibrinolysis; Fibrinolytic Agents; Glycoproteins; Goals; Hemorrhage; Hemostatic Agents; Hour; Human; Immobilization; Impairment; Inflammation; Inflammatory; Injections; Intravenous infusion procedures; Kinetics; Lead; Leg; Life; Lung; Mediating; Mediator; Medical; Microfluidics; Modeling; Modification; Molecular; Mus; Operative Surgical Procedures; Outcome; Pathologic; Patients; Perfusion; Phagocytes; Pharmaceutical Preparations; Physicians; Pilot Projects; Plasminogen Activator; Postoperative Period; Primates; Prophylactic treatment; Regimen; Regional Perfusion; Regulation; Rhesus; Risk; Safety; Site; Source; Structure; Surgical Hemostasis; System; Testing; Therapeutic; Thrombin; Thrombomodulin; Thromboplastin; Thrombosis; Thrombus; Time; Transgenic Mice; Transgenic Organisms; Trauma; Urokinase; Vascular Diseases; Vascular Endothelium; Venous Thrombosis; Work; activated Protein C; biomaterial compatibility; biomechanical test; clinical application; clinical translation; design; high risk; in vivo; in vivo Model; insight; interest; invention; mouse model; mutant; novel; novel strategies; off-target site; pre-clinical; preservation; prevent; prophylactic; prototype; risk/benefit ratio; targeted agent; thromboinflammation; thrombotic; translational study; uptake; Abnormal Red Blood Cell; Acute; Address; Adhesions; Adhesiveness; Animal Model; Animals; Antiinflammatory Effect; Benign; Binding; Biodistribution; Biological Products; Biomechanics; Blood; Blood Cells; Blood Platelets; Blood Vessels; Brain Injuries; COVID-19; Cells; Circulation; Clinic; Clinical; Clinical Research; Coagulation Process; Complement Activation; Complex; Coupled; Coupling; Cytolysis; Data; Development; Disease; Dose; Drug Delivery Systems; Drug Kinetics; Embolism; Endothelium; Endotoxemia; Epitopes; Erythrocytes; Fibrinolysis; Fibrinolytic Agents; Glycoproteins; Goals; Hemorrhage; Hemostatic Agents; Hour; Human; Immobilization; Impairment; Inflammation; Inflammatory; Injections; Intravenous infusion procedures; Kinetics; Lead; Leg; Life; Lung; Mediating; Mediator; Medical; Microfluidics; Modeling; Modification; Molecular; Mus; Operative Surgical Procedures; Outcome; Pathologic; Patients; Perfusion; Phagocytes; Pharmaceutical Preparations; Physicians; Pilot Projects; Plasminogen Activator; Postoperative Period; Primates; Prophylactic treatment; Regimen; Regional Perfusion; Regulation; Rhesus; Risk; Safety; Site; Source; Structure; Surgical Hemostasis; System; Testing; Therapeutic; Thrombin; Thrombomodulin; Thromboplastin; Thrombosis; Thrombus; Time; Transgenic Mice; Transgenic Organisms; Trauma; Urokinase; Vascular Diseases; Vascular Endothelium; Venous Thrombosis; Work; activated Protein C; biomaterial compatibility; biomechanical test; clinical application; clinical translation; design; high risk; in vivo; in vivo Model; insight; interest; invention; mouse model; mutant; novel; novel strategies; off-target site; pre-clinical; preservation; prevent; prophylactic; prototype; risk/benefit ratio; targeted agent; thromboinflammation; thrombotic; translational study; uptake

No anti-thrombotic agent (ATA) is safe and effective in the many patients at a combined risk of acute thrombosis and bleeding, e.g., in the early post-surgery period. To address this unmet need, we develop drug delivery systems (DDS) executing two main functions: A) Block access of ATA to off- target sites, e.g., hemostatic plugs formed after surgery, while B) Optimize pharmacokinetics and deliver ATA into subsequent thrombi, where ATA is activated by thrombin. ATA fused with single- chain fragments (scFv) targeted to red blood cells (RBC) bind to these carriers that execute dual blocking/delivering function. Proof-of-concept is emerging in models of pre-existing and nascent clots in animals. Here we devise humanized scFv/ATA targeted to human RBC and will test them in a humanized microfluidic system (HMF), in transgenic (TG) mice expressing humanized target epitopes on blood cells, and in the perfusion of isolated human lungs. We will pursue three aims. Aim 1. RBC loading. We will characterize scFv/ATA loading onto RBC: A) Binding (copies/cell, on/off kinetics); B) Effect on RBC functionality, biocompatibility and biomechanics; and, B) Regulation of distribution of scFv/ATA between RBC in circulation. We also will characterize biomechanical factors modulating RBC/ATA delivery and effect on clot dynamics and structure, in particular, impact of RBC rigidification, caused by either drug loading or by intrinsic pathophysiological changes in patient's blood. Aim 2. Mechanistic insights. We will interrogate previously unrecognized yet critically aspects of the RBC/ATA workings, in particular their interaction with vascular endothelium and transfer of the drug cargo to these and other vascular cells. In this Aim we will use standard mouse in vivo models, microfluidic model and perfusion of isolated human lungs model. Aim 3. Appraisal of benefit/risk ratio. We are developing TM mice expressing human RBC determinants in mouse EBC, in order to study scFv/ATA loaded on "human RBC" in vivo: A) PK/BD, complement activation, phagocyte uptake and vascular adhesion of RBC/ATA in TG mice; B) Define the time window/extent of anti-thrombotic effect of human RBC/ATA in models of arterial vs venous thrombosis in TG mice; B) Affirm the safety of RBC/ATA. We will detect adversities of scFv/ATA including abnormalities of RBC. To defuse potential issues, if necessary, we will use more benign loading regimen. Together, these studies will advance mechanistic insights and clinical translation of a novel way to mitigate thrombosis in currently unprotected patients by providing a new and tractable approach to understanding thrombus development and a rational approach to deliver cell-directed therapeutics

在许多患者中，没有抗栓性剂（ATA）是安全有效的 急性血栓形成和出血，例如在手术后早期。为了满足这个未满足的需求，我们 开发执行两个主要功能的药物输送系统（DDS）：a）ATA访问到外 靶位部位，例如手术后形成的止血塞，而b）优化药代动力学和 将ATA输送到随后的血栓中，其中ATA被凝血酶激活。 ATA与单人融合 针对红细胞（RBC）的链片段（SCFV）与这些执行双重的载体结合 阻止/交付功能。概念证明正在出现在现有和新生凝块的模型中 在动物中。在这里，我们设计了针对人RBC的人源化SCFV/ATA，并将在 人源化微流体系统（HMF），在表达人源化靶位表位的转基因（TG）小鼠中 在血细胞上，并灌注分离的人肺。我们将追求三个目标。目标1。RBC 加载中。我们将表征加载到RBC上的SCFV/ATA：a）绑定（副本/单元格，开/关化动力学）； b） 对RBC功能，生物相容性和生物力学的影响； b）调节分布的调节 RBC循环中的SCFV/ATA。我们还将表征调节的生物力学因子 RBC/ATA传递以及对凝块动力学和结构的影响，特别是RBC的影响 严格化，是由药物加载或患者内在的病理生理变化引起的 血。目标2。机械见解。我们将询问以前未被认可但批判性的 RBC/ATA工作的各个方面，特别是它们与血管内皮和 将货物转移到这些和其他血管细胞。在此目的中，我们将使用标准鼠标 分离的人肺模型的体内模型，微流体模型和灌注。目标3。评估 利益/风险比率。我们正在开发在小鼠EBC中表达人RBC决定因素的TM小鼠， 为了研究体内“人RBC”上加载的SCFV/ATA：A）PK/BD，补体激活， TG小鼠中RBC/ATA的吞噬细胞摄取和血管粘附； b）定义时间窗口/范围 人类RBC/ATA在TG小鼠中动脉血栓形成模型中人类RBC/ATA的抗栓性作用； b）确认RBC/ATA的安全。我们将检测到SCFV/ATA的逆境，包括异常 RBC。为了减轻潜在问题，如有必要，我们将使用更多的良性加载方案。一起， 这些研究将推动一种新型方法的机械见解和临床翻译 目前未受保护的患者的血栓形成，通过提供一种新的可行方法 了解血栓发展和一种合理的方法来提供细胞指导的治疗剂