BC-mediated delivery of thromboprophylaxis

BC 介导的血栓预防

基本信息

批准号：
10475755
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
项目类别：
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/10475755
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 Circulation Blood Platelets Blood Vessels Brain Injuries COVID-19 Cell physiology Cells 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 of activation protein Medical Microfluidics Modeling Modification Molecular Mus Operative Surgical Procedures Outcome Pathologic Patients 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 base biomaterial compatibility clinical application clinical translation design high risk in vivo in vivo Model insight interest mouse model mutant novel novel strategies off-target site pre-clinical preservation prevent prophylactic prototype risk benefit ratio targeted agent thromboinflammation translational study uptake

项目摘要

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

对于许多面临以下综合风险的患者来说，没有一种抗血栓药物 (ATA) 是安全有效的。急性血栓形成和出血，例如在术后早期。为了解决这个未满足的需求，我们开发药物输送系统 (DDS)，执行两个主要功能： A) 阻止 ATA 访问非目标部位，例如手术后形成的止血塞，同时 B) 优化药代动力学和将 ATA 递送至随后的血栓中，其中 ATA 被凝血酶激活。 ATA 融合单靶向红细胞 (RBC) 的链片段 (scFv) 与这些执行双重功能的载体结合阻塞/传递功能。概念验证正在现有和新生血栓模型中出现在动物中。在这里，我们设计了针对人类红细胞的人源化 scFv/ATA 并将在人源化微流体系统（HMF），在表达人源化靶表位的转基因（TG）小鼠中血细胞以及离体人肺的灌注中。我们将追求三个目标。目标 1. 红细胞加载中。我们将表征 scFv/ATA 加载到 RBC 上的特征：A) 结合（副本/细胞，开/关动力学）；二）对红细胞功能、生物相容性和生物力学的影响；以及，B) 分配的监管循环中红细胞之间的 scFv/ATA。我们还将表征调节生物力学因素 RBC/ATA 输送及其对血块动力学和结构的影响，特别是 RBC 的影响僵化，由药物负荷或患者内在病理生理变化引起血。目标 2. 机制见解。我们将审问以前未被认识但批判性的 RBC/ATA 运作的各个方面，特别是它们与血管内皮的相互作用以及将药物货物转移到这些和其他血管细胞。在此目标中，我们将使用标准鼠标体内模型、微流体模型和离体人肺灌注模型。目标 3. 评估效益/风险比。我们正在开发在小鼠 EBC 中表达人类红细胞决定簇的 TM 小鼠，为了研究体内“人红细胞”上负载的 scFv/ATA：A) PK/BD、补体激活， TG小鼠RBC/ATA的吞噬细胞摄取和血管粘附； B) 定义时间窗口/范围 TG 小鼠动脉与静脉血栓形成模型中人 RBC/ATA 的抗血栓作用； B) 确认RBC/ATA的安全性。我们将检测 scFv/ATA 的逆境，包括异常红细胞。为了化解潜在的问题，如有必要，我们将使用更良性的加载方案。一起，这些研究将推进一种新的缓解方法的机制见解和临床转化通过提供一种新的、易于处理的方法来治疗目前未受保护的患者的血栓形成了解血栓的形成和提供细胞定向治疗的合理方法