Platelet-inspired Delivery System for Targeted Thrombolytic Therapy

用于靶向溶栓治疗的血小板启发输送系统

• 批准号: 9127360
• 负责人: Anirban Sen Gupta
• 金额: $ 46.5万
• 依托单位: CASE WESTERN RESERVE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-09-01 至 2019-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9127360
• 关键词: Adverse effects; Affect; Aftercare; Age; Albumins; Alteplase; Binding; Biocompatible; Blood Platelets; Blood Vessels; Blood flow; Caliber; Carotid Artery Thrombosis; Catheters; Charge; Chromogenic Substrates; Clinic; Clinical; Clinical Research; Coagulation Process; Cues; Drug Delivery Systems; Drug effect disorder; Drug vehicle; Embolism; Engineering; Excision; Fibrinogen; Fibrinolytic Agents; Fluorescence; Fluorescence Microscopy; Goals; Half-Life; Health; Hemorrhage; Imagery; In Vitro; Injectable; Injury; Integrins; Ischemic Stroke; Isoelectric Point; Kinetics; Label; Ligands; Mechanics; Mediating; Microscopy; Modeling; Morbidity - disease rate; Mus; Myocardial Infarction; P-Selectin; P-selectin ligand protein; Particulate; Pathology; Peptides; Perfusion; Peripheral arterial disease; Pharmaceutical Preparations; Pharmacologic Substance; Physiological; Plasma; Plasminogen Activator; Platelet aggregation; Procedures; Property; Research; Risk; Shapes; Site; Spectrophotometry; Stroke; Surface; System; Technology; Testing; Therapeutic; Thrombolytic Therapy; Thrombus; Tissues; Treatment Efficacy; Vascular Diseases; aqueous; base; blood flow measurement; controlled release; density; design; drug distribution; flexibility; hemodynamics; high risk; in vitro testing; in vivo; innovation; interest; intravital microscopy; microscopic imaging; mimetics; mortality; mouse model; particle; prevent; receptor; restoration; stem; targeted treatment; tegrin; thrombolysis

 DESCRIPTION: In occlusive vascular diseases, dissolution of thrombus and rapid restoration of blood flow is critical in preventing tissue morbidity and mortality. To this end, current clinicl `clot-busting' strategies using intravascular systemic administration of thrombolytic drugs like tPA present issues of plasma-induced drug deactivation, short plasma half-life of drugs, rapid drug washout, reduced drug availability at target site, and systemic hemorrhagic side effects. These issues can be potentially resolved by controlled release of the drugs from injectable delivery vehicles that can selectively accumulate at the clot site under hemodynamic flow. For this, it is necessary for the vehicles to (i) actively bind to the clot site, (ii) stay retained uner flow, and (iii) allow localized drug release for site-selective action. For designing such a vehicl, natural platelets provide an excellent paradigm, since platelets have the innate capability to marginate towards the vascular wall, bind actively to clot via specific ligand-receptor interactions, and release cytoplasmic contents to modulate clot activities. The margination of platelets towards the vascular wall is facilitated by their discoid shape, ~2 µ diameter size and ~25-50 kPa modulus, while their active binding at the thrombus site is mediated via ligands fibrinogen (Fg) and PSGL-1 binding to platelet surface integrin GPIIb-IIIa and P-selectin respectively. Inspired by these physico-mechanical and biointeractive cues of natural platelets, we propose to engineer vehicles that will have (i) platelet-mimetic shape, size and modulus to facilitate enhanced margination towards the vascular wall, and (ii) heteromultivalent surface-decoration with GPIIb-IIIa- and P-selectin-binding peptide ligands to enable thrombus-selective anchorage and retention under hemodynamic flow. Our central hypothesis is that controlled release of thrombolytic drugs from such platelet-inspired vehicles anchoring at the thrombus site will enhance the site-specific therapeutic efficacy, while minimizing systemic drug distribution and side-effects. To test this, we will fabricate albumin-based platelet-inspired discoid flexible vehicles, modify their surface heteromultivalently with the active platelet-targeting ligands and evaluate their thrombus anchoring capability in vitro and in vivo (Aim1). In parallel, we will cationically modify tPA (mtPA) to facilitate its loading into the albumin particles since albumin i negatively charged at physiological pH, and characterize the loading and release of mtPA from these particles in vitro (Aim 2). After optimizing the platelet-inspired thrombus- anchoring mechanisms and the mtPA loading/release kinetics via these two synergistic yet independent aims, we will integrate them to create mtPA-loaded platelet-inspired vehicles which will be evaluated for targeted thrombolysis efficacy in vitro and in vivo (Aim 3). Our principal innovation is the design of a delivery vehicle that combines key physico-mechanical and biointeractive parameters inspired by platelets for enhanced thrombus-anchoring ability. The current application will test this technology for targeted thrombolytic therapy. The technology also holds the promise to become a platform for other targeted therapies in vascular diseases.

 描述：在闭塞性血管疾病中，血栓的溶解和血流的快速恢复对于防止组织发病率和死亡率至关重要。为此，使用血管内溶解药物（如TPA）的血管内全身给药的当前临床“凝块破坏”策略提出了血浆诱导的药物失活的问题，较短的药物血浆半衰期，快速药物冲洗，降低目标部位的药物可用性以及全身性出血副作用。这些问题可以通过从可注射的输送车中受控释放药物来解决这些问题，这些释放可以在血流动力学流动下选择性地积累在凝块部位。为此，车辆有必要（i）积极地与凝块部位结合，（ii）保持释放的河流，（iii）允许局部药物释放以进行现场选择性作用。为了设计这种车辆，天然血小板提供了极好的范式，因为血小板具有天生的能力，可以通过特定的配体 - 受体相互作用积极地与凝块结合，并释放细胞质含量以调节凝块活性。血小板向血管壁的边缘由它们的盘状形状（直径约为2 µ和〜25-50 kPa模量）支撑，而它们在血栓部位的积极结合是通过配体纤维蛋白原（FG）（FG）和PSGL-1介导的，PSGL-1与Platelet Surface Interafer interical Enteribib-iib-iiiA和P-iia和p-iia和P链蛋白均相应地介导。 Inspired by these physical-mechanical and biointeractive cues of natural platelets, we propose to engineer vehicles that will have (i) platelet-mimetic shape, size and modulus to facilitate enhanced margination towards the vascular wall, and (ii) heteromultivalent surface-decoration with GPIIb-IIIa- and P-selectin-binding pepper ligands to enable thrombus-selective anchorage and retention under血流动力学流动。我们的中心假设是，从锚定在血栓部位的这种血小板启发的车辆中释放了溶栓药物将提高特定部位的治疗有效性，同时最大程度地减少全身药物分布和副作用。为了测试这一点，我们将制造基于专辑的血小板启发的盘式柔性车辆，使用活跃的血小板靶向配体对其表面异质化，并评估其在体外和体内的血栓锚固能力（AIM1）。同时，我们将在阳离子上修改TPA（MTPA），以促进其从白蛋白I产生pH值的白蛋白颗粒的负载，并表征体外这些颗粒中MTPA的负载和释放（AIM 2）。通过这两个协同的但独立的目标优化了血小板启发的血栓锚定机制和MTPA载荷/释放动力学，我们将整合它们以创建由MTPA负载的血小板启发的车辆，这些车辆将评估针对靶向血栓溶液效率的靶向血栓化效率，并在体外和In Vivo中（AIM 3）。我们的主要创新 是通过由血小板增强的血小板锚定能力启发的关键物理机械和生物互动参数的运输工具的设计。当前的应用将测试该技术的靶向溶栓疗法。该技术还承诺成为血管疾病其他靶向疗法的平台。