FcRn-enabling strategies for improved thrombolytic therapy

改善溶栓治疗的 FcRn 启用策略

• 批准号: 10406291
• 负责人: Patrick McKenna Glassman
• 金额: $ 15.29万
• 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: 2021-06-01 至 2022-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10406291
• 关键词: Acute; Adverse effects; Affect; Affinity; Albumins; Alteplase; Area; Binding; Biodistribution; Biological Response Modifier Therapy; Biomedical Research; Blood; Blood Cells; Blood Circulation; Catabolism; Charge; Chemicals; Clinical; Coagulation Process; Coupling; Cytolysis; Data; Development; Diffusion; Disease; Dose; Drug Delivery Systems; Drug Kinetics; Drug usage; Engineering; Erythrocytes; Event; Eye; Faculty; Fc Receptor; Filtration; Functional disorder; Genetic; Grant; Half-Life; Hemorrhage; Hemostatic Agents; Hemostatic function; Immunoglobulin G; Individual; Infusion procedures; Injections; Intervention; Ischemic Stroke; Kidney; Kinetics; Knockout Mice; Knowledge; Label; Libraries; Life; Life Extension; Ligand Binding; Ligands; Mediating; Mentors; Modeling; Molecular Conformation; Mus; Mutation; Myocardial Infarction; Nature; Pathologic; Pathway interactions; Patient-Focused Outcomes; Penetration; Pennsylvania; Pharmaceutical Preparations; Pharmacodynamics; Pharmacological Treatment; Pharmacology; Physiological; Plasma; Plasma Proteins; Plasminogen Activator; Polymers; Property; Proteins; Pulmonary Embolism; Recycling; Research; Resources; Risk; Role; Safety; Serum Proteins; Structure; Testing; Therapeutic; Therapeutic Uses; Thrombin; Thrombolytic Therapy; Thrombosis; Thrombus; Time; Transcend; Universities; Urokinase; antibody engineering; base; biological systems; blood fractionation; compliance behavior; cost; design; improved; in silico; in vivo; inhibitor; insight; mathematical model; member; mouse model; mutant; nanobodies; neonatal Fc receptor; novel; predictive modeling; prophylactic; receptor; risk benefit ratio; side effect; therapeutic protein; thrombolysis; thrombotic; treatment strategy; uptake

PROJECT SUMMARY/ABSTRACT While clinical use of therapeutic proteins has grown exponentially in recent decades, utility is often limited by unfavorable pharmacokinetics (PK), mediated by rapid elimination. One approach to overcome this limitation is half-life extension (HLE), achieved by attachment of biotherapeutics to polymers (PEG), plasma proteins (albumin, IgG), and blood cells. There is a paucity of data describing the impact of properties of cargo proteins and HLE-conferring ligands on PK of proteins tested using these strategies. One area where HLE has the potential to advance clinical therapy is acute, life-threatening, thrombosis (pulmonary embolism, ischemic stroke, etc.). The only pharmacologic treatment available for treatment of this condition is infusion of plasminogen activators, which have an unfavorable pharmacologic profile, mediated by a half-life of minutes, rapid inactivation in plasma, and severe adverse effects (e.g. hemorrhagic transformation). Pilot data demonstrates that conjugation of a derivative of urokinase selectively activated in thrombin-rich pathological thrombi (scuPA-T) to an albumin-binding nanobody (Nb) leads to PK that is identical to RBC-binding scuPA-T, an approach pioneered by Drs. Muzykantov and Cines, albeit with distribution in the plasma rather than the cellular fraction of blood. The central hypothesis of this proposal is that coupling of scuPA-T to albumin-binding ligands will provide prolonged circulation, mediated by the neonatal Fc receptor (FcRn), and selective thrombolysis of pathological thrombi, sparing hemostatic plugs from lysis. In the mentored stage, the impact of properties of the cargo drug on FcRn-enabled HLE will be identified, using FcRn knockout mice to directly elucidate the FcRn-mediated component of HLE conferred by albumin binding (Aim 1; K99). Based on pilot data, safety and efficacy of albumin-binding scuPA-T will be studied in a mouse model of pulmonary embolism. These results will be used to develop a predictive modeling platform that will be used to for further engineering and PK optimization of HLE-scuPA-T constructs (Aim 2; K99). With an eye on mechanistic and translational advancement of this strategy, the role of albumin-binding affinity on the PK of scuPA-T will be defined, using a newly developed library of nanobodies (Aim 3; R00). Additionally, thrombin-cleavable HLE ligands will be devised, permitting selective release of scuPA-T in thrombi, improving diffusion into clots and lysis (Aim 3; R00). Overarching themes of this proposal include identification of critical features of albumin-mediated HLE and mechanism-based modeling to guide optimization and reengineering of protein therapeutics. A mentoring team has been identified with expertise spanning the areas of research in this grant, namely, thrombosis and hemostasis, antibody engineering, and mathematical modeling of biological systems. Mentored research will be conducted at the University of Pennsylvania, which has a highly collegial and collaborative faculty and extensive resources available to conduct the proposed research. This proposal is geared towards gaining the expertise necessary to be successful as a tenured faculty member conducting high quality biomedical research.

项目摘要/摘要 尽管近几十年来治疗性蛋白质的临床应用呈指数级增长，但其实用性往往受到以下因素的限制 不良药代动力学(PK)，由快速消除介导。克服这一限制的一种方法是 半衰期延长(HLE)，通过将生物治疗药物附着到聚合物(PEG)、血浆蛋白上实现的 (白蛋白、免疫球蛋白)和血细胞。描述货运蛋白性质的影响的数据很少。 以及使用这些策略测试的蛋白质的PK上的HLE授权配体。HLE在一个领域拥有 促进临床治疗的潜力是急性的、危及生命的血栓形成(肺栓塞、缺血性中风、 等)。治疗这种疾病的唯一有效药物治疗方法是输注纤溶酶原。 激活剂，具有不利的药理学特征，由几分钟的半衰期介导，迅速失活 在血浆中，和严重的不良反应(如出血性转化)。试点数据表明， 富凝血酶病理性血栓中选择性激活的尿激酶衍生物(scuPA-T)与 白蛋白结合纳米体(NB)导致与RBC结合的scuPA-T相同的PK，这是一种首创的方法 Muzykantov博士和Cines博士提出的，尽管分布在血浆中，而不是血液的细胞部分。 这一提议的中心假设是scuPA-T与白蛋白结合配体的偶联将 提供由新生儿Fc受体(FcRN)介导的延长循环和选择性溶栓 防止病理性血栓的形成，避免止血塞溶解。在指导阶段， 将利用FcRN基因敲除小鼠直接检测FcRN基因敲除的HLE上的药物性质 阐明FcRN介导的白蛋白结合所致HLE的成分(目标1；K99)。基于飞行员 白蛋白结合scuPA-T的数据、安全性和有效性将在小鼠肺栓塞模型中进行研究。 这些结果将被用于开发预测建模平台，该平台将用于进一步的工程 和HLE-scuPA-T结构的PK优化(目标2；K99)。着眼于机械化和平移化 在这一策略的推进下，白蛋白结合亲和力在scuPA-T的PK上的作用将被定义，使用 新开发的纳米体库(目标3；R00)。此外，凝血酶可切割的HLE配体将是 设计，允许scuPA-T在血栓中选择性释放，改善扩散到血栓和溶解(目标3；R00)。 这项建议的主要主题包括确定白蛋白介导的HLE的关键特征和 基于机理的建模，以指导蛋白质疗法的优化和重组。一个指导团队 已被认定为拥有这笔赠款研究领域的专业知识，即血栓形成和 止血、抗体工程和生物系统的数学建模。有指导的研究将是 在宾夕法尼亚大学进行，该校拥有高度合作的教职员工和广泛的 可用于开展拟议研究的资源。这项建议旨在获得专业知识。 作为一名从事高质量生物医学研究的终身教员，成功所必需的。