FcRn-enabling strategies for improved thrombolytic therapy

改善溶栓治疗的 FcRn 启用策略

• 批准号: 10684075
• 负责人: Patrick McKenna Glassman
• 金额: $ 24.9万
• 依托单位: TEMPLE UNIV OF THE COMMONWEALTH
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-01 至 2025-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10684075
• 关键词: Acute; Adverse effects; Affect; Affinity; Albumins; Area; Binding; Biodistribution; Biological Response Modifier Therapy; Biomedical Research; Blood Cells; Blood Circulation; Catabolism; Charge; Circulation; Clinical; Coagulation Process; Coupling; Cytolysis; Data; Development; Diffusion; Disease; Dose; Drug Delivery Systems; Drug Kinetics; Drug usage; Endosomes; 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; Physiological; Plasma; Plasma Proteins; Plasminogen Activator; Polymers; Property; Protein Engineering; Proteins; Pulmonary Embolism; Recycling; Research; Resources; Risk; Role; Safety; Serum Proteins; Structure; Testing; Therapeutic; Therapeutic Uses; Thrombin; Thrombolytic Therapy; Thrombosis; Thrombus; Time; Tissues; Transcend; Universities; Urokinase; antibody engineering; biological systems; blood fractionation; chemical conjugate; compliance behavior; cost; design; improved; in silico; in vivo; inhibitor; insight; mathematical model; member; mouse model; mutant; nanobodies; neonatal Fc receptor; novel; pharmacologic; 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 虽然最近几十年的临床使用治疗蛋白的临床使用呈指数增长，但公用事业通常受到限制 不利的药代动力学（PK），由快速消除介导。克服这一限制的一种方法是 半衰期延伸（HLE），通过将生物疗法固定到聚合物（PEG），血浆蛋白上实现 （白蛋白，IgG）和血细胞。描述货物蛋白特性影响的数据很少 以及使用这些策略测试的蛋白质PK上的HLE限制配体。 HLE具有的一个区域 促进临床治疗的潜力是急性，威胁生命，血栓形成（肺栓塞，缺血性中风， ETC。）。可用于治疗此疾病的唯一药理治疗方法是输注纤溶酶原 激活剂具有不利的药理学特征，由半衰期介导，快速失活 在血浆和严重的不良反应（例如出血转化）中。飞行员数据表明 在富含凝血酶的病理血栓（SCUPA-T）中选择性激活尿激酶的衍生物的结合 白蛋白结合纳米机（NB）导致PK与RBC结合Scupa-T相同，这是一种率先进行的方法 由Drs。 Muzykantov和Cines，尽管在血浆中分布而不是血液的细胞分数。 该提议的核心假设是Scupa-T与白蛋白结合配体的耦合将 提供长时间的循环，由新生儿FC受体（FCRN）和选择性溶栓介导 病理性血栓，从裂解中保留止血塞。在指导阶段， 将使用FCRN基因敲除小鼠直接确定货物药物在支持FCRN的HLE上的特性 阐明由白蛋白结合赋予的HLE的FCRN介导的成分（AIM 1; K99）。基于飞行员 将在肺栓塞的小鼠模型中研究白蛋白结合SCUPA-T的数据，安全性和功效。 这些结果将用于开发一个预测建模平台，该平台将用于进一步的工程 和HLE-SCUPA-T构建体的PK优化（AIM 2; K99）。着眼于机械和翻译 该策略的进步，将使用一个 新开发的纳米生物库（AIM 3; R00）。此外，凝血酶可裂的HLE配体将是 设计，允许在血栓中选择性释放Scupa-T，从而改善了凝块和裂解的扩散（AIM 3; R00）。 该提案的总体主题包括识别白蛋白介导的HLE和 基于机制的建模，以指导蛋白质治疗剂的优化和重新设计。指导团队 已经确定了跨越该赠款领域的专业知识，即血栓形成和血栓形成 生物系统的止血，抗体工程和数学建模。指导的研究将是 在宾夕法尼亚大学举办，该大学拥有高度合作和协作的教师 可用于进行拟议研究的资源。该建议旨在获得专业知识 作为一名终身教师进行高质量生物医学研究的成功，必不可少。