Bispecific immunotherapeutic delivery system for lung diseases

项目概要/摘要 现代医学已经创造出精确的药物，以高亲和力阻断TGF-β等单一治疗靶点 和特异性。然而，治疗肺部疾病仍然具有挑战性，部分原因是肺微血管内皮细胞 是有效药物输送的关键限制性障碍。目前的全身治疗仅依赖于 对流和扩散以被动地外渗到疾病靶点和细胞所在的组织中 可以容易地到达并直接处理。本研究提案的目标是设计、开发和测试一种 本发明涉及一种用于免疫治疗的新型药物递送系统，其通过靶向小窝来克服这一关键障碍， 促进静脉注射后主动和特异性转胞吞入肺。理想的是提供整个 肺组织内的治疗剂量，所有其他组织最低限度地暴露。我们试图接近这个 理想的方法是实现稳健的经内皮泵送精确进入肺组织， 治疗靶点TGF-β，其调节患病组织中的炎症和重塑。因为TGF-β 也在许多器官中发挥各种稳态作用，当系统靶向其 功能正在尝试。本文提出的精确肺靶向将通过以下方式最大化疗效和治疗指数： 最小化剂量、消除毒性和降低治疗成本。为此，我们从基因上 设计了第一种“双精度”免疫治疗剂，即具有一个双特异性抗体的四重格式的双特异性抗体。 臂对介导经由小窝泵送的精确结合/递送和穿透肺组织， 另一对构成阻断TGF-β效应子功能的精确治疗模式。活性 与标准被动转运相比，经内皮递送将精确的肺靶向提高了100倍。 在1小时内将大部分注射剂量递送到肺部，在一个实施例中将治疗效力增强>1000倍。 大鼠肺炎模型。现在，我们的目标是扩大这一有希望的初步工作，并进一步改善和 严格测试这种药物输送系统，以治疗从早期急性 炎症到慢性和进行性纤维化。我们将优化我们的双精度肺靶向 免疫治疗剂，并研究其特异性肺递送，渗透，积累，定位， 使用多种成像技术（SPECT-CT、IVM、EM和IHC）在大鼠中的治疗影响。治疗 将在大鼠博来霉素模型中评估效果，该模型再现了许多致命的人类肿瘤的病理学特征。 包括ALI、ARDS、COVID、肺炎和纤维化的疾病。我们的具体目标是：1）工程师和 评价不同的小窝靶向抗体构建体用于精确活性递送到正常肺组织中，2） 为了量化肺部疾病中双特异性免疫治疗剂的靶向和优化递送，3）为了测试功效 双特异性免疫治疗剂改善肺部疾病和阻断TGF-β通路。这项工作为 为靶向治疗奠定了基础，并可能开始从被动药物到主动药物的范式转变 为许多疾病提供治疗。

Project summary/abstract Modern medicine has created precision drugs blocking a single therapeutic target like TGF-β with high affinity and specificity. Yet treating lung diseases remains challenging in part because lung microvascular endothelium represents a key restrictive barrier to effective drug delivery. Current systemic therapeutics rely solely on convection and diffusion to extravasate passively into the tissue interstitium where disease targets and cells can readily be reached and directly treated. The goal of this research proposal is to design, develop and test a novel drug delivery system for immunotherapeutics that overcomes this key barrier by targeting caveolae to facilitate active and specific transcytosis into lungs after intravenous injection. The ideal is to deliver the entire therapeutic dose inside the lung tissue with all other tissues minimally exposed. We attempt to approach this ideal by achieving robust transendothelial pumping precisely into lung tissue to comprehensively block the therapeutic target TGF-β, which regulates inflammation and remodeling in diseased tissues. Because TGF-β also exerts various homeostatic effects in many organs, caution is necessary when systemic targeting of its function is attempted. Precision lung targeting proposed here will maximize efficacy and therapeutic indices by minimizing dosages, eliminating toxicities, and reducing cost of treatment. To that end, we have genetically engineered the first “dual precision” immunotherapeutics, namely bispecific antibodies in quad format with one arm pair mediating precise binding/delivery to and penetration of lung tissue via caveolae pumping and the other pair constituting the precision therapeutic modality that blocks TGF-β effector function. Active transendothelial delivery improved precision lung targeting by 100-fold over standard passive transport. Delivering most of the injected dose into lungs within 1 hour enhanced therapeutic potency by >1000-fold in a rat pneumonitis model. Now our goal is to expand this promising preliminary work and further improve and rigorously test this drug delivery system to treat key lung diseases at distinct stages ranging from early acute inflammation to chronic and progressive fibrosis. We will optimize lung targeting of our dual precision immunotherapeutics and study their specific lung delivery, penetration, accumulation, localization, and therapeutic impact in rats using multiple imaging techniques (SPECT-CT, IVM, EM, and IHC). Therapeutic effects will be assessed in a rat bleomycin model that reproduces pathological hallmarks of many fatal human diseases including ALI, ARDS, COVID, pneumonias, and fibrosis. Our specific aims are: 1) to engineer and evaluate distinct caveolae-targeted antibody constructs for precision active delivery into normal lung tissue, 2) to quantify targeting and optimize delivery of bispecific immunotherapeutics in lung disease, 3) to test efficacy of bispecific immunotherapeutics to ameliorate lung disease and block TGF-β pathways. This work sets a foundation for caveolae-targeted therapies and could begin a paradigm shift from passive to active drug delivery for many diseases.