Protease Activated Bi-Specific Antibody Prodrugs for Efficient Transportation Across Blood-Brain Barrier

蛋白酶激活的双特异性抗体前药可有效跨血脑屏障运输

• 批准号: 10300356
• 负责人: Xin Ge
• 金额: $ 42.9万
• 依托单位: UNIVERSITY OF TEXAS HLTH SCI CTR HOUSTON
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-01 至 2023-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10300356
• 关键词: Alzheimer' s Disease; Amyloid; Amyloid beta-Protein; Amyloid deposition; Antibodies; Antibody-drug conjugates; Back; Behavioral; Binding; Biochemical; Biochemistry; Biological; Biological Assay; Biological Models; Bispecific Antibodies; Blood; Blood - brain barrier anatomy; Blood Circulation; Blood Vessels; Brain; Cathepsins; Central Nervous System Agents; Cephalic; Cerebral Amyloid Angiopathy; Cerebrovascular system; Cleaved cell; Cognitive; Data Analyses; Disease; Drug Delivery Systems; Drug Kinetics; ERBB2 gene; Enzyme Kinetics; Epithelial Cells; Equilibrium; Fluorescent Antibody Technique; Foundations; Funding Mechanisms; Glioblastoma; Goals; Human; I125 isotope; Immunoglobulin G; In Situ; Interleukin-1 alpha; Label; Location; Malignant neoplasm of brain; Mammalian Cell; Matrix Metalloproteinases; Mediating; Modeling; Monoclonal Antibodies; Mus; N-terminal; Neuraxis; Neurodegenerative Disorders; Outcome; Penetration; Peptide Hydrolases; Peptides; Pharmaceutical Preparations; Pharmacodynamics; Process; Prodrugs; Protein Engineering; Proteomics; Radiolabeled; Renal clearance function; Research; Side; Site; TFRC gene; TNF gene; Technology; Therapeutic; Therapeutic Monoclonal Antibodies; Therapeutic antibodies; Transportation; Treatment Efficacy; Tumor Antigens; Vascular Endothelial Growth Factors; abeta deposition; base; blood-brain barrier crossing; blood-brain barrier penetration; cerebral amyloidosis; cognitive function; cognitive testing; conditioned fear; design; design and construction; effective therapy; epidermal growth factor receptor VIII; experimental study; gamma secretase; high risk; improved; in vivo; in vivo two-photon imaging; innovation; large scale data; mouse model; nervous system disorder; novel; prevent; protease E; receptor; receptor binding; side effect; transcytosis

PROJECT SUMMARY The blood-brain barrier (BBB) poses the greatest challenge for developing effective therapies for neurological diseases. Inspired by receptor-mediated transcytosis, bi-specific antibodies (bsAbs) against transferrin receptor (TfR) have demonstrated significant improvements of CNS delivery. However, the overall brain penetration was still modest, with large majority of administrated bsAbs remain in blood. As transcytosis at BBB is a bi-directional process and inevitably leads anti-TfR bsAb reaching a concentration equilibrium between the blood and the brain sides, we hypothesize that by minimizing abluminal-to-luminal efflux, the concentration equilibrium can be shifted toward BBB penetration. Our design principle is to fuse the variable fragment (Fv) of anti-TfR to the N-terminal of a therapeutic IgG, via cleavable linker(s) specific to disease-associated protease present in the brain. Once delivered to the brain by TfR-mediated transcytosis, therapeutic IgG will be activated and stays at the brain side because it loses binding ability to TfR. Released anti-TfR Fv will transport back to the blood side then be eliminated by renal clearance. TfR-bound prodrugs will be further transcytosed and thus forming a net flow of therapeutic Ab penetration from blood to brain. Our long-term goal is to develop a highly efficient BBB delivery approach that enables effective treatments of neurological disorders such as brain cancer and neurodegenerative diseases. The objective of this MPI R21 project is to prove the concept of this novel BBB delivery technology based on protease-activated prodrug designs. We will use cathepsin S activated anti-amyloid β (Aβ) for Alzheimer’s disease (AD) as the model system in this study. Building on our collective expertise on protein engineering, protease biochemistry, BBB transportation and AD, we will, Aim 1: design, construct and optimize protease-activated bi-specific antibody prodrugs; and Aim 2: validate BBB penetration and therapeutic efficacy of antibody prodrugs using mouse models of cerebral amyloid angiopathy (CAA). The approaches are innovative, because the protease-cleavable prodrug designs can prevent the reverse transcytosis, shift the concentration equilibrium, and thus promote therapeutics penetration from blood to brain. The proposed research is significant because it develops a platform technology enabling to (1) improve BBB penetration of biologics including monoclonal antibodies (mAbs) and antibody-drug conjugates (ADCs), (2) greatly reduce off-site on- target side effects by in situ activation in brain, and (3) treat a variety of neurological disorders currently non- targetable.

项目摘要 血脑屏障（BBB）是开发有效治疗神经系统疾病的最大挑战。 疾病受受体介导的转胞吞作用的启发，针对转铁蛋白受体的双特异性抗体（bsAb） (TfR)已经证明了CNS递送的显著改善。然而，整体的大脑渗透率是 仍然是适度，大部分施用的bsAb保留在血液中。由于血脑屏障处的转胞吞作用是双向的， 这一过程不可避免地导致抗TfR bsAb在血液和大脑之间达到浓度平衡 另一方面，我们假设通过最小化腔外到腔外的外排，可以改变浓度平衡 向血脑屏障渗透我们的设计原则是将抗TfR的可变片段（Fv）融合到N-末端， 通过对脑中存在的疾病相关蛋白酶具有特异性的可切割接头，一旦 通过TfR介导的转胞吞作用递送至脑，治疗性IgG将被激活并停留在脑侧 因为它失去了与转铁蛋白受体结合的能力。释放的抗-TfR Fv将转运回血液侧，然后被 通过肾清除消除。TfR结合的前药将被进一步转胞吞，从而形成TfR结合的前药的净流动。 治疗性抗体从血液渗透到大脑。我们的长期目标是开发高效的BBB交付 这种方法能够有效治疗神经系统疾病，如脑癌， 神经退行性疾病这个MPI R21项目的目的是证明这种新型BBB的概念 基于蛋白酶激活的前药设计的递送技术。我们将使用组织蛋白酶S激活的抗淀粉样蛋白 β（Aβ）为阿尔茨海默病（AD）的模型系统。利用我们的集体专长， 蛋白质工程、蛋白酶生物化学、血脑屏障运输和AD，我们将，目标1：设计，构建和 优化蛋白酶活化的双特异性抗体前药;和目的2：验证BBB渗透和治疗 使用脑淀粉样血管病（CAA）小鼠模型的抗体前药的功效。这些方法是 创新的，因为蛋白酶可切割的前药设计可以防止反向转胞吞作用，将 浓度平衡，从而促进治疗剂从血液渗透到脑。拟议研究 重要的是，它开发了一种平台技术，能够（1）改善生物制剂的BBB渗透 包括单克隆抗体（mAb）和抗体-药物偶联物（ADC），（2）大大减少了非位点的在- 通过在脑中原位激活靶向副作用，和（3）治疗目前非- 有针对性