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

基本信息

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
项目类别：
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&apos 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

项目摘要

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.

项目概要血脑屏障（BBB）对开发神经系统疾病的有效疗法提出了最大的挑战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) 已证明中枢神经系统输送有显着改善，但总体大脑渗透率却较低。仍然是适度的，因为 BBB 的转胞吞作用是双向的，因此大部分施用的 bsAb 仍保留在血液中。过程并不可避免地导致抗 TfR bsAb 在血液和大脑之间达到浓度平衡侧面，我们重新认识到通过近腔到腔外流的最小化，浓度平衡可以改变我们的设计原则是将抗TfR的可变片段（Fv）融合到N端。治疗性 IgG，通过大脑中存在的疾病相关蛋白酶特异性的可裂解接头。通过 TfR 介导的转胞吞作用输送到大脑，治疗性 IgG 将被激活并停留在大脑一侧因为它失去了与TfR的结合能力，释放的抗TfR Fv会转运回血液侧。通过肾脏清除，TfR结合的前药将被进一步转胞吞，从而形成净流。我们的长期目标是开发高效的血脑屏障输送。能够有效治疗脑癌等神经系统疾病的方法 MPI R21 项目的目标是证明这种新型 BBB 的概念。基于蛋白酶激活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。阿尔茨海默病 (AD) β (Aβ) 作为本研究的模型系统，以我们的集体专业知识为基础。蛋白质工程，蛋白酶生物化学，BBB运输和AD，我们将，目标1：设计，构建和优化蛋白酶激活的双特异性抗体前药；目标 2：验证 BBB 渗透和治疗使用脑淀粉样血管病（CAA）小鼠模型研究抗体前药的功效。创新，因为蛋白酶可切割的前药设计可以防止反向转胞吞作用，改变浓度平衡，从而促进治疗药物从血液渗透到大脑。意义重大，因为它开发了一种平台技术，能够 (1) 提高生物制剂的 BBB 渗透率包括单克隆抗体（mAb）和抗体药物偶联物（ADC），（2）大大减少异地现场通过大脑中的原位激活来靶向副作用，以及（3）治疗目前非治疗的各种神经系统疾病可针对的。