Engineering a unique antibody for patients with RA

为 RA 患者设计独特的抗体

基本信息

批准号：
10005747
负责人：
Toshi Maruyama
金额：
$ 30万
依托单位：
ABWIZ BIO, INC.
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-04-16 至 2022-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10005747
关键词：
Achievement Affect Affinity Agonist Anti-Inflammatory Agents Anti-Tumor Necrosis Factor Therapy Antibodies Antibody Therapy Antigens Antirheumatic Agents Attenuated Autoimmune Diseases Bacteriophages Binding Blood Cells CCL20 gene Cell Differentiation process Cell Line Cells Chronic Collagen Arthritis Complementarity Determining Regions Data Dependovirus Development Disease Drug Kinetics Effectiveness Engineering Enzyme-Linked Immunosorbent Assay Epitopes Fab Immunoglobulins Flagellin Funding Goals Granulocyte-Macrophage Colony-Stimulating Factor Human Impairment Inflammation Inflammatory Inflammatory Response Injections Interleukin-1 Interleukin-1 beta Interleukin-17 Interleukin-6 Investigational New Drug Application Joints Lead Libraries Ligands Ligation Light Maximum Tolerated Dose Methotrexate Molecular Conformation Mus Mutate Myelogenous Pathway interactions Patients Phage Display Phase Pre-Clinical Model Process Recombinants Reporter Rheumatoid Arthritis Rodent Role Scientist Site-Directed Mutagenesis Synovial Fluid T-Lymphocyte TLR5 gene TNF gene Testing Therapeutic Toxicology alternative treatment antibody libraries antigen binding arthritis therapy base bone bone erosion bone loss cellular transduction commercialization cross reactivity disability humanized mouse in vivo interleukin-22 joint destruction joint inflammation macrophage monocyte mouse model multidisciplinary nonhuman primate novel peripheral blood pre-clinical receptor receptor function safety study synergism

项目摘要

RA is one of the most common chronic autoimmune disorders that can lead to complete joint destruction and severe disability if untreated. There is no cure for RA and up to 50% of RA patients do not respond to anti-TNF therapies as circulating Th-17/IL-17 levels are highly elevated subsequent to TNF blockade. For this subset of RA patients, disruption of a novel pathway that impairs the synergy between TNF (M1 macrophages) and IL-17 (Th-17 cells) cascades may resolve the critical barrier in RA treatment. Hence objective of this project is to develop a therapeutic human TLR5 antibody (Ab) for RA patients whose disease is driven by the cross-talk between the effector macrophages and T cells. We documented that ligation of TLR5 to its natural ligand expressed in the joints, transforms RA peripheral blood (PB) cells into proinflammatory M1 macrophages which produce high levels of TNF, IL-1 and IL-6. In addition, IL-6 produced from TLR5 driven M1 macrophages can differentiate the naïve T cells into inflammatory RA TH-17 cells that secrete IL-17A, IL-17F, IL-22, IL-24, IL-26, CCL20 and GM-CSF. In mice, systemic and local injection of a TLR5 agonist exacerbates joint swelling; conversely anti-TLR5 Ab treatment alleviates collagen induced arthritis (CIA) joint inflammation. To investigate the role of TLR5 Ab as a potential treatment for RA, we have partnered with scientists at Abwiz Inc. Using a TLR5 antigen, a human fragment antigen-binding (Fab) phage display library was screened for TLR5 high affinity binders. Up to 40 positive clones were sequenced and 10 selected TLR5 Fab clones were expressed, purified and assessed by ELISA for TLR5 binding and cross reactivity. Ten Fab clones were examined by the Shahrara lab for TLR5 neutralization capacity in human and murine cells and one was selected based on its superior blocking capacity. The overall goal of this project is to develop an anti-TLR5 Ab for RA therapy. In Phase I, our approach is to enhance the affinity of anti-TLR5 Ab in order to reach a Kd value that is within the range of commercially available Abs. Candidates provided through site directed mutagenesis using phage-display Ab library, will be tested for their TLR5 blocking affinity in human cells. Subsequently, the most promising candidates will be tested for their ability to abrogate RA synovial fluid from promoting inflammatory response in humanized RA mouse model. The long term goal of this project is to generate a safe and completely novel TLR5 Ab for RA patients that do not respond to the current therapies.

RA 是最常见的慢性自身免疫性疾病之一，可导致完全关节损伤 RA 无法治愈，高达 50% 的 RA 患者无法治愈。对抗 TNF 疗法有反应，因为 TNF 治疗后循环 Th-17/IL-17 水平大幅升高对于这部分 RA 患者，破坏了一种新途径，从而损害了 TNF 之间的协同作用。（M1 巨噬细胞）和 IL-17（Th-17 细胞）级联可能解决 RA 治疗中的关键障碍。该项目的目标是为患有 RA 疾病的患者开发一种治疗性人类 TLR5 抗体 (Ab) 由效应巨噬细胞和 T 细胞之间的串扰驱动。我们记录了 TLR5 与其在关节中表达的天然配体的连接，改变了 RA 外周血 (PB) 细胞转化为促炎性 M1 巨噬细胞，产生高水平的 TNF、IL-1 和此外，TLR5 驱动的 M1 巨噬细胞产生的 IL-6 可以将幼稚 T 细胞分化为小鼠中分泌 IL-17A、IL-17F、IL-22、IL-24、IL-26、CCL20 和 GM-CSF 的炎性 RA TH-17 细胞。局部注射 TLR5 激动剂，全身关节肿胀恶化；反之，抗 TLR5 抗体治疗；减轻胶原蛋白诱导的关节炎 (CIA) 关节炎症。为了研究 TLR5 Ab 作为 RA 潜在治疗方法的作用，我们与以下机构的科学家合作： Abwiz Inc. 使用 TLR5 抗原筛选人类片段抗原结合 (Fab) 噬菌体展示文库对于 TLR5 高亲和力结合物，对多达 40 个阳性克隆进行了测序，并选择了 10 个 TLR5 Fab 克隆。表达、纯化并通过 ELISA 评估 10 个 Fab 克隆的 TLR5 结合和交叉反应性。 Shahrara 实验室检查了人类和小鼠细胞中的 TLR5 中和能力，其中之一是基于其卓越的阻塞能力而选择。该项目的总体目标是开发一种用于 RA 治疗的抗 TLR5 抗体，这是我们的方法。是增强抗TLR5抗体的亲和力，以达到商业化范围内的Kd值使用噬菌体展示抗体库通过定点诱变提供的可用抗体将是随后，最有希望的候选者将在人类细胞中测试其 TLR5 阻断亲和力。测试了它们消除 RA 滑液促进人源化 RA 炎症反应的能力该项目的长期目标是为 RA 生成安全且全新的 TLR5 Ab。对当前疗法没有反应的患者。