Engineering a unique antibody for patients with RA

为 RA 患者设计独特的抗体

• 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
• 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

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.

类风湿关节炎是一种最常见的慢性自身免疫性疾病，可导致完全性关节炎， 如果不治疗，会造成严重的破坏和残疾。RA无法治愈，高达50%的RA患者无法治愈 对抗TNF治疗有反应，因为循环Th-17/IL-17水平在TNF治疗后高度升高 封锁对于RA患者的这一亚组，破坏一种新的途径，损害TNF-α和TNF-α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治疗中的关键屏障。因此 本项目的目的是开发用于RA患者的治疗性人TLR5抗体（Ab）， 是由效应巨噬细胞和T细胞之间的相互作用驱动的。 我们证明，TLR5与其在关节中表达的天然配体的连接， 外周血（PB）细胞转化为促炎性M1巨噬细胞，其产生高水平的TNF、IL-1和 IL-6。此外，由TLR5驱动的M1巨噬细胞产生的IL-6可以使幼稚T细胞分化为 炎性RA TH-17细胞分泌IL-17 A、IL-17 F、IL-22、IL-24、IL-26、CCL 20和GM-CSF。在小鼠中， 全身和局部注射TLR5激动剂加剧关节肿胀;相反，抗TLR5 Ab治疗 缓解胶原蛋白诱导性关节炎（CIA）关节炎症。 为了研究TLR5 Ab作为RA潜在治疗方法的作用，我们与来自以下机构的科学家合作： 阿布维兹公司以TLR5为抗原，筛选人源Fab噬菌体展示文库 用于TLR5高亲和力结合剂。对多达40个阳性克隆进行测序，并选择10个TLR5 Fab克隆 表达、纯化并通过ELISA评估TLR5结合和交叉反应性。10个Fab克隆是 Shahrara实验室检查了人和鼠细胞中的TLR5中和能力， 选择基于其优越的阻断能力。 本项目的总体目标是开发用于RA治疗的抗TLR5抗体。在第一阶段，我们的方法 增强抗TLR5 Ab的亲和力，以达到在商业上可接受的范围内的Kd值。 可用Abs.通过使用噬菌体展示Ab文库的定点诱变提供的候选物将被 测试了它们在人类细胞中的TLR5阻断亲和力。随后，最有希望的候选人将是 测试它们消除RA滑液促进人源化RA中的炎症反应的能力 小鼠模型该项目的长期目标是产生一种安全且完全新颖的用于RA的TLR5 Ab。 对目前的治疗没有反应的患者。