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.

RA是最常见的慢性自身免疫性疾病之一，可能导致联合关节 如果未治疗，破坏和严重的残疾。 RA无法治愈，多达50％的RA患者无法治愈 响应抗TNF疗法，因为TNF之后循环TH-17/IL-17水平高度升高 glocade。对于RA患者的这一子集，破坏了TNF之间协同作用的新途径的破坏 （M1巨噬细胞）和IL-17（TH-17细胞）级联反应可能解决RA处理中的关键障碍。因此 该项目的目的是为疾病的RA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）细胞进入促炎的M1巨噬细胞，产生高水平的TNF，IL-1和 IL-6。另外，由TLR5驱动的M1巨噬细胞产生的IL-6可以将幼稚的T细胞区分为 炎症性RA Th-17细胞，该细胞秘密IL-17A，IL-17F，IL-22，IL-24，IL-26，IL-26，CCL20和GM-CSF。在老鼠中， TLR5激动剂的全身和局部注射加剧了关节旋转；相反的抗TLR5 AB处理 减轻胶原蛋白诱导关节炎（CIA）关节炎症。 为了调查TLR5 AB作为RA的潜在治疗的作用，我们与科学家合作 Abwiz Inc.使用TLR5抗原，筛选人类碎片抗原结合（FAB）噬菌体展示库 对于TLR5高亲和力粘合剂。测序多达40个阳性克隆，并选择了10个TLR5 Fab克隆 通过ELISA表示，纯化并评估TLR5结合和交叉反应性。十个工厂克隆是 由Shahrara实验室检查的人和鼠细胞中的TLR5神经化能力，一个是 根据其出色的阻塞能力选择。 该项目的总体目标是开发用于RA治疗的抗TLR5 AB。在第一阶段，我们的方法 是为了达到商业范围内的KD值，以增强抗TLR5 AB的亲和力 可用的腹肌。通过使用噬菌体 - 播放AB库通过网站提供的候选人将是 测试了其TLR5阻断人类细胞中的亲和力。随后，最有前途的候选人将是 测试了它们从人源化RA中促进炎症反应消除RA滑液的能力 鼠标模型。该项目的长期目标是为RA生成安全且完全新颖的TLR5 AB 对当前疗法不反应的患者。