Probes to target the 3-way hotspot of IL1RacP to abolish aberrant interleukin inflammation

靶向 IL1RacP 3 路热点以消除异常白细胞介素炎症的探针

基本信息

批准号：
9085212
负责人：
Lance Allen Liotta
金额：
$ 56.88万
依托单位：
GEORGE MASON UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2015
资助国家：
美国
起止时间：
2015-07-01 至 2018-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9085212
关键词：
Acute Affinity Agonist American Amino Acids Animal Model Arginine Arthritis Asthma Autoimmune Process Binding Binding Proteins Biodistribution Biological Assay Biological Availability Cell Culture Techniques Cell Line Cell membrane Cell surface Chemistry Clinical Complex Cytokine Activation Degenerative polyarthritis Deuterium Disease Dose Drug Targeting Engineering Family Family member Health Hot Spot In Vitro Inflammation Inflammatory Interleukin Therapy Interleukin-1 Interleukin-1 beta Interleukins Intra-Articular Injections Kineret Lead Ligand Binding Ligands Maps Mediating Methods Modeling Modification Molecular Monoclonal Antibodies Neonatal Obstructive Lung Diseases Peptide Hydrolases Peptides Pharmaceutical Preparations Play Positioning Attribute Protein Chemistry Proteins Psoriasis Publishing Receptor Signaling Recruitment Activity Role Signal Transduction Site Specificity Surface Surface Plasmon Resonance Tail Technology Testing Transmembrane Domain Traumatic Arthropathy Validation Work anakinra base crosslink cytokine design domain mapping in vitro Assay in vivo inhibitor/antagonist interleukin-1 receptor accessory protein member mouse model neoplastic new technology new therapeutic target next generation novel osmotic minipump peptidomimetics prevent protein complex protein folding protein protein interaction receptor small molecule targeted treatment

项目摘要

DESCRIPTION (provided by applicant): Aberrant interleukin family cytokine activation of inflammation characterizes a wide spectrum of autoimmune, inflammatory, neoplastic, and neonatal disorders, including arthritis, obstructive lung disease, psoriasis and asthma. We created a novel technology, protein painting, and used it to discover, and create, a novel peptide inhibitor, called Arg286p, that abolishes the necessary interaction of IL1-RAcP with the IL-1β-IL1Receptor complex. Arg286p constitutes a novel specific and potent approach to treat interleukin mediated inflammatory diseases. The binding interface contact points between interacting protein partners are important drug targets for the next generation of therapies that block such interactions. Employing a new class of small molecule affinity chemistry, protein painting has a positive hit specificity of 93% and yield up to ten fold higher compared to conventional cross linking or deuterium exchange methods. Using our technology to study the 3-way interaction of interleukin-1β (IL-1β, IL-1 receptor I (IL-1RI), and IL-1 receptor accessory protein (IL-1RAcP), we identified a highly conserved beta-loop Arg286 region of the IL-1RAcP protein that participates in a multivalent interaction with the ligand and the receptor. We validated this novel target to be necessary for IL1β-mediated signaling. We created a folded protein peptide Arg286p that corresponds to the Arg286 beta loop domain, and a monoclonal antibody that recognizes the IL-1RAcP Arg286 surface domain. Our Arg286 peptide compound abolished interleukin signaling in a cell culture model in a dose-dependent manner, compared to a control peptide lacking the arginine in the 286 position. The Arg286 peptide and the anti-Arg286 mAb also abolished formation of the IL-1β, IL-1RI, and IL-1RAcP 3-way complex, in vitro, in a dose-dependent manner. IL1-RAcP is an accessory protein that is recruited after the interleukin ligand binds to the receptor, and signaling requires all members of this three-way complex formation. Our new inhibitor Arg286p mimics a small key multivalent interaction region at the arginine 286 beta loop of IL1-RacP, thereby abolishing complex formation and preventing downstream inflammatory signaling. Since Arg286p acts downstream of receptor-ligand binding, it can massively amplify the potency for interleukin therapy even in the face of excess ligand, and may be superior to, or synergistic with, existing IL-1 therapies that competitively target the ligand. Under the Aims, we will create new structural modifications in this lead compound to optimize its affinity and stability for IL-1 and we will use it as a basis to create novel inhibitos specific for IL33 and IL36, for which clinical inhibitors do not exist. We will employ our protein painting technology in a novel iterative workflow to guide the design of peptide modulators (agonists and antagonists) that are either optimized in potency for IL-1 or specific for IL-33 and IL-36 in vitro, in three types of assays. The optimized version of our lead compound will be studied in a well-established animal model of osteoarthritis. Osteoarthritis causes suffering for 27 million Americans.

描述（由应用提供）：炎症的异常白细胞介素家族细胞因子激活表征了各种自身免疫性，炎症，肿瘤和新生儿疾病，包括关节炎，阻塞性肺部疾病，牛皮癣和哮喘。我们创建了一种新型的技术，蛋白质绘画，并用它来发现并创建了一种称为Arg286p的新型肽抑制剂，该肽抑制剂废除了IL1-RACP与IL-1β-IL1受体复合物的必要相互作用。 ARG286P构成了治疗白介素介导的炎症性疾病的新型特异性和潜在方法。相互作用蛋白质伴侣之间的结合界面接触点是阻断这种相互作用的下一代疗法的重要药物靶标。与常规的交叉链接或氘交换方法相比，蛋白质绘画采用新的小分子亲和力化学化学，其正命中率为93％，高达十倍。使用我们的技术研究白细胞介素1β（IL-1β，IL-1受体I（IL-1RI）和IL-1受体附件的3向相互作用蛋白质（IL-1RACP），我们确定了IL-1RACP蛋白的高度组成的β环ARG286区，该区域与配体和接收器进行了多价相互作用。我们验证了这一新型目标是IL1β介导的信号传导所必需的。我们创建了与ARG286β环域相对应的折叠蛋白肽ARG286P，以及一种识别IL-1RACP ARG286表面域的单克隆抗体。我们的ARG286肽化合物以剂量依赖性的方式在细胞培养模型中取消了白介素信号传导，与在286位缺乏精氨酸的对照肽相比。 ARG286肽和抗ARG286 MAB也以剂量依赖性方式消除了体外IL-1β，IL-1RI和IL-1RACP 3向复合物的形成。 IL1-RACP是一种辅助蛋白，在白介素配体与受体结合后募集，信号传导需要这种三向复合物形成的所有成员。我们的新抑制剂ARG286P模仿了IL1-RACP的精氨酸286β环的小钥匙多价相互作用区域，从而消除了复合物的形成并防止下游炎症信号传导。由于ARG286P在受体配体结合的下游起作用，因此即使面对过量配体，它也可以大大扩大白介素治疗的效力，并且可能优于竞争性靶向配体的现有IL-1疗法。在此目的下，我们将在该铅化合物中创建新的结构修饰，以优化其对IL-1的亲和力和稳定性，我们将使用它作为创建针对IL33和IL36的新型抑制剂的基础，为此临床抑制剂不存在。我们将在新型的迭代工作流程中采用蛋白质绘画技术来指导肽调节剂（激动剂和拮抗剂）的设计，这些肽调节剂（激动剂和拮抗剂）在IL-1的效力中进行了优化，或者在三种类型的测定中对IL-33和IL-33和IL-36的特定于体外进行了优化。我们的铅化合物的优化版本将在骨关节炎的动物模型中进行研究。骨关节炎会造成2700万美国人的痛苦。