Development of Chemical Probes to Investigate Adhesion GPCR Tethered Agonism

开发用于研究粘附 GPCR 系链激动作用的化学探针

• 批准号: 9917826
• 负责人: Gregory Gordon Tall
• 金额: $ 48.77万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-07-01 至 2022-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9917826
• 关键词: ADGR1 gene; Adhesions; Adult; Affinity; Agonist; Bilateral; Binding; Biochemical; Biological Assay; Brain; Brain Injuries; Cell Adhesion; Cell Adhesion Molecules; Cell Proliferation; Cells; Chemicals; Chemistry; Collaborations; Collection; Databases; Deposition; Development; Disease; Dissociation; Engineering; Enzymes; Event; Exhibits; Fractionation; Functional disorder; G-Protein Signaling Pathway; G-Protein-Coupled Receptors; G13 Protein; GTP-Binding Proteins; Genomics; Goals; Growth; Heterotrimeric GTP-Binding Proteins; Human; Informatics; Injections; Investigation; Laboratories; Libraries; Ligands; Luc Gene; Luciferases; Measures; Mediating; Michigan; Microgyria; Modeling; Multiple Sclerosis; Mutation; Myelin; Natural Products; Nerve; Neuraxis; Neurodevelopmental Disorder; Oligodendroglia; Pathway interactions; Patients; Peptides; Pharmaceutical Preparations; Physiological; Physiology; Positioning Attribute; Preparation; Procedures; Process; Proteins; Receptor Activation; Reporter Genes; Research; Resources; Rodent; Role; Schizophrenia; Scientist; Screening Result; Serum Response Element; Signal Transduction; System; Testing; Therapeutic; Universities; Work; base; brain malformation; counterscreen; efficacy testing; follow-up; high throughput screening; inhibitor/antagonist; malignant neurologic neoplasms; myelination; nervous system disorder; novel; oligodendrocyte precursor; peptidomimetics; polypeptide; precursor cell; programs; prospective; protein activation; protein aminoacid sequence; protein reconstitution; receptor; receptor binding; receptor function; reconstitution; repaired; restoration; scaffold; screening; small molecule; small molecule libraries; synthetic peptide; therapeutic development; therapeutic target; tool

Summary/Abstract: GPR56/ADGRG1 is an Adhesion G protein Coupled Receptor (AGPCR) that is required for brain development and function by acting as a critical regulator of the cellular pathway responsible for myelin deposition in the brain. GPR56 mutations cause a debilitating human brain malformation disease. Our recent work unveiled the general AGPCR activation mechanism. AGPCRs are two-fragment receptors that arise from a single polypeptide following a defined self-cleavage event. The physical arrangement of the two bound receptor fragments conceals a peptide sequence, that when revealed by fragment dissociation, serves as a tethered-peptide self-activator (agonist) of the GPCR-like fragment. We used information from our delineation of the AGPCR mechanism to engineer model GPR56 receptors in high and low activity states. These receptors will be used to find small molecule compounds, through screening diverse chemical libraries, that inhibit the high-activity receptors or activate the low-activity receptors. The immediate application of our work will be to develop chemical probes that bind GPR56 for use in investigative studies of physiological AGPCR activation mechanism(s). In the longer term, our chemical scaffolds may be developed as therapeutics to correct defective brain myelination in a variety of disease contexts, including myelin-deficient schizophrenia patients. GPR56 promotes nerve myelination by controlling the growth of oligodendrocyte precursor cells (OPCs). OPCs become mature oligodendrocytes, which are the cells that wrap processes around nerves in the brain to produce the protective myelin coat. Past studies in which OPCs were injected into myelin-deficient rodent brains promoted nerve re-myelination and partial restoration of brain functions. OPC injection into human brains would not be a tenable therapy at present, but use of a new GPR56 activating drug to stimulate growth of natural OPCs might be a more realistic approach. The prospect of directing re- myelination programs in damaged brains by artificially activating GPR56 is a highly appealing goal for patients afflicted with diseases including schizophrenia or multiple sclerosis. We are confident that our endeavors to develop probes for GPR56 will be successful, as our pilot screening efforts with small compound libraries (5346 total compounds from four small libraries) identified a low-affinity, first-in-class GPR56 antagonist and a partial agonist. It is the combination of our well-honed screen and counter-screen assays and our exquisitely tuned G protein reconstitution secondary assays that made our pilot work successful. We are eager to apply our established assays to the large chemical libraries and natural product extract collections at the University of Michigan Center for Chemical Genomics to fulfill the critical, unmet need for AGPCR modulatory compounds.

摘要/摘要：GPR56/ADGRG1是一种必需的粘附性G蛋白偶联受体(AGPCR) 作为细胞通路的关键调节器，负责 脑内髓鞘沉积。GPR56突变会导致一种使人衰弱的大脑畸形疾病。我们的 最近的工作揭示了AGPCR的一般激活机制。AGPCRs是两个片段的受体， 产生于定义的自切割事件之后的单个多肽。两者的物理排列 结合的受体片段隐藏了一个肽序列，当片段解离时，该序列起作用 作为GPCR样片段的栓系多肽自激活剂。我们使用了来自我们的 描述AGPCR机制以设计处于高和低活性状态的GPR56受体模型。 这些受体将被用来寻找小分子化合物，通过筛选不同的化学库， 抑制高活性受体或激活低活性受体。立即应用我们的 工作将是开发结合GPR56的化学探针，用于生理研究 AGPCR激活机制(S)。从长远来看，我们的化学支架可能会发展为 用于纠正包括髓鞘缺陷在内的各种疾病背景下的脑髓鞘形成缺陷的治疗方法 精神分裂症患者。GPR56通过控制少突胶质细胞的生长促进神经髓鞘形成 前体细胞(OPC)。OPC成为成熟的少突胶质细胞，也就是包裹过程的细胞 围绕着大脑中的神经，产生保护性的髓鞘外衣。注射OPC的过去研究 进入髓鞘缺乏的啮齿类动物的大脑，促进了神经的重新髓鞘形成和大脑功能的部分恢复。 目前，将OPC注射到人脑中并不是一种站得住脚的治疗方法，而是使用一种新的GPR56激活 用药物刺激天然OPC的生长可能是更现实的方法。重新定向的前景-- 通过人工激活GPR56来修复受损大脑的髓鞘形成计划对患者来说是一个非常有吸引力的目标 患有精神分裂症或多发性硬化症等疾病的。我们相信，我们将努力 为GPR56开发探针将是成功的，因为我们利用小型化合物文库进行了试点筛选工作 (来自四个小文库的5346个化合物)鉴定了一个低亲和力、一流的GPR56拮抗剂和一个 部分激动剂。这是我们精心打磨的筛查和反筛查分析的结合，以及我们精致的 调整了G蛋白重组二次分析，使我们的试点工作取得成功。我们渴望申请 我们建立了对大学大型化学图书馆和天然产品提取物收藏的分析 密歇根化学基因组学中心以满足关键的、未得到满足的AGPCR调控需求 化合物。