Cell-Based Screening for Multi-Functional Chemokine Receptor Modulators

基于细胞的多功能趋化因子受体调节剂筛选

• 批准号: 8881070
• 负责人: Peter Krutzik
• 金额: $ 55.3万
• 依托单位: PRIMITY, INC.
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-06-02 至 2017-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8881070
• 关键词: Antigens; Asthma; Autoimmunity; Biological Assay; Biology; CCL3 gene; CCR1 gene; CCR5 gene; Calcium; Calcium Signaling; Calcium-Sensing Receptors; Cell Count; Cell Line; Cells; Chemotaxis; Complex; Data; Data Set; Databases; Disease; Disease model; Dose; Drug Targeting; Event; FDA approved; Flow Cytometry; Fluorescence; GTP-Binding Proteins; Goals; Grant; Health; Human; Imagery; Immune; Immune System Diseases; Immune response; Inflammation; Inflammatory; Kinetics; Lead; Life; Ligand Binding; Ligands; Malignant Neoplasms; Maps; Marketing; Measurable; Measurement; Measures; Mediating; Membrane; Mus; Nature; Outcome; Pattern; Pharmaceutical Preparations; Phase; Phenotype; Population; Positioning Attribute; Property; Proteins; Receptor Activation; Recycling; Relative (related person); Signal Transduction; System; Techniques; Technology; Testing; Therapeutic; Time; Treatment Efficacy; Validation; Virus Diseases; base; chemokine; chemokine receptor; commercialization; cost; desensitization; design; drug discovery; falls; inhibitor/antagonist; insight; meetings; new technology; novel; phase 1 study; protein activation; receptor; receptor binding; receptor internalization; receptor recycling; release of sequestered calcium ion into cytoplasm; response; screening; small molecule; success; tool

DESCRIPTION (provided by applicant): Chemokine receptors belong to the most druggable class of receptors and modulate immune responses central to a wide variety of disease states ranging from inflammation and autoimmunity to viral infection, asthma and cancer, making these receptors prime drug targets. In spite of their apparent therapeutic tractability, numerous clinica trials of seemingly promising compounds have met with limited success. A unique challenge in chemokine drug discovery is the complexity of ligand- receptor interactions, with many chemokine ligands binding to multiple receptors and many receptors responding to numerous chemokines. These overlapping features make it difficult to clearly identify single receptors or ligands as drug targets, and to achieve therapeutic efficacy by targeting a single protein. Furthermore, it is now appreciated that although several chemokines may activate the same receptor, the nature of the transduced signals can vary dramatically between ligands. Thus, not only is the ability to define the selectivity of the ligands across the range of chemokine receptor vitally important, but also the ability to measure the intensity, duration, and final outcome of th signals that are produced. With 20+ chemokine receptors, 45+ ligands, and multiple downstream readouts, measuring the matrix of possible phenotypes with standard techniques is time and cost-prohibitive. In Phase I studies, we developed a functioning multiplex assay platform for receptor internalization that enabled nine chemokine receptors to be screened simultaneously in a single assay well. In this Phase II proposal we will create two multiplex panels containing 18 human or murine chemokine receptors and expand the number of assay readouts to include calcium signaling, receptor desensitization, and chemotaxis. The system will be used to create a comprehensive ligand-receptor interaction database of all naturally-occurring chemokines and to discover synthetic chemokine ligands with non-natural profiles of selectivity and signaling. These results will provide the basis for commercialization of this technology as a drug discovery platform for the identification of compounds with unique selectivity profiles for treatment of inflammation, autoimmunity and other chemokine-mediated diseases.

说明书(申请人提供)：趋化因子受体属于最容易下药的受体类别，调节免疫反应，对多种疾病状态至关重要，从炎症和自身免疫到病毒感染、哮喘和癌症，使这些受体成为主要的药物靶点。尽管它们明显具有治疗效果，但许多看似有希望的化合物的临床试验却取得了有限的成功。趋化因子药物开发中的一个独特挑战是配体-受体相互作用的复杂性，许多趋化因子配体与多个受体结合，许多受体对多种趋化因子做出反应。这些重叠的特征使得很难清楚地识别单一受体或配体作为药物靶点，也很难通过靶向单一蛋白质来实现治疗效果。此外，现在人们意识到，尽管几种趋化因子可以激活同一受体，但不同配体之间转导信号的性质可能会有很大不同。因此，不仅定义整个趋化因子受体范围内配体的选择性的能力至关重要，而且测量产生的TH信号的强度、持续时间和最终结果的能力也是至关重要的。使用20+趋化因子受体、45+配体和多个下游读数，用标准技术测量可能的表型矩阵既费时又费钱。在第一阶段的研究中，我们开发了一个用于受体内化的功能性多重分析平台，使9种趋化因子受体能够在一次良好的测试中同时进行筛选。在这个第二阶段的提案中，我们将创建两个包含18个人或小鼠趋化因子受体的多重面板，并扩大检测读数的数量，以包括钙信号、受体脱敏和趋化作用。该系统将用于创建所有自然产生的趋化因子的全面配体-受体相互作用数据库，并发现具有非自然选择性和信号特征的合成趋化因子配体。这些结果将为这项技术的商业化提供基础，作为一个药物发现平台，用于鉴定具有独特选择性的化合物，用于治疗炎症、自身免疫和其他趋化因子介导的疾病。