Cell-Based Screening for Multi-Functional Chemokine Receptor Modulators

描述（由申请人提供）：趋化因子受体属于最具药物药性的受体类别，可调节多种疾病状态的免疫反应，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。尽管它们具有明显的治疗效果，但许多看似有希望的化合物的临床试验却收效甚微。趋化因子药物发现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，许多受体对许多趋化因子作出反应。这些重叠的特征使得很难清楚地识别单个受体或配体作为药物靶点，也很难通过靶向单个蛋白质来达到治疗效果。此外，现在认识到，尽管几种趋化因子可能激活相同的受体，但转导信号的性质在配体之间可能会发生巨大变化。因此，不仅定义配体在趋化因子受体范围内的选择性的能力至关重要，而且测量所产生的信号的强度、持续时间和最终结果的能力也至关重要。使用20+趋化因子受体，45+配体和多个下游读数，使用标准技术测量可能表型的矩阵是时间和成本过高的。在I期研究中，我们开发了一个多功能的受体内化检测平台，可以在一个检测井中同时筛选9种趋化因子受体。在这个II期提案中，我们将创建两个包含18个人类或小鼠趋化因子受体的多重面板，并扩大检测读数的数量，包括钙信号，受体脱敏和趋化性。该系统将用于创建所有自然发生的趋化因子的综合配体-受体相互作用数据库，并发现具有选择性和信号传导的非自然特征的合成趋化因子配体。这些结果将为该技术的商业化提供基础，作为药物发现平台，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。

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.