Molecular and in-silico interrogation of novel modes of binding at G protein-coupled receptors (GPCRs)

GPCRs是存在于细胞膜上的重要信号转导蛋白，是许多动态平衡过程的重要调节因子，超过30%的药物以GPCRs为靶标。β-肾上腺素能受体(A家族GPCRs)是心血管和呼吸系统的关键调节因子，也是公认的药物靶点。Beta1、Beta2和Beta3亚型具有不同的组织分布和药理活性，但很少有已知的配体具有选择性，这使得研究它们不同的功能变得困难。我们最近通过先进的分子模拟方法确定了一种可能的新模式，即实验亚型选择性配体与β受体的结合。这种结合模式可以解释原本看起来相互矛盾的结构-活性关系数据。了解这些典型的GPCRs如何与配体相互作用可能与更广泛的GPCRs家族的其他成员高度相关。这一多学科项目的目标将是设计、合成和药理学表征一个配体文库，以更严格地检验这一假说。同时，将进行最终将与药理学数据相关的分子模拟研究。目标：-设计和合成一个β受体配体文库。-使用基于细胞的分析方法对这些配体进行药理学表征。-使用先进的分子模拟方法，包括灵活的对接和分子动力学模拟，预测新的β受体配体与蛋白质的结合模式。-计算结合亲和力，并确定结构-活性关系。主要方法：-合成有机化学和结构阐明。-细胞培养和配基亲和力和亚型选择性的药理学评价。-配体和蛋白质复合体的分子模拟和结构-活性数据分析。

GPCRs are important signal transduction proteins residing in the cell membrane and are essential regulators of many homeostatic processes and targeted by great than 30% of drugs. The beta-adrenoceptors (family A GPCRs), are key regulators of the cardiovascular and respiratory systems and well-established drug targets. beta1, beta2, and beta3 subtypes have distinct tissue distributions and pharmacological activity, but few known ligands show selectivity, making it difficult to research their distinct functions. We have recently identified, through advanced molecular modelling methods, a possible novel mode of binding of experimental subtype-selective ligands to beta adrenoceptors. This mode of binding may explain structure-activity relationship data that otherwise appears contradictory. An understanding of how these prototypical GPCRs interact with ligands is likely to be highly relevant to other members of the wider GPCR family.The aim of this multidisciplinary project will be to design, synthesise and pharmacologically characterise a library of ligands to test this hypothesis more rigorously. In parallel, molecular modelling studies will be performed that eventually will be correlated with pharmacological data.Objectives: -Design and synthesise a library of beta adrenoceptor ligands.-Characterise these ligands pharmacologically, using cell-based assays.-Use advanced molecular modelling methods, including flexible docking and molecular dynamics simulations, to predict the mode of binding of new beta adrenoceptor ligands to the proteins.-Make calculations of the binding affinities, and identify structure-activity relationships.Major methods:-Synthetic organic chemistry and structural elucidation.-Cell culture and pharmacological evaluation of ligand affinity and subtype selectivity.-Molecular modelling of ligand-protein complexes and analysis of structure-activity data.