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

GPCR是存在于细胞膜中的重要信号转导蛋白，并且是许多稳态过程的必需调节剂，并且被超过30%的药物靶向。β-肾上腺素受体（A家族GPCR）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。β 1、β 2和β 3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显示出选择性，使得难以研究它们的独特功能。我们最近已经确定，通过先进的分子模拟方法，一种可能的新模式的实验亚型选择性配体β肾上腺素受体的结合。这种结合模式可以解释结构-活性关系数据，否则似乎矛盾。了解这些原型GPCR如何与配体相互作用可能与更广泛的GPCR家族的其他成员高度相关。这个多学科项目的目的是设计，合成和构建配体库，以更严格地验证这一假设。与此同时，将进行分子模拟研究，最终将与药理学数据相关联。目标：-设计和合成β肾上腺素受体配体库。使用基于细胞的测定来表征这些配体的特性。使用先进的分子建模方法，包括灵活的对接和分子动力学模拟，以预测新的β肾上腺素受体配体与蛋白质的结合模式。计算化合物的结合亲和力，并确定构效关系。主要方法：-有机合成化学和结构鉴定。-配体亲和力和亚型选择性的细胞培养和药理学评价。配体-蛋白质复合物的分子模拟与构效数据分析。

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.