New chemical biology approaches to study P2Y2 receptors in health and disease

细胞交流的过程是所有生命至关重要和不可或缺的一部分，并控制着生物体的内部运作，使它们能够做出反应、适应和生存。G蛋白偶联受体(GPCRs)是一大类位于细胞表面的相关蛋白。受体可以检测细胞外的分子，并通过一个称为细胞信号传递的过程激活细胞内的细胞反应。这个项目将解决基本问题，因为我们将能够通过合成包含荧光基团的分子来可视化和微调生物受体的反应，从而使我们能够可视化这一关键细胞信号传递过程中的关键过程。我们将制造能够开启(痛苦)或开启(对抗)受体发出信号的能力的分子。膜结合的P2受体介导小分子(核苷酸)在细胞间信号转导中的作用，而P2Y受体属于GPCRs家族。值得注意的是，对该受体的阻滞剂(拮抗剂)和激动剂(激动剂)的阐明和开发都落后于许多其他GPCR。因此，缺乏有效和选择性的化学P2Y2R探针，作为药理学工具来阐明受体在未得到满足的医疗需求领域的生理作用。这项研究提案的引人注目的目标在于应用一种基于荧光的合成化学平台技术，同时偶然发现抑制这一关键受体作用的新分子。我们将合成新的高亲和力的P2Y2R拮抗剂和激动剂探针，用于未来的药物发现研究计划，以证明这种未被研究的生物受体的临床益处。我们将建立新的方法来研究受体的功能，通过建立新的生化分析和可视化技术，使我们能够更好地了解激活P2Y2R的结构要求，并研究受体所在的细胞群体。这项研究横跨合成和计算化学与药理学，在医疗保健、科学就业创造和培训领域具有广泛的重要性。新出现的证据表明，对P2Y2R的对抗在未得到满足的医疗需求的关键领域发挥了作用。最近的研究表明，P2Y2R在某些癌细胞中表达，并可能在肿瘤细胞的增殖中发挥重要作用。癌症是世界范围内主要的死亡原因，是一大类疾病的总称，这些疾病可以影响身体的任何部位，如肺、胃、肝、结肠和乳房。癌症的一个决定性特征是迅速产生异常细胞，这些细胞超出它们通常的边界生长，然后可以侵袭身体的邻近部分，并扩散到其他器官。这个被称为转移的过程是癌症死亡的主要原因。干扰G蛋白偶联的P2Y受体信号是治疗侵袭性和难治性癌症的一种有前途的替代疗法。此外，还证实了P2Y2R在纤维化肺疾病中的关键中介作用。特发性肺纤维化(IPF)是一种肺部结疤和呼吸变得越来越困难的疾病。这是一种毁灭性的疾病，几乎没有可用的治疗选择。总之，这一应用将确保高亲和力和稳定的化学荧光探针的合成，从而使未来的药物发现拨款申请成为可能。在本项目中发现的高亲和力P2Y2R拮抗剂和激动剂配体的合成将用于探索未检测疾病状态下P2Y2R的生物潜力，而荧光高亲和力P2Y2R化学探针将用于建立新的高通量生物检测方法，以便将来对这一重要受体进行筛选。

The process in which cells communicate is a vital and integral part of all life and controls the inner workings of organisms allowing them to respond, adapt and survive. G protein-coupled receptors (GPCRs) are a large group of related proteins that are located on the surface of cells. The receptors can detect molecules outside the cell and activate cellular responses within the cells through a process called cell signalling. This project will address fundamental questions as we will be able to visualise and finely tune a biological receptor's response through the synthesis of molecules that contain a fluorescent group allowing us to visualise the key processes in this critical cell signalling process. We will make molecules that can turn on (agonise) or turn of (antagonise) the ability of the receptor to signal. Membrane-bound P2-receptors mediate the actions of small molecules (nucleotides) in cell-to-cell signalling and P2Y-receptors belong to the family of GPCRs. The P2Y2 receptor (P2Y2R) is notable that elucidation and exploitation of the both blockers (antagonists) and activators (agonists) of this receptor has lagged behind that of many other GPCRs. As such, there is a lack of potent and selective chemical P2Y2R probes, required as pharmacological tools to elucidate the physiological roles of the receptor in areas of un-met medical need. The compelling goal of this research proposal lies in the application of a fluorescence-based synthetic chemistry platform technology in combination with a serendipitous discovery of the discovery of new molecules that inhibit the action of this key receptor. We will synthesise new high-affinity P2Y2R antagonist and agonist probes for use in future drug discovery research programmes in order to prove the clinical benefit of this understudied biological receptor. We will establish new ways to study the receptor function, through the establishment of novel biochemical assays and visualisation techniques enabling us to understand better the structural requirements for activation of the P2Y2R and for studying cell populations where the receptor is located. The research spanning synthetic and computational chemistry with pharmacology, has widespread importance in the areas of healthcare, scientific job creation and training. Emerging evidence suggests antagonism of the P2Y2R has roles in key areas of unmet medical need. Recent disclosures have demonstrated that P2Y2Rs are expressed in certain cancer cells and could play a vital role in tumour cell proliferation. Cancer is a leading cause of death worldwide and is a generic term for a large group of diseases that can affect any part of the body such as; lung, stomach, liver, colon and breast. One defining feature of cancer is the rapid creation of abnormal cells that grow beyond their usual boundaries, and which can then invade adjoining parts of the body and spread to other organs. This process known as metastasis is the major cause of death from cancer. Interfering with G protein-coupled P2Y receptor signalling is a promising therapeutic alternative to treat aggressive and difficult-to-manage cancers. In addition, the role of the P2Y2R as a key mediator in fibrotic lung diseases was demonstrated. Idiopathic pulmonary fibrosis (IPF) is a condition in which the lungs become scarred and breathing becomes increasingly difficult. It is a devastating disease with few available treatment options. In conclusion, this application will secure the synthesis of the high affinity and stable chemical fluorescent probes to enable future drug discovery grant applications. The synthesis of high affinity P2Y2R antagonist and agonist ligands discovered in this programme will be used to probe the biological potential of P2Y2R in unmet disease states and the fluorescent high affinity P2Y2R chemical probes will be used to establish novel high-throughput biological assays to enable future screening of this important receptor.