Development of GPR30-Selective Ligands

说明(申请人提供)：雌激素是人体内的一种关键荷尔蒙，调节许多组织的生长、发育和动态平衡。对雌激素的生理反应包括对哺乳动物生殖和乳房功能、中枢神经和免疫系统、骨骼生理和血管功能的调节。我们最近描述了七个跨膜G蛋白偶联雌激素受体GPR30的新功能。该受体被经典雌激素受体ER1和ER2的激动剂和拮抗剂激活。直到最近，还没有已知的GPR30的特定配体，这使得传统的药理学方法研究这种受体变得困难。然而，我们最近的研究结合了虚拟筛选和生物分子筛选，发现了第一个GPR30选择性激动剂G-1。本申请的具体目的是：1.进行虚拟和生物分子筛选的组合，以基于目前已知结合和激活GPR30的化合物来鉴定额外的GPR30特异性配体。将进行结构-活性分析，以确定与经典雌激素受体相比，GPR30结合选择性和活性的关键分子决定因素。2.基于目标1的生物分子筛选结果和构效关系分析，合理设计和合成新型G-1配体的小文库(每个周期最多20个化合物)。这一目标的目的是分离配体内的激活性和拮抗性，并通过有针对性的合成化学来进一步评估新的GPR30配体的SAR。3.鉴定AIMS 1和AIMS 2中鉴定和合成的化合物的生物学功能。将使用一系列功能生物测定来表征显示活性的化合物的生物学效应。这些分析将包括细胞内信号分析，如钙动员、ERK和EGFR磷酸化以及PI3K激活；更复杂的细胞分析，如转录激活、细胞迁移和增殖；以及使用小鼠模型的体内研究。了解与GPR30结合的配体的药理学特征和构效关系，对于发现针对该受体的新药至关重要，目的是揭示该受体的潜在生理学，并开发改善雌激素依赖性癌症治疗的方法。公共卫生相关性：雌激素在正常和疾病生物学中发挥着重要作用。我们已经鉴定了一种新的细胞膜雌激素受体，它可能在雌激素生物学中发挥作用。这项工作的目标是开发新的化合物，能够特异性地靶向这种新的受体，在不影响其他雌激素受体的情况下激活或抑制其活性。

DESCRIPTION (provided by applicant): Estrogen is a critical hormone in the human body that regulates the growth, development and homeostasis of many tissues. Physiological responses to estrogen include the regulation of mammalian reproduction and breast function, central nervous and immune systems, skeletal physiology and vascular function. We have recently described novel functions of the seven transmembrane G protein-coupled estrogen receptor, GPR30. This receptor is activated by both agonists and antagonists of the classical estrogen receptors, ER1 and ER2. Until recently there were no known specific ligands for GPR30, making traditional pharmacological approaches to the study of this receptor difficult. Our recent studies however have combined both virtual and biomolecular screening to discover the first GPR30-selective agonist, G-1. The specific aims of this application are: 1. Perform a combination of virtual and biomolecular screening to identify additional GPR30-specific ligands based on compounds presently known to bind and activate GPR30. Structure-activity analyses will be carried out to determine the critical molecular determinants for GPR30 binding selectivity and activity as compared to classical estrogen receptors. 2. Based on the biomolecular screening results and structure-activity analyses of Aim 1, rationally design and synthesize small libraries (up to 20 compounds per cycle) of novel G-1-based ligands. The goal of this aim is to separate agonism from antagonism within ligands, and to further evaluate the SAR of novel GPR30 ligands through targeted synthetic chemistry. 3. Characterize the biological functions of the compounds identified and synthesized in Aims 1 and 2. A collection of functional bioassays will be employed to characterize the biological effects of the compounds displaying activity. These assays will include intracellular signaling assays such as calcium mobilization, ERK and EGFR phosphorylation and PI3K activation; more complex cellular assays such as transcriptional activation, cell migration and proliferation; and in vivo studies using mouse models. Understanding the pharmacological profile and structure-activity relationships for ligand binding to GPR30 will be critical to the discovery of novel drugs that target this receptor for the purposes of revealing the underlying physiology of the receptor and developing therapeutic approaches for the improved treatment of estrogen-dependent cancers. PUBLIC HEALTH RELEVANCE: Estrogen plays an important role in normal and disease biology. We have characterized a novel membrane estrogen receptor that likely plays a role in estrogen biology. The goal of this work is to develop novel compounds that can specifically target this new receptor to either activate or inhibit its activity without affecting other estrogen receptors.