Selective Targeting of G Protein beta gamma Subunits with Small Molecules

小分子选择性靶向 G 蛋白 β γ 亚基

• 批准号: 8051989
• 负责人: Alan V. Smrcka
• 金额: $ 4.03万
• 依托单位: UNIVERSITY OF ROCHESTER
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-05-01 至 2011-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8051989
• 关键词: Absence of pain sensation; Address; Adenylate Cyclase; Affect; Animal Model; Binding; Binding Sites; Biological Assay; Cardiovascular system; Cell Line; Cells; Characteristics; Chemistry; Chemotactic Factors; Chemotaxis; Computer software; Coupled; Crystallography; Data; Development; Disease; Dissociation; Docking; Equilibrium; Family; Fostering; G Protein-Coupled Receptor Signaling; G-Protein-Coupled Receptors; G-protein Beta gamma; GTP Binding; GTP-Binding Proteins; HL60; Heart failure; Human; IL8 gene; In Vitro; Individual; Inflammation; Investigation; Kinetics; Knowledge; Laboratories; Lead; Ligands; Measures; Mediating; Methods; Modeling; Modification; Molecular; Morphine; Mutagenesis; Pharmacologic Substance; Phosphotransferases; Play; Process; Proteins; Receptor Mediated Signal Transduction; Regulation; Role; Screening procedure; Signal Pathway; Signal Transduction; Site; Site-Directed Mutagenesis; Specificity; Surface Plasmon Resonance; System; Testing; Therapeutic; Therapeutic Agents; X-Ray Crystallography; base; cofactor; improved; inhibitor/antagonist; inward rectifier potassium channel; neutrophil; novel; novel therapeutic intervention; novel therapeutics; protein protein interaction; public health relevance; receptor; research study; small molecule

DESCRIPTION (provided by applicant): G proteins 23 subunits play a central role in G-protein coupled receptor (GPCR)- mediated signal transduction. They act as cofactors in the receptor-mediated activation process as well as playing direct roles in signal transfer to downstream targets. Considerable data has accumulated in number of systems that excess 23 signaling has pathological consequences and that manipulation of 23 subunit signaling could be an effective therapeutic strategy in heart failure as well as other diseases. We have developed a novel targeting strategy for selective manipulation of G protein 23 subunit signaling pathways by selectively blocking 23-subunit binding interactions with functional protein partners using small molecules. In our preliminary data we demonstrate these compounds are efficacious in animal models of heart failure, inflammation and morphine-dependent analgesia. In the proposed experiments we will explore the fundamental mechanisms underlying binding and selectivity of these 23 binding compounds. In Specific aim 1 the experiments will characterize the selectivity of compounds for a broad range of G23 targets and develop new assays for predicting compound selectivity. Specific aim 2 will combine biophysical ligand interaction methods, mutagenesis and x-ray crystallography to develop a detailed understanding of the mechanisms for selectivity. Specific aim3 will exploit the information from specific aim 2 to refine our computational screening approach to allow for more efficient compound identification that may have unique mechanisms of action. Specific aim 4 will apply our knowledge of selectivity to GPCR signaling in neutrophil functions. Successful completion of the proposed experiments will lead to a thorough understanding of a the mechanism of action of a new family of molecules that target G23 signaling that have potential uses in dissecting the mechanisms of action of GPCR stimulated signaling and providing the basis for novel therapeutic approaches. G protein coupled receptors (GPCRs) are a major class of transmembrane receptors responsible for recognition of a large class of diverse ligands. Here we propose investigation of selective small molecule inhibitors of G protein 23 subunits identified in our laboratory which could be used to inhibit multiple GPCRs and modify actions of existing GPCR directed pharmaceuticals. Public Health Relevance: Results of these experiments will help to validate this alternate approach to modification of signaling pathways downstream of GPCRs that could ultimately lead to development of novel therapeutics.

描述（由申请人提供）：G蛋白23亚基在G蛋白偶联受体（GPCR）介导的信号转导中发挥核心作用。它们在受体介导的激活过程中充当辅助因子，并在信号传递至下游靶点中发挥直接作用。许多系统中积累的大量数据表明，超过 23 信号传导会产生病理后果，并且操纵 23 亚基信号传导可能是心力衰竭和其他疾病的有效治疗策略。我们开发了一种新颖的靶向策略，通过使用小分子选择性阻断 23 亚基与功能蛋白伴侣的结合相互作用，选择性操纵 G 蛋白 23 亚基信号通路。在我们的初步数据中，我们证明这些化合物在心力衰竭、炎症和吗啡依赖性镇痛的动物模型中有效。在拟议的实验中，我们将探索这 23 种结合化合物的结合和选择性的基本机制。在具体目标 1 中，实验将表征化合物对广泛 G23 靶标的选择性，并开发用于预测化合物选择性的新测定方法。具体目标 2 将结合生物物理配体相互作用方法、诱变和 X 射线晶体学，以详细了解选择性机制。具体目标 3 将利用具体目标 2 的信息来完善我们的计算筛选方法，以便更有效地识别可能具有独特作用机制的化合物。具体目标 4 将应用我们对中性粒细胞功能中 GPCR 信号传导选择性的了解。成功完成拟议的实验将有助于彻底了解以 G23 信号传导为目标的新分子家族的作用机制，这些分子在剖析 GPCR 刺激信号传导的作用机制方面具有潜在用途，并为新的治疗方法提供基础。 G 蛋白偶联受体 (GPCR) 是一类主要的跨膜受体，负责识别一大类不同的配体。在这里，我们建议研究我们实验室鉴定的 G 蛋白 23 亚基的选择性小分子抑制剂，该抑制剂可用于抑制多种 GPCR 并改变现有 GPCR 导向药物的作用。 公共卫生相关性：这些实验的结果将有助于验证这种修改 GPCR 下游信号通路的替代方法，最终可能导致新型疗法的开发。