Sulfotyrosine-guided discovery of small molecule chemokine inhibitors

磺基酪氨酸引导的小分子趋化因子抑制剂的发现

• 批准号: 8639582
• 负责人: Brian F Volkman
• 金额: $ 28.29万
• 依托单位: MEDICAL COLLEGE OF WISCONSIN
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-04-01 至 2016-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8639582
• 关键词: Accounting; Affinity; Agonist; Amino Acids; Animals; Anti-Inflammatory Agents; Anti-inflammatory; Autoimmune Diseases; Autoimmune Process; Binding; Binding Sites; Biological; Biological Assay; Bone Marrow; CXCL12 gene; CXCR4 Receptors; CXCR4 gene; Cardiovascular Diseases; Cell surface; Cells; Chemicals; Chronic; Cleaved cell; Complex; Computer Simulation; Data; Development; Diagnostic; Disseminated Malignant Neoplasm; Disulfides; Docking; Drug Targeting; Embryonic Development; Epitopes; Evaluation; Extracellular Domain; Extracellular Space; Family; Funding; G Protein-Coupled Receptor Genes; G Protein-Coupled Receptor Signaling; G-Protein-Coupled Receptors; GTP-Binding Proteins; Goals; Golgi Apparatus; HIV Infections; Health; Homing; Human; Immune; Immune response; Infection; Inflammation; Inflammatory; Integral Membrane Protein; Laboratories; Lead; Ligand Binding; Ligands; Liver; Lung; Malignant Neoplasms; Measures; Methods; Modeling; Modification; Molecular; Molecular Conformation; Multiple Sclerosis; N-terminal; Neoplasm Metastasis; Pharmacologic Substance; Post-Translational Protein Processing; Prevalence; Process; Proteins; Receptor Activation; Research; Role; Scheme; Side; Signal Transduction; Signaling Molecule; Site; Stem cells; Structure; Surface; System; Testing; Therapeutic Agents; Time; Tissues; Titrations; Transmembrane Domain; Tyrosine; United States National Institutes of Health; Validation; Work; Wound Healing; base; cancer type; cell motility; chemokine; combinatorial; conformer; design; drug development; drug discovery; extracellular; flexibility; human disease; improved; inhibitor/antagonist; innovation; lymph nodes; member; molecular recognition; novel; preclinical study; prevent; receptor; receptor binding; receptor coupling; response to injury; screening; small molecule; sulfation; trafficking; tyrosine O-sulfate

DESCRIPTION (provided by applicant): The goal of this project is the development of an innovative platform for discovery of small molecule chemokine inhibitors. Chemokines direct cell migration primarily by binding and activating a family of G protein-coupled receptors that are post-translationally modified by tyrosine sulfation. Sulfation is required for chemokine function, suggesting that receptor sulfotyrosines participate directly in specific binding of the chemokine ligand. We recently revealed the molecular basis for sulfotyrosine recognition in the chemokine family by solving the NMR structure of a complex between the chemokine SDF1/CXCL12 and a sulfotyrosine-containing fragment of its receptor CXCR4. The N-terminal extracellular domain of CXCR4 contains three potential sulfation sites, and each sulfotyrosine occupies a unique cleft on the CXC12 surface. We hypothesize that because sulfotyrosine recognition sites are essential for high affinity receptor binding, each one represents a target for inhibition using small molecules directed at the chemokine ligand. In specific aim 1 we propose to define additional sulfotyrosine recognition sites on the CXCL12 surface and use a combination of high-throughput docking calculations, validation of site-specific ligand binding by 2D NMR, and functional assays to test for chemokine inhibition. We will refine our in silico screening methods in aim 2 using the structure of the CXCL12/CXCR4 complex as a test case for evaluation of flexible side chain docking and fragment screening against the ensemble of NMR conformers. Hit-to-lead and lead optimization will be performed for affinity and selectivity in specific aim 3 using SAR analysis and by designing and synthesizing novel compounds that combine ligands from adjacent sulfotyrosine recognition sites. Compounds that bind CXCL12 and function as potent and selective inhibitors in cell-based chemokine assays will be made available to existing collaborators for preclinical studies in animals. To assess the broader potential of sulfotyrosine- directed inhibition, we will develop a streamlined approach to site identification in specific aim 4 and apply our screening method to a representative set of chemokine targets. Structure-based drug discovery guided by sulfotyrosine recognition is a conceptual advance that can be applied to all 50 members of the chemokine family and other complexes that require this protein modification to function in the extracellular space. New treatments for inflammatory and autoimmune disease or cancer that emerge from application of this strategy will have a major impact on human health.

描述（由申请人提供）：该项目的目标是开发一个创新平台，用于发现小分子趋化因子抑制剂。趋化因子主要通过结合和激活G蛋白偶联受体家族来引导细胞迁移，这些受体在翻译后被酪氨酸硫酸化修饰。趋化因子的功能需要硫酸化，这表明受体硫代酪氨酸直接参与趋化因子配体的特异性结合。最近，我们通过解析趋化因子SDF1/CXCL12与其受体CXCR4中含有硫代酪氨酸片段的复合物的核磁共振结构，揭示了趋化因子家族中硫代酪氨酸识别的分子基础。CXCR4的n端胞外结构域包含三个潜在的硫酸化位点，每个硫代酪氨酸在CXC12表面占据一个独特的间隙。我们假设，因为硫代酪氨酸识别位点对于高亲和力受体结合是必不可少的，每个位点都代表了使用小分子直接抑制趋化因子配体的目标。在具体目标1中，我们建议在CXCL12表面定义额外的硫代酪氨酸识别位点，并结合高通量对接计算，通过二维核磁共振验证位点特异性配体结合，以及功能分析来测试趋化因子的抑制作用。我们将在目标2中完善我们的硅筛选方法，使用CXCL12/CXCR4复合物的结构作为评估柔性侧链对接和针对核磁共振构象集合的片段筛选的测试案例。将使用SAR分析和设计和合成结合邻近巯基酪氨酸识别位点配体的新化合物，对特定目标3的亲和力和选择性进行命中-导联和导联优化。结合CXCL12并在基于细胞的趋化因子分析中作为有效和选择性抑制剂的化合物将提供给现有的合作者用于动物临床前研究。为了评估磺基酪氨酸定向抑制的更广泛潜力，我们将开发一种简化的方法来识别特定目标4的位点，并将我们的筛选方法应用于一组具有代表性的趋化因子靶点。以巯基酪氨酸识别为指导的基于结构的药物发现是一种概念上的进步，可以应用于趋化因子家族的所有50个成员和其他需要这种蛋白质修饰才能在细胞外空间发挥作用的复合物。应用这一策略产生的针对炎症和自身免疫性疾病或癌症的新疗法将对人类健康产生重大影响。