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: 8245006
• 负责人: Brian F Volkman
• 金额: $ 41.04万
• 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 2015-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8245006
• 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 Delivery Systems; 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; Screening procedure; 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; 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）：本项目的目标是开发一个用于发现小分子趋化因子抑制剂的创新平台。趋化因子主要通过结合和激活由酪氨酸硫酸化修饰的G蛋白偶联受体家族来指导细胞迁移。硫酸化是趋化因子功能所必需的，表明受体磺基酪氨酸直接参与趋化因子配体的特异性结合。我们最近揭示了在趋化因子家族中磺基酪氨酸识别的分子基础，通过解决趋化因子SDF 1/CXCL 12和其受体CXCR 4的含磺基酪氨酸片段之间的复合物的NMR结构。CXCR 4的N-末端胞外结构域包含三个潜在的硫酸化位点，并且每个磺基酪氨酸占据CXC 12表面上的独特裂缝。我们假设，因为磺基酪氨酸识别位点是必不可少的高亲和力受体结合，每一个代表了一个目标的抑制使用小分子针对趋化因子配体。在具体目标1中，我们建议在CXCL 12表面上定义额外的磺基酪氨酸识别位点，并使用高通量对接计算、通过2D NMR验证位点特异性配体结合和功能测定的组合来测试趋化因子抑制。我们将在目标2中使用CXCL 12/CXCR 4复合物的结构作为测试案例来改进我们的计算机筛选方法，以评估针对NMR构象异构体的集合的柔性侧链对接和片段筛选。将使用SAR分析并通过设计和合成结合来自相邻磺基酪氨酸识别位点的联合收割机配体的新型化合物，对特定目标3中的亲和力和选择性进行命中-引导和引导优化。结合CXCL 12并在基于细胞的趋化因子测定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鉴定特定目标4中的位点，并将我们的筛选方法应用于一组代表性的趋化因子靶标。由磺基酪氨酸识别指导的基于结构的药物发现是一个概念上的进步，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。应用这种策略产生的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。