CHEMICAL SYNTHESIS & TARGET IDENTIFICATION OF CERATOSPONGAMIDE

化学合成

• 批准号: 8363735
• 负责人: Jack Taunton
• 金额: $ 0.21万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-06-01 至 2012-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8363735
• 关键词: Affinity Chromatography; Asthma; Autoimmune Diseases; Binding; Cell Surface Receptors; Cells; Complex; Cyclization; Cysteine; Funding; Gene Expression; Grant; Immune; Inflammation Mediators; Inflammatory; Inhibitory Concentration 50; Mass Spectrum Analysis; Measurement; Methodology; National Center for Research Resources; Pattern; Peptides; Principal Investigator; Protein Kinase; Proteins; Radiolabeled; Research; Research Infrastructure; Resources; Rheumatoid Arthritis; Side; Signal Pathway; Signaling Molecule; Source; TNF gene; Thiazoles; Threonine; Tissues; United States National Institutes of Health; Vertebral column; cellular targeting; chemical synthesis; cost; cytokine; marine natural product; radiotracer; response

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. Inflammatory and autoimmune diseases, such as asthma and rheumatoid arthritis, are characterized by the accumulation of immune cells that cause tissue damage in response to a complex array of secreted signaling molecules. Two cytokines thought to be critical mediators of inflammation are IL-1b and TNF-a. These cytokines exert distinct yet overlapping effects on target tissues, primarily by altering patterns of gene expression. IL-1b and TNF-a control gene expression by binding to distinct cell surface receptors, whereupon multiple protein kinase cascades are activated. We propose a pharmacological approach to study these complex signaling pathways. Our idea derives from the recent discovery that a marine natural product, ceratospongamide, potently inhibits gene expression induced by IL-1b with an IC50 of 32 nM. We seek to elucidate its mechanism of action. Ceratospongamide is a macrocyclic heptapeptide that is conformationally constrained by two additional rings (thiazole and oxazoline) formed by the cyclization of cysteine and threonine side chains onto the peptide backbone. We will synthesize ceratospongamide and derivatives that will enable the identification of its cellular targets. both radiolabeled and immobilized derivatives for affinity purification will be synthesized using modern synthetic methodology. Mass spectrometry measurements obtained at the UCSF Facility will be essential for two reasons: (1) characterizing synthetic intermediates and (2) identifying ceratospongamide's protein target.

该子项目是利用资源的众多研究子项目之一 由 NIH/NCRR 资助的中心拨款提供。子项目的主要支持 并且子项目的主要研究者可能是由其他来源提供的， 包括其他 NIH 来源。 子项目可能列出的总成本 代表子项目使用的中心基础设施的估计数量， NCRR 赠款不直接向子项目或子项目工作人员提供资金。 炎症和自身免疫性疾病，例如哮喘和类风湿性关节炎，其特征是免疫细胞积聚，这些细胞响应一系列复杂的分泌信号分子而导致组织损伤。 IL-1b 和 TNF-a 被认为是炎症关键介质的两种细胞因子。 这些细胞因子主要通过改变基因表达模式对靶组织产生独特但重叠的影响。 IL-1b 和 TNF-a 通过与不同的细胞表面受体结合来控制基因表达，从而激活多个蛋白激酶级联。 我们提出了一种药理学方法来研究这些复杂的信号通路。 我们的想法源自最近的发现，即海洋天然产物角海绵酰胺可有效抑制 IL-1b 诱导的基因表达，IC50 为 32 nM。 我们试图阐明其作用机制。 角海绵酰胺是一种大环七肽，其构象受到两个附加环（噻唑和恶唑啉）的限制，这两个环是由半胱氨酸和苏氨酸侧链环化到肽主链上形成的。 我们将合成角海绵酰胺及其衍生物，从而能够识别其细胞靶点。 用于亲和纯化的放射性标记和固定化衍生物将使用现代合成方法合成。 在加州大学旧金山分校设施获得的质谱测量至关重要，原因有两个：(1) 表征合成中间体；(2) 识别角海绵酰胺的蛋白质靶标。