Novel Small-Molecule Inhibitors of the Tie2/Angiopoietin Interactions and Signali

Tie2/血管生成素相互作用和信号的新型小分子抑制剂

• 批准号: 7426768
• 负责人: DIMITAR B NIKOLOV
• 金额: $ 18.8万
• 依托单位: SLOAN-KETTERING INST CAN RESEARCH
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-09-30 至 2009-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7426768
• 关键词: Angiopoietin-2; Angiopoietins; Animals; Antineoplastic Agents; Binding; Biochemical; Biological; Biological Assay; Blood Vessels; Cells; Combined Modality Therapy; Complex; Development; Disruption; Fluorescence; Goals; Growth; In Vitro; Libraries; Ligands; Lymphatic System; Maintenance; Measures; Mediating; Phosphotransferases; Principal Investigator; Property; Protein Tyrosine Kinase; Research; Role; Screening procedure; Signal Transduction; Structure; TIE-2 Receptor; Therapeutic Agents; Time; Validation; angiogenesis; anti-cancer therapeutic; antiangiogenesis therapy; base; cancer type; clinical application; cytotoxicity; design; extracellular; high throughput screening; in vivo; inhibitor/antagonist; novel; programs; receptor; small molecule; small molecule libraries; tumor

DESCRIPTION (provided by applicant): The goal of this study is to develop a robust and sensitive assay and to implement it for the high throughput screening (HTS) of chemical libraries for compounds that block the Tie2/Angiopoietin interactions and signaling. The Tie2 receptor tyrosine kinase and its angiopoietin ligands regulate both developmental and tumor-induced angiogenesis and, therefore, our proposed research will facilitate the development of novel anti-tumor strategies based on the targeted disruption of tumor-induced blood vessel formation. Our preliminary structural, biophysical and biochemical characterization of the angiopoietins, Tie2, and of their interactions, suggests that small-molecules would be able to disrupt the Tie2/Ang complex formation and the initiation of Tie2-mediated signaling. Furthermore, we are currently developing a Homogeneous Time Resolved Fluorescence (HTRF) ligand-receptor interactions assay, which we have documented is suitable for implementation in a high-throughput screen format. We have also identified suitable secondary- and counter- screening assays for "hit" validation and prioritization. We propose to further optimize the primary HTRF assay, configure it in HTS format and perform a validation screen of ~2700 selected compounds. Subsequently, we propose to implement this assay to screen a complete compound library for small molecules that disrupt respectively the Tie2/Ang1 and the Tie2/Ang2 complex formation. Finally, we will use a variety of in vitro and in vivo biochemical, biophysical, as well as cell- and animal-based assays, to validate the HTS hits and to characterize their biological properties, cytotoxicity and potential as anti-angiogenesis-based anti-cancer agents.

描述（由申请人提供）：本研究的目标是开发一种强大而敏感的检测方法，并将其用于化学文库的高通量筛选（HTS），以筛选阻断Tie2/血管生成素相互作用和信号传导的化合物。Tie2受体酪氨酸激酶及其血管生成素配体调节发育和肿瘤诱导的血管生成，因此，我们提出的研究将促进基于靶向破坏肿瘤诱导的血管形成的新型抗肿瘤策略的发展。我们对血管生成素、Tie2及其相互作用的初步结构、生物物理和生化表征表明，小分子能够破坏Tie2/Ang复合物的形成和Tie2介导的信号传导的启动。此外，我们目前正在开发一种均匀时间分辨荧光（HTRF）配体-受体相互作用测定，我们已经证明它适合在高通量屏幕格式下实施。我们还确定了用于“命中”验证和优先排序的合适的二次筛选和反筛选试验。我们建议进一步优化初级HTRF分析，将其配置为HTS格式，并对约2700个选定的化合物进行验证筛选。随后，我们建议实施该实验来筛选一个完整的化合物文库，以分别破坏Tie2/Ang1和Tie2/Ang2复合物形成的小分子。最后，我们将使用各种体外和体内生化、生物物理以及基于细胞和动物的分析来验证HTS命中，并表征其生物学特性、细胞毒性和作为基于抗血管生成的抗癌药物的潜力。