Novel High-throghput Assay:Angiogenesis Inhibitors (RMI)

新型高通量检测：血管生成抑制剂 (RMI)

• 批准号: 6879886
• 负责人: Sunil A David
• 金额: $ 7.2万
• 依托单位: UNIVERSITY OF KANSAS LAWRENCE
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-09-30 至 2006-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6879886
• 关键词: angiogenesis inhibitors; biotechnology; cell proliferation; chemical binding; chemical registry /resource; chorioallantoic membrane; drug discovery /isolation; fibroblast growth factor; heparan sulfate; heparin; high throughput technology; intermolecular interaction; neoplasm /cancer chemotherapy; small molecule; spectrometry; technology /technique development; vascular endothelial growth factors; vascular endothelium

DESCRIPTION (provided by applicant): The vascularization (angiogenesis) of tumors plays an important role in the growth and hematogenous dissemination of neoplasms, and the inhibition of angiogenesis is being increasingly appreciated as a novel and effective therapeutic strategy for limiting tumor growth and spread. Endothelial growth factors play a dominant role in angiogenesis, many of which are dependent on the presence of heparan for binding to their cognate cell-surface receptors and subsequent initiation of downstream signaling cascades that lead to endothelial cell proliferation. We hypothesize that the sequestration of heparan would prevent the binding of heparan-dependent growth factors to endothelial cells and inhibit their mitogenic effects. Based on existing knowledge on the structural requisites for the molecular recognition of heparan sulfate, we have constructed a preliminary pharmacophore for heparan sulfate binding, and propose to screen a carefully chosen focused library of compounds. We will first test the hypothesis that small molecules possessing the pharmacophore will bind heparan sulfate, using heparin as a model heparin compound. We will then examine if the binding will be manifested in inhibition of endothelial growth factor activity using a small subset of compounds. The utilization of a novel dye-displacement spectrophotometric assay as the primary high-throughput screen has already yielded provisional "hits", one of which has been shown to inhibit both FGF and VEGF-induced angiogenic activity in human umbilical endothelial cells. Promising compounds will be examined in greater detail with respect to details of the physical interactions with heparin. Further biological activity will be ascertained in chicken chorioallantoic membrane assays.

描述（由申请人提供）：肿瘤的血管化（血管生成）在肿瘤的生长和血行播散中起着重要作用，并且抑制血管生成作为限制肿瘤生长和扩散的新颖且有效的治疗策略越来越受到重视。 内皮生长因子在血管生成中起主导作用，其中许多依赖于乙酰肝素的存在来与其同源细胞表面受体结合，并随后启动导致内皮细胞增殖的下游信号级联。 我们假设乙酰肝素的隔离会阻止乙酰肝素依赖性生长因子与内皮细胞的结合并抑制其促有丝分裂作用。 基于硫酸乙酰肝素分子识别结构要求的现有知识，我们构建了硫酸乙酰肝素结合的初步药效团，并建议筛选精心选择的重点化合物库。 我们将首先使用肝素作为模型肝素化合物来测试具有药效团的小分子将结合硫酸乙酰肝素的假设。 然后，我们将检查这种结合是否会通过使用一小部分化合物抑制内皮生长因子活性而表现出来。 利用新型染料置换分光光度测定作为主要高通量筛选已经取得了暂时的“成功”，其中一种已被证明可以抑制人脐带内皮细胞中 FGF 和 VEGF 诱导的血管生成活性。 将更详细地研究有前景的化合物与肝素的物理相互作用的细节。 进一步的生物活性将通过鸡绒毛尿囊膜测定来确定。