Novel Approaches to Identify Anti-Angiogenesis Drugs

鉴定抗血管生成药物的新方法

• 批准号: 6792527
• 负责人: Joseph M Colacino
• 金额: $ 18.82万
• 依托单位: PTC THERAPEUTICS, INC.
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-04-01 至 2004-09-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6792527
• 关键词: angiogenesis inhibitors; athymic mouse; biomarker; cell line; chemical structure function; drug screening /evaluation; enzyme linked immunosorbent assay; fibroblast growth factor; genetically modified animals; human tissue; hypoxia; immunocytochemistry; laboratory mouse; messenger RNA; neoplasm /cancer blood supply; neoplastic growth; nucleic acid sequence; pharmacokinetics; posttranscriptional RNA processing; retinopathy of prematurity; ribosomal RNA; vascular endothelial growth factors

DESCRIPTION (provided by applicant): Up-regulation of vascular endothelial growth factor (VEGF), a key factor for angiogenesis, is an important contributor to the pathogenesis of cancers, diabetic retinopathy, and exudative macular degeneration. The abundance of VEGF is in large part controlled at the post-transcriptional level by sequences in both the 5'- and 3'-untranslated regions (UTRs) of its mRNA. The 5'-UTR contains an internal ribosomal entry site (IRES) that mediates a unique, cap-independent mode of translation initiation. Under hypoxic conditions, cap-dependent translation is dramatically impaired and the translation of the VEGF mRNA occurs through its cap-independent IRES. Thus, even under severe hypoxic conditions, cells are capable of producing large amounts of VEGF resulting in angiogenesis to support further tumor growth or aberrant neovascularization as occurs in ocular diseases. In addition, the 3'-UTR harbors multiple AU-rich stability determinants that have been previously shown to regulate VEGF mRNA turnover rates. Using PTC's proprietary technology platforms, we have successfully identified a collection of compounds that inhibit the expression of VEGF post transcriptionally. The initial results from medicinal chemistry efforts are very encouraging, since all three lead series identified contain compounds with low nanomolar activity for the inhibition of VEGF expression. Preliminary selectivity studies demonstrated that there is a subset of compounds that specifically inhibit VEGF production, while a second set of compounds inhibits VEGF expression as well as other angiogenesis factors such as FGF-2 expression. Based on these results, the aims of this proposal are to further characterize these compounds identified from HTS and preliminary SAR studies in a battery of selectivity assays and cytotoxicity assays; initiate structure-activity studies around the lead compounds to enhance potency, efficacy and selectivity; and develop end-point assays to measure the therapeutic efficacies of compounds for future studies in animals.

描述（由申请人提供）： 血管内皮生长因子 (VEGF) 是血管生成的关键因素，其上调是癌症、糖尿病视网膜病变和渗出性黄斑变性的重要发病机制。 VEGF 的丰度在很大程度上在转录后水平上由其 mRNA 5' 和 3' 非翻译区 (UTR) 中的序列控制。 5'-UTR 包含一个内部核糖体进入位点 (IRES)，可介导独特的、与帽无关的翻译起始模式。在缺氧条件下，帽依赖性翻译显着受损，并且 VEGF mRNA 的翻译通过其帽非依赖性 IRES 进行。因此，即使在严重缺氧条件下，细胞也能够产生大量 VEGF，导致血管生成，以支持肿瘤的进一步生长或眼部疾病中发生的异常新血管形成。此外，3'-UTR 包含多个富含 AU 的稳定性决定簇，先前已证明这些决定簇可调节 VEGF mRNA 周转率。利用 PTC 的专有技术平台，我们成功鉴定了一系列可抑制 VEGF 转录后表达的化合物。药物化学工作的初步结果非常令人鼓舞，因为所有三个已鉴定的先导系列均含有具有低纳摩尔活性的化合物，可抑制 VEGF 表达。初步选择性研究表明，有一组化合物特异性抑制 VEGF 产生，而第二组化合物则抑制 VEGF 表达以及其他血管生成因子（例如 FGF-2 表达）。基于这些结果，该提案的目的是进一步 在一系列选择性测定和细胞毒性测定中表征从 HTS 和初步 SAR 研究中鉴定出的这些化合物；围绕先导化合物启动结构活性研究，以增强效力、功效和选择性；并开发终点测定法来测量化合物的治疗效果，以供未来的动物研究使用。