Substrate specific ERK docking domain inhibitors

底物特异性 ERK 对接域抑制剂

• 批准号: 7076716
• 负责人: PAUL S SHAPIRO
• 金额: $ 21.09万
• 依托单位: UNIVERSITY OF MARYLAND BALTIMORE
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-04-01 至 2011-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7076716
• 关键词: cell proliferation; lead; neoplastic cell; phosphorylation; proteins

DESCRIPTION (provided by applicant): The extracellular signal regulated kinase (ERK1 and ERK2) signalling pathway plays a critical role in cell proliferation. Activation of the ERK proteins either through increased expression of growth factor receptors or genetic mutations of upstream proteins is thought to be involved in the pathogenesis of many human cancers. Thus, inhibition of ERK signaling is viewed as a potential approach to prevent cancer cell proliferation. Currently, no specific inhibitors of the ERK proteins exist. Moreover, most kinase inhibitors lack specificity because they target ATP binding sites, which are well conserved among the protein kinase families. Taking advantage of recently identified ERK2 docking domains, which are reported to facilitate substrate interactions, we have used computer-aided drug design (CADD) to identify novel low molecular weight ERK inhibitors. We hypothesize that low molecular weight compounds that bind to the ERK2 docking domains will selectively block ERK interactions with substrate proteins and inhibit cell proliferation. Following CADD screening, potential compounds were selected and tested for activity in biological assays. Several compounds identified inhibited ERK-specific phosphorylation of ribosomal S6 kinase-1 (RSK-1) and the transcription factor, ELK-1, both of which promote cell proliferation. In addition, active compounds showed a dose-dependent reduction in the proliferation of cancer cell lines as measured by colony survival assays and bind directly to ERK2 as indicated by fluorescence spectroscopy. Aim 1 of the current application will extend the use of CADD to identify additional compounds with a high probability of binding to nine different putative binding pockets on the ERK2 docking domains. MAP kinase substrate phosphorylation assays will be used to characterize the biological activity of the test compounds. The specific interactions between the active compounds and ERK2 will be tested by fluorescence spectroscopy and X-ray crystallography in Aim 2. Lead compounds that inhibit ERK-substrate phosphorylation and bind to specific docking domains will then be tested for their ability to inhibit cancer cell proliferation in cell and animal models. At the end of this application, it is expected that a collection of lead compounds will be identified that block specific ERK-substrate interactions. These compounds will be appropriate for research tools and the development into novel chemotherapeutic agents aimed at treating cancers associated with elevated ERK pathway activity.

描述（申请人提供）：细胞外信号调节激酶（ERK1和ERK2）信号通路在细胞增殖中发挥着关键作用。通过增加生长因子受体的表达或上游蛋白的基因突变来激活 ERK 蛋白被认为与许多人类癌症的发病机制有关。因此，抑制 ERK 信号传导被视为预防癌细胞增殖的潜在方法。目前，不存在 ERK 蛋白的特异性抑制剂。此外，大多数激酶抑制剂缺乏特异性，因为它们靶向 ATP 结合位点，而这些位点在蛋白激酶家族中非常保守。据报道，最近发现的 ERK2 对接域可促进底物相互作用，利用最近发现的 ERK2 对接域，我们使用计算机辅助药物设计 (CADD) 来鉴定新型低分子量 ERK 抑制剂。我们假设与 ERK2 对接结构域结合的低分子量化合物将选择性阻断 ERK 与底物蛋白的相互作用并抑制细胞增殖。 CADD 筛选后，选择了潜在的化合物并测试了生物测定中的活性。鉴定出的几种化合物可抑制核糖体 S6 激酶-1 (RSK-1) 和转录因子 ELK-1 的 ERK 特异性磷酸化，这两种因子均能促进细胞增殖。此外，通过集落存活测定法测量，活性化合物显示出剂量依赖性的癌细胞系增殖减少，并且通过荧光光谱法显示，活性化合物直接与 ERK2 结合。当前申请的目标 1 将扩展 CADD 的使用，以识别其他化合物，这些化合物很可能与 ERK2 对接域上的九个不同的推定结合袋结合。 MAP 激酶底物磷酸化测定将用于表征测试化合物的生物活性。在目标 2 中，将通过荧光光谱和 X 射线晶体学测试活性化合物和 ERK2 之间的特异性相互作用。然后将测试抑制 ERK 底物磷酸化并与特定对接域结合的先导化合物抑制癌细胞增殖的能力在细胞和动物模型中。在本申请结束时，预计将鉴定出一系列可阻断特定 ERK-底物相互作用的先导化合物。这些化合物将适用于研究工具和开发新型化疗药物，旨在治疗与 ERK 通路活性升高相关的癌症。