Substrate specific ERK docking domain inhibitors

底物特异性 ERK 对接域抑制剂

基本信息

批准号：
7578927
负责人：
PAUL S SHAPIRO
金额：
$ 23.03万
依托单位：
UNIVERSITY OF MARYLAND BALTIMORE
依托单位国家：
美国
项目类别：
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/7578927
关键词：
Animal Model Binding Binding Sites Biological Biological Assay Breast Cancer cell line Cell Proliferation Cells Clinical Collection Complex Computer Assisted Development Docking Dose Drug Design Drug Kinetics ELK1 gene Effectiveness Extracellular Signal Regulated Kinases Family Fluorescence Spectroscopy Future Gene Mutation Goals Growth Factor Receptors Human Investigation Large Intestine Carcinoma Lead MAPK1 gene MAPK3 gene Malignant Neoplasms Measures Mitogen-Activated Protein Kinase 3 Mitogen-Activated Protein Kinases Molecular Weight Mus Pathogenesis Pathway interactions Phosphorylation Physiological Play Probability Prostate Protein Kinase Protein-Serine-Threonine Kinases Proteins Reporting Research Research Personnel Ribosomal Protein S6 Kinase Role Screening procedure Signal Pathway Signal Transduction Site Specificity Substrate Interaction Testing Therapeutic Therapeutic Agents Toxic effect X-Ray Crystallography Xenograft Model base cancer cell chemotherapeutic agent inhibitor/antagonist kinase inhibitor novel novel therapeutics prevent programs ribosomal protein S6 kinase 1 success tool tool development transcription factor tumor growth

项目摘要

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 (GADD) 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 showeda 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 1of the current proposal will extend the use 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 assayswill 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 proposal, 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.

细胞外信号调节激酶（ERK1和ERK2）信号通路在细胞增殖。通过增加生长因子受体的表达来激活 ERK 蛋白或上游蛋白质的基因突变被认为与许多人类疾病的发病机制有关癌症。因此，抑制 ERK 信号被视为预防癌细胞的潜在方法。增殖。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中非常保守家庭。利用最近发现的 ERK2 对接域，据报道可以促进底物相互作用；我们使用计算机辅助药物设计（GADD）来识别新型低分子药物重量 ERK 抑制剂。我们假设与 ERK2 对接结合的低分子量化合物结构域将选择性地阻断 ERK 与底物蛋白的相互作用并抑制细胞增殖。下列的 CADD 筛选，选择潜在化合物并测试其生物测定活性。一些鉴定出的化合物可抑制核糖体 S6 激酶-1 (RSK-1) 的 ERK 特异性磷酸化和转录因子 ELK-1，两者均促进细胞增殖。此外，活性化合物显示通过集落存活测定测量，癌细胞系增殖的剂量依赖性减少荧光光谱表明直接与 ERK2 结合。当前提案的目标 1 将延长使用 CADD 来识别很可能与九种不同的假定结合的其他化合物 ERK2 对接域上的结合口袋。将使用 MAP 激酶底物磷酸化测定表征测试化合物的生物活性。活性物质之间的具体相互作用目标 2 中的化合物和 ERK2 将通过荧光光谱和 X 射线晶体学进行测试。抑制 ERK 底物磷酸化并与特定对接结构域结合的化合物将被在细胞和动物模型中测试了它们抑制癌细胞增殖的能力。在本提案的最后，预计将鉴定出一系列可阻断特定 ERK 底物的先导化合物互动。这些化合物将适用于研究工具和开发新型化合物化疗药物旨在治疗与 ERK 通路活性升高相关的癌症。