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：
7495874
负责人：
PAUL S SHAPIRO
金额：
$ 2.88万
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/7495874
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 Thinking Toxic effect X ray spectroscopy 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）信号通路在细胞中起关键作用增殖。通过增加生长因子受体的表达来激活ERK蛋白或上游蛋白的遗传突变被认为与许多人的发病机理有关癌症。因此，抑制ERK信号被视为预防癌细胞的潜在方法增殖。当前，尚无对ERK蛋白的特定抑制剂。而且，大多数激酶抑制剂缺乏特异性，因为它们靶向ATP结合位点，这些位点在蛋白质激酶中是充分保守的家庭。利用最近确定的ERK2对接域，据报道底物相互作用；我们已经使用计算机辅助药物设计（GADD）来识别新颖的低分子重量ERK抑制剂。我们假设低分子量的化合物与ERK2对接结合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。下列的选择了CADD筛选，潜在化合物并测试生物测定中的活性。一些鉴定核糖体S6激酶1（RSK-1）的ERK特异性磷酸化的化合物和转录因子ELK-1，两者都会促进细胞增殖。另外，活性化合物展示通过菌落存活分析和如荧光光谱法所示，直接与ERK2结合。目前的提案将延长目标1 使用CADD来识别具有与九种不同推定的高概率的其他化合物 ERK2对接域上的绑定口袋。将使用MAP激酶底物磷酸化测定法表征测试化合物的生物学活性。活动之间的特定相互作用 AIM 2中的荧光光谱和X射线晶体学测试化合物和ERK2。抑制ERK覆盖磷酸化并结合特定对接结构域的化合物将是测试了它们在细胞和动物模型中抑制癌细胞增殖的能力。在此提案的结尾，预计将确定一系列铅化合物的集合互动。这些化合物适用于研究工具和发展为新颖的化合物旨在治疗与ERK途径升高相关的癌症的化学治疗剂。