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：
7215142
负责人：
PAUL S SHAPIRO
金额：
$ 20.48万
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/7215142
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

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（由申请人提供）：细胞外信号调节激酶（ERK1和ERK2）信号通路在细胞增殖中起关键作用。人们认为，通过增加生长因子受体的表达或上游蛋白的遗传突变的激活被认为与许多人类癌的发病机理有关。因此，抑制ERK信号传导被视为预防癌细胞增殖的潜在方法。当前，尚无对ERK蛋白的特定抑制剂。此外，大多数激酶抑制剂都缺乏特异性，因为它们靶向ATP结合位点，而ATP结合位点在蛋白激酶家族中均能保守。利用最近鉴定出的ERK2对接结构域，据报道，这些结构域促进了底物相互作用，我们已经使用了计算机辅助药物设计（CADD）来识别新型的低分子量ERK 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有选择地阻断ERK与底物蛋白的相互作用并抑制细胞增殖。 CADD筛选后，选择了潜在化合物并测试生物测定中的活性。几种化合物鉴定了核糖体S6激酶-1（RSK-1）和转录因子ELK-1的ERK特异性磷酸化，这两者都促进细胞增殖。此外，通过菌落存活分析测量的癌细胞系增殖的剂量依赖性降低，并直接与ERK2结合，如荧光光谱法所示。当前应用程序的目标1将扩展使用CADD以识别具有高概率与ERK2对接域上9个不同推定的结合口袋结合的其他化合物。 MAP激酶底物磷酸化测定法将用于表征测试化合物的生物学活性。活性化合物与ERK2之间的特定相互作用将通过AIM 2中的荧光光谱和X射线晶体学测试。铅化合物抑制ERK-SUBSTRATE磷酸化并与特定对接结构域结合，然后测试其在细胞和动物模型中抑制癌细胞增殖的能力。在此应用程序的结尾，预计将确定铅化合物的集合，以阻止特定的ERK-SUBSTRATE相互作用。这些化合物将适用于研究工具和发展为旨在治疗与ERK途径升高相关的癌症的新型化学治疗剂的发展。