Evaluation of novel substrate specific inhibitors of ERK1/2 in the treatment of asthma

新型ERK1/2底物特异性抑制剂治疗哮喘的评价

基本信息

批准号：
9293245
负责人：
Deepak A Deshpande
金额：
$ 19.41万
依托单位：
UNIVERSITY OF MARYLAND BALTIMORE
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-06-10 至 2019-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9293245
关键词：
Address Affect Allergens Animal Model Anti-Inflammatory Agents Anti-inflammatory Apoptosis Arthritis Asthma Atherosclerosis Binding Binding Sites Breathing Bronchoconstriction Carcinogens Catalytic Domain Cell Count Cell Cycle Progression Cell Proliferation Cell Size Cell Survival Cell secretion Cells Chemicals Chronic Disease Chronic Obstructive Airway Disease Clinical Data Clinical Trials Complex Computer Assisted Data Development Disease Disease Progression Down-Regulation Drug Design Drug Targeting Drug resistance Drug toxicity EGF gene Effectiveness Epithelial Cells Evaluation Extracellular Matrix Extracellular Signal Regulated Kinases FOS Protein FOS gene FOXP3 gene Family Fibrosis Fos-Related Antigens Gene Expression Growth Growth Factor Health Care Costs Hepatitis Human Hyperplasia Immune Inflammation Inflammation Mediators Inflammatory Interleukin-13 Interleukin-4 Interleukin-5 Lead Lung MAPK14 gene MAPK8 gene Malignant Neoplasms Mediating Mediator of activation protein Methods Mitogen-Activated Protein Kinases Mitogens Modeling Molecular Morbidity - disease rate Mucous body substance Muscle Cells Normal Cell Parkinson Disease Pathogenesis Pathologic Pharmaceutical Preparations Pharmacology Phenotype Phosphorylation Phosphotransferases Platelet-Derived Growth Factor Production Proteins Psoriasis Pyroglyphidae Recruitment Activity Regulation Role Safety Severity of illness Signal Pathway Signal Transduction Signaling Molecule Site Smooth Muscle Myocytes Specificity Stimulus Testing Therapeutic Intervention Time Tissues Toxic effect Transcription Factor AP-1 Transcriptional Activation Validation activator 1 protein airway hyperresponsiveness airway inflammation airway remodeling angiogenesis asthmatic cell growth cell type chemokine cytokine in vivo inhibitor/antagonist kinase inhibitor mortality mouse model novel novel strategies pre-clinical protein function receptor respiratory smooth muscle response scaffold small molecule inhibitor success targeted agent therapeutic target transcription factor validation studies

项目摘要

PROJECT SUMMARY: Airway inflammatory diseases, such as asthma, are characterized by inflammation, airway remodeling and hyperresponsiveness resulting in severe bronchoconstriction. Asthma pathogenesis is driven by inflammatory mediators that act on immune cells and resident airways cells through receptors that regulate signaling molecules and gene expression. Transcriptional activation is a common end point in the signaling elicited by multiple cytokines in target cells. The transcription factor, Activator Protein (AP)-1 is activated by mitogen-activated protein (MAP) kinases, such as ERK1/2, and is known to mediate airway inflammation, airway smooth muscle (ASM) remodeling, and mucus production, and therefore is an attractive anti-asthma therapeutic target. Current kinase inhibitors used as anti-inflammatory or anti-oncogenic agents target ATP binding or catalytic sites and block all enzymatic activity. These drugs invariably lead to drug resistance or toxicity due to lack of specificity. Using advanced computer-aided drug design methods we have identified a novel thienyl benzenesulfonate chemical scaffold that selectively inhibits ERK1/2-mediated AP-1 activity taking advantage of the structural requirements of ERK1/2 and specific substrates. These novel substrate- specific inhibitors of ERK1/2 bind the F-recruitment site (FRS) of ERK1/2, which mediates interactions with AP-1 substrates containing an F-site or FXF motif. Functional validation studies demonstrate that the compounds inhibit specific kinase functions associated with disease while preserving ERK1/2 functions in normal cells. In this proposal we aim at evaluating the efficacy of newly developed small molecule inhibitors of ERK1/2 in mitigating features of asthma. Aim 1 studies will establish the effect of substrate-specific inhibitors of ERK1/2 on human ASM cell growth and secretion of extracellular matrix, chemokines and cytokines. Aim 2 studies will ascertain specificity and signaling mechanisms modulated by these compounds. Finally, we will employ an integrated mouse model of asthma to determine the in vivo efficacy of ERK1/2 inhibitors in mitigating allergen-induced airway inflammation, remodeling, mucus production and airway hyperresponsiveness. Preliminary data suggest that the compounds inhibit mitogen-induced ASM cell growth and AP-1 activation. Importantly, the compounds did not inhibit growth factor-induced phosphorylation of ERK1/2 or Akt further confirming the specificity of targeting downstream ERK1/2 substrates in ASM. Considering the ubiquitous expression and functional role of ERK1/2-induced activation of AP-1 in asthma pathogenesis, we predict that novel substrate specific inhibitors of ERK1/2 mitigate multiple features of asthma in the animal model. These studies will provide important pre-clinical data to advance a novel class of small molecule inhibitors targeting a versatile signaling molecule as anti-asthma therapies.

项目摘要：气道炎症性疾病（例如哮喘）的特征是炎症，气道重塑和反应性过高，导致严重的支气管收缩。哮喘发病机理是由对免疫细胞和常驻气道细胞作用的炎症介质驱动的调节信号分子和基因表达的受体。转录激活是靶细胞中多个细胞因子引起的信号传导中的常见终点。转录因子，激活蛋白（AP）-1通过有丝分裂原激活蛋白（MAP）激酶激活，例如 ERK1/2，众所周知会介导气道炎症，气道平滑肌（ASM）重塑和粘液产生，因此是一个有吸引力的抗窒息治疗靶标。当前的激酶用作抗炎或抗疾病剂的抑制剂靶向ATP结合或催化位点，以及阻止所有酶活性。由于缺乏特异性。使用先进的计算机辅助药物设计方法，我们已经确定了一种新颖的硫烯基苯磺酸盐化学支架有选择地抑制ERK1/2介导的AP-1活性 ERK1/2和特定底物的结构要求的优势。这些新颖的底物 - ERK1/2的特异性抑制剂结合了ERK1/2的F恢复位点（FRS），该位点介导了相互作用与含有F位置或FXF基序的AP-1底物。功能验证研究表明这些化合物抑制了与疾病相关的特异性激酶功能，同时保存ERK1/2 正常细胞的功能。在此提案中，我们旨在评估新开发的小型的功效 ERK1/2的分子抑制剂缓解哮喘的特征。 AIM 1研究将确定效果 ERK1/2对人ASM细胞生长的底物特异性抑制剂和细胞外分泌基质，趋化因子和细胞因子。 AIM 2研究将确定特异性和信号传导机制由这些化合物调节。最后，我们将采用哮喘的综合鼠标模型确定ERK1/2抑制剂在减轻过敏原炎症的体内功效，重塑，粘液产生和气道高反应性。初步数据表明化合物抑制有丝分裂原诱导的ASM细胞生长和AP-1激活。重要的是，化合物没有抑制生长因子诱导的ERK1/2或AKT的磷酸化进一步确认 ASM中靶向下游ERK1/2底物的特异性。考虑无处不在 ERK1/2诱导的AP-1在哮喘发病机理中的表达和功能作用，我们预测ERK1/2的新型底物特异性抑制剂减轻了哮喘的多种特征动物模型。这些研究将提供重要的临床前数据，以推动一类新的小型小型靶向多功能信号分子作为抗哮喘疗法的分子抑制剂。