Optimizing function-selective ERK1/2 inhibitors for reducing AP-1-mediated airway pathology in asthma.

项目摘要 哮喘的发病机制以呼吸道炎症、重塑和高反应性为特征。 导致严重的支气管收缩。变应原诱导的炎症介质作用于免疫细胞 和结构呼吸道细胞，并激活细胞内信号。激活蛋白-1(AP-1) 转录因子复合体是一种中央调节因子，它对由 细胞因子、生长因子等炎症信号在气道重塑中的作用 哮喘。因此，上调的AP-1参与了哮喘发病机制的多个特征。 一个有吸引力的抗哮喘治疗靶点。细胞外信号调节蛋白激酶 (ERK1/2)是AP-1活性的关键调节因子，在气道平滑肌(ASM)、肺成纤维细胞(LF)和 参与哮喘病理的其他肺细胞。利用ERK1/2结构 与特定底物的相互作用，我们发现了一种新的化合物，它与一个独特的ERK1/2结合 介导与AP-1复合蛋白相互作用并抑制ERK1/2的底物对接部位 介导的AP-1活性。靶向选择激酶功能在减少获得性药物方面具有优势 靶向三磷酸腺苷结合部位并阻断的当前激酶抑制剂的耐药性和毒性 所有的酶活性。我们证明了功能选择性ERK1/2抑制剂抑制ASM细胞 在小鼠模型中，AP-1的增殖、AP-1活性和减轻过敏性哮喘的多种特征。 考虑到ERK1/2活性上调与哮喘的发病机制有关，我们假设 通过AP-1选择性抑制ERK1/2信号将缓解ASM和LF细胞 哮喘具有细胞外基质(ECM)过度分泌、肥大、肥大等特点。这个 R61阶段将包括两个目标。目标1将使用计算机辅助药物设计和化学合成 为了产生针对AP-2调节的铅功能选择性ERK1/2抑制剂的优化类似物 1蛋白质。目的2将评估新化合物在调节AP-1介导的增殖，ECM分泌， 正常肺和哮喘肺原代ASM和LF细胞中的炎性介质。目标 3在R33阶段将使用集成的哮喘小鼠模型来评估最有效的 化合物在缓解过敏性哮喘的多种特征方面。此外，R33阶段将与 Accelerator合作伙伴Gen1E，生命科学，执行TOP的临床前测试和开发 3个化合物侧重于药代动力学评价、激酶选择性、靶外效应和毒性。 这些研究将提供重要的临床前数据，以推进有效抑制 参与哮喘病理的一个主要效应靶点(如AP-1)。

Project Summary Asthma pathogenesis is characterized by airway inflammation, remodeling and hyperresponsiveness resulting in severe bronchoconstriction. Allergen-induced inflammatory mediators act on immune cells and structural airways cells and activate intracellular signaling. The Activator Protein-1 (AP-1) transcription factor complex is a central regulator that responds to signaling pathways activated by cytokines, growth factors and other inflammatory signals in airway cells to mediate airway remodeling in asthma. Therefore, upregulated AP-1, which contributes to multiple features of asthma pathogenesis, is an attractive anti-asthma therapeutic target. The Extracellular signal‑Regulated protein Kinases (ERK1/2) are key regulators of AP-1 activity in airway smooth muscle (ASM), lung fibroblasts (LF), and other lung cells that contribute to the pathology of asthma. Taking advantage of ERK1/2 structural interactions with specific substrates, we identified a novel compound that binds to a unique ERK1/2 substrate docking site that mediates interactions with AP-1 complex proteins and inhibits ERK1/2- mediated AP-1 activity. Targeting select kinase functions offers advantages in reducing acquired drug resistance and toxicity observed with the current kinase inhibitors that target ATP binding sites and block all enzymatic activity. We demonstrate that function-selective ERK1/2 inhibitors inhibit ASM cell proliferation, AP-1 activity, and mitigate multiple features of allergic asthma in a murine model. Considering that upregulated ERK1/2 activity contributes to the pathogenesis of asthma, we hypothesize that function-selective inhibition of ERK1/2 signaling through the AP-1 will mitigate ASM and LF cell hyperplasia, hypertrophy, extracellular matrix (ECM) hypersecretion, and other features of asthma. The R61 phase will consist of two aims. Aim 1 will use computer-aided drug design and chemical synthesis to generate optimized analogs of a lead function-selective ERK1/2 inhibitor that targets regulation of AP- 1 proteins. Aim 2 will evaluate new compounds in regulating AP-1 mediated hyperplasia, ECM secretion, and inflammatory mediators in primary ASM and LF cells obtained from normal and asthmatic lungs. Aim 3 in the R33 phase will employ an integrated mouse model of asthma to assess the most potent compounds in mitigating multiple features of allergic asthma. In addition, R33 phase will collaborate with an Accelerator Partner, Gen1E, Life Sciences, to perform pre-clinical testing and development of the top 3 compounds focusing on pharmacokinetic evaluation, kinase selectivity, off-target effects, and toxicity. These studies will provide important pre-clinical data to advance a novel therapy that effectively inhibits a major effector target (e.g., AP-1) involved in the pathology of asthma.