Non-catalytic FAK inhibitors as novel therapeutics for lung fibrosis

项目摘要 特发性肺纤维化（IPF）是一种持续进行性和致命性的纤维化肺病， 对男性和老年人的影响尤为严重。尽管最近有两种药物（吡非尼酮和尼达尼布） 获得FDA批准治疗IPF和结缔组织疾病相关的进行性纤维化肺病 （类风湿性关节炎和硬皮病，在年轻女性中更常见），这些药物绝不是治愈性的。 事实上，这些疗法仅显示出肺功能下降率的适度降低，并没有改善 生活质量不幸的是，纤维化管道中的几种潜在疗法未能达到其终点 在最近的审判中。因此，我们只剩下次优的治疗方法和肺移植作为唯一的当前治疗方法。 治疗IPF患者。重要的是，没有可用的疗法“逆转”纤维化。粘着斑激酶（FAK）是 一种调节肺促纤维化表型的非受体酪氨酸激酶和支架蛋白 成纤维细胞，包括分泌细胞外基质蛋白（纤连蛋白和胶原蛋白），肌成纤维细胞 分化、细胞迁移和抗凋亡。在最近的肺组织基因表达分析中， 在IPF患者中，与健康对照相比，FAK在早期IPF和晚期IPF中均高度上调。 此外，FAK的粘着斑靶向（FAT）结构域的支架功能已经被证实。 在体外和体内证实对肺纤维化的发展至关重要。然而，FAK 迄今为止开发的抑制剂仅针对其激酶，而忽略了FAK作为支架蛋白的作用。 因为目前的FAK激酶抑制剂不能抑制肺成纤维细胞中关键的FAT结构域相互作用， 高脱靶毒性，开发靶向非催化支架功能的新型FAK抑制剂 或FAK的FAT结构域仍然是显著未满足的临床需求。FAKnostics有限责任公司发现了一个 一系列基于钉合肽的FAK抑制剂，直接靶向FAK的FAT结构域。我们有初步的 前导肽FN-2023在肺成纤维细胞（IMR 90）中引起有效的抗纤维化作用的数据，包括减少 α-SMA、纤连蛋白和胶原蛋白的蛋白水平。第一阶段STTR的目标是证明- 使用这些新型FAK FAT抑制剂作为肺纤维化治疗方法的概念。目标1： 将优化前导肽FN-2023以改善ADMET性质。在目标2中，我们将描述优化的 对肺成纤维细胞和精密切割肺切片（PCLS）具有抗纤维化作用的肽。在目标3中，我们 评价最优化的肽的体内药代动力学，并使用肺的体内功效模型进行测试 纤维化（博来霉素损伤模型）。最终，该项目将产生优化的FAK FAT肽， 改善ADMET在肺纤维化小鼠模型中的特性和功效，支持进一步的临床前研究。 在未来的第二阶段项目中。

PROJECT ABSTRACT Idiopathic pulmonary fibrosis (IPF) is a relentlessly progressive and fatal fibrotic lung disorder which disproportionately affects men and the elderly. Although two drugs (pirfenidone and nintedanib) have recently gained FDA-approval for IPF and progressive fibrosing lung disorders related to connective tissue diseases (rheumatoid arthritis and scleroderma, more common in younger women), these drugs are by no means curative. In fact, these therapies show only a modest reduction in the rate of lung function decline and do not improve quality of life. Unfortunately, several potential therapies in the fibrosis pipeline have failed to meet their endpoints in recent trials. Hence, we are left with suboptimal treatments and lung transplantation as the only current treatment for IPF patients. Importantly, no available therapies ‘reverse’ fibrosis. Focal Adhesion Kinase (FAK) is a non-receptor tyrosine kinase and scaffolding protein that regulates the pro-fibrotic phenotype of lung fibroblasts, including secretion of extracellular matrix proteins (fibronectin and collagen), myofibroblast differentiation, cell migration, and resistance to apoptosis. In recent analyses of gene expression in lung tissue from IPF patients, FAK is highly upregulated in both early IPF and advanced IPF compared to health controls. Moreover, the scaffolding function of the Focal Adhesion Targeting (FAT) domain of FAK has been demonstrated to be critical for the development of lung fibrosis in vitro and in vivo. However, the FAK inhibitors developed to date only target its kinase enzyme and ignore FAK’s role as a scaffolding protein. Because current FAK-kinase inhibitors do not inhibit key FAT domain interactions in lung fibroblasts and show high off-target toxicity, the development of novel FAK inhibitors that target the non-catalytic scaffolding function or FAT domain of FAK remains a significant unmet clinical need. FAKnostics, LLC has identified a first-in-class series of stapled peptide-based FAK inhibitors that directly target the FAT domain of FAK. We have preliminary data that lead peptide FN-2023 causes potent anti-fibrotic effects in lung fibroblasts (IMR90), including reduction in protein levels of α-SMA, fibronectin, and collagen. The goal of this Phase I STTR is to demonstrate proof- of-concept for the use of these novel FAK FAT inhibitors as therapeutics of lung fibrosis. In Aim 1, we will optimize lead peptide FN-2023 to improve ADMET properties. In Aim 2, we will characterize optimized peptides for anti-fibrotic effects on lung fibroblasts and precision cut lung slices (PCLS). In Aim 3, we will evaluate in vivo pharmacokinetics of top optimized peptides and test using an in vivo efficacy model of lung fibrosis (bleomycin injury model). Ultimately, this project will result in optimized FAK FAT peptides that show improved ADMET properties and efficacy in a mouse model of lung fibrosis, supporting further preclinical development in a future Phase II project.