The development of a transcriptional inhibitor for lung fibrosis.

摘要/摘要 特发性肺纤维化(IPF)在美国影响10万人，总发病率每年增加5% 年。IPF患者的5年生存率在20%-40%之间。Esbriet和Nintdanib是 临床批准的治疗特发性肺纤维化的方法。然而，Esbriet将疾病进展的风险降低了50% 其作用机制目前尚不清楚。Inetedanib是一种血管激酶抑制剂，靶向超过30 包括血小板衍生生长因子受体、成纤维细胞生长因子受体和血管 内皮生长因子受体和临床试验中，宁替达尼使IPF下降50% 治疗的一年。尽管Esbriet和Nintdanib是用来治疗IPF的，但所有患者都取得了进展 治疗，发展为需要氧气的晚期疾病，要么死于这种疾病，要么接受肺部手术 移植。因此，寻找既能预防又能逆转纤维化组织的新疗法是非常必要的。 进展性特发性肺纤维化患者组织中的转录谱显示MASTER显著激活 转录调控因子STAT3是由SH2-二聚结构域(pSTAT3-)的磷酸化增加决定的 Y705)。IPF中激活的STAT3与炎症和ECM导致的患者生存不良相关。此外， 遗传学证据表明，STAT3激活剂IL6基因的多态与疾病独立相关 进步。我们假设靶向STAT3将阻断多条促纤维化通路并减少 肺部炎症和胶原蛋白堆积。虽然像STAT3这样的转录因子是有吸引力的治疗靶点， 它们很难以小分子为靶标，因为它们缺乏清晰的结合口袋，表面很大 对蛋白质相互作用很重要的区域，包含大的内在无序的结构域。在阿勒泰 治疗公司，我们开发了一个平台，能够识别内在的小捆绑口袋 先前不可用药的TF中的无序结构域，使靶向STAT3的新的可用药方法成为可能 具有特异性的和开发有效和高度特异性的STAT3抑制剂(STAT3i)。我们完成了In-Silico 筛选和鉴定减少STAT3DNA结合的抑制剂。重要的是，这些STAT3i具有最小 STAT1抑制活性，低细胞毒性，并显示出对STAT3靶点和纤维化的有效抑制 基因。我们提出三个目标，以确定和描述最有希望的线索，并继续努力 开发一种基于抑制STAT3的治疗IPF的可行方法。在目标1中，我们将测量细胞毒性。 一组正常细胞以及使用Altay的新STAT3 IS测量STAT3靶基因抑制。在目标2中， 我们将利用我们的领先的STAT3进行博莱霉素诱导的IPF小鼠的研究。在目标3中，我们将确定 特发性肺纤维化患者肺切片中STAT3i的抗纤维化活性及细胞因子的检测 分泌物。拟议的研究将建立针对STAT3治疗IPF的潜力，并指导新的 在这种情况下的治疗策略。然后，我们将寻求SBIR第二阶段赠款，其中将包括 化学方面的努力、更多的动物研究，以及我们针对IPF的STAT3抑制剂的最终商业化。

Abstract/Summary Idiopathic Pulmonary Fibrosis (IPF) affects 100,000 people in the US with total incidences to increase 5% every year. The 5-year survival rate of patients suffering from IPF is between 20-40%. Esbriet and Nintedanib are clinically approved treatments for IPF. Esbriet reduces risk of disease progression by 50% however, the mechanism of action is currently unknown. Nintedanib is an angiokinase inhibitor that targets more than 30 kinases including platelet-derived growth factor receptor, fibroblasts growth factor receptor and vascular endothelial growth factor receptor and in clinical trials Nintedanib reduced the decline of IPF by 50% after one year of treatment. Although Esbriet and Nintedanib are marketed to treat IPF, all patients progress despite therapy, develop advanced disease requiring oxygen and either succumb to the disease or undergo lung transplantation. Therefore, finding new treatments that both prevent and reverse fibrotic tissue are in great need. Transcriptional profiles in progressive IPF patient tissues have demonstrated significant activation of the master transcriptional regulator STAT3 determined by increased phosphorylation in SH2-dimerization domain (pSTAT3- Y705). Active STAT3 in IPF correlated with poor patient survival driven by inflammation and ECM. Furthermore, genetic evidence shows polymorphisms within IL6, STAT3 activator, was independently associated with disease progression. We hypothesize that targeting STAT3 will block multiple profibrogenic pathways and reduce inflammation and collagen accumulation in the lung. Although TFs like STAT3 are attractive therapeutic targets, they are challenging to target with small molecules because they lack clear binding pockets, have large surface areas important for protein-protein interactions and contain large intrinsically disordered domains. At Altay Therapeutics, we developed a platform that enables identification of small binding pockets within intrinsically disordered domains in previously undruggable TFs, allowing a novel druggable approach for targeting STAT3 with specific and development of potent and highly specific STAT3 inhibitors (STAT3i). We completed in-silico screening and identified inhibitors that reduced STAT3 DNA binding. Importantly, these STAT3i had minimal STAT1 inhibitory activity, low cytotoxicity and demonstrated potent inhibition of STAT3 targets and fibrosis genes. We propose three aims to identify and characterize the most promising lead and continue our efforts to develop a viable treatment for IPF based on inhibiting STAT3. In Aim 1, we will measure cellular cytotoxicity in a panel of normal cells as well as measure STAT3 target gene inhibition with Altay’s novel STAT3is. In Aim 2, we will carry out bleomycin induced IPF mouse studies with our lead STAT3is. In Aim 3, we will determine antifibrotic activity of STAT3i in precision cut lung slices isolated from IPF patients as well as measure cytokine secretion. The proposed studies will establish the potential for targeting STAT3 in treating IPF and guide new therapeutic strategies in this setting. We will then pursue an SBIR phase 2 grant that will include medicinal chemistry efforts, additional animal studies and ultimately commercialization of our STAT3 inhibitor for IPF.