An oxysterol drug candidate, Oxy210, for inhibition of pulmonary fibrosis through targeting hedgehog and TGFβ signaling

摘要 特发性肺纤维化(IPF)是一种慢性间质性肺疾病，其特征是进行性、 肺组织不可逆转的疤痕形成和肺功能下降，最终导致呼吸衰竭。中位数患者 存活时间限制为确诊后3-5年。目前可用的两种疗法是吡非尼酮和宁替达尼， 只是延缓了疾病的发展，并没有显著降低IPF的死亡率或显著改善 IPF患者自2014年获得批准以来的生活质量。因此，新的和改进的IPF候选药物 都是迫切需要的。IPF疾病的进展在很大程度上是通过不适当的重新激活 胚胎细胞信号通路，如转化生长因子β(β)和刺猬 (Hh)信号通路。事实上，Pirfenidone和Nintdanib的作用部分是通过适度阻断这些信号来实现的 小路。在Max BioPharma，我们正在开发新型的、专有的、基于氧固醇的ABABRANT拮抗剂 在人类肺间质纤维化和其他纤维化疾病中可能具有疾病修改特性的细胞信号。这 应用涵盖了一种基于氧固醇的候选药物Oxy210，它是HH和转化生长因子β的双重抑制剂，发出信号 改善非脂肪性肝炎(NASH)人源化小鼠模型的肝纤维化和炎症。在……里面 在这一应用中，我们提供了Oxy210体外研究的令人鼓舞的初步数据，证明了它 抑制正常肺成纤维细胞系增殖和促纤维化基因表达的研究 90)和IPF患者来源的细胞系(LL97A)。Oxy210在脂多糖治疗中的抗炎作用 通过直接抑制Toll样受体(TLR4)、TLR2和AP-1而不干扰巨噬细胞 转化生长因子β的抗炎作用。初步研究结果表明，Oxy210具有潜在的抑制作用 博莱霉素诱导的小鼠体内肺部病理。我们建议进一步描述Oxy210在相关的 体外和体内的特发性肺纤维化模型。具体而言，我们建议处理以下具体目标：目标1。 Oxy210对原代肺肌成纤维细胞分化和激活作用的研究 体外培养的正常肺和IPF患者肺成纤维细胞。目的2.鉴定Oxy210对血管紧张素转换酶的影响 不同疾病进展阶段的肺纤维化、成纤维细胞活化和巨噬细胞表型 博莱霉素处理的小鼠的RNA测序。Aim3.用体外Ames法检测Oxy210的安全性， 细胞色素P450抑制及靶外活性研究。我们预计来自拟议研究的数据 将评估Oxy210作为进一步开发和靶向IPF的候选药物的潜力。相比较 与现有的治疗方法相比，Oxy210作为一种单独的治疗方法或在 与现有治疗IPF的方法相结合。

ABSTRACT Idiopathic Pulmonary Fibrosis (IPF) is a chronic interstitial lung disease, characterized by progressive, irreversible scarring of lung tissue and declining lung function, terminating in respiratory failure. Median patient survival is limited to 3-5 years post diagnosis. Two currently available therapies, Pirfenidone and Nintedanib, only slow the progression of the disease and have not greatly reduced IPF mortality or significantly improved quality of life in IPF patients since they were approved in 2014. Hence, new and improved IPF drug candidates are urgently needed. IPF disease progression is driven in significant part through inappropriate reactivation of embryonic cellular signaling pathways, such as the Transforming Growth Factor beta (TGFβ) and Hedgehog (Hh) signaling pathways. In fact, Pirfenidone and Nintedanib act in part through modestly blocking these signaling pathways. At MAX BioPharma, we are developing novel, proprietary, oxysterol-based antagonists of aberrant cellular signaling that may have disease modifying properties in human IPF and other fibrotic diseases. This application covers an oxysterol-based drug candidate, Oxy210, a dual inhibitor of Hh and TGFβ signaling that ameliorates hepatic fibrosis and inflammation in a humanized mouse model of non-steatohepatitis (NASH). In this application, we present encouraging preliminary data from in vitro studies with Oxy210, demonstrating its inhibitory effects on proliferation and pro-fibrotic gene expression in a normal pulmonary fibroblast cell line (IMR- 90) and an IPF patient-derived cell line (LL97A). Oxy210 has anti-inflammatory effects in LPS-treated macrophages through direct inhibition of Toll-Like Receptor (TLR4), TLR2, and AP-1, and does not interfere with the anti-inflammatory effects of TGFβ. Preliminary findings show the promising potential of Oxy210 to inhibit bleomycin-induced pulmonary pathology in vivo in mice. We propose to further characterize Oxy210 in relevant in vitro and in vivo models of IPF. Specifically, we propose to address the following specific aims: Aim 1. Elucidation of the effects of Oxy210 on myofibroblast differentiation and activation of primary pulmonary fibroblasts from normal lungs and from IPF patients in vitro. Aim 2. Identification of the effects of Oxy210 on lung fibrosis, fibroblast activation, and macrophage phenotypes at different stages of disease progression using RNA sequencing in bleomycin-treated mice. Aim3. Examination of Oxy210 safety using in vitro Ames, Cytochrome P450 inhibition, and off-target activity studies. We anticipate that data from the proposed studies will evaluate the potential of Oxy210 as a drug candidate for further development and targeting of IPF. Compared to existing therapies, Oxy210 may be more effective and safer as a standalone therapy or when used in combination with existing therapies for treating IPF.