Development, Formulation and Inhalational Delivery of a New Peptide for ILD

间质性肺病（ILD）的特征是进行性肺瘢痕形成。特发性肺 肺纤维化（IPF）是ILD最常见的形式之一，是一种致死性肺病，发病率为60 例/100，000人在美国每年。IPF的中位5年生存率仅为20%。目前存在 无法治愈IPF的发病机制以肺泡上皮细胞（AEC）衰老和凋亡为特征， 活化的肌成纤维细胞和纤维化肺成纤维细胞（fLfs）的增殖和积累以及细胞外 基体沉积这些特征导致进行性肺功能障碍。形态学变化包括空间和 时间异质性包括与含有凋亡的病变区域相邻的正常肺区域 AEC和fLfs。最近，药物治疗已被证明可以减缓IPF的进展，这表明， 可以开发有效的治疗方法。我们发现p53的增加有助于AEC凋亡和成纤维细胞的形成。 激活，并且其fLfs的下降促进肌成纤维细胞扩增。我们还发现，Caveolin-1（Cav 1） 在受损的AEC中增加，而在IPF患者肺的增殖fLfs中其水平显著降低 我们鉴定了一个20聚体的Cav 1支架结构域肽（CSP）， 7-mer片段，CSP 7，其抑制p53、TGF-β、CTGF、AEC凋亡和fLf扩增。这些肽阻断了 单次（1X）或多次（8X）使用博来霉素（BLM）、腺病毒TGF-β1（Ad-TGF-β1）和硅胶后小鼠的PF 导致肺损伤。由于fLfs中mdm 2表达增加，CSP 7抑制p53的降解，并阻断其表达。 增殖CSP 7还抑制AEC衰老和凋亡，否则它们在纤维化肺中增加， 包括IPF。CSP 7在小鼠中耐受良好，并且可以通过气道有效地递送。我们的工作 验证了Cav 1作为PF的治疗靶点。我们的出版物为开发气道CSP 7提供了前提 通过雾化（neb）或干粉吸入（DPI）给药作为一种新的、安全的和更有效的治疗PF的方法。 目的一、明确CSP 7的细胞表面结合/作用模式，优化CSP 7的结构，确定CSP 7的作用范围 有效气道（雾化吸入或DPI）给药，并评估CSP 7的耐受性和解决PF的能力 在4种PF模型中（1X BLM在年轻和老年小鼠中，Ad-TGF-β1和8X BLM），我们将选择最多的 目标I中确定的有效、耐受性良好、优化形式的CSP 7（CSP 7 OP），然后评价非GLP 药代动力学、毒理学和评估气道递送后的全身生物分布。在Aim III中，我们将 阐明CSP 7 OP与吡非尼酮或尼达尼布联合作用影响损伤的分子机制 AEC和fLfs解决现有的PF。用CSP 7靶向Cav 1代表了一种新的和有前途的方法， 通过气道治疗IPF。该项目成功的可能性很高，要生成的数据是 用于IND启用工作的谓词。我们的团队拥有细胞/分子生物学、临床前 建模、药物发现、制剂和气道药物递送。该项目可能会定义新的和潜在的 对IPF或其他ILD患者更有效的治疗，对他们来说，有效的治疗选择仍然有限。

Interstitial lung diseases (ILDs) are characterized by progressive pulmonary scarring. Idiopathic pulmonary fibrosis (IPF) is one of the most common forms of ILDs and is a fatal lung disease with an incidence of 60 cases/100,000 individuals in the US annually. IPF has a median five-year survival of only 20%. There is presently no cure. The pathogenesis of IPF is characterized by alveolar epithelial cell (AEC) senescence and apoptosis, proliferation and accumulation of activated myofibroblasts and fibrotic lung fibroblasts (fLfs) and extracellular matrix deposition. These features lead to progressive lung dysfunction. Morphologic changes include spatial and temporal heterogeneity incorporating areas of normal lung adjacent to diseased areas containing apoptotic AECs, and fLfs. Recently, pharmacotherapy has been shown to slow progression of IPF, suggest that even more effective treatment can be developed. We found that increased p53 contributes to AEC apoptosis and fibroblast activation, and that its decline in fLfs promotes myofibroblast expansion. We also found that caveolin-1 (Cav1) is increased in injured AECs while its level is markedly reduced in proliferating fLfs from the lungs of IPF patients and mice with established PF. We identified a 20-mer Cav1 scaffolding domain peptide (CSP), and its truncated 7-mer fragment, CSP7, that inhibits p53, TGF-β, CTGF, AEC apoptosis and fLf expansion. These peptides block PF in mice after single (1X) or multi-hit (8X) bleomycin (BLM), adenoviral TGF-β1 (Ad-TGF-β1)- and silica- induced lung injury. CSP7 inhibits degradation of p53 due to increased mdm2 expression in fLfs and blocks their proliferation. CSP7 also inhibits AEC senescence and apoptosis, which are otherwise increased in fibrotic lungs, including in IPF. CSP7 is well-tolerated in mice and can effectively be delivered via the airways. Our work validates Cav1 as a therapeutic target in PF. Our publications offer the premise for developing CSP7 for airway delivery by nebulization (neb) or dry power inhalation (DPI) as a new, safe and more effective therapy for PF. In Aim I, we will define cell surface binding/mode action of CSP7, optimize the structure of CSP7, identify the range of dosing for effective airway (neb or DPI) delivery and assess the tolerability and ability of CSP7 to resolve PF in 4 (1X BLM in young and aged mice, Ad-TGF-β1 and 8X BLM) models of PF. In Aim II, we will select the most effective, well-tolerated, optimized form CSP7s (CSP7OPs) identified in Aim I, then evaluate non-GLP pharmacokinetics, toxicology and evaluate systemic biodistribution after airway delivery. In Aim III, we will elucidate the molecular mechanisms by which CSP7OP combined with pirfenidone or nintedanib, affects injured AECs and fLfs to resolve existing PF. Targeting Cav1 with CSP7 represents a novel and promising approach for treatment of IPF via the airway. This project has a high probability of success and the data to be generated is a predicate for IND-enabling work. Our team has all the requisite expertise in cellular/molecular biology, preclinical modeling, drug discovery, formulation and airway drug delivery. This project will likely define novel and potentially more effective therapy for patients with IPF or other ILDs, for whom effective treatment options remain limited.