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

间质性肺疾病(ILDS)的特征是进行性肺瘢痕形成。特发性肺 纤维化(IPF)是ILDS最常见的形式之一，是一种致命的肺部疾病，发病率为60% 美国每年的病例/10万人。IPF的五年存活率中值只有20%。目前有一种 没有解药。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(FLf)及细胞外的增殖和积聚 基质沉积。这些特征会导致进行性肺功能障碍。形态变化包括空间和 时间异质性包括正常肺区域与含有细胞凋亡的病变区相邻 AECS和fLFS。最近，药物治疗已被证明可以延缓IPF的进展，提示更多 可以开发出有效的治疗方法。我们发现P53的增加促进了血管内皮细胞的凋亡和成纤维细胞 它在fLf中的减少促进了肌成纤维细胞的扩张。我们还发现了小窝蛋白-1(Cav1) 在受损的血管内皮细胞中，其水平升高，而在IPF患者肺组织中增殖的fLf中，其水平显著降低。 以及已建立PF的小鼠。我们鉴定了一个20聚体的Cav1支架结构域肽(CSP)，并将其截短 7-聚体片段CSP7，可抑制P53、转化生长因子-β、结缔组织生长因子、血管内皮细胞的凋亡和FLF的增殖。这些多肽可以阻断 单次(1倍)或多次(8倍)博莱霉素、腺病毒转化生长因子-β-1(Ad-转化生长因子-β1)和二氧化硅对小鼠肺功能的影响 致肺损伤。CSP7抑制FLf中MDM2表达增加所致的P53降解，并阻断其 扩散。CSP7还抑制AEC的衰老和凋亡，而在纤维化的肺中，AEC的衰老和凋亡会增加。 包括在IPF中。CSP7在小鼠体内耐受性良好，可以通过呼吸道有效地给药。我们的工作 验证Cav1作为PF的治疗靶点。我们的出版物为开发用于呼吸道的CSP7提供了前提 雾化吸入(NEB)或干粉吸入(DPI)是治疗PF的一种新的、安全和有效的治疗方法。在……里面 目的一、确定CSP7的细胞表面结合/模式作用，优化CSP7的结构，确定CSP7的作用范围 评估CSP7的耐受性和分解肺泡灌洗液的能力 4只(青年和老年小鼠1x BLm，Ad-转化生长因子-β1和8x BLm)建立PF模型。在AIM II中，我们将选择最多 在AIM I中确定有效、耐受性良好、优化的CSP7(CSP7OPs)，然后评估非GLP 药物动力学、毒理学和评价呼吸道给药后的全身生物分布。在AIM III中，我们将 阐明CSP7OP联合吡非尼酮或9tedanib影响损伤的分子机制 AECS和fLFS来解析现有的PF。使用CSP7靶向Cav1代表了一种新的、有前途的方法 经呼吸道治疗特发性肺纤维化。这个项目成功的概率很高，要生成的数据是 支持IND的工作的谓词。我们的团队拥有所有必要的细胞/分子生物学专业知识，临床前 建模、药物发现、配方和呼吸道给药。这个项目可能会定义新的和潜在的 对IPF或其他ILDS患者更有效的治疗，他们的有效治疗选择仍然有限。

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.