Nanotechnology Approach for Inhalation Treatment of Pulmonary Fibrosis

描述（由申请人提供）：吸入治疗肺纤维化的纳米技术方法特发性肺纤维化（IPF）是一种慢性、进行性且通常致命的间质性肺病，通常导致患者发病和死亡。然而，IPF的治疗是一个重大的临床挑战，因为这种疾病没有可靠的治疗选择。前列腺素E2（PGE 2）是一种环加氧酶衍生的脂质介质，由于其在IPF的发生和进展中的作用以及作为限制免疫炎症反应、抑制特定肺成纤维细胞功能、其增殖和基质蛋白（如胶原蛋白）合成的可能治疗剂而引起了相当大的关注。然而，使用PGE 2治疗IPF的主要挑战是其对肺部的低效递送和对其他器官的严重不良副作用。为了验证PGE 2可以成功地用于治疗IPF，我们通过吸入将脂质体形式的PGE 2递送至患有IPF的小鼠，并且发现PGE 2的局部递送具有高治疗潜力。PGE 2的作用与导致IPF的主要蛋白质表达的正常化有关。然而，并非所有靶向蛋白都被有效抑制，IPF的一些体征（最明显的是间质性肺水肿、炎症和过度胶原蛋白生成）没有完全消除。基于这些观察结果，我们假设IPF治疗结果的成功可能通过PGE 2和负责炎症、细胞外基质降解和缺氧损伤的蛋白质的抑制剂的组合局部肺递送来增强。因此，本研究的主要目标是开发和体内测试一种专门设计的用于吸入的基于纳米载体的药物递送系统（DDS），其含有PGE 2和靶向基质金属蛋白酶（MMP 3）、趋化因子（CCL 12）和缺氧诱导因子1 α（HIF 1A）的siRNA。该提案主要集中在（1）用于肺部递送PGE 2和siRNA的纳米结构脂质载体（NLC）的合成和表征;（2）鉴定参与IPF发展的最重要蛋白质;（3）siRNA序列的选择;（4）纳米颗粒气溶胶化的表征和优化，包括雾化器性能的优化，气载DDS浓度的测定，DDS的动态稳定性分析;（5）测定递送的DDS在体内的身体分布;和（6）评价所提出的治疗方法在IPF小鼠模型中的治疗效率（博莱霉素的阴道内施用）。计划中的研究解决了IPF治疗中的关键问题-治疗有效性低和严重不良副作用。本申请进行了一种新的纳米颗粒策略的概念验证实验，用于同时局部肺递送PGE 2和主要负责炎症、细胞外基质降解和缺氧损伤的蛋白质的siRNA抑制剂。该项目提出了切实实现这一概念的创新办法和方法。

DESCRIPTION (provided by applicant): Nanotechnology Approach for Inhalation Treatment of Pulmonary Fibrosis Idiopathic pulmonary fibrosis (IPF) is a chronic, progressive and often fatal form of interstitial lung disease often resulting in patient morbidity and mortality. However, treatment of IPF represents a major clinical challenge since this disorder does not have reliable therapeutic options. Prostaglandin E2 (PGE2), a cyclooxygenase-derived lipid mediator, has attracted considerable attention for its role in the development and progression of IPF and as a possible therapeutic agent for limitation of the immune-inflammatory response, inhibition of specific lung fibroblast functions, their proliferation and synthesis of matrix proteins such as collagen. However, the major challenge in the use of PGE2 for treatment of IPF is its inefficient delivery to the lungs and severe adverse side effects on other organs. To verify that PGE2 can be successfully used for treatment of IPF, we delivered liposomal form of PGE2 via inhalation to the mice with IPF and found that local delivery of PGE2 has a high therapeutic potential. The effect of PGE2 was related to the normalization of the expression of major proteins responsible for the IPF. However, not all targeted proteins were effectively suppressed and some signs of IPF (most notably interstitial lung edema, inflammation, and excessive collagen production) were not completely eliminated. Based on these observations, we hypothesize that the success in the treatment outcome of IPF might be enhanced by combinatorial local lung delivery of PGE2 and suppressors of proteins responsible for inflammation, extracellular matrix degradation, and hypoxic damage. Consequently, the major goal of this study is to develop and test in vivo a specially designed for inhalation nanocarrier-based drug delivery system (DDS) containing PGE2 and siRNA targeted to matrix metalloproteinase (MMP3), chemokine (CCL12) and hypoxia inducible factor one alpha (HIF1A). The proposal is focused mainly on the (1) synthesis and characterization of nanostructured lipid carrier (NLC) for a pulmonary delivery of PGE2 and siRNA; (2) identification of most important proteins involved in the development of IPF; (3) selection of siRNA sequences; (4) characterization and optimization of nanoparticles aerosolization, including optimization of nebulizer performance, determination of airborne DDS concentration, analysis of dynamic stability of DDS; (5) determination of body distribution of delivered DDS in vivo; and (6) evaluation of therapeutic efficiency of the proposed therapeutic approach in a mouse model of IPF (intratracheal administration of bleomycin). The planned research addresses critical problems in treatment of IPF - low effectiveness of therapy and severe adverse side effects. The application performs proof-of-concept experiments of a novel nanotherapeutic strategy for simultaneous local lung delivery of PGE2 and siRNA - suppressors of proteins primarily responsible for inflammation, extra cellular matrix degradation, and hypoxic damage. The project proposes an innovative approach and methodology for the practical realization of this concept.