Nanotechnology Approach for Inhalation Treatment of Pulmonary Fibrosis

特发性肺纤维化（IPF）是一种慢性、进行性和经常致命的间质性肺疾病，通常导致患者发病率和死亡率。然而，治疗IPF是一个主要的临床挑战，因为这种疾病没有可靠的治疗选择。前列腺素E2 （PGE2）是一种环氧化酶衍生的脂质介质，因其在IPF的发生和进展中所起的作用而引起了人们的广泛关注，并可能成为一种限制免疫炎症反应、抑制特定肺成纤维细胞功能、它们的增殖和胶原等基质蛋白合成的治疗药物。然而，使用PGE2治疗IPF的主要挑战是其向肺部的输送效率低下以及对其他器官的严重不良副作用。为了验证PGE2可以成功用于治疗IPF，我们通过吸入将PGE2脂质体形式传递给IPF小鼠，发现局部传递PGE2具有很高的治疗潜力。PGE2的作用与负责IPF的主要蛋白的表达正常化有关。然而，并不是所有的靶蛋白都被有效抑制，一些IPF的迹象（最明显的是间质性肺水肿、炎症和过多的胶原蛋白产生）没有完全消除。基于这些观察结果，我们假设IPF的成功治疗结果可能通过联合局部肺递送PGE2和负责炎症、细胞外基质降解和缺氧损伤的蛋白质抑制因子而增强。因此，本研究的主要目标是开发和体内测试一种专门设计用于吸入的基于纳米载体的药物递送系统（DDS），该系统含有PGE2和siRNA，靶向基质金属蛋白酶（MMP3）、趋化因子（CCL12）和缺氧诱导因子α (HIF1A)。该提案主要集中在(1)用于肺递送PGE2和siRNA的纳米结构脂质载体（NLC）的合成和表征；(2)鉴定IPF发生过程中最重要的蛋白；(3) siRNA序列的选择；(4)纳米颗粒雾化的表征与优化，包括雾化器性能优化、机载DDS浓度测定、DDS动态稳定性分析；(5)测定给药DDS的体内分布；(6)在IPF小鼠模型（气管内给药博莱霉素）中评估所提出的治疗方法的治疗效果。计划的研究解决了治疗IPF的关键问题-治疗效果低和严重的不良反应。该应用程序执行了一种新型纳米治疗策略的概念验证实验，该策略可同时局部肺递送PGE2和siRNA -主要负责炎症，细胞外基质降解和缺氧损伤的蛋白质抑制因子。该项目提出了一种创新的方法和方法来实际实现这一概念。

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.