Nanotechnology Approach for Inhalation Treatment of Pulmonary Fibrosis

描述(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、进行性的、通常是致命的间质性肺部疾病，通常会导致患者的发病率和死亡率。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(PGE2)是一种环氧合酶衍生的脂质介质，因其在IPF的发生发展中的作用而备受关注，并可能作为一种治疗药物来限制免疫炎症反应，抑制特定的肺成纤维细胞功能，抑制其增殖和合成胶原等基质蛋白。然而，使用PGE2治疗IPF的主要挑战是它对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可以成功地用于IPF的治疗，我们通过吸入的方式将PGE2脂质体输送到IPF小鼠身上，发现局部给药具有很高的治疗潜力。PGE2的作用与参与IPF的主要蛋白表达正常化有关。然而，并不是所有的靶蛋白都被有效地抑制，一些IPF的迹象(最明显的是间质性肺水肿、炎症和过度的胶原蛋白产生)并没有完全消除。基于这些观察，我们假设肺内联合应用前列腺素E_2和抑制炎症、细胞外基质降解和低氧损伤的蛋白可以提高IPF治疗结果的成功率。因此，本研究的主要目标是开发并在体内测试一种专门设计的吸入型纳米载体给药系统(DDS)，该系统包含针对基质金属蛋白酶(MMP3)、趋化因子(CCL12)和低氧诱导因子1α(HIF1a)的PGE2和siRNA。该建议主要集中在(1)用于PGE2和siRNA肺部递送的纳米结构脂质载体(NLC)的合成和表征；(2)确定参与IPF发展的最重要的蛋白质；(3)siRNA序列的选择；(4)纳米颗粒雾化的表征和优化，包括雾化性能的优化、DDS气载浓度的测定、DDS动态稳定性的分析；(5)DDS体内分布的测定；以及(6)对所提议的治疗方法在IPF(气管内给药)小鼠中的治疗效率的评估。这项计划中的研究针对IPF治疗中的关键问题--治疗效率低和严重的副作用。该应用程序对一种新的纳米治疗策略进行了概念验证实验，该策略可同时在肺内局部递送PGE2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抑制物。该项目为实际实现这一概念提出了一种创新的办法和方法。

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.