Lung Delivery of Nebulized Tissue Plasminogen Activator

描述（由申请人提供）：在美国，急性呼吸窘迫综合征（ARDS）每年折磨数万人，并与显著的死亡率相关。目前，没有有效的治疗方法，目前正在研究的前瞻性治疗方法也不乐观。在肺部炎症动物模型中，一种独特的抗氧化剂（蛋白质）可减少中性粒细胞活性氧（ROS）介导的损伤，并已被鉴定为ARDS的潜在可行疗法。因此，这项研究和药物开发计划的广泛，长期目标是开发一种独特的，局部的（肺靶向）治疗，将有效降低与ARDS相关的死亡率。本I期申请的目的是通过开发一种能够维持雾化过程并充分分布到人体呼吸道中的药物制剂，证明该策略在人体中的原理。为此，本项目的具体目标是：1）配制药剂，使其适合通过雾化进行局部肺部给药，同时保持活性。通过添加不同浓度的表面活性剂将得到最佳配方。制剂活动将包括蛋白质稳定性、空气动力学粒度和生物学功能的评估。将通过总蛋白回收率和结构完整性测量蛋白稳定性。将使用级联冲击器测定粒度，并通过试剂抑制嗜中性粒细胞ROS产生的能力来定量生物活性; 2）表征试剂的雾化制剂在人肺复制模型中的呼吸沉积。该目的旨在生成额外的支持性证据，以证明该制剂在人体中的原理。为了实现该目的，将根据特定目的#1的可行性标准确定的每种可行制剂雾化到人肺复制品中。将使用由人尸体制成的主模型复制的高保真人肺复制品，其中包括五至九代支气管。通过流经上游气道并收集在泡沫过滤器上的材料模拟铸件小气道和肺泡区域中的沉积。在这两个具体目标中进行的工作的结果将确定这种新的和新颖的药剂的雾化制剂的可行性，以及作为II期应用和随后的产品商业化的基础。

DESCRIPTION (provided by applicant): Acute respiratory distress syndrome (ARDS) afflicts tens of thousands of individuals annually in the United States and is associated with significant mortality. Presently, there is no effective treatment and little optimism exists for prospective therapies currently under investigation. A unique antiinflammatory agent (protein) that reduces neutrophil reactive oxygen species (ROS)-mediated injury in an animal model of lung inflammation has been identified and represents a potentially viable therapy for ARDS. Therefore, the broad, long-term objectives of this research and drug development program are to develop a unique, localized (lung-targeted) therapy that will be effective in reducing the mortality associated with ARDS. The purpose of this phase I application is to prove the principle of this strategy in man by developing a drug formulation that will sustain the nebulization process and distribute adequately into the human respiratory tract. To this end, the specific aims of this project are to: 1) formulate the agent to make it suitable for local pulmonary delivery via nebulization while retaining activity. Optimal formulation will be derived by the addition of varying concentrations of surfactant. The formulation activities will include an assessment of protein stability, aerodynamic particle size, and biological function. Protein stability will be measured by total protein recovery and structural integrity. Particle size will be determine using a cascade impactor and biological activity will be quantified by the ability of the agent to inhibit neutrophil ROS production; 2) characterize the respiratory deposition of a nebulized formulation of the agent in a human lung replica model. This aim is designed to generate additional supportive evidence to prove the principle of the formulation in man. To accomplish this aim, each viable formulation, as determined by the feasibility criteria of specific aim #1, will be nebulized into a human lung replica. A high-fidelity human lung replica reproduced from a master cast made from a human cadaver which includes five to nine generations of bronchi will be utilized. Deposition in the small airways and alveoli regions of the cast is simulated by material that passes through the upstream airways and is collected on foam filters. The results of the work performed in both specific aims will determine the feasibility of the nebulized formulation of this new and novel agent as well as serve as the basis for a phase II application and the subsequent commercialization of the product