S-Nitrosothiol-Based Rinse/Aerosol Solutions for Treatment/Prevention of Rhinosinusitis (Phase II)

摘要： 据估计，慢性鼻窦炎（CRS）影响13%的美国人口，每年的医疗保健 每年的直接成本近100亿美元，但CRS的有效疗法很少。检测 鼻窦粘膜中的金黄色葡萄球菌和/或铜绿假单胞菌生物膜与 严重CRS表现。鼻腔通道内的健康细胞通常会释放出大量的 一氧化氮（NO），一种有效的天然抗菌分子，刺激粘膜纤毛清除， 减少细菌生物膜的形成。CRS患者呼出鼻气中NO水平降低 这表明提高鼻窦内局部NO浓度可能是非常有益的。 在该项目的第一阶段，NOTA实验室，有限责任公司，清楚地表明，一个自然发生的， 廉价的NO释放剂，S-亚硝基谷胱甘肽（GSNO），可以掺入到典型的 生理盐水/碳酸氢盐窦冲洗溶液，并产生足够的NO水平，以显著降低 两种生物膜形成CRS微生物（S.金黄色葡萄球菌和铜绿假单胞菌）。在 在这些实验中，细菌生物膜生长在人上气道上皮细胞的表面上， 细胞我们进一步证明了GSNO和伴随的NO释放水平不会损害细胞的功能。 通过监测毒性相关细胞因子（IL-8）的水平来评估。在 此外，NOTA表明，NO从GSNO的释放速率可以通过 抗坏血酸盐（Asc;维生素C）的存在，并且所得的GSNO/Asc的粉末制剂可以 储存超过6个月（在37 °C下），同时保留>80%的NO释放能力。 通过第二阶段SBIR资金，NOTA将：1）进一步优化GSNO/Asc混合物的浓度 在等渗NaCl/NaHCO 3/EDTA溶液中， S.金黄色葡萄球菌和铜绿假单胞菌在人上气道上皮细胞上形成的生物膜 2）探索商业包装工艺（在聚合物薄膜内衬的铝袋中）并测试 干燥包装的GSNO/Asc/NaCl/NaHCO 3/EDTA混合粉末的1年稳定性， 临用前用240 mL纯水在塑料鼻窦冲洗瓶中复溶， 3）在兔子中进行动物研究以确定NOTA是否重构 RSNO/Asc/NaCl/NaHCO 3/EDTA制剂，以气雾剂喷雾剂形式给药，每日两次，持续7天， 与对照组相比，显著降低感染家兔（CRS）窦腔中的细菌计数 (only NaCl/NaHCO 3/EDTA）;以及4）通过获得以下方面的专家评估开始监管过程： FDA在批准新的GSNO制剂作为潜在的非处方产品方面的作用 CRS市场。在第二阶段完成这些工作将使国家海洋和海洋事务管理局能够规划第一个 作为III期研究的一部分，在密歇根大学鼻窦中心进行的人类受试者临床试验。

Abstract: Chronic rhinosinusitis (CRS) is estimated to affect 13% of the US population, with annual health care direct costs of nearly $10 billion per year, yet there are few effective therapies for CRS. Detection of Staphylococcus aureus and/or Pseudomonas aeruginosa biofilm in the sinus mucosa is associated with severe CRS presentation. Healthy cells that line the nasal passages normally emit significant levels of nitric oxide (NO), a potent and natural antimicrobial molecule that stimulates mucociliary clearance and reduces bacterial biofilm formation. Reduced levels of NO in the exhaled nasal breath of CRS patients suggest that boosting local NO concentrations within the sinuses may be quite beneficial. In Phase I of this project, NOTA Laboratories, LLC, clearly demonstrated that a naturally occurring and inexpensive NO releasing agent, S-nitrosoglutathione (GSNO), can be incorporated into a typical saline/bicarbonate sinus rinse solution and generate adequate levels of NO to significantly reduce levels of live bacteria counts for two biofilm forming CRS microbes (S. aureus and P. aeruginosa). In these experiments, the bacterial biofilms were grown on the surface of human upper airway epithelial cells. We further demonstrated that the levels of GSNO and concomitant NO released do not harm the underlying human cells, as assessed by monitoring the levels of a toxicity-associated cytokine (IL-8). In addition, NOTA demonstrated that the release rates of NO from GSNO can be enhanced by the presence of ascorbate (Asc; vitamin C) and that the resulting powder formulations of GSNO/Asc can be stored for more than 6 months (at 37 °C) while retaining >80% of their NO release capability. Via Phase II SBIR funding, NOTA will: 1) Further optimize the concentration of the GSNO/Asc mixture within an isotonic NaCl/NaHCO3/EDTA solution and frequency of application for the in vitro disruption of S. aureus and P. aeruginosa biofilms formed on human upper airway epithelial cells over 3 d test periods; 2) Explore a commercial packaging process (in polymer film lined aluminum pouches) and test the resulting 1-year stability of a dry packaged GSNO/Asc/NaCl/NaHCO3/EDTA mixed powder that will be reconstituted in plastic sinus flush bottles with 240 mL of pure water immediately before use by patients; 3) Conduct animal studies in rabbits to determine if NOTA's reconstituted RSNO/Asc/NaCl/NaHCO3/EDTA formulation, given as an aerosol spray twice daily over a 7 d period, significantly decreases bacterial counts in the sinus cavities of infected rabbits (with CRS) vs. controls (only NaCl/NaHCO3/EDTA); and 4) Begin the regulatory process by obtaining expert assessments of the role of the FDA in approval of the new GSNO formulations as a potential over-the-counter product for the CRS market. The completion of these efforts during Phase II will enable NOTA to plan the first human subject clinical trials at the University of Michigan Sinus Center as part of Phase III studies.