Li H-Y, Seville P C, Williamson I J, Birchall J C
Gene Delivery Research Group, Welsh School of Pharmacy, Cardiff University, Cardiff CF10 3XF, Wales, UK.
J Gene Med. 2005 Mar;7(3):343-53. doi: 10.1002/jgm.654.
Pulmonary delivery of gene therapy offers the potential for the treatment of a range of lung conditions, including cystic fibrosis, asthma and lung cancer. Spray-drying may be used to prepare dry powders for inhalation; however, aerosolisation of such powders is limited, resulting in poor lung deposition and biological functionality. In this study, we examine the use of amino acids (arginine, aspartic acid, threonine, phenylalanine) to enhance the aerosolisation of spray-dried powders containing model non-viral gene vectors.
Lipid/polycation/pDNA (LPD) vectors, in the presence or absence of amino acids, were dispersed in lactose solutions, and spray-dried to produce appropriately sized dry powders. Scanning electron microscopy and laser diffraction were used to determine particle morphology and diameter, respectively. Gel electrophoresis was used to examine the influence of amino acids on the structural integrity of the LPD complex. In vitro cell (A549) transfection was used to determine the biological functionality of the dry powders, and the in vitro aerosolisation performance was assessed using a multistage liquid impinger (MSLI).
Both gel electrophoresis and in vitro cell transfection indicated that certain amino acids (aspartic acid, threonine) can adversely affect the integrity and biological functionality of the LPD complex. All amino acids significantly increased the aerosolisation of the powder, with the arginine and phenylalanine powders showing optimal deposition in the lower stages of the MSLI.
Amino acids can be used to enhance the aerosolisation of spray-dried powders for respiratory gene delivery, allowing the development of stable and viable formulations for pulmonary gene therapy.
基因治疗的肺部给药为一系列肺部疾病的治疗提供了潜力，包括囊性纤维化、哮喘和肺癌。喷雾干燥可用于制备吸入用干粉；然而，此类粉末的雾化受限，导致肺部沉积和生物功能不佳。在本研究中，我们考察了使用氨基酸（精氨酸、天冬氨酸、苏氨酸、苯丙氨酸）来增强含有模型非病毒基因载体的喷雾干燥粉末的雾化效果。
在有或没有氨基酸存在的情况下，将脂质/聚阳离子/pDNA（LPD）载体分散在乳糖溶液中，并进行喷雾干燥以产生尺寸合适的干粉。分别使用扫描电子显微镜和激光衍射来确定颗粒形态和直径。凝胶电泳用于考察氨基酸对LPD复合物结构完整性的影响。体外细胞（A549）转染用于确定干粉的生物功能，并且使用多级液体撞击器（MSLI）评估体外雾化性能。
凝胶电泳和体外细胞转染均表明某些氨基酸（天冬氨酸、苏氨酸）会对LPD复合物的完整性和生物功能产生不利影响。所有氨基酸均显著增加了粉末的雾化效果，精氨酸和苯丙氨酸粉末在MSLI的较低阶段显示出最佳沉积。
氨基酸可用于增强喷雾干燥粉末用于呼吸道基因递送的雾化效果，从而开发出用于肺部基因治疗的稳定且可行的制剂。
