Pillai R, Petrak K, Blezinger P, Deshpande D, Florack V, Freimark B, Padmabandu G, Rolland A
GENE MEDICINE, INC., The Woodlands, Texas 77381-4248, USA.
Pharm Res. 1998 Nov;15(11):1743-7. doi: 10.1023/a:1011964813817.
This study relates to the development of gene therapies for the treatment of lung diseases. It describes for the first time the use of ultrasonic nebulization for administration of plasmid/lipid complexes to the lungs to transfect lung epithelial cells.
Plasmid complexed to cationic liposomes at a specific stoichiometric ratio was nebulized using an ultrasonic nebulizer. We assessed: (i) the stability of plasmid and plasmid/lipid complexes to ultrasonic nebulization, (ii) the in vitro activity of plasmid in previously nebulized plasmid/lipid complex, (iii) the in vivo transgene expression in lungs following intratracheal instillation of nebulized plasmid/lipid formulations compared to un-nebulized complexes, (iv) the emitted dose from an ultrasonic nebulizer using plasmid/lipid complexes of different size, and (v) the transgene expression in lungs following oral inhalation of aerosolized plasmid/lipid complex generated using an ultrasonic nebulizer.
Integrity of plasmid formulated with cationic lipids, and colloidal stability of the plasmid/lipid complex were maintained during nebulization. In contrast, plasmid alone formulated in 10% lactose was fragmented during nebulization. The efficiency of transfection of the complex before and after nebulization was comparable. Nebulization produced respirable aerosol particles. Oral exposure of rodents for 10 minutes to aerosol produced from the ultrasonic nebulizer resulted in transgene expression in lungs in vivo.
The performance characteristics of the ultrasonic nebulizer with our optimized plasmid/lipid formulations suggests that this device can potentially be used for administering gene medicines to the airways in clinical settings for the treatment of respiratory disorders.
本研究涉及用于治疗肺部疾病的基因疗法的开发。首次描述了使用超声雾化将质粒/脂质复合物给药至肺部以转染肺上皮细胞。
将以特定化学计量比与阳离子脂质复合的质粒用超声雾化器进行雾化。我们评估了：（i）质粒和质粒/脂质复合物对超声雾化的稳定性，（ii）先前雾化的质粒/脂质复合物中质粒的体外活性，（iii）与未雾化的复合物相比，经气管内滴注雾化的质粒/脂质制剂后肺部的体内转基因表达，（iv）使用不同大小的质粒/脂质复合物时超声雾化器的发射剂量，以及（v）经超声雾化器产生的雾化质粒/脂质复合物经口服吸入后肺部的转基因表达。
在雾化过程中，用阳离子脂质配制的质粒的完整性以及质粒/脂质复合物的胶体稳定性得以维持。相比之下，单独在10%乳糖中配制的质粒在雾化过程中发生了片段化。雾化前后复合物的转染效率相当。雾化产生了可吸入的气溶胶颗粒。将啮齿动物经口暴露于超声雾化器产生的气溶胶10分钟导致体内肺部的转基因表达。
超声雾化器与我们优化的质粒/脂质制剂的性能特征表明，该装置有可能在临床环境中用于将基因药物给药至气道以治疗呼吸系统疾病。
