The use of absorption enhancers to enhance the dispersibility of spray-dried powders for pulmonary gene therapy.
BACKGROUND
Pulmonary gene therapy requires aerosolisation of the gene vectors to the target region of the lower respiratory tract. Pulmonary absorption enhancers have been shown to improve the penetration of pharmaceutically active ingredients in the airway. In this study, we investigate whether certain absorption enhancers may also enhance the aerosolisation properties of spray-dried powders containing non-viral gene vectors.


METHODS
Spray-drying was used to prepare potentially respirable trehalose-based dry powders containing lipid-polycation-pDNA (LPD) vectors and absorption enhancers. Powder morphology and particle size were characterised using scanning electron microscopy and laser diffraction, respectively, with gel electrophoresis used to assess the structural integrity of the pDNA. The biological functionality of the powders was quantified using in vitro cell (A549) transfection. Aerosolisation from a Spinhaler dry powder inhaler into a multistage liquid impinger (MSLI) was used to assess the in vitro dispersibility and deposition of the powders.


RESULTS
Spray-dried powder containing dimethyl-beta-cyclodextrin (DMC) demonstrated substantially altered particle morphology and an optimal particle size distribution for pulmonary delivery. The inclusion of DMC did not adversely affect the structural integrity of the LPD complex and the powder displayed significantly greater transfection efficiency as compared to unmodified powder. All absorption enhancers proffered enhanced powder deposition characteristics, with the DMC-modified powder facilitating high deposition in the lower stages of the MSLI.


CONCLUSIONS
Incorporation of absorption enhancers into non-viral gene therapy formulations prior to spray-drying can significantly enhance the aerosolisation properties of the resultant powder and increase biological functionality at the site of deposition in an in vitro model.
Li
H-Y
HY
Gene Delivery Research Group, Welsh School of Pharmacy, Cardiff University, Cardiff CF10 3XF, UK.
Seville
P C
PC
Williamson
I J
IJ
Birchall
J C
JC
D000042:Absorption / Q000502:physiology*; D045744:Cell Line, Tumor; D004247:DNA / Q000737:chemistry; D005434:Flow Cytometry; D015316:Genetic Therapy / Q000379:methods*; D006801:Humans; D066298:In Vitro Techniques; D008171:Lung Diseases / Q000628:therapy*; D008954:Models, Biological; D010316:Particle Size; D011208:Powders / Q000008:administration & dosage* / Q000737:chemistry / Q000627:therapeutic use*; D014162:Transfection
D011208:Powders; D004247:DNA