TPP-dendrimer nanocarriers for siRNA delivery to the pulmonary epithelium and their dry powder and metered-dose inhaler formulations

被引:55
作者
Bielski, Elizabeth [1 ,3 ,4 ]
Zhong, Qian [1 ,3 ,4 ]
Mirza, Hamad [1 ]
Brown, Matthew [1 ]
Molla, Ashura [1 ]
Carvajal, Teresa [2 ]
da Rocha, Sandro R. P. [1 ,3 ,4 ]
机构
[1] Wayne State Univ, Dept Chem Engn & Mat Sci, Detroit, MI 48202 USA
[2] Purdue Univ, Dept Agr & Biol Engn, W Lafayette, IN 47907 USA
[3] Virginia Commonwealth Univ, Sch Pharm, Dept Pharmaceut, Med Coll Virginia Campus, Richmond, VA 23284 USA
[4] Virginia Commonwealth Univ, Sch Engn, Dept Chem & Life Sci Engn, Med Coll Virginia Campus, Richmond, VA 23284 USA
基金
美国国家科学基金会;
关键词
PAMAM dendrimers; Dendriplexes; Triphenylphosphonium (TPP) ion; Short interfering RNA (siRNA); Inhalation therapy; Pressurized metered dose inhalers (pMDIs); Dry powder inhalers (DPIs); IN-VIVO; TARGETED DELIVERY; GENE DELIVERY; EFFICIENT; DESIGN; CARRIER; VITRO; CORE; RNAI; TRIPHENYLPHOSPHONIUM;
D O I
10.1016/j.ijpharm.2017.05.046
中图分类号
R9 [药学];
学科分类号
1007 ;
摘要
The regulation of genes utilizing the RNA interference (RNAi) mechanism via the delivery of synthetic siRNA has great potential in the treatment of a variety of lung diseases. However, the delivery of siRNA to the lungs is challenging due to the poor bioavailability of siRNA when delivered intraveneously, and difficulty in formulating and maintaining the activity of free siRNA when delivered directly to the lungs using inhalation devices. The use of non-viral vectors such as cationic dendrimers can help enhance the stability of siRNA and its delivery to the cell cytosol. Therefore, in this work, we investigate the ability of a triphenylphosphonium (TPP) modified generation 4 poly(amidoamine) (PAMAM) dendrimer (G4NH(2)-TPP) to enhance the in vitro transfection efficiency of siRNA in a model of the pulmonary epithelium and their aerosol formulations in pressurized metered dose inhalers (pMDIs) and dry powder inhalers (DPIs). Complexes of siRNA and G4NH(2)-TPP were prepared with varying TPP densities and increasing N/P ratios. The complexation efficiency was modulated by the presence of the TPP on the dendrimer surface, allowing for a looser complexation compared to unmodified dendrimer as determined by gel electrophoresis and polyanion competition assay. An increase in TPP density and N/P ratio led to an increase in the in vitro gene knockdown of stably green fluorescent protein (eGFP) expressing lung alveolar epithelial (A549) cells. G4NH(2)-12TPP dendriplexes (G4NH(2) PAMAM dendrimers containing 12 TPP molecules on the surface complexed with siRNA) at N/P ratio 30 showed the highest in vitro gene knockdown efficiency. To assess the potential of TPP-dendriplexes for pulmonary use, we also developed micron particle technologies for both pMDIs and DPIs and determined their aerosol characteristics utilizing an Andersen Cascade Impactor (ACI). Mannitol microparticles encapsulating 12TPP-dendriplexes were shown to be effective in producing aerosols suitable for deep lung deposition for both pMDI formulations (fine particle fraction of 50-53%) and DPI formulations (fine particle fraction of 39%) with no impact on the in vitro gene knockdown efficiency of the siRNA. This work demonstrates the potential benefits of utilizing TPP-conjugated dendrimers in the formation of dendriplexes for siRNA delivery to the pulmonary epithelium and their aerosol formulation for local delivery to the lungs using portable inhalers. (C) 2017 Elsevier B.V. All rights reserved.
引用
收藏
页码:171 / 183
页数:13
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