Nebulised siRNA encapsulated crosslinked chitosan nanoparticles for pulmonary delivery.

PURPOSE: To explore the potential of crosslinked chitosan nanoparticles as carriers for delivery of siRNA using a jet nebuliser. MATERIALS AND METHODS: Nanoparticles encapsulating siRNA were prepared using an ionic crosslinking technique at chitosan to siRNA weight/weight ratios of 10:1, 30:1 and 50:1. Particles were characterised for their size, charge, morphology, pH stability and siRNA encapsulation efficiency. Gel electrophoresis was used to assess the association and stability of siRNA with nanoparticles, including after aerosolisation using a Pari LC Sprint jet nebuliser. The aerosolisation properties of FITC labelled chitosan nanoparticles were investigated using a two-stage impinger. Cell viability was performed with H-292 cells using a WST-1 assay. RESULTS: Positively charged spherical nanoparticles were produced with mean diameters less than 150 nm, at all chitosan to siRNA ratios. Nanoparticles were non-aggregated at the pH of the airways and showed high siRNA encapsulation efficiency (>96%). Complete binding of siRNA to chitosan nanoparticles was observed when the w/w ratio was 50:1. Nebulisation produced fine particle fractions of 54±11% and 57.3±1.9% for chitosan and chitosan:siRNA (10:1 w/w) nanoparticles respectively. The stability of chitosan-encapsulated siRNA was maintained after nebulisation. Cell viability was high (>85%) at the highest chitosan concentration (83 µg/ml). CONCLUSION: The results suggest that crosslinked chitosan nanoparticles have potential for siRNA delivery to the lungs using a jet nebuliser.

Challenges with developing in vitro dissolution tests for orally inhaled products (OIPs).

The purpose of this article is to review the suitability of the analytical and statistical techniques that have thus far been developed to assess the dissolution behavior of particles in the respirable aerodynamic size range, as generated by orally inhaled products (OIPs) such as metered-dose inhalers and dry powder inhalers. The review encompasses all analytical techniques publicized to date, namely, those using paddle-over-disk USP 2 dissolution apparatus, flow-through cell dissolution apparatus, and diffusion cell apparatus. The available techniques may have research value for both industry and academia, especially when developing modified-release formulations. The choice of a method should be guided by the question(s) that the research strives to answer, as well as by the strengths and weaknesses of the available techniques. There is still insufficient knowledge, however, for translating the dissolution data into statements about quality, performance, safety, or efficacy of OIPs in general. Any attempts to standardize a dissolution method for compendial inclusion or compendial use would therefore be premature. This review reinforces and expands on the 2008 stimulus article of the USP Inhalation Ad Hoc Advisory Panel, which "could not find compelling evidence suggesting that such dissolution testing is kinetically and/or clinically crucial for currently approved inhalation drug products."

Crosslinked chitosan nanoparticle formulations for delivery from pressurized metered dose inhalers.

Crosslinked chitosan nanoparticles, prepared using ionic gelation, have been successfully formulated into pressurized metered dose inhalers (pMDIs) with potential for deep lung delivery of therapeutic agents. Nanoparticles were prepared from crosslinked chitosan alone and incorporating PEG 600, PEG 1000 and PEG 5000 for dispersion in aerosol propellant, hydrofuoroalkane (HFA) 227. Spherical, smooth-surfaced, cationic particles of mean size less than 230 nm were produced. Nanoparticles were positively charged and non-aggregated at the pH of the airways. Crosslinked chitosan-PEG 1000 nanoparticles demonstrated greatest dispersibility and physical stability in HFA-227, whereas other formulations readily either creamed or sedimented. Following actuation from pMDIs, the fine particle fraction (FPF) for crosslinked chitosan-PEG 1000 nanoparticles, determined using a next generation impactor, was 34.0±1.4% with a mass median aerodynamic diameter of 4.92±0.3 µm. The FPFs of crosslinked chitosan, crosslinked chitosan-PEG 600 and crosslinked chitosan-PEG 5000 nanoparticles were 5.7±0.9%, 11.8±2.7% and 17.0±2.1%, respectively. These results indicate that crosslinked chitosan-PEG 1000-based nanoparticles are promising candidates for delivering therapeutic agents, particularly biopharmaceuticals, using pMDIs.