Enhanced Lung Delivery of mRNA Using Nebulized Lipid Nanoparticles
Ricardo Velez1, Susana Ramalhete2, Rute Mota1,2,3,4 & Luís Marques1
1Hovione FarmaCiencia S.A., R&D Inhalation & Advanced Drug Delivery, Estrada do Lumiar, Campus do Lumiar, Edifício R, 1649-038 Lisbon, Portugal
2Hovione FarmaCiencia SA, R&D Analytical Development, Estrada do Lumiar, Campus do Lumiar, Edifício R, 1649-038 Lisbon, Portugal
3CICECO – Aveiro Institute of Materials, Department of Chemistry, University of Aveiro, 3810-193 Aveiro, Portugal
4LAQV-REQUIMTE, Department of Chemistry, NOVA School of Science and Technology, NOVA University Lisbon, 2829-516 Caparica, Portugal
Summary
The present study successfully achieved the development and formulation of messenger RNA (mRNA) encapsulated in lipid nanoparticles (LNPs) for lung delivery through nebulization. tRNA-LNPs and mRNA-LNPs were produced via microfluidics and were dialyzed prior to nebulization. The formulation screening stage used transfer RNA (tRNA) as a model molecule and different formulations containing sodium chloride (NaCl), phosphate buffered saline (PBS), Poloxamer 188, Tween 80, sucrose, and/or arginine were evaluated. Among the tested conditions, the formulation with PBS and Tween 80 demonstrated the maintenance of tRNA-LNPs colloidal stability upon nebulization when a dilution ratio of 1:10 was applied. The previous formulation was used for mRNA-LNPs production and it was reported no loss of mRNA integrity throughout the product’s lifecycle, effectively enduring mechanical stress during production, dialysis, and nebulization. The nebulized aerosol droplets, generated using a vibrating mesh nebulizer, displayed appropriate aerodynamic properties, with particle sizes ranging from 1 to 5 µm, thus ensuring effective lung deposition. Moreover, the nebulization of the formulation resulted in a Fine Particle Fraction of Emitted Dose (FPFED) of 51% and an Emitted Dose (ED) of 92% of the encapsulated mRNA.