Design and scalable manufacturing of mucus-penetrating liposomal systems for enhanced pulmonary delivery of ciclesonide and indacaterol maleate

According to the 2025 GINA Guidelines, the management of asthma remains fundamentally dependent on the combination of an inhaled corticosteroid (ICS) and a long-acting β2-agonist (LABA). Clinical practice is increasingly shifting toward molecules with a balanced hydrophilic/lipophilic profile, such as budesonide and formoterol.This balance facilitates rapid dissolution in the airway lining fluid and efficient penetration across the mucus barrier to access their molecular receptors. While the high lipophilicity of newer ICS and LABA molecules enables convenient once-daily dosing regimens, this property renders them more susceptible to mucociliary clearance and phagocytosis by alveolar macrophages, thereby compromising their ability to reach the target cells. To address these pulmonary delivery limitations, this study focuses on the formulation development and production scale-up of an advanced liposomal carrier system for a new ICS-LABA combination: ciclesonide (CIC) and indacaterol maleate (IND). Initially, we focused on the design of a liposome architecture that allows for mucus penetration, stable drug encapsulation, evasion of macrophage phagocytosis, and enhanced epithelial uptake. By meticulously optimizing the type and concentration of PEGylated lipids and phospholipids, an optimal formulation (DLPC/Cholesterol/DSPE-PEG) containing CIC and IND was identified. This formulation significantly improved the attenuation of airway hyperresponsiveness and inflammation in an in vivo acute asthma mouse model compared to non-liposomal drug formulation. Furthermore, the liposomal delivery of CIC and IND exhibited a substantial dose-sparing potential for both therapeutic agents. Building upon the proof-of-concept for this optimized formulation, the second objective aimed to facilitate its production at a pilot scale without using organic solvents. This scale-up effort required a comprehensive application of Design of Experiments principles to characterize multiple critical process parameters, such as dispersion volume, pressure, agitation speed, and depressurization rate, within a pilot-scale, GMP-like environment. The rigorous optimization process resulted in the generation of liposomes exhibiting a mean particle size of 152 ± 8 nm and a PdI of 0.36 ± 0.02, while maintaining high drug encapsulation efficiencies (>90%). Furthermore, the detailed characterization of the liposomes manufactured at the pilot scale confirmed that the chosen method did not induce statistically significant increases in lipid oxidation or hydrolysis, confirming the chemical stability and industrial suitability of the process. In conclusion, this study successfully met its primary goals. We first validated a highly effective liposomal platform that significantly enhances the anti-asthmatic efficacy of the CIC and IND combination in vivo. Second, we demonstrated the feasibility of transitioning this delivery system to a pilot manufacturing scale using the advanced, organic solvent-free PGSS process. These results provide a robust and highly viable platform for developing next-generation inhaled therapies for chronic respiratory diseases.