Abstract
Inhalation therapy has advantages for the treatment of multidrug resistant bacterial lung infections with high drug concentrations at the infection sites in the airways and reduced systemic exposure. We have developed liposomal formulations for pulmonary delivery of synergistic ciprofloxacin (Cipro) and colistin (Col) as the potential candidate for treatment of lung infections caused by multidrug resistant Gram-negative bacteria. This study aims to: (1) further optimize the powder formulation by adding drying stabilizers (polyvinyl pyrrolidone or poloxamer) to protect the liposomes during spray-freeze-drying; (2) evaluate the transport and cellular uptake of drugs in a human lung epithelial Calu-3 cell model. The liposomal powder formulations were produced using the ultrasonic spray-freeze-drying technique. The optimal formulation (F5) used mannitol (8% w/v) and sucrose (2% w/v) as the internal lyoprotectants. Adding external lyoprotectants/aerosolization enhancers (i.e. 8% w/v mannitol, 2% w/v sucrose and 1%, w/w PVP 10) produced the superior rehydrated EE values of ciprofloxacin and colistin (50.2 ± 0.9% for Cipro and 37.8 ± 1.2% for Col) as well as satisfactory aerosol performance (FPF: 34.2 ± 0.8% for Cipro and 33.6 ± 0.9% for Col). The cytotoxicity study indicated that F5 with the colistin concentration at 50 μg/mL and ciprofloxacin at 200 μg/mL was not cytotoxic to human lung epithelial Calu-3 cells. The intracellular uptake of ciprofloxacin was concentration-dependent in Calu-3 cells and the uptake of A-B was more than that of B-A for all samples (p < 0.05). This study demonstrates that co-delivery of ciprofloxacin and colistin in a single liposome can lower the transport capability of both drugs across the Calu-3 cell monolayer and their accumulation in the cells. These findings indicate that co-loaded liposomal powder of ciprofloxacin and colistin is a promising potential treatment for respiratory infections caused by multidrug resistant Gram-negative bacteria.