Formulation Optimization of Freeze-Dried Long-Circulating Liposomes and In-Line Monitoring of the Freeze-Drying Process Using an NIR Spectroscopy Tool

Abstract

The effect of lyoprotectant type and concentration on the stability of freeze-dried prednisolone sodium phosphate-loaded long-circulating liposomes was investigated. Trehalose at a 5:1 carbohydrate to lipid molar ratio proved to be superior in maintaining the structural integrity and the permeability properties of the liposome bilayers, assuring the desired characteristics of the final product: a cake with a porous structure and easy to reconstitute, a similar size to the liposomes before freeze-drying, a high percent of encapsulated drug, and a low residual moisture content. Further on, the study demonstrated the possibility of near-infrared spectroscopy to provide valuable insights for detecting critical changes in acyl chain packing of the liposome bilayer. By visualizing the spectra after principal component analysis, one can predict if any harm has occurred to liposome integrity during the process. Moreover, near-infrared spectroscopy enabled us to determine the end points of primary and secondary drying without disturbing the normal freeze-drying procedure, which allowed us to gain a better understanding of the process and to improve process efficiency by optimizing the primary and secondary drying time.

Keywords: encapsulation; freeze drying/lyophilization; liposomes; near-infrared spectroscopy; principal component analysis; process analytical technology (PAT).