Influences of copolymers (Copovidone, Eudragit RL PO and Kollicoat MAE 30 DP) on stability and bioactivity of spray-dried and freeze-dried lysozyme

Abstract

Protein stability is the most crucial factor in protein pharmaceutical preparations. Various techniques were applied for producing stable protein formulations such as spray-drying and freeze-drying. However, heating and freezing stresses are disadvantages for proteins using these methods, respectively. Accordingly, excipients have been used to preserve therapeutic effects of proteins during processing and for long period of time. Therefore, influences of Copovidone, Eudragit® RL-PO and Kollicoat® MAE-30 DP (as excipients) on stability and integrity of lysozyme (as a model protein) in spray-dried and freeze-dried forms were investigated. Protein formulations in both dried forms were prepared without and with the addition of mentioned excipients at different concentrations. Protein formulations were characterised for yield determination, morphology using scanning electron microscopic (SEM), thermal analysis by Differential Scanning Calorimetry (DSC), secondary structure stability using Fourier transform infrared (FT-IR) spectroscopy and biological activity. All protein formulations were subjected to a stability study as solid protein formulations for 3 weeks at 24 °C/76% relative humidity and aqueous protein samples were stored at 50°C for 30 minutes in a water bath. Results showed that Copovidone successfully preserved integrity and biological activity of lysozyme before and after storage in both spray-dried and freezedried forms with more advantage for using higher concentration of the same excipient.Smooth spheres of spray-dried lysozyme formulations with Copovidone were smaller than spray-dried lysozyme without and with Kollicoat® MAE-30 DP, which affected %yield produced. Copovidone has demonstrated valuable protection ability for lysozyme.

Progress in spray-drying of protein pharmaceuticals: Literature analysis of trends in formulation and process attributes

“…[101] Another study showed that poly(vinylpyrrolidone-co-vinyl acetate) (PVP-VA) was successful in stabilizing lysozyme and trypsin upon spray-drying. [50] 4.6.2. Lipids and lipophilic molecules With respect to lipid excipients, dipalmitoyl phosphatidylcholine (DPPC), an endogenous phospholipid to the lung, has been frequently applied to protein formulations, particularly, inhaled ones.…”

Effect of Spray-Drying and Electrospraying as Drying Techniques on Lysozyme Characterisation

“…Increasing the percentage yield can be achieved by introducing polymeric excipients with high glass transition temperature in addition to optimise the drying condition used during the spray-drying [37]. A largescale production is feasible in pharmaceutical industries for spray-drying formulations by using a large scalable spray-drier that would generate the highest possible yield [38]. This is also feasible for electrospraying by using, for example, a multi-tip emitter to improve the potential upscale electrospraying [39].…”