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  • Title: Compare and contrast the effects of surfactants (PluronicF-127 and CremophorEL) and sugars (β-cyclodextrin and inulin) on properties of spray dried and crystallised lysozyme.
    Author: Haj-Ahmad RR, Elkordy AA, Chaw CS, Moore A.
    Journal: Eur J Pharm Sci; 2013 Jul 16; 49(4):519-34. PubMed ID: 23712374.
    Abstract:
    The stabilisation of proteins using different excipients in dried forms for possible therapeutic use is extensively studied. However, the effects of excipients on proteins in crystallised forms are sparsely documented. Therefore, the influences of PluronicF-127 and CremophorEL (as surfactants) and β-cyclodextrin and inulin (as sugars) on stability and biological activity of lysozyme, a model protein, in spray dried and crystallised forms were investigated. Spray dried and crystallised lysozyme were prepared in absence and presence of the mentioned excipients in a concentration of 0.05% w/v. The protein formulations were characterised in both solution state (using biological assay, particle size analysis and protein concentration determination) and solid state (employing yield determination, scanning electron microscopic (SEM) examination, Fourier transform infrared (FT-IR) spectroscopy for secondary structure analysis and Differential Scanning Calorimetry (DSC) for thermal study). Also, protein samples were assayed for their biological activities after exposing to storage stability study for 20 weeks in solid states at 24 °C/76% relative humidity (RH) and in aqueous states at 24 °C. The results showed that lysozyme crystals with CremophorEL, PluronicF-127, β-cyclodextrin and inulin maintained protein thermal stability (as indicated by DSC) to greater extent compared with spray dried protein formulations. Also, PluronicF-127 was competent to recover 100% lysozyme from crystallisation protein solutions (as confirmed by yield determination); this surfactant was able to prevent aggregate formation within spray dried lysozyme (as demonstrated by particle size analysis). The presence of PluronicF-127, β-cyclodextrin and inulin preserved the protein biological activity in freshly prepared spray dried and crystallised samples. PluronicF-127 was competent to protect lysozyme in both spray dried and crystallised forms after storage. PluronicF-127 has proved to be a promising protectant of proteins. The improved stability of the spray dried and crystallised protein containing PluronicF-127 shows promise for delivery of proteins via inhalation (in a spray dried form which has particle size range suitable for inhalation as revealed by particle size analysis and SEM) and injectable routes (in spray dried and crystallised forms). The way excipients react with proteins is different in the case of spray drying and crystallisation techniques, hence the choice of the additives and the processing techniques play a great role in controlling protein properties, activity and stability as shown in this study.
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