New Insight into the Role of Polyethylene Glycol Acting as Protein Release Modifier in Lipidic Implants
It has recently been shown that the addition of polyethylene glycol 6000 (PEG) to lipidic implants fundamentally affects the resulting protein release kinetics and moreover, the underlying mass transport mechanisms (Herrmann, Winter, Mohl, F. Siepmann, & J. Siepmann, J. Control. Release, 2007). However, it is yet unclear in which way PEG acts. It was the aim of this study to elucidate the effect of PEG in a mechanistic manner.
Materials and Methods
rh-interferon α-2a (IFN-α)-loaded, tristearin-based implants containing various amounts of PEG were prepared by compression. Protein and PEG release was monitored in phosphate buffer pH 4.0 and pH 7.4. IFN-α solubility and stability were assessed by reverse phase and size exclusion HPLC, SDS PAGE, fluorescence and FTIR.
Importantly, in presence of PEG IFN-α was drastically precipitated at pH 7.4. In contrast, at pH 4.0 up to a PEG concentration of 20% no precipitation occurred. These fundamental effects of PEG on protein solubility were reflected in the release kinetics of IFN-α from the tristearin implants: At pH 7.4 the protein release rates remained nearly constant over prolonged periods of time, whereas at pH 4.0 high initial bursts and continuously decreasing release rates were observed. Interestingly, it could be shown that IFN-α release was governed by pure diffusion at pH 4.0, irrespective of the PEG content of the matrices. In contrast, at pH 7.4 both—the limited solubility of the protein as well as diffusion through tortuous liquid-filled pores—are dominating.
For the first time it is shown that the release of pharmaceutical proteins can be controlled by an in-situ precipitation within inert matrices.
Key wordslipid polyethylene glycol protein release mechanism solubility
This study was financially supported by the Centre de Coopération Franco-Bavarois. We further express our grateful thanks to Roche Diagnostics (Penzberg, Germany) for the supply of interferon α-2a and Sasol (Witten, Germany) for the donation of the lipids.
- 1.S. Herrmann, G. Winter, S. Mohl, F. Siepmann, and J. Siepmann. Mechanisms controlling protein release from lipidic implants: effects of PEG addition. J. Control. Release (2007). http://www.sciencedirect.com/science/journal/01683659.
- 11.S. P. Schwendeman, M. Cardamone, A. Klibanov, R. Langer, and M. R. Brandon. Stability of proteins and their delivery from biodegradable polymer microspheres. Drugs Pharm. Sci. 77:1–49 (1996).Google Scholar
- 13.A. Maschke, A. Lucke, W. Vogelhuber, C. Fischbach, T. Appel, T. Blunk, and A. Goepferich. Lipids: an alternative material for protein and peptide release. In A.C.S. Symp. Ser. 79 (Carrier-Based Drug Delivery), 2004, pp. 176–196.Google Scholar
- 23.I. J. Castellanos, R. Crespo, and K. Griebenow. Poly(ethylene glycol) as stabilizer and emulsifying agent: a novel stabilization approach preventing aggregation and inactivation of proteins upon encapsulation in bioerodible polyester microspheres. J. Control. Release 88:135–145 (2003).PubMedCrossRefGoogle Scholar
- 24.S. Mohl. The development of a sustained and controlled release device for pharmaceutical proteins based on lipid implants. PhD thesis, LMU Munich, Munich (2004). Document available from http://edoc.ub.uni-muenchen.de/.
- 25.J. M. Vergnaud. Controlled Drug Release of Oral Dosage Forms. Ellis Horwood Limited, Chichester (1993).Google Scholar
- 26.J. M. Harris. Poly(ethylene glycol) Chemistry: Biotechnical and Biomedical Applications. Plenum, New York (1992).Google Scholar
- 34.O. L. Johnson, W. Jaworowicz, J. L. Cleland, L. Bailey, M. Charnis, E. Duenas, C. Wu, D. Shepard, S. Magil, T. Last, A. J. S. Jones, and S. D. Putney. The stabilization and encapsulation of human growth hormone into biodegradable microspheres. Pharm. Res. 14:730–735 (1997).PubMedCrossRefGoogle Scholar
- 41.Dustrup. Interferon formulations. US Pat. Appl. Publ. 41 (2003).Google Scholar