Release of PEGylated granulocyte-macrophage colony-stimulating factor from chitosan/glycerol films.
We have prepared a new formulation for mucosal delivery of GM-CSF or PEGylated GM-CSF based on a chitosan carrier plus added glycerol to control the rate of release of the protein. Thin dry films comprised of various weight ratios of chitosan to glycerol and containing either granulocyte-macrophage colony-stimulating factor (GM-CSF) or PEGylated GM-CSF, PEG-(GM-CSF), were prepared. The amount of GM-CSF or PEG-(GM-CSF) released from the chitosan/glycerol films was determined using size exclusion high performance liquid chromatography (HPLC-SEC). The amount of PEG-(GM-CSF) released from the films decreased with an increase in the amount of glycerol present in the film. In parallel with this, films with higher glycerol content exhibited a lower degree of equilibrium swelling when immersed in release media. pH measurements of the release media and analysis of the dried films by Fourier-transform infrared spectroscopy (FTIR) suggested that the amount of residual acetic acid in the dry films decreased as the glycerol content increased. This indicates that glycerol may act by displacing and releasing bound acetic acid from the chitosan molecules, resulting in chitosan--glycerol hydrogen bond formation as the film dries. Further, it was found that the release rate and the amount of PEG-(GM-CSF) released decreased with increasing molecular weight of the conjugated PEG. This effect was not observed with films containing physical mixtures of PEG and GM-CSF. The decrease in the fraction of PEG-(GM-CSF) released with increasing PEG molecular weight is believed to be due to the increased steric hindrance of the PEGylated protein molecule during its diffusion out of the swollen chitosan/glycerol film.
Brown
C D
CD
University of Washington, Department of Bioengineering, Box 352255, Seattle, WA 98195, USA.
Kreilgaard
L
L
Nakakura
M
M
Caram-Lelham
N
N
Pettit
D K
DK
Gombotz
W R
WR
Hoffman
A S
AS
D001672:Biocompatible Materials; D002021:Buffers; D002686:Chitin / Q000031:analogs & derivatives; D048271:Chitosan; D005990:Glycerol; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000008:administration & dosage* / Q000737:chemistry; D006863:Hydrogen-Ion Concentration; D008567:Membranes, Artificial; D014677:Pharmaceutical Vehicles; D011092:Polyethylene Glycols / Q000737:chemistry; D011994:Recombinant Proteins; D017550:Spectroscopy, Fourier Transform Infrared
D001672:Biocompatible Materials; D002021:Buffers; D008567:Membranes, Artificial; D014677:Pharmaceutical Vehicles; D011994:Recombinant Proteins; D002686:Chitin; D011092:Polyethylene Glycols; D016178:Granulocyte-Macrophage Colony-Stimulating Factor; D048271:Chitosan; D005990:Glycerol