Copolymers of 2-(dimethylamino)ethyl methacrylate with ethoxytriethylene glycol methacrylate or N-vinyl-pyrrolidone as gene transfer agents.
Random copolymers of 2-(dimethylamino)ethyl methacrylate (DMAEMA) with ethoxytriethylene glycol methacrylate (triEGMA) or N-vinylpyrrolidone (NVP) of different molecular weights and compositions were synthesized, characterized and evaluated as polymeric transfectants in vitro. All synthesized copolymers (comonomer fraction up to 66 mol%) were able to bind to DNA, yielding polymer-plasmid complexes (polyplexes). However, the polymer-plasmid ratio at which small complexes (size 0.2-0.3 microm) were formed, increased with increasing mole fraction of the comonomer. zeta-Potential measurements revealed that the polymer-plasmid ratio where charge neutralization of DNA occurred, increased with increasing mole fraction of triEGMA. The cytotoxicity of the copolymers, either complexed with DNA or in the free form, decreased with increasing mole fraction of both comonomers (triEGMA and NVP). This reduction was even more than what could be expected based on the DMAEMA mole fraction in the copolymer. The copolymers with a molecular weight up to 170¿ omitted¿000 had the same transfection capability as a homopolymer of comparable molecular weight. However, higher molecular weight copolymers showed a reduced transfection capability compared to the homopolymer, which was ascribed to the reduced capability to condense the size of plasmid. Transfection efficiency of polyplexes composed of copolymers with a low triEGMA content increased with increasing molecular weight. Although the copolymers with 50 mol% triEGMA were also better transfectants than the homopolymer, the transfection efficiency did not increase further with increasing molecular weight. Interestingly, NVP-DMAEMA copolymers synthesized by polymerization to high conversion showed both excellent DNA binding and condensing characteristics (polyplex size <0.3 microm) and transfection capabilities. This is ascribed to a synergistic effect of DMAEMA-rich copolymers and NVP-rich copolymers present in this system on the complex formation with plasmid DNA.
van de Wetering
P
P
Department of Pharmaceutics, Utrecht Institute for Pharmaceutical Sciences (UIPS), Faculty of Pharmacy, Utrecht University, P.O. Box 80.082, 3508, TB Utrecht, The Netherlands.
Schuurmans-Nieuwenbroek
N M
NM
van Steenbergen
M J
MJ
Crommelin
D J
DJ
Hennink
W E
WE
D002851:Chromatography, High Pressure Liquid; D004247:DNA / Q000378:metabolism; D018736:DNA Adducts; D004337:Drug Carriers / Q000737:chemistry*; D018014:Gene Transfer Techniques; D008689:Methacrylates / Q000737:chemistry*; D010957:Plasmids / Q000378:metabolism; D011108:Polymers / Q000138:chemical synthesis / Q000737:chemistry*; D011760:Pyrrolidinones / Q000737:chemistry*; D014162:Transfection
D018736:DNA Adducts; D004337:Drug Carriers; D008689:Methacrylates; D011108:Polymers; D011760:Pyrrolidinones; C005044:hydroxyethyl methacrylate; D004247:DNA; C049840:2-(dimethylamino)ethyl methacrylate