A combination of poloxamers increases gene expression of plasmid DNA in skeletal muscle.
Intramuscular administration of plasmid DNA is a promising strategy to express therapeutic genes, however, it is limited by a relatively low level of gene expression. We report here that a non-ionic carrier, SP1017, composed of two amphiphilic block copolymers, pluronics L61 and F127, also known as poloxamers, significantly increases intramuscular expression of plasmid DNA. Two reporter genes, luciferase and beta-galactosidase, and one therapeutic gene, erythropoietin, were injected intramuscularly with and without SP1017 into C57Bl/6 and Balb/C mice and Sprague-Dawley rats. SP1017 increased gene expression by about 10-fold and maintained higher gene expression compared with naked DNA. Comparison of SP1017 with polyvinyl pyrrolidone (PVP) showed that SP1017 exhibited a significantly higher efficacy and its optimal dose was 500-fold lower. Experiments with beta-galactosidase using X-gal staining suggested that SP1017 considerably increased plasmid DNA diffusion through the tissue. SP1017 also improved expression of the erythropoietin gene leading to an increase in its systemic level and hematocrits. Previous toxicity studies have suggested that SP1017 has over a 1000-fold safety margin. Poloxamers used in SP1017 are listed in the US Pharmacopeia as inactive excipients and are widely used in a variety of clinical applications. We believe that the described system constitutes a simple and efficient gene transfer method to achieve local or systemic production of therapeutic proteins.
Lemieux
P
P
Supratek Pharma Inc., Laval, Quebec, Canada.
Guérin
N
N
Paradis
G
G
Proulx
R
R
Chistyakova
L
L
Kabanov
A
A
Alakhov
V
V
D000818:Animals; D004337:Drug Carriers; D004921:Erythropoietin / Q000235:genetics; D005260:Female; D015870:Gene Expression; D018014:Gene Transfer Techniques; D015316:Genetic Therapy / Q000379:methods*; D006400:Hematocrit; D007273:Injections, Intramuscular; D008156:Luciferases / Q000235:genetics; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D018482:Muscle, Skeletal / Q000378:metabolism*; D010957:Plasmids / Q000008:administration & dosage*; D020442:Poloxamer / Q000008:administration & dosage*; D051381:Rats; D017207:Rats, Sprague-Dawley; D018709:Statistics, Nonparametric; D001616:beta-Galactosidase / Q000235:genetics
D004337:Drug Carriers; C488695:SP1017; D020442:Poloxamer; D004921:Erythropoietin; D008156:Luciferases; D001616:beta-Galactosidase