RRC ID |
41668
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Author |
Wang J, Zhang PC, Lu HF, Ma N, Wang S, Mao HQ, Leong KW.
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Title |
New polyphosphoramidate with a spermidine side chain as a gene carrier.
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Journal |
J Control Release
|
Abstract |
A new cationic polymer (PPA-SP), polyphosphoramidate bearing spermidine side chain, was prepared as a non-viral vector for gene delivery. PPA-SP was synthesized from poly(1,2-propylene H-phosphonate) by the Atherton-Todd reaction. The weight average molecular weight of PPA-SP was 3.44x10(4) with a number average degree of polymerization of 90, as determined by GPC/LS/RI method. The average net positive charge per polymer chain was 102. PPA-SP was able to condense plasmid DNA efficiently and formed complexes at an N/P ratio (free amino groups in polymer to phosphate groups in DNA) of 2 and above, as determined by agarose gel electrophoresis. This new gene carrier offered significant protection to DNA against nuclease degradation at N/P ratios above 2, and showed lower cytotoxicity than PLL and PEI in cell culture. The LD(50) of PPA-SP was 85 microg/ml in COS-7 cells, in contrast to 20 and 42 microg/ml for PLL and PEI, respectively. The complexes prepared in saline at N/P ratios of 5 approximately 10 had an average size of 250 nm and zeta-potential of 26 mV. PPA-SP mediated efficient gene transfection in a number of cell lines, and the transfection protocol was optimized in HEK293 cells using a luciferase plasmid as a marker gene. Gene expression mediated by PPA-SP was greatly enhanced when chloroquine was used in conjunction at a concentration of 100 microM. Under the optimized condition, PPA-SP/DNA complexes yield a luciferase expression level closed to PEI/DNA complexes or Transfast mediated transfection. In a non-invasive CNS gene delivery model, PPA-SP/DNA complexes yielded comparable bcl-2 expression as PEI/DNA complexes in mouse brain stem following injection of the complexes in the tongue.
|
Volume |
83(1)
|
Pages |
157-68
|
Published |
2002-9-18
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DOI |
10.1016/s0168-3659(02)00180-3
|
PII |
S0168365902001803
|
PMID |
12220847
|
MeSH |
Animals
COS Cells / drug effects
COS Cells / metabolism
Chlorocebus aethiops
Drug Carriers / administration & dosage*
Drug Carriers / chemistry
Drug Carriers / pharmacokinetics
Female
Genetic Therapy / methods*
Humans
Mice
Mice, Inbred BALB C
Polymers / administration & dosage*
Polymers / chemistry
Polymers / pharmacokinetics
Spermidine / administration & dosage*
Spermidine / chemistry
Spermidine / pharmacokinetics
Tumor Cells, Cultured / drug effects
Tumor Cells, Cultured / metabolism
|
IF |
7.727
|
Times Cited |
100
|
WOS Category
|
PHARMACOLOGY & PHARMACY
CHEMISTRY, MULTIDISCIPLINARY
|
Resource |
Human and Animal Cells |
HuH-7(RCB1366) |