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Novel biodegradable poly(disulfide amine)s for gene delivery with high efficiency and low cytotoxicity
- Source :
- Bioconjugate chemistry. 19(3)
- Publication Year :
- 2008
-
Abstract
- Novel biodegradable poly(disulfide amine)s with defined structure, high transfection efficiency, and low cytotoxicity were designed and synthesized as nonviral gene delivery carriers. Michael addition between N, N'-cystaminebisacrylamide (CBA) and three N-Boc protected diamines ( N-Boc-1,2-diaminoethane, N-Boc-1,4-diaminobutane, and N-Boc-1,6-diaminohexane) followed by N-Boc deprotection under acidic condition resulted in final cationic polymers with disulfide bonds, tertiary amine groups in main chains, and pendant primary amine groups in side chains. Polymer structures were confirmed by 1H NMR, and their molecular weights were in the range 3.3-4.7 kDa with narrow polydispersity (1.12-1.17) as determined by size exclusion chromatography (SEC). Acid-base titration assay showed that the poly(disulfide amine)s possessed superior buffering capacity to branched PEI 25 kDa in the pH range 7.4-5.1, which may facilitate the escape of DNA from the endosomal compartment. Gel retardation assay demonstrated that significant polyplex dissociation was observed in the presence of 5.0 mM DTT within 1 h, suggesting rapid DNA release in the reduction condition such as cytoplasm due to the cleavage of disulfide bonds. Genetic transfections mediated by these poly(disulfide amine)s were side-chain spacer length dependent. The poly(disulfide amine) with a hexaethylene spacer, poly(CBA-DAH), had comparable transfection efficiency to bPEI 25 kDa in the tested cell lines, i.e., 293T cells, Hela cells, and NIH3T3 cells. This same poly(disulfide amine) mediated 7-fold higher luciferase expression than bPEI 25 kDa in C2C12 cells (mouse myoblast cell line), a cell line difficult to transfect with many cationic polymers. Furthermore, MTT assay indicated that all three poly(disulfide amine)s/pDNA polyplexes were significantly less toxic than bPEI/pDNA complexes.
- Subjects :
- Magnetic Resonance Spectroscopy
Tertiary amine
Light
Cell Survival
Size-exclusion chromatography
Biomedical Engineering
Pharmaceutical Science
Tetrazolium Salts
Bioengineering
Gene delivery
Buffers
Transfection
Article
Mice
Drug Delivery Systems
Genes, Reporter
Cell Line, Tumor
Polymer chemistry
Side chain
Polyamines
Organic chemistry
Animals
Humans
Scattering, Radiation
Particle Size
Luciferases
Pharmacology
Chemistry
Organic Chemistry
Cationic polymerization
Gene Transfer Techniques
Nuclear magnetic resonance spectroscopy
3T3 Cells
DNA
Thiazoles
Michael reaction
Chromatography, Gel
Amine gas treating
Indicators and Reagents
Biotechnology
HeLa Cells
Subjects
Details
- ISSN :
- 10431802
- Volume :
- 19
- Issue :
- 3
- Database :
- OpenAIRE
- Journal :
- Bioconjugate chemistry
- Accession number :
- edsair.doi.dedup.....2a5d7a1dc6298bb40e01008b8afd427a