A biodegradable gene transfer vector, poly(ethylenimine)-grafted-poly[(aspartic acid)-co-lysine] has been developed by thermal polycondensation of aspartic acid and lysine, and branch poly(ethylenimine) (Mw less than 600) was grafted to the backbone. The polymer was characterized by ^1H NMR. It appeared lower cytotoxity compared to poly(ethylenimine) (25KDa),which was quantified by MTT assay. Electrophoresis indicated that the polymer could retardate DNA at N/P ratio 1.2-1.8 (w/w). Transfection efficiency of the complexes was studied in NT2 cell lines. It was 1.5 fold higher than molecular weight PEI (Mw=25KDa). 相似文献
A biocompatible complex has been prepared as gene carrier via electrostatic interaction, which is composed of a polycation, that is, poly[(dimethylamino)ethyl methacrylate] end-capped with cholesterol moiety (Chol-PDMAEMA30), along with a polyanion named poly(aspartic acid)-grafted-poly(ethylene glycol) (PASP-g-PEG). The complexes have less cytotoxicity compared to the case of alone Chol-PDMAEMA30 or branched polyethylenimine (PEI) system. In the present study, biocompatible complexes have been prepared as gene carrier via electrostatic interaction, which is composed of a polycation, that is, poly[(dimethylamino)ethyl methacrylate] end-capped with cholesterol moiety (Chol-PDMAEMA30), along with a polyanion named poly(aspartic acid)-grafted-poly(ethylene glycol) (PASP-g-PEG). We first synthesized polysuccinimide (PSI) via condensation polymerization of aspartic acid, and then used PEG-NH2 to react with the partial pentacyclic rings of PSI to yield a kind of graft copolymer polysuccinimide-grafted-poly(ethylene glycol) (PSI-g-PEG). After hydrolysis of the residual succinimide units, a new biodegradable and biocompatible graft copolymer PASP-g-PEG was prepared successfully. Chol-PDMAEMA30 was synthesized via oxyanion-initiated polymerization, as reported in our previous literature. We investigated the interactions between every pair among calf thymus DNA, Chol-PDMAEMA30, and PASP-g-PEG by agarose gel retardation assay. The results indicate that the prepared complexes could completely bind DNA and may become more stable during systemic circulation. The complexes have less cytotoxicity compared to the case of alone Chol-PDMAEMA30 or branched polyethylenimine (PEI) system. Furthermore, the physicochemical properties of the complexes were also investigated by zeta potential, transmission electron microscopy (TEM) and dynamic light scattering (DLS) measurements. These biodegradable and biocompatible polymeric carriers have potential applications in gene delivery. 相似文献
Integrin-mediated delivery of genes is evaluated using a synthetic vector formed by self-assembly of DNA with an oligolysine- peptide sequence containing RGD (referred to as K16-RGD). The RGD peptide binds plasmid DNA effectively and inhibits ethidium bromide/DNA fluorescence at N-to-P ratios of less than 1.0. At N:P ratio 1.0, peptide/DNA complexes formed show a mixture of normal DNA migration and retention at the origin when analysed by agarose electrophoresis. At N:P ratio of 1.2, the complexes have a slight positive surface charge (5 mV) and in the absence of serum they show 10-fold increase uptake into 293 cells, compared with control poly(
-lysine)/DNA vectors, together with a 100-fold increase in transfection. In the presence of serum, RGD-mediated uptake is decreased about 3-fold, but the targeted vectors achieve over 150 times greater transfection than poly(
-lysine)/DNA controls. Transfection could be inhibited by addition of competing RGD, and to a lesser extent RGE, peptides. The targeted vector is believed to achieve cell uptake and transfection by binding av integrins in the cell surface, and the approach could be employed to promote internalisation of vectors following their binding to other, high affinity, receptors, in a system analogous to adenovirus entry. 相似文献
Poly(lactic acid)(PLA)is one of the most important bio-plastics,and chemical modification of the already-polymerized poly(lactic acid)chains may enable optimization of its material properties and expand its application areas.In this study,we demonstrated that poly(lactic acid)can be readily dissolved in acrylic acid at room temperature,and acrylic acid can be graft-polymerized onto poly(lactic acid)chains in solution with the help of photoinitiator benzophenone under 254 nm ultraviolet(UV)irradiation.Similar photo-grafting polymerization of acrylic acid(PAA)has only been studied before in the surface modification of polymer films.The graft ratio could be controlled by various reaction parameters,including irradiation time,benzophenone content,and monomer/polymer ratios.This photo-grafting reaction resulted in high graft ratio(graft ratio PAA/PLA up to 180%)without formation of homopolymers of acrylic acid.When the PAA/PLA graft ratio was higher than 100%,the resulting PLA-g-PAA polymer was found dispersible in water.The pros and cons of the photo-grafting reaction were also discussed. 相似文献
Dendritic poly(amidoamine)s (PAMAM)s were introduced into the side chains of disulfide‐containing poly(amidoamine)s via repetitive Michael addition and amidation. The bioreducible poly(amidoamine)s grafted with dendritic polyamidoamines showed high buffer capacity, low cytotoxicity and strong DNA binding ability at low N/P ratio. They were able to condense DNA into small sized polycation/DNA complexes, which degraded and released the incorporated DNA under reductive conditions. In comparison to the original disulfide‐containing poly(amidoamine) with aminoethyl side chain, the grafting of the bioreducible poly(amidoamine) with dendrimer greatly improved the transfection efficiencies of 293T and HeLa cells with foreign DNA at various N/P ratios. The structure–gene delivery property relations of dendrimer‐grafted polycations will provide valuable insight into the design of highly efficient and less toxic polycationic gene carriers.
A novel amine‐functionalized polycarbonate was synthesized and its excellent gene transfection ability in vitro is demonstrated. In the framework of adapting the cationic polycarbonate for in vivo gene delivery applications, here the design and synthesis of biodegradable block copolymers of poly(ethylene glycol) (PEG) and amine‐functionalized polycarbonate with a well‐defined molecular architecture and molecular weight is achieved by metal‐free organocatalytic ring‐opening polymerization. Copolymers in triblock cationic polycarbonate‐block‐PEG‐block‐cationic polycarbonate and diblock PEG‐block‐cationic polycarbonate configurations, in comparison with a non‐PEGylated cationic polycarbonate control, are investigated for their influence on key aspects of gene delivery. Among the polymers with similar molecular weights and N content, the triblock copolymer exhibit more favorable physicochemical (i.e., DNA binding, size, zeta‐potential, and in vitro stability) and biological (i.e., cellular uptake and luciferase reporter gene expression) properties. Importantly, the various cationic polycarbonate/DNA complexes are biocompatible, inducing minimal cytotoxicities and hemolysis. These results suggest that the triblock copolymer is a more useful architecture in future cationic polymer designs for successful systemic therapeutic applications. 相似文献
A new poly(aminoester) (EPAE-FA) containing folic acid and amino groups in the backbone and side chain was synthesized. EPAE-FA self-assembled readily with the plasmid DNA (pCMV-βgal) in HEPES buffer and was characterized by dynamic light scattering, zeta potential, fluorescence images, and XTT cell viability assays. To evaluate the transfection effect of graft ratio of FA on the EPAE system, EPAE-FA polymers with two different graft ratios (EPAE-FA12k and EPAE-FA14k) were also prepared. This study found that all EPAE-FA polymers were able to bind plasmid DNA and yielded positively charged complexes with nano-sized particles ( < 200 nm). To assess the transfection efficiency mediated by EPAE and EPAE-FA polymers, we performed in vitro transfection activity assays using FR-negative (COS-7) and FR-positive (HeLa) cells. The EPAE-FA12k/DNA and EPAE-FA14k/DNA complexes were able to transfect HeLa cell in vitro with higher transfection efficiency than PEI25k/DNA at the similar weight ratio. These results demonstrated that the introduction of FA into EPAE system had a significant effect on transferring ability for FR-positive cells (HeLa). Examination of the cytotoxicity of PEI25k and EPAE-FA system revealed that EPAE-FA system had lower cytotoxicity. In this paper, EPAE-FA seemed to be a novel cationic poly(aminoester) for gene delivery and an interesting candidate for further study. 相似文献
Abstract Chemical modification of poly(oxyethylene)s (MW from 600 Da to 3000 Da), leading to the obtaining of poly(oxyethylene)s with (2-benzoxazolon-3-yl)acetyl endgroups, is described. The corresponding diesters were synthesized in high yields-70—84% and degree of esterification-82—92% under mild conditions in the presence of dicyclohexylcarbodiimide and 4-dimethylaminopyridine as catalyst. The diesters were characterized by 1H-NMR, 13C-NMR and IR spectra, and SEC analysis. The relatively narrow molecular weight distribution was retained in the process of chemical modification. The complex formation between polyacrylic or polymeth-acrylic acids and (2-benzoxazolon-3-yl)acetyl derivatives of poly -(oxyethylene)s was studied in dilute aqueous solutions by potentio-metric titration and viscometry. The presence of (2-benzoxazolon-3-yl)acetyl end groups attached to the poly(oxyethylene)s chains results in a significant decrease of the critical poly(oxyethylene) chain length necessary for the formation of stable polymer-polymer complexes. 相似文献
