Reduction sensitive polyethyleneimine derivative as well as preparation method and application thereof

A polyethylenimine and sensitive technology, which is applied in the field of reduction-sensitive polyethyleneimine derivatives and their preparation and application, can solve the problem of reducing the ability of polymer binding and protecting DNA, low yield, complicated process, etc. question

Inactive Publication Date: 2012-07-04
WUHAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Recently Hong Kong scholars reported that the degree of cross-linking of PEI derivatives synthesized by this method is not easy to control, the solubility of the obtained polymer is not good, and the contained microgel must be removed with a column, the process is complicated, and the yield is low[ R.Deng, Y.N.Yue, F.Jin, Y.C.Chen, H.F.Kung, M.C.M.Lin, C.Wu, Revisit the complexation of PEI and DNA-How to make low cytotoxic and highly efficient PEI gene transfection non-viral vectors with a controllable chain length and structure? J.Control.Release 140(2009)40-46.], they found that excessive cross-linking will greatly reduce the ability of the polymer to bind and protect DNA, and the gene transfection efficiency will be low, so controlling the appropriate polymer Structure is very important for polymer gene carriers

Method used

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  • Reduction sensitive polyethyleneimine derivative as well as preparation method and application thereof
  • Reduction sensitive polyethyleneimine derivative as well as preparation method and application thereof
  • Reduction sensitive polyethyleneimine derivative as well as preparation method and application thereof

Examples

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preparation example Construction

[0065] The above-mentioned reduction-sensitive micro-crosslinked modified polyethyleneimine derivatives of the present invention (using the simplified formula PEI-(ASSA-PEI) n indicated that PEI 1800 -(ASSA-PEI 1800 ) n , representing that the weight-average molecular weight that it uses PEI is 1800, also expresses with PEI-SS-C) preparation method, comprises following three steps:

[0066]

[0067]In step a) above, a disulfide bond-containing chain extender (crosslinking agent) can be prepared: compound 1 propargyl carbonylimidazole and cystamine dehydrochloride are added in a molar ratio of 1-4:1 Proton-free organic solvents such as chloroform, dichloromethane, acetone, tetrahydrofuran, DMF, stirred at 20-60 ° C for 2-48 hours, purified to obtain bis (allyl carbamate ethyl) disulfide ( bis[(propargyl carbamate) ethyl] disulfide, BPPA-Cyst, compound 2) (see Example 1), can also adjust the feed molar ratio of compound 1 and cystamine to be 0.5-1.5: 1 to prepare (allylami...

Embodiment 1

[0088] Example 1 Synthesis of two (allyl carbamate ethyl) disulfide (bis[(propargyl carbamate) ethyl] disulfide, BPPA-Cyst, 2):

[0089] According to the literature [X.L.Jiang, M.C.Lok, W.E.Hennink, Degradable-BrushedpHEMA-pDMAEMA synthesized via ATRP and click chemistry for gene delivery, Bioconjugate Chem.18(2007) 2077-2084] through propargyl alcohol and 1,1'-carbonyldiimidazole Synthesis of propargyl ester of carbonyl-imidazole (PPA-CI, 1). First desalt cystamine hydrochloride, weigh 4.40 g of dehydrochlorated cystamine and 6.93 g of propargyl carbonylimidazole in 50 ml of dichloromethane, stir and react at room temperature for 24 hours, remove the solvent, add 100 ml of phosphoric acid Sodium dihydrogen solution (pH 4.0) was dissolved, and then extracted 3 times with ether to obtain BPPA-Cyst with a yield of 80%. 1 H-NMR in CDCl 3 : δ(ppm) 2.48(s, 2H, CH≡C), 2.82(t, 4H, S-CH 2 -C), 3.51 (app.quartet, 4H, NH-CH 2 -C), 4.69(s, 4H, O-CH 2 -C≡C), 5.31 (s, 2H, NH-C=O). Inf...

Embodiment 2

[0090] Embodiment 2 (allylcarbamate ethyl) dithioethyl 1-carboxamide imidazole ((propargylcarbamate) ethyl disulfide ethyl 1-carbamide-imidazole, PPA-Cyst-CI, 7) is first synthesized (ene Propyl carbamate ethyl) dithioethylamine ((propargyl carbamate) ethyl disulfideethylamine, PPA-Cyst, 6): Propargyl carbamate carbonylimidazole (PPA-CI, 1) was synthesized according to the steps in Example 1. First desalt cystamine hydrochloride, weigh 4.40 grams of cystamine and 3.47 grams of propargyl carbonylimidazole in 50 milliliters of chloroform, stir and react at room temperature for 24 hours, remove the solvent, add 80 milliliters of sodium dihydrogen phosphate solution (pH 4.0 ) was dissolved, extracted 3 times with diethyl ether, adjusted the pH of the obtained lower aqueous phase to 9.0, then extracted 3 times with 40 ml of ethyl acetate, combined the organic phases, washed once with 40 ml of pH=9 sodium phosphate solution, and obtained The organic phase was dried with anhydrous ma...

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Abstract

The invention discloses a reduction sensitive polyethyleneimine derivative containing modified disulfide bonds, having a chemical formula: PEI-(ASSA-PEI)n-(ASSB-PEI)m, wherein PEI is polyethyleneimine, the component of A is C10H14O4N4, the component of B is C3H5ON, n=0-30; m=0-30; and n and m are not equal to zero simultaneously. The weight-average molecular weight of the reduction sensitive polyethyleneimine derivative is 5,000-80,000. Through simple and special modification and click chemistry, the polyethyleneimine is connected into modified polyethyleneimine with high molecular weight through reductive and degradable disulfide bonds. The invention has the advantages of simple method, easy structure control, less amino loss for reaction and higher amino utilization ratio. The obtained product has the characteristics of bio-reduction and degradation, has less cytotoxicity, can well bind and compound gene DNA under a physiology condition, enters cells to release carried gene because the reduction environment in the cells can be broken more rapidly, and has higher gene expression transfection efficiency. Results show that the modified polyethyleneimine derivative has higher transfection efficiency and lower cytotoxicity than 25kDa PEI.

Description

technical field [0001] The invention relates to a reduction-sensitive cationic polymer, its preparation method and application. Specifically, it relates to a degradable derivative of polyethyleneimine, its preparation method and its application as a gene carrier. Background technique [0002] Gene therapy refers to inserting genes that can produce specific proteins into patient cells to correct genetic defects or treat acquired diseases, including cancer, AIDS, diabetes, etc. It is a new treatment method that has emerged in the past two decades and has a good development prospect . Gene carrier is an important part of gene medicine, and it is also the key and bottleneck of current gene therapy. Gene vectors mainly include viral vectors and non-viral vectors. Among them, viral vectors were firstly used in clinical research of gene therapy because of their high transfection efficiency; however, the potential safety hazards of viral vectors and the complicated and expensive ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C08G73/04A61K48/00A61K47/48A61K47/58
Inventor 蒋序林刘佳徐丽卓仁禧
Owner WUHAN UNIV
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