Reducible and biodegradable comb type high polymer gene vector and preparation method of same

A comb-shaped polymer and gene carrier technology, applied in the field of comb-shaped polymer gene carrier and its preparation, can solve the problems of low yield, complicated process, poor polymer solubility and the like

Inactive Publication Date: 2011-09-07
WUHAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Similar studies have been discussed in our previous invention patent [Chinese patent application publication number: CN101812178A]. The main disadvantage of this type of PEI derivatives is that the degree of crosslinking is not easy to control, and the resulting polymer has poor solubility and contains The microgel must be removed with a column, the process is complicated and the yield is 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.]

Method used

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  • Reducible and biodegradable comb type high polymer gene vector and preparation method of same
  • Reducible and biodegradable comb type high polymer gene vector and preparation method of same
  • Reducible and biodegradable comb type high polymer gene vector and preparation method of same

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0061] Embodiment 1 (allyl carbamate ethyl) dithioethylamine ((propargyl carbamate) ethyldisulfide ethylamine, PPA-cyst) and (allyl carbamate ethyl) dithioethyl 1-carboxamide Synthesis of imidazole ((propargyl carbamate) ethyl disulfide ethyl 1-carbamide-imidazole, PPA-cyst-CI)

[0062] (Allyl carbamate ethyl) dithioethylamine ((propargyl carbamate) ethyl disulfideethylamine, PPA-cyst) synthesis: According to the literature [X.L.Jiang, M.C.Lok, W.E.Hennink, Degradable-Brushed pHEMA-pDMAEMA synthesized via ATRP and click chemistry for gene delivery, Bioconjugate Chem.18 (2007) 2077-2084] synthesize propargyl ester of carbonyl-imidazole (PPA-CI) by propargyl alcohol and 1,1'-carbonyldiimidazole. 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 ) wa...

Embodiment 2

[0064] Example 2 Mono-azide terminated poly(2-(dimethylamino)ethyl methacrylate), L-PDE-N 3 )Synthesis:

[0065] 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.] first synthesized the initiator 2-bromo-isobutyric acid - 3-Azidopropyl ester (BiBAP). 6.29 grams of monomer methacrylate-2-N, N-dimethylaminoethyl ester, 260 mg of ligand 1,1,4,7,7,-pentamethyldiethylenetriamine (PMDETA) and 250 mg of initiator BiBAP was dissolved in 15 mL of 1,2-dichlorobenzene, 210 mg of cuprous bromide was added under nitrogen, and then the oxygen in the system was removed through three vacuum-nitrogen cycles, and the mixture was heated in an oil bath at 50°C for reaction 2 Hour. Add 200 ml of n-hexane to stop the reaction and precipitate the product, then repeat the precipitation twice for purification, and dry in a vacuum oven to obtain the product L-...

Embodiment 3

[0067] The synthesis of the polyethylenimine of embodiment 3 azide end group functionalization

[0068] Azide-end functionalized polyethyleneimine PEI 800 -(N 3 ) 1 Synthesis of (subscript number 800 means PEI weight average molecular weight is 800, and subscript number 1 means that the average number of azide end groups per polyethyleneimine is about 1): reference [2008-033s Chem Comm, 20071219, N2 , 190-192, Biodegradable microcapsules designed via 'click' chemistry, Bruno G. De Geest; Wim Van Camp, Filip E. Du Prez; Stefaan C. De Smedt; Jo Demeester; Wim E. Hennink] via azidopropanol and 1, 1'-Carbonyldiimidazole Synthesis of liquid azidopropyl ester of carbonylimidazole (3-azidopropyl ester of carbonylimidazole, AP-CI). 0.30 gram of propyl azidocarbonylimidazole (AP-CI) was dissolved in 60 milliliters of chloroform, and 1.23 grams of polyethyleneimine with a weight average molecular weight of 800 was dissolved in 20 milliliters of chloroform, and they were mixed and rea...

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Abstract

The invention discloses a reducible and biodegradable comb type high polymer gene vector and a preparation method of the same. The structure of the vector is as shown in the following formula, wherein CP is a cationic polymer with low molecular weight between 600 and 30000; F1 and F2 are groups capable of interaction; F1 or F2 contains a disulfide bond; M is a main chain unit structure of the polymer; n equals to 10-500; m is smaller than n; m equals to 0-400; l equals to 0-800; and l, m and n are integers. In the invention, the comb type high polymer gene vector has small cytotoxicity, gene DNA can be well bound and compounded under physical conditions so as to enter cells; and the carried genes can be released as the reducing environment in the cells can rupture quickly. The result showthat the reducible and biodegradable comb type high polymer gene vector has higher transfection efficiency and lower cytotoxicity compared with 25kDa PEI (polyether imide); moreover, the preparation method is simple, and the structure is easy to control.

Description

technical field [0001] The invention relates to a reductively degradable comb-shaped polymer gene carrier and a preparation method thereof, belonging to the field of gene therapy. Background technique [0002] The completion of the human genome project draft has enabled people to better understand the relationship between genes and diseases. As of 2008, there have been more than 1,400 clinical cases, using genes to correct genetic defects or treat acquired diseases (http: / / www.wiley.co.uk / genmed / clinical / ). The field of genetics has identified many genes with the potential to treat and prevent diseases, but the progress of gene delivery systems is limited, which is the bottleneck of gene therapy at present. Gene delivery vectors mainly include viral vectors and non-viral vectors. Because of the high transfection efficiency, viral vectors were first applied in the clinical research of gene therapy; however, their application and promotion were limited due to safety hazards ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/87C08G81/02
Inventor 蒋序林刘佳张光彦杨奇志卓仁禧
Owner WUHAN UNIV
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