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A kind of amphipathic comb-shaped sma-pei graft copolymer used as non-viral gene carrier and its preparation method and application

A graft copolymer and amphiphilic technology are applied to the amphiphilic comb-shaped SMA-PEI graft copolymer used as a non-viral gene carrier and the fields of preparation and use thereof, and the materials are cheap and easy to obtain and the preparation method is simple. , the effect of mild reaction conditions

Inactive Publication Date: 2016-02-10
SHANGHAI INST OF MATERIA MEDICA CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, SMA has been used in chemical preparations and polymeric prodrugs to improve drug solubility and stability, prolong plasma half-life, improve drug pharmacokinetics and pharmacodynamics parameters, and show high potential for clinical application. Reports of SMA used in gene delivery

Method used

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  • A kind of amphipathic comb-shaped sma-pei graft copolymer used as non-viral gene carrier and its preparation method and application
  • A kind of amphipathic comb-shaped sma-pei graft copolymer used as non-viral gene carrier and its preparation method and application
  • A kind of amphipathic comb-shaped sma-pei graft copolymer used as non-viral gene carrier and its preparation method and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0050] Example 1 Polystyrene maleic anhydride (SMA, molecular weight of 5500Da, molar ratio of styrene unit to maleic anhydride unit of 1:1)-polyethyleneimine (molecular weight of 800Da, PEI800) graft copolymer non-viral Synthesis of gene vector

[0051] Weigh 792 mg of PEI800 and dissolve it in 20 mL of dimethyl sulfoxide. Under stirring conditions, slowly add the SMA dimethyl sulfoxide solution (100 mg, 20 mL) dropwise at a rate of 1 to 2 drops per second. When SMA was added dropwise, the reaction mixture was stirred at room temperature for 12 hours. After the reaction, the reaction solution was transferred to a dialysis bag and dialyzed with water for 5 days to remove dimethyl sulfoxide and unreacted PEI. Then, most of the water is removed under reduced pressure and dried in vacuum to obtain the amphiphilic comb-shaped polystyrene maleic anhydride-polyethyleneimine graft copolymer (SMA-PEI800).

Embodiment 2

[0052] Example 2 Confirmation of the structure of SMA-PEI800

[0053] 1) 1H-NMR

[0054] Dissolve 8 mg of SMA-PEI800 prepared in Example 1 in 500 μL of a mixed solution of heavy water and deuterated methanol (the volume ratio of heavy water to deuterated methanol is 1:1), measure its 1H-NMR spectrum at 400 MHz, and perform Resolve. The result is Figure 1A As shown, the 1H-NMR spectrum analysis result is consistent with its structure.

[0055] 2)FT-IR

[0056] The SMA-PEI800 powder prepared in Example 1 and kBr were pressed into tablets, and the infrared spectrum (FT-IR) was measured by a Fourier transform infrared spectrometer, and analyzed. The result is Figure 1B As shown, the FT-IR spectrum analysis result is consistent with its structure.

Embodiment 3

[0057] Example 3 Determination of molecular weight of SMA-PEI800

[0058] Dissolve dextran with a molecular weight of 4400-401000Da in an aqueous solution containing 0.2M acetic acid / sodium acetate and 0.1% sodium azide to prepare a 1mg / mL solution as a standard product containing 0.2M acetic acid / sodium acetate and 0.1% sodium azide. % Sodium azide aqueous solution is the mobile phase, and a standard curve is established on a gel permeation chromatograph with a differential refractive index detector. Dissolve the SMA-PEI800 prepared in Example 1 in the mobile phase solution to prepare a 2mg / mL solution. Use an aqueous solution containing 0.2M acetic acid / sodium acetate and 0.1% sodium azide as the mobile phase. The molecular weight is measured by a refractive index detector on the instrument. The results are shown in Table 1 below:

[0059] Table 1: The weight average molecular weight (Mw) and dispersion (Mw / Mn) of SMA-PEI800 measured in Example 3

[0060]

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Abstract

The invention discloses an amphiphilic comb SMA (Styrene Maleic Anhydride)-PEI (Polyethylenimine) graft copolymer which is used as a non-viral gene vector as well as a preparation method and a use thereof. The amphiphilic comb styrene maleic anhydride-polyethylenimine graft copolymer is formed by carrying out aminolysis onto acid anhydride of the styrene maleic anhydride with primary amine relative molecular weight of 2KDa-1000KDa of low-molecular weight polyethylenimine with molecular weight of 0.2 KDa-10KDa. The mphiphilic comb styrene maleic anhydride-polyethylenimine graft copolymer can be used for conveying nucleic acid molecules into a cell and capable of enabling the nucleic acid molecules to express gene coding products or express silent target genes. The non-viral gene vector of the amphiphilic comb copolymer has the characteristics of being gentle in reaction condition, cheap and easily available in material, simple in synthesizing and post-processing method, high in recovery rate, simple in compound preparation method, high in transfection efficiency of carried nucleic acid, low in biological toxicity and the like.

Description

Technical field [0001] The invention belongs to the field of biotechnology, and specifically relates to an amphiphilic comb-shaped polystyrene maleic anhydride-polyethyleneimine (SMA-PEI) graft copolymer used as a non-viral gene carrier and a preparation method thereof And use, the polystyrene maleic anhydride-polyethyleneimine graft copolymer can deliver nucleic acid molecules into cells, and make them express gene coding products or silence target gene expression in the cells. Background technique [0002] Gene therapy is to transport the target gene into specific tissue cells through a suitable carrier and make it express therapeutic protein or replace or repair defective genes in the cell, so as to achieve the purpose of treating diseases. It has become one of the most active fields of biotechnology. One. After entering body fluids, naked nucleic acid molecules will be rapidly degraded under the action of ribozymes in the serum, making it difficult to stably exist in body fl...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C08F257/02C08F267/04C08F212/08C08F222/08C08F226/02C12N15/87C08L51/00C08L99/00
Inventor 李亚平段晓品肖计生顾王文张志文
Owner SHANGHAI INST OF MATERIA MEDICA CHINESE ACAD OF SCI
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