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Gene carrier system and preparation method thereof

A gene carrier and gene technology, applied in other methods of inserting foreign genetic materials, sulfonamide preparation, organic chemistry, etc., can solve the problems of low transfection efficiency, low efficiency of carrier system and target cell tissue combination, and achieve protection transport effect

Active Publication Date: 2011-12-14
CHANGZHOU INST OF ENERGY STORAGE MATERIALS &DEVICES
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Although the gene carrier system containing the targeting shielding system can avoid the non-specific adsorption of the gene carrier, it can also successfully reach the target tissue cells, but the negative charge on the surface of the shielding system and the negative charge on the cell surface will generate repulsion, affecting the carrier. The binding efficiency of the system to the target cell tissue is still low, that is, its in vivo transfection efficiency is low

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  • Gene carrier system and preparation method thereof

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preparation example Construction

[0057] The present invention also provides a method for preparing a gene carrier system, comprising the following steps:

[0058] The aqueous solution of the gene carrier and the aqueous solution of the gene material are mixed and incubated to obtain a complex, the mass ratio of the gene carrier to the gene material is 0.5:1 to 50:1, and the gene carrier is polyethyleneimine, dendrimer Amide or polylysine;

[0059] Adding an aqueous solution of a pH-sensitive material to the complex to obtain a gene carrier system, the mass ratio of the pH-sensitive material to the gene carrier is 1:1 to 80:1, and the pH-sensitive material is an overrun Copolymers of polyethyleneimine, polylysine and polyglutamic acid, or copolymers of polyethylene glycol, hyperbranched polyethyleneimine, polylysine and polyglutamic acid, or oligomeric Sulfonamide, or the condensation product of polyethylene glycol and oligomeric sulfonamide, wherein the molecular weight of the hyperbranched polyethyleneimine...

Embodiment 1～8

[0095] The preparation of embodiment 1~8 pH value sensitive material

[0096] According to the ratio of raw materials shown in Table 1, the pH value sensitive material composed of hyperbranched PEI, PLL and PGlu links was prepared according to the following method:

[0097] Dissolve hyperbranched polyethyleneimine (PEI) in chloroform, then add ε-benzyloxycarbonyl-L-lysine N-carboxylic acid anhydride (Lys(z)-NCA) in chloroform and react at 30°C for 72 hours Afterwards, a chloroform solution of γ-benzyl-L-glutamic acid-N-carboxylic acid anhydride (BLG-NCA) was added continuously, and reacted at 30° C. for 72 hours. Settling and drying the obtained reaction product with ether to obtain a solid product, dissolving the solid product in trifluoroacetic acid, adding hydrogen bromide in acetic acid solution for deprotection, continuing to settle and dry with ether, and drying the obtained dry product with The dialysis bag was dialyzed for 72 hours, the water was changed 6 times durin...

Embodiment 9～16

[0103] Embodiment 9~16 preparation of pH value sensitive material

[0104] According to the proportion of raw materials shown in Table 3, the pH value sensitive material composed of PEG, hyperbranched PEI, PLL and PGlu link was prepared according to the following method:

[0105] Dissolve polyethylene glycol with carboxyl groups in water, add N-hydroxysuccinimide (NHS) and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide (EDC), and react After 24 hours, the aqueous solution of the PEI-PLL-PGlu pH sensitive material prepared in Examples 1 to 8 was added respectively, reacted for 24 hours, and the obtained reaction product was dialyzed for 72 hours with a dialysis bag, and the water was changed 6 times during the dialysis, and PEG- PEI-PLL-PGlu pH sensitive material.

[0106] The pH-sensitive material was dissolved in an aqueous solution, and its particle size and surface potential were measured. See Table 4 for the results.

[0107] Raw material and ratio of table 3 embodiment 9...

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Abstract

The invention provides a gene vector system. The gene vector system comprises a pH value sensitive material, wherein the pH value sensitive material is a hyperbranched polyethylenimine-polylysine-polyglutamic acid copolymer, a polyethylene glycol-hyperbranched polyethylenimine-polylysine-polyglutamic acid copolymer, low molecular weight sulfanilamide, or a polyethylene glycol-low molecular weightsulfanilamide condensation compound, and a complex comprising gene vectors and a gene substance, wherein a mass ratio of the gene vectors to the gene substance is in a range of (0.5: 1) to (50: 1); the pH value sensitive material is bonded to the surface of the complex; and a mass ratio of the pH value sensitive material to the gene vectors of the complex is in a range of (1: 1) to (80: 1). The gene vector system provided by the invention can guarantee effectively transportation of gene vectors and a gene substance in normal cells and promote the gene vectors and the gene substance to bond with tumor cells thereby improving transfection efficiency.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a gene carrier system and a preparation method thereof. Background technique [0002] With the continuous advancement of biological science and technology, gene therapy will occupy an important position in the process of overcoming chronic diseases such as cancer and genetic diseases, and will gradually become a common and effective method. Gene therapy refers to a new biomedical technology that introduces human normal genes or genes with therapeutic effects into human target cells in a certain way to correct gene defects or exert therapeutic effects, so as to achieve the purpose of treating diseases. [0003] Introducing gene material into cells for expression is a key step in realizing gene therapy, and successful gene therapy depends on effective gene carriers. Common vectors are divided into viral vectors and non-viral vectors. Viral vectors include retrovirus, adeno...

Claims

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Application Information

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IPC IPC(8): C12N15/87C08G69/10C08G81/00C08G65/00C07C311/39C07C303/40
Inventor 田华雨陈学思郭兆培林琳陈杰焦自学董璇
Owner CHANGZHOU INST OF ENERGY STORAGE MATERIALS &DEVICES
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