A kind of gene carrier system and its preparation method

A gene carrier and system technology, applied in the field of biological carriers, can solve the problems of low transfection efficiency, affecting the binding efficiency of the carrier system and cells, etc., and achieve the effects of reducing cytotoxicity, improving binding efficiency, and improving turnover ability

Active Publication Date: 2016-07-06
CHANGCHUN INST OF APPLIED CHEMISTRY - CHINESE ACAD OF SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the negative charge on the surface of the introduced shielding system will generate repulsion with the negative charge on the cell surface, affecting the binding efficiency of the carrier system and cells, resulting in low transfection efficiency

Method used

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  • A kind of gene carrier system and its preparation method
  • A kind of gene carrier system and its preparation method
  • A kind of gene carrier system and its preparation method

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preparation example Construction

[0051] ③ The preparation method of the polymer α is: dissolve PEI-PPLG-g-(MEA-BOC / MAA), add acid for deprotection, then settle and drain, and obtain the polymer α through dialysis purification. The reaction has no requirement on the dissolved solvent, preferably chloroform or DMF, more preferably DMF; the reaction has no requirement on the type of acid added, preferably trifluoroacetic acid and hydrochloric acid; the reaction has no requirement on temperature, preferably 20-40°C . The molecular cutoff during dialysis is preferably 1,000 to 5,000.

[0052] In the amino acid polymer of the present invention, when the amino acid polymer is the small molecule alkylamine, the ring-opening polymerization of γ-propynyl-L-glutamic acid-N-internal carboxylic acid anhydride is initiated to obtain polyglutamate and 2- When tert-butoxycarbonyl aminoethanethiol and mercaptoacetic acid react and take off the polymer obtained by tert-butoxycarbonyl protection, i.e. polymer β, it is preferab...

Embodiment 1-8

[0081] The ring-opening polymerization of γ-propynyl-L-glutamic acid-N-internal carboxylic acid anhydride was initiated by polyethyleneimine to obtain polyglutamate, and then reacted with 2-tert-butoxycarbonylaminoethanethiol and thioglycolic acid to remove The polymer obtained by tert-butoxycarbonyl protection, i.e. polymer α, its preparation method is as follows:

[0082] ①Preparation of PLG: Disperse 20g of L-glutamic acid in 30mL of propynyl alcohol, and add 8mL of concentrated sulfuric acid drop by drop under stirring in an ice bath. After half an hour of reaction, the reaction was carried out at room temperature of 28°C for 12h. The product was neutralized with 28 g of sodium bicarbonate in ice water and stirred until a solid precipitated. Filter and dry. The optimized experiment requires recrystallization and purification: heating and dissolving in water once, then cooling to precipitate, suction filtration, and drying. Gamma-propynyl-L-glutamate (PLG) was obtained. ...

Embodiment 9-16

[0097] The ring-opening polymerization of γ-propynyl-L-glutamic acid-N-internal carboxylic acid anhydride is initiated by a small molecule alkylamine to obtain polyglutamate, which is then reacted with 2-tert-butoxycarbonylaminoethanethiol and thioglycolic acid to remove The polymer obtained by tert-butoxycarbonyl protection, i.e. polymer β, its preparation method is as follows:

[0098] ①Preparation of PLG: Disperse 20g of L-glutamic acid in 30mL of propynyl alcohol, and add 8mL of concentrated sulfuric acid drop by drop under stirring in an ice bath. After half an hour of reaction, the reaction was carried out at room temperature of 28°C for 12h. The product was neutralized with 28 g of sodium bicarbonate in ice water and stirred until a solid precipitated. Filter and dry. The optimized experiment requires recrystallization and purification: heating and dissolving in water once, then cooling to precipitate, suction filtration, and drying. Gamma-propynyl-L-glutamate (PLG) ...

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Abstract

The invention provides a gene carrier system, comprising: a pH-sensitive masking system, a cationic carrier, and a genetic material; The obtained polymer is protected by tert-butoxycarbonyl, and the polyglutamate is obtained by ring-opening polymerization of γ-propynyl-L-glutamic acid-N-internal carboxylic acid anhydride initiated by polyethyleneimine or small molecule alkylamine; The mass ratio of the pH-sensitive masking system to the cationic carrier is (2-100):1; the mass ratio of the pH-sensitive masking system to the genetic material is (5-80):1; the genetic material is plasmid DNA or siRNA. The invention also provides a preparation method of the gene carrier system and an amino acid polymer. The gene carrier provided by the invention introduces more charged groups, reduces the dosage of the shielding system, and reduces the cytotoxicity caused by the shielding system during application.

Description

technical field [0001] The invention relates to the field of biological carriers, in particular to a gene carrier system and a preparation method thereof. Background technique [0002] As a new treatment mode, gene therapy has made great progress. It plays an important role in the process of overcoming persistent diseases such as genetic diseases and cancers, and has gradually become a common and effective method. Gene therapy refers to a new biomedical technology that introduces exogenous normal genes into target cells to correct diseases caused by gene defects and abnormalities, so as to achieve therapeutic purposes. Using vectors to introduce foreign genes into target cells is an effective method for gene therapy, and successful gene therapy relies heavily on gene carriers. Common vectors include viral vectors and non-viral vectors. However, viral vectors have great potential safety hazards in clinical application. In the history of gene therapy, there are many patient d...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/63C12N15/87C08G69/48
Inventor 田华雨焦自学陈学思林琳郭兆培陈杰
Owner CHANGCHUN INST OF APPLIED CHEMISTRY - CHINESE ACAD OF SCI
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