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Genetic vector system and preparation method thereof

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

Active Publication Date: 2014-01-08
CHANGCHUN INST OF APPLIED CHEMISTRY - CHINESE ACAD OF SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

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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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  • Genetic vector system and preparation method thereof
  • Genetic vector system and preparation method thereof
  • Genetic vector system and preparation method thereof

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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 genetic vector system. The genetic vector system comprises a pH sensitive masking system, a cation carrier and a genetic material, wherein the pH sensitive masking system is a polymer obtained by reaction of polyglutamate, 2-(Boc-amino)ethanethiol and mercaptoacetic acid and with removal of t-butyloxycarboryl protection, the polyglutamate is obtained by gamma-propinyl-L-glutamic acid-N-carboxyanhydride initiated by polyethyleneimine or micromolecule alkyl amine through ring opening polymerization, the mass ratio of the pH sensitive masking system and the cation carrier is (2-100):1, the mass ratio of the pH sensitive masking system and the genetic material is (5-80):1, and the genetic material is plasmid DNAs or siRNAs. The invention also provides a preparation method of the genetic vector system and an amino acid polymer. The genetic vector can introduce relatively multiple electrified groups, the using amount of the masking system is reduced, and the cytotoxicity generated by the masking system can be reduced when the genetic vector system is used.

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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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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