Non-viral gene transfection vector material based on cationic helical peptide

A technology of gene transfection carrier and cation, which is applied in the field of non-viral gene transfection carrier materials and its preparation, can solve the problems of large acidity changes, low efficiency, and many steps, and achieve the effect of low cytotoxicity and high transfection efficiency

Active Publication Date: 2017-04-26
GUANGZHOU MEDICAL UNIV
View PDF3 Cites 10 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

There are many steps in this preparation process, and two consecutive functional group coupling reactions are used for side chain modification, and the efficiency is low
The third step of side chain oxidation and the fourth step of side chain reduction amine reaction take a long time, high temperature, and large changes in acidity. These variable factors may lead to the degradation of the polypeptide main chain.

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Non-viral gene transfection vector material based on cationic helical peptide
  • Non-viral gene transfection vector material based on cationic helical peptide
  • Non-viral gene transfection vector material based on cationic helical peptide

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0071] Embodiment 1: the synthesis of PALG

[0072] Weigh 5 g (0.034 mol) of L-glutamic acid and 8.90 g (0.153 mol) of allyl alcohol, mix them uniformly in the flask, and slowly add 4.0 g (0.041 mol) of concentrated sulfuric acid dropwise under ice-bath conditions, and the dropwise addition is completed within 30 minutes. After cooling for 1 h, the ice bath was removed and the temperature was raised to room temperature, and the reaction was continued for 48 h. After the reaction is finished, add enough triethylamine to the system for neutralization, then add enough acetone to stir, and filter out the precipitate. The precipitate was dried overnight in a vacuum oven at room temperature, and the crude product was recrystallized from isopropanol / water, filtered, washed with a sufficient amount of cold acetone, and dried in vacuo. The product γ-allyl-L-glutamate was white flaky crystal, and the yield was 47%.

[0073] Suspend 1.87g (0.01mol) of γ-allyl-L-glutamate in 30mL of anh...

Embodiment 2

[0076] Embodiment 2: the synthesis of PPAETH

[0077] Dissolve 1.41g (0.01mol) of N-propyl-4-piperidone in 120mL of anhydrous methanol, and then add 1.70g (0.015mol) of cysteamine hydrochloride to fully dissolve it. At this point, add 0.94g (0.015mol) sodium cyanoborohydride, and after it is completely dissolved, add 1.5-2.0mL acetic acid as a catalyst dropwise, and react at room temperature for 72 hours. After the reaction was completed, the solvent was removed by rotary evaporation, quenched by adding 4 mL of concentrated HCl and stirred for 1 h, and most of the water was removed by rotary evaporation. Pour hot ethanol solution into the mixture, filter off the inorganic salt while it is hot, remove ethanol by rotary evaporation, and recrystallize the crude product in methanol. The obtained hydrochloride of thiol-type small molecule amine is white granular, named as PPAETH, and the yield is 34%. 1 H NMR diagram see figure 2 , and its chemical structure is shown in the fol...

Embodiment 3

[0079] Embodiment 3: the synthesis of PPALG

[0080] Weigh 0.1g (0.59mmol C=C bond) of polymer PALG obtained in Example 1 and dissolve it in 3mL DMF to make solution 1; then dissolve 0.18g (0.65mmol-SH) of PPAETH obtained in Example 2 In the least amount of methanol / water mixed solution, make solution two; add photoinitiator Irgacure 651 accounting for 5% of the total mass of all reactants to solution one, and turn on the flashlight Wood Burner's lamp, then slowly add solution 2 dropwise, and the dropwise addition is completed within 1h. Continue to illuminate for 30 minutes, then add 3 mL of DMF, and continue to illuminate for 30 minutes to end the reaction, and dialyze the final mixed solution with a dialysis bag with a molecular weight cut-off of 3500. The distilled water was changed every 4 hours on the first day, and the ethanol solution was changed every 12 hours on the second day to remove the photoinitiator, and then continued to be dialyzed with distilled water for 1...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

No PUM Login to view more

Abstract

The invention belongs to the field of biomedical macromolecular materials and discloses a non-viral gene transfection vector material based on cationic helical peptide, and a preparation method and application thereof. The structure of the vector material is shown by general formulas (1) to (6) in the description, wherein R1 is C1-6 alkyl, benzyl, methoxypolyethylene glycol, polyethylene glycol or polypropylene oxide, R2 is C1-3 alkyl, benzyl or phenethyl, R3 is methyl or ethyl, R4 is hydroxy, and A is H, C1-3 alkyl, alkoxy, halogen or nitryl; in the general formulas (1) to (4), x represents polymerization degree of polypeptide and is not less than 5; in the general formulas (5) and (6), x represents the ratio of side chain repeating units, with 0<x<1. The cationic helical polypeptide main chain is of Alpha-helical conformation and effectively forms complex micelles with DNA and siRNA, and the material has good transfection efficiency, low cell toxicity and applicable to the transfer of pDNA and siRNA.

Description

technical field [0001] The invention belongs to the field of biomedical polymer materials, and in particular relates to a non-viral gene transfection carrier material based on a cationic helical polypeptide and its preparation method and application. Background technique [0002] Gene therapy is one of the most promising technologies emerging today for the treatment of tumors and various immune diseases. It regards genes as a "drug" to treat diseases, which is an advanced treatment method. This therapy can treat a variety of serious diseases, including various congenital genetic diseases and acquired diseases, such as tumors, acquired immunodeficiency syndrome, cardiovascular diseases, cystic fibrosis, etc., among which the treatment of tumors is current research hotspot. There are two ways of gene therapy, one is to replace the disordered gene in the cell with exogenous gene, or to correct the problem gene with exogenous gene, which belongs to DNA therapy; the other is to...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
Patent Type & Authority Applications(China)
IPC IPC(8): C08G69/48C07K7/06A61K47/64A61K47/69A61K31/713A61P35/00A61P37/02
CPCA61K31/713C07K7/06C08G69/48
Inventor 黄玉刚易玲叶国东黄珺珺
Owner GUANGZHOU MEDICAL UNIV
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap