Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Cationic polymer gene carrier and preparation method and application thereof

A cationic polymer, gene carrier technology, applied in the field of medicine, can solve the problems of non-targeting, high production cost of viral vectors, and immunogenicity, etc. Effect of Transfection Efficiency

Active Publication Date: 2017-06-27
JIANGSU CANCER HOSPITAL
View PDF8 Cites 5 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Viral vectors are expensive to produce and have many disadvantages after entering the body (such as no targeting, immunogenicity, etc.), these factors limit their practical use [19]

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
  • Cationic polymer gene carrier and preparation method and application thereof
  • Cationic polymer gene carrier and preparation method and application thereof
  • Cationic polymer gene carrier and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0079] Cationic polymer carrier of embodiment 1 gene medicine and its preparation

[0080] (1) Preparation of poly DHPMA polymer:

[0081] DHPMA (1.0430g, 7.0mmol) was dissolved in 1.0mLmL distilled water, and then added to a 5mLmL polymerization vial, dithiobenzoic acid (4-cyanovaleric acid) ester (0.0336g, 0.12mmol), ACVA (0.0167g, 0.06 mmol) was dissolved in 0.5 mL of 1,4-dioxane and added to the polymerization vial. The system was ventilated with dry nitrogen for 1 h, and the polymerization vial was sealed with wax. Put the polymerization bottle into an oil bath preheated to 70°C for 24 hours. The polymerization tube was taken out and cooled in a cold pump at 0°C to terminate the polymerization reaction and obtain the product. The product was added to ice acetone to precipitate, centrifuged, and dissolved / precipitated repeatedly three times, and the product was dried in a vacuum oven at 50° C. to obtain 0.0362 g (yield: 72%) of the product poly-DHPMA polymer (PDHPMA). ...

Embodiment 2

[0092] The cationic polymer carrier of embodiment 2 gene medicine and its preparation

[0093] (1) Preparation of poly DHPMA polymer:

[0094] With the first (1) step of embodiment one

[0095] (2) Preparation of polyDA polymer:

[0096] With the first (2) step of embodiment one

[0097] (3) Synthesis of guanidinated DG polymer:

[0098] Take DA polymer (1.78g, 0.1mmol) and dissolve in water, add guanidinating reagent 1-H-pyrazole-1-carboxamidine hydrochloride (0.059g, 0.4mmol), and adjust the pH of the reaction solution with saturated sodium carbonate solution To 9.0, react at room temperature for 24h. The reaction solution was added to an ultrafiltration tube (10 kd), washed with ultrapure water three times, and 1.23 g of DG polymer was freeze-dried (yield: 67%).

[0099] Table 3 shows the polymer compositions and average molecular weights of polymers DG1, DG2, and DG3 prepared according to the above method with different segment ratios.

[0100] Table 3 Composition an...

Embodiment 3

[0105] Example 3 Preparation and Characterization of mDG / siRNA Complex

[0106] (1) Preparation of mDG / siRNA complex:

[0107] Dissolve quantitative siRNA stock solution in enzyme-free water, prepare mDG / siRNA complex according to the molar ratio of amino nitrogen in mDG polymer to the molar ratio of phosphate in siRNA (siRNA1, siRNA2, siRNA3 and siRNA-NC), and take Add siRNA (any one of siRNA-1, siRNA-2, siRNA-3, siRNA-NC) stock at the N / P ratio of 1:1, 1:4, and 1:8 to mDG1, mDG2, and mDG3 polymers The solution was mixed and incubated at room temperature for 30 min to prepare a series of DHPMA-b-GPMA / siRNA complexes (i.e. complexes of any two of mDG1, mDG2, mDG3 and siRNA-1, siRNA-2, siRNA-3, siRNA-NC), and then The complex solution was sonicated and passed through a 0.22 μm filter membrane and stored at 4°C for later use.

[0108] (2) Agarose gel electrophoresis of mDG / siRNA complex

[0109] Mix mDG polymer and siRNA according to the charge ratio of 1:4, 1:8 and 1:16, and...

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

PropertyMeasurementUnit
particle diameteraaaaaaaaaa
particle sizeaaaaaaaaaa
diameteraaaaaaaaaa
Login to View More

Abstract

Provided is a cationic polymer gene carrier. N-3-aminopropyl methacrylamide and N-(1,3-dihydroxyl-2-propyl) methacrylamide serve as monomers to synthesizea block polymer N-(1,3-dihydroxyl-2-propyl) methacrylamide-b-N-3-aminopropyl methacrylamide polymer, then a guanidine group is coupled to a chain segment of the N-3-aminopropyl methacrylamide to obtain the N-(1,3-dihydroxyl-2-propyl) methacrylamide-b-N-3-guanidinomethacrylamide polymer; or methionine having tumor targeting property is grafted and a primaquine group is guanidinated to obtain N-(1,3-dihydroxyl-2-propyl) methacrylamide-b(N-3-aminopropyl methacrylamide-g-methionine)-b-guanidinomethacrylamide polymer. The invention further provides a preparation method of the gene carrier. The gene carrier can carry a gene drug, has good biocompatibility and tumor targeting property and is large in application prospect.

Description

technical field [0001] The invention belongs to the field of medicine, and in particular relates to a cationic polymer gene carrier and its preparation method and application. Background technique [0002] Cationic polymer is a new type of non-viral gene carrier, which has good physical and chemical properties, high molecular diversity, low immunogenicity, easy structure modification, and unlimited gene size and type that virus-like gene carriers do not have. Etc [1] . Polyethyleneimine (PEI) is currently the most widely used cationic polymer gene carrier. In 1995, Boussif et al. reported for the first time that PEI can be used as a cationic polymer gene carrier to carry negatively charged DNA molecules to form PEI / DNA complexes [2] . After the PEI / DNA complex enters the lysosome of the cell, the nitrogen atom on the PEI molecular chain is protonated, which increases the pH of the lysosome and causes the influx of Cl ions, thereby allowing the complex to escape from the l...

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): C08F293/00A61K48/00
CPCA61K47/32A61K48/0041C08F8/30C08F293/00C08F293/005C08F2438/03C12N15/87
Inventor 吴旸唐金海吴建中马蓉张薇孙春龙
Owner JIANGSU CANCER HOSPITAL
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

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

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com