Eureka AIR delivers breakthrough ideas for toughest innovation challenges, trusted by R&D personnel around the world.

Lipid-cationic polymer siRNA gene transfection agent and preparation method thereof

A cationic polymer and gene transfection technology, applied in the field of compounds, can solve the problems of low efficiency of cationic polymers, low affinity between cationic polymers and cell membranes, etc., to overcome poor biocompatibility, save experimental time, and facilitate arrangement and adjustment Effect

Inactive Publication Date: 2007-08-15
ZHONGSHAN HOSPITAL XIAMEN UNIV
View PDF0 Cites 9 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The purpose of the present invention is to aim at the low affinity between cationic polymer and cell membrane, and cationic polymer is too strong to small molecule siRNA binding force, so that siRNA is difficult to be separated from polymer in the cell and play a role, causes cationic polymer in small molecule For problems such as low efficiency in siRNA delivery, a degradable lipid-cationic polymer siRNA gene transfection reagent containing brasilenoic acid side chains that can be used as an siRNA transfection reagent and its preparation method are provided

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

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Example 1 Preparation of lipid-cationic polymer siRNA gene transfection reagent

[0026] 1) synthetic lipid-cationic polymer: 1. add 200ml CH in reaction vessel 2 Cl 2 , 0.3ml N,N-dimethylformamide and 28.4g brazilenic acid (C22:1, trans-13), stirred to dissolve. Place in an ice bath to cool to 0°C, maintain this temperature, slowly add 20ml of oxalyl chloride dropwise, stir for 20min, heat up to 25°C in a water bath, add 200ml of toluene, then heat in a 56°C water bath, keep the constant temperature at 56°C, reduce the temperature under vacuum The solvent was evaporated by rotary evaporation until no liquid flowed out. 2. Dissolve the residue in 200ml CH 2 Cl 2 , and cooled to 0°C, added 90ml diethanolamine, 15g 4-dimethylaminopyridine and 28ml ethylamine, stirred at 0°C for 30min and then stirred at room temperature overnight. 3. Add 100ml CH 2 Cl 2 Dilute, stir for 5min, wash with 200ml 0.1mmol / L hydrochloric acid, and wash the organic layer with 200ml 0.02mol...

Embodiment 2

[0029] Example 2 Preparation of lipid-cationic polymer siRNA gene transfection reagent

[0030] 1) synthetic lipid-cationic polymer: 1. add 200ml CH in reaction vessel 2 Cl 2 , 0.6ml of N,N-dimethylformamide and 37.2g of brassenoic acid, stirred to dissolve it. Place in an ice bath to cool to 0°C, maintain this temperature, slowly add 40ml of oxalyl chloride dropwise, stir for 20min, then heat up to 25°C in a water bath, add 300ml of toluene, then heat in a 56°C water bath, keep the constant temperature at 56°C, vacuum reduce The solvent was evaporated by rotary evaporation until no liquid flowed out. 2. Dissolve the residue in 200ml CH 2 Cl 2 , and cooled to 0°C, added 120ml diethanolamine, 21g 4-dimethylaminopyridine and 40ml ethylamine, stirred at 0°C for 30min and then stirred at room temperature overnight. 3. Add 100ml CH 2 Cl 2 Dilute, stir for 5min, wash with 300ml 0.1mmol / L hydrochloric acid, and wash the organic layer with 300ml 0.02mol / L NaHCO 3 Wash, and fin...

Embodiment 3

[0033] Example 3 Preparation of lipid-cationic polymer siRNA gene transfection reagent

[0034] 1) synthetic lipid-cationic polymer: 1. add 200ml CH in reaction vessel 2 Cl 2 , 0.8ml of N,N-dimethylformamide and 34g of brassenoic acid, stirred to dissolve it. Place in an ice bath to cool to 0°C, maintain this temperature, slowly add 30ml of oxalyl chloride dropwise, stir for 20 minutes, then heat up to 25°C in a water bath, add 250ml of toluene, then heat in a 56°C water bath, keep the constant temperature at 56°C, vacuum The solvent was evaporated by rotary evaporation until no liquid flowed out. 2. Dissolve the residue in 200ml CH 2 Cl 2 , and cooled to 0°C, added 100ml diethanolamine, 18g 4-dimethylaminopyridine and 35ml ethylamine, stirred at 0°C for 30min and then stirred overnight at room temperature. 3. Add 100ml CH 2 Cl 2 Dilute, stir for 5min, wash with 250ml 0.1mmol / L hydrochloric acid, and wash the organic layer with 250ml 0.02mol / L NaHCO 3 Wash, and finally...

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 discloses a lipoid-cationic polymer siRNA gene diverting-dye agent and preparing method, which is characterized by the following: supplying a lipoid-cationic polymer and siRNA gene inverting-dye agent hydraulic fluid; making the polymer as brassidic acid structure with branch lateral chain and cationic polymer with molecular weight at 50KD; setting the hydraulic liquid incorporates normal saline, dimethyl sulfoxide and glycerol; synthesizing lipoid-cationic polymer and hydraulic liquid; mixing the lipoid-cationic polymer and hydraulic liquid; getting the product. This invention can overcome defects of poor compatibility and high toxicity, which product can be added into cell culture medium.

Description

technical field [0001] The invention relates to a compound, in particular to a degradable lipid-cationic polymer containing brassenoic acid side chains which can be used as siRNA transfection reagent. Background technique [0002] Short interfering RNA (small interfering RNA, siRNA) is a gene expression silencing technology caused by double-stranded RNA. In 1998, Andrew Fire et al. (Fire A, Xu S, Montgomery MK, Kostas SA, Driver SE, and MelloCC. Potent and specific genetic interference by double-stranded RNA in Caenorhabditis elegans. Nature 1998.391: 806-811) first described a Gene expression silencing technology caused by double-stranded RNA has become RNA-mediated interference technology. RNAi is stimulated by siRNA. One siRNA molecule can cut about 1000 mRNA molecules. RNAi is by far the most effective technique for inhibiting gene expression. Recent studies have found that chemically synthesized siRNA can inhibit the expression of hepatitis B virus in mice, which show...

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): C12N15/63
Inventor 杨天赐
Owner ZHONGSHAN HOSPITAL XIAMEN 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
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