Polycation lipesome telomere enzyme antiseuse oligonucleotide complex and preparation

An antisense oligonucleotide and polycation technology, which is applied in the field of preparation of polycation liposome telomerase antisense oligonucleotide complex, can solve the problem of cell toxicity, high cytotoxicity, and no transfection Efficiency and other issues, to achieve the effect of improving antisense curative effect, simple preparation process, and broad application prospects

Active Publication Date: 2007-03-28
ZHEJIANG PHARMA COLLEGE +1
View PDF0 Cites 13 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The former has good biocompatibility, simple preparation, but poor stability, and has certain toxicity to cells; the latter has a wide variety, and polyethyleneimine (PEI) is a commonly used one, with linear and branched types and different molecular

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
  • Polycation lipesome telomere enzyme antiseuse oligonucleotide complex and preparation
  • Polycation lipesome telomere enzyme antiseuse oligonucleotide complex and preparation
  • Polycation lipesome telomere enzyme antiseuse oligonucleotide complex and preparation

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] Take 5 mg of polyethyleneimine-cholesterol (PEI-Chol) and 5 mg of dioleoylphosphatidylethanolamine (DOPE), dissolve it in an appropriate amount of chloroform, prepare it by thin film dispersion method, remove the chloroform by evaporation under reduced pressure, and use deionized Water and dilute to 5mL. PEI-Chol / DOPE liposomes were obtained by probe ultrasonication (power 400W, working time 1 second, gap time 2 seconds, 120 cycles, ice bath) and extruded polycarbonate membrane (0.1 μm). The polycationic liposome and anti-hTERT were properly diluted with serum-free medium (RPMI 1640), mixed in equal volumes at an N / P ratio of 1.9:1 to 9.5:1, and incubated at room temperature for 20 minutes. The total volume of the complex was 100 μL, and the final concentration of anti-hTERT was 2.0 μmol / L.

Embodiment 2

[0020] Take 3 mg of polyethyleneimine-cholesterol (PEI-Chol), 10 mg of soybean lecithin (SPC), and 10 mg of cholesterol (Chol) and dissolve them in an appropriate amount of chloroform, prepare by thin film dispersion method, remove chloroform by evaporation under reduced pressure, and use Hydrate with deionized water and dilute to 10 mL. PEI-Chol / SPC / Chol liposomes were obtained by probe ultrasonication (power 400W, working time 1 second, gap time 2 seconds, 120 cycles, ice bath) and extruded polycarbonate membrane (0.1 μm). The polycationic liposome and anti-hTERT are properly diluted with serum-free medium (RPMI1640), mixed in equal volumes according to the N / P ratio of 0.5:1-2.5:1, and incubated at room temperature for 20 minutes. The total volume of the complex was 100 μL, and the final concentration of anti-hTERT was 2.0 μmol / L.

Embodiment 3

[0022] Take 3 mg of polyethyleneimine-cholesterol (PEI-Chol), 10 mg of egg yolk lecithin (EPC), and 10 mg of cholesterol (Chol) dissolved in an appropriate amount of chloroform, prepare by thin film dispersion method, evaporate under reduced pressure to remove chloroform, and use Hydrate with deionized water and dilute to 10 mL. PEI-Chol / EPC / Chol liposomes were obtained by probe ultrasonication (power 400W, working time 1 second, gap time 2 seconds, 120 cycles, ice bath) and extruded polycarbonate membrane (0.1 μm). The polycationic liposome and anti-hTERT are properly diluted with serum-free medium (RPMI1640), mixed in equal volumes according to the N / P ratio of 0.5:1-2.5:1, and incubated at room temperature for 20 minutes. The total volume of the complex was 100 μL, and the final concentration of anti-hTERT was 2.0 μmol / L.

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 supplies a polycation liposome telomere enzyme antisense oligonucleotide compound that is made up of polycation liposome and antisense oligonucleotide. It uses low molecular weight PEI and acyl chloride cholesterol as raw material to compound PEI-Chol, uses phospholipid, PEI-Chol and cholesterin as film material. The polycation liposome would be made through film dispersion method and reverse evaporation method. The compound could obviously improve ability of cell to absorb anti-hTERT and have higher inhibition ratio than that of cation polymer. The method is simple technology and widely application prospect.

Description

technical field [0001] The invention relates to the carrier of gene medicine, in particular to the preparation of polycationic liposome telomerase antisense oligonucleotide complex. technical background [0002] With the completion of the Human Genome Project, people have a deeper understanding of the genetic material basis of diseases, and gene therapy (gene therapy) has become a promising treatment method, which is considered to treat hereditary congenital diseases, malignant tumors, etc. , infectious diseases, etc., is one of the most promising treatment options. [0003] Antisense oligonucleotides are oligodeoxynucleotides with a specific sequence, which can specifically block the replication, transcription and translation of target genes, and are a method of gene therapy. Telomerase is a special nucleoproteinase that can use its own RNA sequence as a template to add the telomeric repeat sequence TTAGGG at the 3' end of the chromosome. Two main components of telomerase...

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
IPC IPC(8): C12N15/63A61K48/00
Inventor 胡英陈海靓陈金亮高建青梁文权
Owner ZHEJIANG PHARMA COLLEGE
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