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

peg modified cpg oligonucleotide and its application

An oligonucleotide and cpg-7 technology, which is applied in the field of vaccines and tumor immunotherapy delivery systems, can solve the problems of few modified oligonucleotides and low efficiency, so as to increase in vivo stability, improve response, Increased immune modulating effect

Inactive Publication Date: 2016-06-01
SHANGHAI JIAOTONG UNIV
View PDF2 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] In addition, although PEG modification (PEGylation) has been successfully applied to the modification of various protein molecules and nano-drug delivery systems, which can greatly improve the serum stability and half-life of proteins, it is rarely used to modify oligonucleotides (ODNs). ) molecules, including siRNA, microRNA, CpgODN, etc. The main reason is that most of the targets of ODN are in cells, and the molecular weight of PEG-modified ODN is larger, and the efficiency of entering cells should be lower

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
  • peg modified cpg oligonucleotide and its application
  • peg modified cpg oligonucleotide and its application
  • peg modified cpg oligonucleotide and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0065] Embodiment 1, the impact of different reaction times on the synthesis rate of structure 1

[0066] (1) After the ultrapure water is boiled, N 2 After cooling, to remove oxygen, the ultrapure water was used to prepare pH=7.4, 10 mM Tris-HCl as the reaction solvent. Take 20nmol of CpG-SH and dissolve it in 100ul of 10mM Tris-HCl. After dissolving, blow N 2 5 minutes. Weigh 3.5mg of TCEP, add 4ml of 10mM Tris-HCL (pH=7.4) to obtain 3nmol / μl TCEP, blow with nitrogen for 5min, draw 10ul into the EP tube of CPG-SH, and mix well. Weigh 1.0mg of mPEG 20K -mal (other molecular weight increases or decreases in the same proportion) was dissolved in CpG-SH solution, and nitrogen was protected during the whole process. On a constant temperature oscillator at 25°C and 800rpm, react for 5h, 10h, 15h, and 25h, respectively.

[0067] (2) Use agarose gel electrophoresis to compare the difference in synthesis rate of the samples obtained in (1). The results are shown in figure 1 ,...

Embodiment 2

[0068] Embodiment 2, the impact of the feed ratio of CpG-PEG and CpG on the reaction yield of structure 1

[0069] (1) After the ultrapure water is boiled, N 2 After cooling, to remove oxygen, the ultrapure water was used to prepare pH=7.4, 10 mM Tris-HCl as the reaction solvent. Take 15nmol of CpG-SH and dissolve it in 20ul of 10mM Tris-HCl. After dissolving, blow nitrogen gas for 5 minutes. Weigh 3.5mg of TCEP, add 4ml of 10mM Tris-HCl (pH=7.4) to obtain 3nmol / μl TCEP, blow with nitrogen for 5min, draw 10ul into the ep tube of CPG-SH, and mix well. And divided into 3 tubes, weighed and weighed 0.2mg, 0.4mg, 0.6mg of mPEG 20K -mal (other molecular weight increases or decreases in the same proportion) was dissolved in the CpG-SH solution and vortexed. Nitrogen protection during the whole process. React at 25°C and 800rpm on a constant temperature oscillator for 5h respectively.

[0070] (2) Use agarose gel electrophoresis to compare the difference in synthesis rate of t...

Embodiment 3

[0071] The synthesis and purification method of CpG-PEG shown in embodiment 3, structure 1

[0072] 1. CpG-PEG 20K synthetic method ( image 3 ).

[0073] (1) After the ultrapure water is boiled, N 2 After cooling, to remove oxygen, the ultrapure water was used to prepare pH=7.4, 10 mM Tris-HCL as a reaction solvent. Take 20 nmol of CpG-SH and dissolve it in 100 ul of 10 mM Tris-HCL. After dissolving, blow nitrogen gas for 5 minutes. Weigh 3.5mg of TCEP, add 4ml of 10mM Tris-HCL (pH=7.4) to obtain 3nmol / μl TCEP, blow with nitrogen for 5min, draw 10ul into the EP tube of CPG-SH, and mix well. Weigh 1.6 mg of mPEG20K-mal (other molecular weights increase or decrease in the same proportion) and add to the above solution, vortex to dissolve completely. N throughout the process 2 Protect. React at 25°C and 800rpm on a constant temperature oscillator for 5h respectively. The dosage can be enlarged or reduced in the same proportion for synthesis.

[0074] (2) Extract the s...

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
molecular weightaaaaaaaaaa
Login to View More

Abstract

The invention relates to PEGylated CpG oligonucleotide and application of the PEGylated CpG oligonucleotide in the preparation of a medicament for improving the IL-12 expression quantity in serum, a medicament for inducing the immunogenicity to an endogenous antigen such as a tumor antigen and a medicament for stimulating the immunocompetence of B cells, plasmacytoid dendritic cells, myeloid dendritic cells, macrophages and peripheral mononuclear cells in the field of vaccine and tumor immunotherapy administration systems. The PEGylated CpG oligonucleotide has the effects of homeostasis improvement and immunoregulation, and response to persistent antigens in vivo can be improved.

Description

technical field [0001] The invention belongs to the field of drug delivery systems for vaccines and tumor immunotherapy, and in particular relates to a PEG-modified CpG oligonucleotide and an application thereof. Background technique [0002] CpG oligodeoxyribonucleotides (oligodeoxynucleotides, ODN) are some artificially synthesized oligodeoxynucleotides whose core is the CpG sequence (cytosine-phosphate-guanine) motif with immune activation function. It is 5'-PuPuCPGPyPy-3, an oligonucleotide sequence with two purines or a purine followed by a thymine at the 5' end and two pyrimidines at the 3' end. [0003] In the 1890s, when American oncologist Coley used bacterial crude extracts in the treatment of 900 long-term cancer patients, he found that 40% of the patients' tumors regressed by themselves. At that time, it was believed that the lipopolysaccharide played a role, but it did not attract enough attention. Later, when BCG was used for in vivo experiments in mice, it w...

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 Patents(China)
IPC IPC(8): C12N15/117A61K48/00A61P37/02
Inventor 徐宇虹向小飞岳洋吴彩兴王辉
Owner SHANGHAI JIAOTONG 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