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Folate-receptor-mediated pH-sensitive Decoy-ODN (Decoy-oligodeoxynucleotide) nanoparticle preparation and preparation method thereof

A folic acid receptor and sensitivity technology, applied in the field of medical applications, can solve the problems of low transfection efficiency, immunogenicity and carcinogenicity

Inactive Publication Date: 2011-11-16
SHANDONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The gene vectors currently used all have low transfection efficiency or even high transfection efficiency, but are immunogenic and potentially carcinogenic

Method used

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  • Folate-receptor-mediated pH-sensitive Decoy-ODN (Decoy-oligodeoxynucleotide) nanoparticle preparation and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] Take Decoy-ODN separately, add 0.1ml TE buffer solution to fully dissolve, dilute to a concentration of 5μg / ml Decoy-ODN solution A, take folic acid-coupled trimethyl chitosan, dissolve it with distilled water and dilute to a concentration of 0.01mg / ml The folic acid-coupled trimethyl chitosan solution B of ml (the coupling rate of folic acid is 10%), polyacrylic acid resin S100 is taken, and diluted with phosphate buffer to a concentration of 10 μg / mlS100 solution C; Decoy-ODN solution A Mix 50 μl each with solution C to obtain drug solution D, and 100 μl folic acid-coupled trimethyl chitosan solution B respectively in a water bath at 55°C for 30 minutes, and drip drug solution D dropwise into carrier material solution B under vortex conditions , continue to vortex for 30 seconds, and incubate at room temperature for 30 minutes to obtain the folate receptor-mediated pH-sensitive Decoy-ODN nanoparticle preparation.

[0041] The binding rate of the obtained nanoparticles...

Embodiment 2

[0043] Decoy-ODN solution A, 50 μg / ml polyacrylic acid resin S100 solution C and 0.01mg / ml folic acid coupling trimethyl chitosan solution B of 10 μ g / ml of Decoy-ODN solution A, 50 μ g / ml were configured according to the method of Example 1 (folate coupling rate 30%), mix 50 μl each of Decoy-ODN solution A and 50 μg / ml polyacrylic acid resin S100 solution C to obtain drug solution D, and 200 μl folic acid-coupled trimethyl chitosan solution B respectively in a 55° C. water bath for 30 min, vortex Add the drug solution D dropwise to the carrier material solution B under the condition of swirling, continue to vortex for 30 s, and incubate at room temperature for 1 h to obtain the folate receptor-mediated pH-sensitive Decoy-ODN nanoparticles of the present invention.

[0044] The resulting nanoparticle binding rate was 55%.

Embodiment 3

[0046] Decoy-ODN solution A, 50 μg / ml carboxymethyl chitosan solution C and 1.2mg / ml folic acid coupling trimethyl chitosan solution B (folate coupling The rate is 20%), each 50 μ l of Decoy-ODN solution A and 50 μ g / ml carboxymethyl chitosan solution C are mixed to obtain drug solution D, and 300 μ l folic acid-coupled trimethyl chitosan solution B respectively at 55 ℃ Water bath for 20 minutes, under vortex conditions, drop the drug solution D into the carrier material solution B drop by drop, continue to vortex for 1 minute, and incubate at room temperature for 3 hours to obtain the folic acid receptor-mediated pH-sensitive trimethyl chitosan of the present invention sugar nanoparticles.

[0047] Take an appropriate amount of the suspension, drop it on the copper grid, carry out negative staining with 2% phosphotungstic acid solution, and observe it under the transmission electron microscope (TEM) after natural drying. Between 100-200nm, according to the size and shape of ...

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Abstract

The invention relates to a folate-receptor-mediated pH-sensitive Decoy-ODN (Decoy-oligodeoxynucleotide) nanoparticle preparation and a preparation method thereof. A nanoparticle suspension is prepared from 5-200 mu g / ml of the Decoy-ODN used as a generic medicament, 0.01-6.0 mg / ml of folate-conjugated trimethyl chitosan used as a medicament carrier and 10-500 mu g / ml of pH-sensitive material by a complex coacervation method, wherein the folate conjugation rate of the folate-conjugated trimethyl chitosan is 5-30%, and the ratio of the total volume of the Decoy-ODN solution and the pH-sensitive material solution to the volume of the folate-conjugated trimethyl chitosan solution is 1:(1-4). The folate-receptor-mediated pH-sensitive Decoy-ODN nanoparticle preparation provided by the invention has the advantages of good shaping property, uniform particle size distribution, high DNA (deoxyribonucleic acid) binding rate, better stability, low cytotoxicity and high in-vitro transfection efficiency.

Description

technical field [0001] The invention relates to a folic acid receptor-mediated pH-sensitive Decoy-ODN nanoparticle preparation and a preparation method thereof, belonging to the technical field of medical applications. Background technique [0002] In recent years, with the maturity of gene transfer technology, gene therapy has become one of the research hotspots in biological science and clinical medicine, and has gradually become a new means of treating cancer. The so-called gene therapy refers to the introduction of human normal genes or therapeutic DNA sequences into human target cells through gene transfer technology, and by controlling the expression of target genes, inhibiting, correcting, replacing or compensating for defective or abnormal genes to change human genetics. Substance-based biomedical treatments. [0003] However, the occurrence and development of tumors is a complex process involving multiple genes, multiple links, multiple stages, and multiple steps. ...

Claims

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Application Information

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IPC IPC(8): A61K9/14A61K47/22A61K47/36A61K48/00A61P35/00
Inventor 翟光喜陈焕蕾张建赵丽霞李涛
Owner SHANDONG UNIV
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