Hybrid nucleic acid drug carrier of DNA and polymer, and preparation method and application thereof

A nucleic acid drug and polymer technology, used in drug combinations, pharmaceutical formulations, and non-active ingredients medical preparations, etc., can solve the problems of off-target effect of nucleic acid fragments, poor tissue penetration, and lack of targeting, and achieve simple equipment. , Easy functionalization, convenient and applicable effect of operation process

Active Publication Date: 2019-11-05
TIANJIN UNIV
View PDF2 Cites 1 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Second, degraded nucleic acid fragments are likely to have off-target effects or cause immune responses
Finally, the nucleic acid drug itself lacks targeting, poor tissue penetration, and a large number of surface negative charges make it difficult to be taken up by cells near the cell membrane

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
  • Hybrid nucleic acid drug carrier of DNA and polymer, and preparation method and application thereof
  • Hybrid nucleic acid drug carrier of DNA and polymer, and preparation method and application thereof
  • Hybrid nucleic acid drug carrier of DNA and polymer, and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047]Prepared by the above method, using N-isopropylacrylamide, 4-acrylamidophenylboronic acid, N,N-methylene bisacrylamide, and acrylamide functionalized DNA as monomer models to prepare polymer / DNA hybrids chemicalized nanoparticles.

[0048] (1) Pipet 467.5 μL of N-isopropylacrylamide (140mM) aqueous solution, 20 μL of N,N-methylenebisacrylamide (130mM) aqueous solution, and 20 μL of APS (5wt%) aqueous solution into a 5mL chicken heart bottle, and add 7μL 4-AAPBA (50mM) methanol solution and 50μL dsDNA-acrylamide (200μM) aqueous solution, add water to make up to 1mL, stir and mix evenly;

[0049] (2) Nitrogen was passed for 20 minutes, then the device was sealed, and the stirring speed was 200 rpm;

[0050] (3) Place in a constant temperature water bath at 70°C for 30 minutes;

[0051] (4) Obtain white nano-gel emulsion after completion of the reaction, centrifuged at 7000rpm for 10min;

[0052] (5) Remove the upper layer solution, redisperse the lower layer of white pr...

Embodiment 2

[0057] Using the above method, DNA functionalized with N-isopropylacrylamide, 4-acrylamidophenylboronic acid, N,N-methylenebisacrylamide, and acrylamide were used as monomer models, and hairpin DNA H1 and H2 were used as The chain hybridization reaction model uses single-stranded DNA as the nucleic acid drug model to prepare nanoparticles loaded with nucleic acid drugs efficiently. The specific implementation methods are as follows:

[0058] (1) Dissolving acrylamide-functionalized DNAC1 and C2 in TAE / Mg 2+ In the buffer, make C1 and C2 complementary to form double-stranded dsDNA-acrylamide;

[0059] (2) Pipet 467.5 μL of N-isopropylacrylamide (140 mM) aqueous solution, 20 μL of N,N-methylenebisacrylamide (130 mM) aqueous solution, and 20 μL of LAPS (5wt%) aqueous solution into a 5 mL chicken heart bottle, and simultaneously add 7 μL of 4- AAPBA (50mM) methanol solution and 100μL dsDNA-acrylamide (200μM) aqueous solution, add water to make up to 1mL, stir and mix evenly;

[...

Embodiment 3

[0068] Embodiment 3 adjusts the breast cancer MDA-MB-231 cells that are in logarithmic growth to 1*10 3 One / well was inoculated in a 96-well culture plate, and different concentrations of nanogel (0-400 μg / mL) were added, and each concentration was paralleled to 5 wells. Give 10% bovine serum RPMI1640 or DMEM culture solution, add 100 μL to each well, culture for 24 hours respectively, and use the MTT method to detect the pro-apoptotic effect of the prepared particles on the cells. Experimental results such as Figure 4 shown.

[0069] It can be seen from the experimental results that there is no obvious cytotoxicity relative to the prepared polymer / DNA.

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
Login to view more

Abstract

The invention discloses a hybrid nucleic acid drug carrier of DNA and a polymer, and a preparation method and an application thereof. Firstly, an acrylamide-modified DNA with an initiating chain I ispolymerized with acrylamide to form acrylamide polymer/DNA hybrid nano gel by free radicals, a DNA hairpin structure H1 and H2 are designed, and the terminals of the H1 and H2 can be complementarily paired with each other by a base or covalently linked with nucleic acid drug sequences. The prepared polymer/DNA hybrid nano gel is dispersed in solutions containing the corresponding hairpin structureH1 and H2. By in-situ DNA hybridization chain reaction, nucleic acid drugs can be highly efficiently assembled in the nano gel, so target nucleic acid nano drugs with high stability and high loadingefficiency are prepared. As a unique biopolymer, the DNA has a precise and adjustable structure, is highly efficient and controllable in assembly, and can give the nano gel stimulated responsiveness and controllable drug release properties. The acrylamide polymer has high stability, is easy to functionalize, can improve the in-vivo stability and tumor targeting ability of the nano gel, and achieves highly efficient transfection and expression of the nucleic acid drugs in tumor sites.

Description

technical field [0001] The invention belongs to the technical field of pharmacy, and in particular relates to a DNA and polymer hybrid nucleic acid drug carrier and its preparation method and application. [0002] technical background [0003] In cancer treatment, chemotherapy is one of the most commonly used methods. Traditional chemotherapy drugs have problems such as poor drug stability, lack of tumor targeting, and large side effects. In recent years, with the development of genomics and the elucidation of the genetic mechanism of cancer pathogenesis, nucleic acid drugs have received extensive attention in the field of cancer treatment. [0004] Nucleic acid drugs mainly include DNA, antisense nucleic acid (ASO), small interfering RNA (siRNA) and microRNA (miRNA), etc., which are research hotspots in the field of medical science and technology in various countries. Compared with small molecule drugs and protein drugs, nucleic acid drugs can achieve efficient and precise ...

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): A61K9/51A61K47/32A61K47/26A61K47/54A61K31/7105A61P35/00
CPCA61K9/5138A61K9/5123A61K47/549A61K31/7105A61P35/00
Inventor 李凤仰大勇张姣姣余文婷
Owner TIANJIN 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
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