Hybrid nucleic acid drug carrier of dna and polymer and its preparation method and application

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 effects of nucleic acid fragments, poor tissue penetration, and lack of targeting, and achieve enhanced effects. , easy to functionalize, to achieve specific release effect

Active Publication Date: 2022-01-28
TIANJIN UNIV
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  • 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

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  • Hybrid nucleic acid drug carrier of dna and polymer and its preparation method and application
  • Hybrid nucleic acid drug carrier of dna and polymer and its preparation method and application
  • Hybrid nucleic acid drug carrier of dna and polymer and its preparation method and application

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.

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Abstract

The invention discloses a DNA and polymer hybrid nucleic acid drug carrier and its preparation method and application. Firstly, acrylamide-modified DNA with initiator chain I and acrylamide are used to construct an acrylamide polymer by free radical polymerization / DNA hybrid nanogel, design DNA hairpin structure H1 and H2, the ends of H1 and H2 can be complementary base pairing or covalently linked nucleic acid drug sequence. The prepared polymer / DNA hybrid nanogel is dispersed in the solution containing the corresponding hairpin structures H1 and H2, and the efficient assembly of nucleic acid drugs in the nanogel can be realized through in situ DNA hybridization chain reaction, thereby preparing stable Targeted nucleic acid nanomedicine with high specificity and high loading efficiency. As a unique biopolymer, DNA has precise and adjustable structure, efficient and controllable assembly, and can endow nanogel with stimuli responsiveness and controllable drug release performance; acrylamide polymers have high stability and are easy to functionalize, which can improve nanogel The in vivo stability and tumor targeting ability of the gel enable efficient transfection and expression of nucleic acid drugs at 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

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Application Information

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Patent Type & Authority Patents(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
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