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Anti-coagulation nano gene carrier and preparation method thereof

A gene carrier and nanotechnology, applied in gene therapy, medical preparations with non-active ingredients, medical preparations containing active ingredients, etc., can solve the problem of not being able to penetrate deep into the focus of the disease

Active Publication Date: 2019-10-22
XIN HUA HOSPITAL AFFILIATED TO SHANGHAI JIAO TONG UNIV SCHOOL OF MEDICINE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The coronary microcirculation is the only way for miRNA-loaded nanoparticles to reach the ischemic myocardium from the systemic blood circulation, but it is difficult for traditional nanocarriers to pass through these embolisms in the coronary microcirculation, which makes them unable to penetrate into the lesion.

Method used

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  • Anti-coagulation nano gene carrier and preparation method thereof
  • Anti-coagulation nano gene carrier and preparation method thereof
  • Anti-coagulation nano gene carrier and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Example 1. The antisense sequence AMO-1 of miRNA-1 was co-incubated with DGL at different ratios for 30 minutes to find a suitable ratio for AMO-1 to be fully loaded by DGL.

[0033] miRNA-1: an anti-apoptosis gene, sequence 5'-UGGAAUGUAAAGAAGUGUGUAU-3'

[0034] AMO-1: the antisense sequence of miRNA-1, which can specifically bind to it, is an inhibitor of miRNA-1, the sequence 5'-AUACACACUUCUUUACAUUCCA-3', synthesized by Shanghai Gemma Pharmaceutical Technology Co., Ltd., and sold in each tube 0.5OD aliquot

[0035] DGL: G3 dendritic polylysine, purchased from colcom company, MW: 20000

[0036] (1) Dissolve a tube of 0.5OD AMO-1 with 100ul DEPC water, and use nanodrop2000 to measure gene concentration: 300ng / ul, temporarily store at 4°C for later use

[0037] (2) Weigh 10mg of DGL powder and vortex with 10ml of DEPC water to dissolve it for later use (1mg / ml)

[0038] (3) With 50ngAMO-1 as 1, according to the DGL:AMO-1 mass ratio of 0, 0.5, 1, 2, 4, 6, 8, 10, slowly...

Embodiment 2

[0042] Example 2. Low molecular weight heparin and AMO-1-loaded DGL were co-incubated in different ratios to modify the surface and neutralize part of the positive charges while ensuring that AMO-1 was still loaded.

[0043] Low molecular weight heparin sodium: purchased from Dalian Meilun Biotechnology Co., Ltd., enoxaparin, MW: 4500

[0044] (1) After incubating 100ng of AMO-1 and 150ng of DGL for 30min, slowly add low molecular weight heparin (Hep) dropwise to DGL according to the mass ratio of 0, 0.5, 1, 2, 4, 6. Mix slowly while adding, incubate at room temperature for 10 minutes, temporarily store at 4°C for later use

[0045] (2) Prepare 1.5% agar gel containing gel red sugar, mix naked AMO-1, and 10ul of each sample prepared above and loading buffer, and load the sample

[0046] (3) stop after running with a voltage of 100V for 10 minutes, and take pictures with a gel imaging system, the results are as follows figure 2 shown.

[0047] When the mass ratio of low-mol...

Embodiment 3

[0048] Embodiment 3. the nanoparticle that obtains in embodiment 2 is detected with Malvern particle size analyzer, as image 3 As shown, the resulting particle size is uniform, and the average particle size is about 180nm. The detection steps of the Malvern particle size analyzer can be carried out according to the steps provided in the instrument manual.

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Abstract

The invention provides an anti-coagulation nano gene carrier and a preparation method thereof. The nano gene carrier comprises a gene carrier and low-molecular-weight heparin modified on the surface of the gene carrier. The nano gene carrier and the preparation method thereof have the advantages that anti-coagulation genes are safely and effectively delivered and integrated as a whole, accordingly, the dual effects of anti-coagulation gene treatment are achieved, the penetration of the embolization and high blood coagulation parts in coronary artery microcirculation is improved, and a deliverysystem deeply penetrates into the infarcted myocardium.

Description

technical field [0001] The invention relates to an anticoagulant nanometer gene carrier, and also relates to a preparation method of the anticoagulant nanometer gene carrier, which belongs to the field of biotechnology. Background technique [0002] Myocardial infarction (MI) is the cardiovascular disease with the highest mortality rate in the world, and the long-term mortality rate remains high. miRNA brings new opportunities for the clinical treatment of MI. Although miRNA has a good clinical application prospect in the treatment of MI, the lack of a safe and efficient delivery system restricts its clinical application, and nano gene carriers are expected to solve this obstacle. The introduction of nanocarriers can improve the stability, sustained release, transfection rate, and targeting of miRNA, and has good biological safety compared with viral vectors. [0003] When nanocarriers mediate the treatment of myocardial infarction, there is a bottleneck that is difficult t...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K48/00A61K31/7088A61K31/727A61K47/64A61P7/02A61P9/10
CPCA61K31/727A61K47/64A61K31/7088A61K48/0008A61P7/02A61P9/10A61K2300/00Y02A50/30
Inventor 何斌洪婷薛晓梅郭小瑜韦亚忠
Owner XIN HUA HOSPITAL AFFILIATED TO SHANGHAI JIAO TONG UNIV SCHOOL OF MEDICINE