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Nano microRNA precise diagnosis and treatment system capable of targeting ischemic myocardium

A diagnosis and treatment system, myocardial technology, applied in the field of gene carrier, can solve the problems of non-targeting and inability to solve the problem of targeted treatment of diseased parts, and achieve the effect of less toxic and side effects

Active Publication Date: 2017-03-22
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

[0008] In addition, in the current therapeutic products for cardiovascular diseases, the carrier used to load the gene is often not targeted during treatment, so it cannot solve the problem of targeted therapy in the case of enriched lesion sites

Method used

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  • Nano microRNA precise diagnosis and treatment system capable of targeting ischemic myocardium
  • Nano microRNA precise diagnosis and treatment system capable of targeting ischemic myocardium
  • Nano microRNA precise diagnosis and treatment system capable of targeting ischemic myocardium

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0131] Embodiment 1, the synthesis of DGL-PEG

[0132] a) Weigh 22 mg (1 μmol) of DGL (MW: 25000) and dissolve it in 5 mL of PBS buffer (10 mM; pH=7.4) (conc.

[0133] 0.2μmol / mL) (after weighing, place the solid DGL in a 10mLep tube);

[0134] In this embodiment, DGL with other molecular weights can also be used, such as: MW=10000, 18000, 20000, 27000, 30000, 50000, 80000 or 100000.

[0135] b) Weigh 100 mg (50 μmol) of NHS-PEG2000-MAL (MW: 2000) and dissolve it in 5 mL of DMSO;

[0136] In this embodiment, NHS-PEG2000-MAL of other molecular weights can also be used, such as: MW=1000, 1800, 2100, 2700, 3000, 4500, 6500, 8000 or 10000.

[0137] In this example, during the scientific research on the reaction conditions, the reaction ratio between DGL and NHS-PEG2000-MAL was adjusted according to the different groups to be substituted. For example, the molar ratio of DGL and NHS-PEG2000-MAL is 1:2, 1:5, 1:10, 1:15, 1:25, 1:30, 1:40, 1:45, 1:50, 1:55, 1:60, 1:65, 1:60 70, 1:8...

Embodiment 2

[0152] Embodiment two, the synthesis of DGL-PEG-AT1

[0153] The reaction equation is as follows:

[0154]

[0155] a) Weigh 22 mg (1 μmol) of DGL (MW: 25000) and dissolve it in 5 mL of PBS buffer (10 mM; pH=7.4) (conc.

[0156] 0.2μmol / mL) (after weighing, place the solid DGL in a 10mLep tube);

[0157] In this embodiment, DGL with other molecular weights can also be used, such as: MW=10000, 18000, 20000, 27000, 30000, 50000, 80000 or 100000.

[0158] b) Weigh 100 mg (50 μmol) of NHS-PEG2000-MAL (MW: 2000) and dissolve it in 5 mL of DMSO;

[0159] In this embodiment, NHS-PEG2000-MAL of other molecular weights can also be used, such as: MW=1000, 1800, 2100, 2700, 3000, 4500, 6500, 8000 or 10000.

[0160] In this example, during the scientific research on the reaction conditions, the reaction ratio between DGL and NHS-PEG2000-MAL was adjusted according to the different groups to be substituted. For example, the molar ratio of DGL and NHS-PEG2000-MAL is 1:2, 1:5, 1:10, 1:...

Embodiment 3

[0183] Embodiment 3, loaded antisense oligonucleotide

[0184] In this embodiment, AMO-1 is selected, and nanomaterials (NHS-PEG or NHS-PEG-MAL) and AMO-1 are loaded according to the mass ratio (20:1). Dissolve the nanomaterials and AMO-1 in DEPC water, add AMO-1 first into a 200 μL ep tube, then add the materials, mix by pipetting, vortex for 30 seconds, and incubate in a 47°C incubator for 1 hour. (Principle: electrostatic effect; DGL is positively charged, AMO-1 is negatively charged)

[0185] (Note 1: miRNA-1 increases in cardiomyocyte hypoxia, which promotes the apoptosis of cardiomyocytes and is a damage factor of cardiomyocytes. AMO-1 refers to the antisense oligonucleotide of miRNA-1, which is chemically synthesized The single-stranded RNA complementary to miRNA. It can complement the mature miRNA-1 in the cytoplasm, so that the damage effect of miRNA-1 is weakened.)

[0186] (Note 2: miRNA-1 sequence: 5'-UGGAAUGUAAAAGAAGUGUGUAU-3'

[0187] AMO-1 sequence: 5'-AUACAC...

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Abstract

The invention provides a nano microRNA precise diagnosis and treatment system capable of targeting ischemic myocardium. The precise diagnosis and treatment system is characterized by comprising a nano gene substrate which has a targeting property; the nano gene substrate is prepared from an amino acid polymer, modifying polyethylene glycol and angiotensin II polypeptide, wherein the amino acid polymer and the angiotensin II polypeptide respectively participate in a chemical reaction with modifying genes at two terminals of the modifying polyethylene glycol, so that the nano gene substrate which has the targeting property is obtained. The novel nano gene material provided by the invention has the targeting property on the ischemic myocardium and has a protective effect on genes, and the nano gene material is good in transfection effect and low in toxicity.

Description

technical field [0001] The present invention relates to the field of gene carriers, in particular to a nanometerized microRNA precision diagnosis and treatment system targeting ischemic myocardium and a manufacturing method thereof. Background technique [0002] In recent years, cardiovascular disease has become one of the diseases with the highest morbidity and mortality worldwide. Although gene therapy has achieved satisfactory results, safe and effective delivery vectors are still the bottleneck. Commonly used cardiac transport vectors can be divided into viral vectors and non-viral vectors. Due to the high transfection efficiency, the application of viral vectors still has an absolute advantage. However, due to the strong immune response caused by the virus vector, its clinical application is restricted. As an effective supplement to viral vectors, the research on non-viral vectors has been deepened in recent years. [0003] Non-viral vectors are non-infectious, have...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K47/60A61K47/64A61K31/7105A61K48/00A61P9/10
CPCA61K48/0041A61K48/005A61K31/7105
Inventor 何斌石学银薛晓梅董海青李永勇
Owner XIN HUA HOSPITAL AFFILIATED TO SHANGHAI JIAO TONG UNIV SCHOOL OF MEDICINE
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