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Cationic liposome genetic vector and preparation method and application thereof

A cationic liposome and gene carrier technology, which is applied in the field of biomedicine, can solve the problems of insufficient clinical treatment, and achieve the effects of small particle size, low cytotoxicity, and reduced cytotoxicity.

Inactive Publication Date: 2017-03-29
SHANDONG ANALYSIS & TEST CENT
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although the laboratory research on the application of cationic liposomes in gene therapy has achieved some preliminary results, it is far from being widely used in clinical treatment.

Method used

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  • Cationic liposome genetic vector and preparation method and application thereof
  • Cationic liposome genetic vector and preparation method and application thereof
  • Cationic liposome genetic vector and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] The preparation of embodiment 1 cationic liposome gene carrier

[0047] Weigh DPPC and DODAB, the molar ratio of the two is 1:3, put them in a 500ml ground-mouthed round bottom flask, add 20ml of chloroform-methanol (1:1, v / v) mixed solvent to fully dissolve it, and obtain lipid solution. Place the round-bottomed flask on a water bath with a constant temperature of 40°C, pass through nitrogen, and rotate at a speed of 140r / min to make the lipids form a uniform film on the inner wall of the round-bottomed flask. Continue to pass nitrogen gas for 10 min. The dried film was washed with a certain amount of twice-distilled water on a rotary evaporator, and the obtained hydration solution was transferred into a closed Erlenmeyer flask, and the first ultrasonic treatment was performed on a cup bath ultrasonicator. The treatment time is 30 minutes, and a relatively transparent lipid dispersion is obtained, and then the dispersion is repeatedly frozen and thawed four times (fr...

Embodiment 2

[0048] The preparation of embodiment 2 cationic liposome gene carrier

[0049] Weigh DPPC and DODAB, the molar ratio of the two is 1:1, put them in a 500ml ground-mouth round bottom flask, add 25ml of chloroform-methanol (1:2, v / v) mixed solvent to fully dissolve it, and obtain lipid solution. Place the round-bottomed flask on a water bath with a constant temperature of 37°C, pass through nitrogen, and rotate at a speed of 130r / min to make the lipids form a uniform film on the inner wall of the round-bottomed flask. Continue to pass nitrogen gas for 15min. The dried film was washed with a certain amount of twice-distilled water on a rotary evaporator, and the obtained hydration solution was transferred into a closed Erlenmeyer flask, and the first ultrasonic treatment was performed on a cup bath ultrasonicator. The treatment time is 25 minutes, and a relatively transparent lipid dispersion is obtained, and then the dispersion is repeatedly frozen and thawed four times (the r...

Embodiment 3

[0050] The preparation of embodiment 3 cationic liposome gene carrier

[0051] Weigh DPPC and DODAB, the molar ratio of the two is 1:4, put them in a 500ml ground-mouthed round bottom flask, add 30ml of chloroform-methanol (1:2, v / v) mixed solvent to fully dissolve it, and obtain lipid solution. Place the round-bottomed flask on a water bath at a constant temperature of 35°C, blow in nitrogen gas, and rotate at a speed of 150r / min to make the lipids form a uniform film on the inner wall of the round-bottomed flask. Continue to pass nitrogen gas for 15min. The dried film was washed with a certain amount of twice-distilled water on a rotary evaporator, and the obtained hydration solution was transferred into a closed Erlenmeyer flask, and the first ultrasonic treatment was performed on a cup bath ultrasonicator. The treatment time is 40 minutes, and a relatively transparent lipid dispersion is obtained, and then the dispersion is repeatedly frozen and thawed four times (refrig...

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Abstract

The invention discloses a novel cationic liposome genetic vector and a preparation method and application thereof. Dipalmitoyl phosphatidyl choline and dimethyl dioctadecyl ammonium bromide are dissolved into a chloroform-methanol solution fully, wherein the molar ratio of the dipalmitoyl phosphatidyl choline to the dimethyl dioctadecyl ammonium bromide is 1: 1 to 1: 5, and a lipoid solution is prepared; then a stable nitrogen flow is used for rotationally drying the lipoid solution, so that a uniform film is formed by the lipoid solution; after drying, distilled water is added for hydration treatment, and then ultrasonic treatment is conducted for the first time, so that transparent lipid dispersion liquid is obtained; and the dispersion liquid is subjected to freeze thawing repeatedly, and ultrasonic treatment is conducted for the second time, so that the cationic liposome genetic vector is obtained. The cationic liposome genetic vector obtained through the preparation method is stable in property, and the encapsulation efficiency and transfection efficiency a cationic liposome medicinal preparation prepared from the cationic liposome genetic vector are high.

Description

technical field [0001] The invention belongs to the technical field of biomedicine and relates to a novel non-viral gene delivery carrier, in particular to a novel cationic liposome gene carrier and its preparation method and application. Background technique [0002] With the development of molecular biology and the completion of the Human Genome Project, gene therapy provides a promising treatment method for the prevention and treatment of major diseases that seriously threaten human health, such as hereditary diseases, malignant tumors, AIDS, and cardiovascular and cerebrovascular diseases. This method is to introduce normal genes or genes with therapeutic effects into target cells in a specific way to correct gene defects and finally achieve the purpose of treating diseases. Compared with traditional treatment methods, gene therapy can specifically treat target cells and has long-term efficacy. However, since exogenous genes are easily degraded in whole or in part by nu...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K48/00A61K9/127A61K47/18A61K47/24C12N15/88
CPCA61K48/0033A61K9/1271A61K47/186A61K47/24C12N15/88
Inventor 刘书花魏云波王利红王兴民丁尚志刘静
Owner SHANDONG ANALYSIS & TEST CENT
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