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A sucrose fatty acid ester embedded type cationic liposome genetic vector system, a preparing method thereof and applications of the method

A technology of sucrose fatty acid ester and cationic liposome, which is applied in the field of sucrose fatty acid ester embedded cationic liposome gene carrier system and its preparation, can solve the problem of restricting the development of cationic liposome, unclear gene transfer mechanism, organ Targeting is not obvious and other problems, to achieve the effect of a wide range of hydrophilic and lipophilic balance, good biodegradability, and enhanced compatibility

Inactive Publication Date: 2016-04-27
DALIAN NATIONALITIES UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, it also has certain limitations, such as high cytotoxicity, unclear targeting of organs, and unclear gene transfer mechanism, which limit the development of cationic liposomes.

Method used

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  • A sucrose fatty acid ester embedded type cationic liposome genetic vector system, a preparing method thereof and applications of the method
  • A sucrose fatty acid ester embedded type cationic liposome genetic vector system, a preparing method thereof and applications of the method
  • A sucrose fatty acid ester embedded type cationic liposome genetic vector system, a preparing method thereof and applications of the method

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Experimental program
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Effect test

Embodiment 1

[0055] The preparation of embodiment 1 cationic liposome CDO14S

[0056] Weigh 1mg of peptide-type cationic lipid CDO14 and colipoid sucrose stearate S070 (1mg) in the same mass ratio, dissolve in 1mL methanol and chloroform mixed solvent (mixing ratio is 2:1 volume ratio), and wait for After fully dissolving, blow it into a uniform film under nitrogen, and dry it in vacuum for 5 hours to evaporate all the solvent (vacuum degree is -0.09MPa, normal temperature). After dissolving with 100 μL absolute ethanol, add 900 μL ultrapure water to make the liposome 1mg / mL, hydrated at about 80°C for 1h, ultrasonically oscillated at an ultrasonic frequency of 100Hz until clear and transparent, and prepared sucrose ester-embedded cationic liposomes with a concentration of 1mL / mg.

Embodiment 2

[0057] The preparation of embodiment 2 cationic liposome CTA14S

[0058] Weigh 0.5 mg of Gemini cationic lipid CTA14 and dissolve it in 1 mL of chloroform, add 1 mg of colipoid sucrose stearate S570 (mass ratio of CTA14 to S570 is 1:2), after fully dissolved, blow it under nitrogen Form a uniform film, vacuum dry for 12 hours to volatilize all the solvent (vacuum degree is -0.09MPa, room temperature), soak in 1mL ultrapure water for 2 hours to make the film fall off, and repeat ultrasonic vibration at about 55°C (oscillation frequency: 100Hz) until clear and transparent , to prepare sucrose ester embedded cationic liposomes with a concentration of 0.5 mg / mL.

Embodiment 3

[0059] The preparation of embodiment 3 cationic liposome CPA14P

[0060] Weigh 5 mg of quaternary ammonium cationic lipid CPA14 and 0.625 mg of colipoid sucrose palmitate P170 (the mass ratio of CPA14 to P-170 is 8:1) and dissolve it in 5 mL of chloroform. Blow down to form a uniform film, vacuum dry for 8 hours to evaporate all the solvent (vacuum degree is -0.09MPa, room temperature), add 5mL phosphate buffer, hydrate at 30°C for 8 hours, and ultrasonically oscillate at an ultrasonic frequency of 100Hz until clear and transparent. A concentration of 5 mg / mL sucrose ester-embedded cationic peptide liposomes was obtained.

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Abstract

The invention relates to a sucrose fatty acid ester embedded type cationic liposome genetic vector system, a preparing method thereof and applications of the method. The genetic vector system is a composition comprising genetic materials and liposomes formed from cationic lipoids and sucrose esters adopted as auxiliary lipoids. The cationic lipoids comprise peptide type cationic lipoids, gemini cationic lipoids and quaternary ammonium salt type cationic lipoids. The sucrose esters comprise sucrose stearate, sucrose palmitate, sucrose laurate, sucrose oleate and sucrose erucate. The vector can effectively compress plasmid DNA and siRNA and is capable of efficient transfection in vitro and in vivo and nearly free of toxicity for cells and mice. The genetic vector system has a good research and application prospect in gene therapy.

Description

technical field [0001] The invention belongs to the field of biotechnology, in particular to a sucrose fatty acid ester embedded cationic liposome gene carrier system and a preparation method thereof. technical background [0002] Gene therapy refers to the introduction of exogenous normal genes into target cells to correct or compensate diseases caused by gene defects and abnormalities, so as to achieve therapeutic purposes. In 1990, the National Institutes of Health of the United States used gene therapy to treat adenosine deaminase deficiency, which was the first clinical application of gene therapy and achieved success. The successful treatment of combined immunodeficiency syndrome using gene therapy in 2000 demonstrated the effectiveness of gene therapy strategies. For the further development of gene therapy, three problems must be overcome: first, there must be a target gene for treatment and cells receiving the target gene; second, the controllability of gene express...

Claims

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Application Information

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IPC IPC(8): A61K48/00A61K9/127A61K31/7088A61K47/26A61K47/34A61P35/00
CPCA61K9/127A61K31/7088A61K47/26A61K47/34A61K48/0033A61K48/0041
Inventor 张树彪赵轶男崔诗慧赫泽坤徐宇虹
Owner DALIAN NATIONALITIES UNIVERSITY
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