Cationic liposome containing natural yam sterol, as well as preparation method and application of cationic liposome

A technology of cationic liposome and diosgenol arginine cation, which is applied in the directions of liposome delivery, medical preparations of inactive ingredients, and other methods of inserting foreign genetic materials, etc. problem, to achieve the effect of high siRNA transfection efficiency

Inactive Publication Date: 2015-09-23
SHANGHAI INST OF ORGANIC CHEM CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, most of the above-mentioned synthetic steroidal lipids used as gene delivery carriers are concentrated in the cell transfection of plasmid DNA, and there are few steroidal lipids used for siRNA transfection.

Method used

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  • Cationic liposome containing natural yam sterol, as well as preparation method and application of cationic liposome
  • Cationic liposome containing natural yam sterol, as well as preparation method and application of cationic liposome
  • Cationic liposome containing natural yam sterol, as well as preparation method and application of cationic liposome

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024]Weigh 1mmoL of cationic lipid Dios-Arg containing natural diossterol in a 250mL round bottom flask, and then add dioleoylphosphatidylethanolamine (DOPE) with a molar ratio of 2:1 to mix with it. Then fully dissolve the mixed lipid in 100mL chromatographic grade tetrahydrofuran solution, then add 1mL of anhydrous n-propanol, and then remove the organic solvent in a rotary evaporator at 60°C until a uniform lipid film is formed on the bottom of the bottle , adding 100mL deionized water, ultrasonic hydration for 1.5h to obtain cationic liposome Dios-DOPE 2 / 1 containing natural diossterol.

[0025]

Embodiment 2

[0027] In a 250 mL round bottom flask, 1 mmoL of cationic lipid Dios-Arg containing natural diossterol was weighed, and then dioleoylphosphatidylethanolamine (DOPE) was added in a molar ratio of 1:1 and mixed with it. Then fully dissolve the mixed lipid in 100mL chromatographic grade chloroform solution, then add 1mL of anhydrous tert-butanol, then remove the organic solvent in a rotary evaporator at 70°C, and wait until a uniform lipid film is formed on the bottom of the bottle , adding 100mL deionized water, ultrasonic hydration for 1.5h to obtain cationic liposome Dios-DOPE 1 / 1 containing natural diossterol.

[0028]

Embodiment 3

[0030] Weigh 1mmoL of cationic lipid Dios-Arg containing natural diossterol in a 250mL round bottom flask, and then add dioleoylphosphatidylethanolamine (DOPE) with a molar ratio of 1:2 to mix with it. Then fully dissolve the mixed lipid in 100mL of chromatographic grade acetone solution, add 1mL of absolute ethanol, and then remove the organic solvent in a rotary evaporator at 60°C. After a uniform lipid film is formed on the bottom of the bottle, add 100mL of deionized water was ultrasonically hydrated for 1.5h to obtain cationic liposome Dios-DOPE 1 / 2 containing natural diossterol.

[0031]

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Abstract

The invention relates to the field of cationic liposome small interference RNA (SiRNA) transfection reagents, in particular to cationic liposome containing natural yam sterols, a preparation method of the cationic liposome, and application of the cationic liposome as an SiRNA transfection reagent. The cationic liposome containing natural yam sterols, provided by the invention, is formed by direct ultrasonic hydration after compounding of the cationic liposome containing natural yam sterols and auxiliary lipid dioleoyl phosphatidyl ethanolamine, the preparation method is simple and convenient, and large-scale preparation is easy to realize. Compared with a commercial gene transfection reagent bPEI-25k sold on the market, the cationic liposome as the SiRNA transfection reagent has remarkable lower cytotoxicity, a relatively efficient SiRNA transfection effect, and can be taken as a novel lipoid gene vector functional reagent to be applied to SiRNA transfection.

Description

technical field [0001] The invention relates to the field of synthetic liposome gene delivery functional carrier materials, in particular to a cationic liposome containing natural diossterol, a preparation method and its application as a small molecule interference gene Bcl-2SiRNA transfection reagent. Background technique [0002] In recent years, gene therapy methods based on small interfering RNA (siRNA) technology have aroused great interest. Since siRNA can be directly expressed in the cytoplasm, it has the advantages of rapid onset of action and flexible and controllable time interval of administration. Therefore, the biomedical technology related to gene therapy through siRNA delivery has been developed rapidly. This technology can be used alone or in combination with other treatments to treat a series of gene defect diseases, such as genetic diseases, viral infectious diseases, and Tumor etc. In terms of tumor therapy, the recently studied small interfering RNA is ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K9/127A61K47/28A61K47/24A61K48/00A61P35/00C12N15/88
Inventor 盛瑞隆曹阿民孔令丹史向阳朱晓夏
Owner SHANGHAI INST OF ORGANIC CHEM CHINESE ACAD OF SCI
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