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Polypeptide-modified liposome, mRNA delivery system and dendritic cell vaccine

A technology of liposomes and auxiliary lipids, applied in the field of biomedicine, can solve problems such as the need to improve transfection efficiency, and achieve the effects of enhancing anti-tumor effect, strong immune function, and improving efficiency

Pending Publication Date: 2021-12-31
SICHUAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In comparison, liposome-mediated mRNA entry into DC does less damage to cells, but the transfection efficiency needs to be improved

Method used

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  • Polypeptide-modified liposome, mRNA delivery system and dendritic cell vaccine
  • Polypeptide-modified liposome, mRNA delivery system and dendritic cell vaccine
  • Polypeptide-modified liposome, mRNA delivery system and dendritic cell vaccine

Examples

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preparation example Construction

[0098] For example, in an example of the present invention, liposomes were further made using DP7-C, DOTAP and cholesterol as raw materials. And provide a kind of typical preparation method based on film dispersion method, the step of this method is:

[0099] a, weigh DOTAP and cholesterol and dissolve in the solvent, and remove the solvent by rotary evaporation under reduced pressure;

[0100] b. The aqueous solution of the hydrophobically modified cationic polypeptide is added to the system for hydration and membrane removal to obtain a liposome suspension;

[0101] c. ultrasonically processing the liposome suspension obtained in b to obtain liposomes.

[0102] The solvent used in step a is generally an organic solvent, and commonly used ones include chloroform, ethanol, methanol, dichloromethane, ethyl acetate and the like.

[0103] During sonication, it is generally necessary to control the temperature of the system not to be too high, preferably below 25°C. For example,...

Embodiment 1

[0153] Embodiment 1, preparation and characterization of liposomes modified by DP7-C

[0154] In this example, we adopted different methods (film dispersion method and ethanol injection method) to prepare DP7-C modified DOTAP / cholesterol cationic liposomes, and prepared DP7-C modified DC-chol / DOPE cations by film dispersion method Liposomes.

[0155] 1. Preparation and characterization of DP7-C modified DOTAP liposomes

[0156] 1) Prepare DOTAP liposomes modified by DP7-C by film dispersion method

[0157] Weigh 20mg DOTAP and 20mg cholesterol, dissolve them in 4ml chloroform, and add them to a 100ml eggplant-shaped bottle. The organic solvent was removed by rotary evaporation in a water bath at 37° C. under reduced pressure, and the rotation speed was set at 50 rpm / min. Add 10ml of 300μg / ml DP7-C aqueous solution into the eggplant-shaped bottle for hydration and film removal, set the temperature at 60°C, and the rotation speed at 75rpm / min, and continue hydration for 30min...

Embodiment 2

[0167] Preparation and characterization of the DOTAP liposome complex mRNA of embodiment two, DP7-C modification

[0168] 1. The method and optimal ratio of DOTAP liposomes modified by DP7-C and mRNA

[0169] The DP7-C modified DOTAP liposome prepared in Example 1 was diluted in an aqueous solution to the use concentration. Add mRNA according to the ratio of liposome:mRNA=2:1, mix gently, and incubate at room temperature for 10 minutes to obtain DP7-C modified DOTAP liposomes. Gel retardation electrophoresis detected the combination of DP7-C modified liposomes and mRNAs of different mass ratios (mRNA encoding neoantigens shown in SEQ ID No.4, the same below), and DP7-C modified liposomes The mass ratios of plastid and mRNA were 0:1, 0.25:1, 0.5:1, 1:1, 2:1, respectively.

[0170] The experimental results show that: the particle diameter of the DOTAP liposome composite mRNA modified by DP7-C is 130.45 ± 9.32nm, and the zeta potential is 34.67 ± 7.45mV ( figure 2 a-b). Tran...

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Abstract

The invention belongs to the field of biological medicines and particularly relates to a liposome modified by a hydrophobic-modified polypeptide, an mRNA delivery system and a dendritic cell vaccine. The technical problem to be solved by the invention is to improve the immune effect of mRNA-based vaccines and mRNA-sensitized DCs vaccines. The technical scheme for solving the technical problem is to provide the liposome modified by a hydrophobic-modified cationic polypeptide. The polypeptide-modified liposome disclosed by the invention can be used for efficiently delivering nucleic acid into cells; and especially, mRNA of a coding antigen can be efficiently delivered into DCs, the efficiency of delivering the mRNA to the DCs by the cationic liposome is obviously increased, the immune effect of the mRNA vaccines and the mRNA-sensitized DC vaccines is enhanced, and good clinical application prospects are achieved.

Description

technical field [0001] The invention belongs to the field of biomedicine, and in particular relates to a liposome modified by a hydrophobized modified polypeptide, an mRNA delivery system and a dendritic cell vaccine. Background technique [0002] In recent years, the application of mRNA to gene therapy, immunotherapy and stem cell biomedicine has attracted extensive attention. Compared with other types of gene therapy and tumor immunotherapy, the advantages of mRNA are mainly reflected in: 1) Unlike viral vectors, mRNA does not need to be integrated into the host genome, so possible insertion mutations and abnormal transcription can be avoided; 2) Compared with plasmid DNA, mRNA has a simple structure, requires fewer elements to be translated into protein, and is biodegradable and only works in a short period of time, thereby reducing the risk of side effects; 3) In the process of individualized immunotherapy Compared with peptide vaccines, mRNA has obvious advantages, suc...

Claims

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Application Information

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IPC IPC(8): A61K9/127A61K47/42A61K47/24A61K47/28A61K39/00A61P35/00
CPCA61K9/1271A61K47/42A61K47/24A61K47/28A61K39/0011A61P35/00A61K2039/5154A61K2039/53
Inventor 杨莉张瑞
Owner SICHUAN UNIV
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