Dendritic cell-targeted pH-response type DNA vaccine delivery system and preparation method

A technology of DNA vaccines and dendritic cells, which is applied to medical preparations with non-active ingredients, medical preparations containing active ingredients, and pharmaceutical formulations, etc., which can solve DNA vaccine enzymatic hydrolysis, insufficient antigen presentation, and low transfection efficiency To achieve the effects of mild and controllable reaction conditions, good targeting of dendritic cells, and improved transfection efficiency

Inactive Publication Date: 2017-05-10
CHINA PHARM UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are risks in the delivery process of DNA vaccines such as enzymatic hydro

Method used

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  • Dendritic cell-targeted pH-response type DNA vaccine delivery system and preparation method
  • Dendritic cell-targeted pH-response type DNA vaccine delivery system and preparation method
  • Dendritic cell-targeted pH-response type DNA vaccine delivery system and preparation method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Embodiment 1: the preparation of DNA carrier

[0035] Dissolve 2 mg of DSPE-G2 in 100 μL of chloroform / methanol (v / v=4:1), inject 1 mL of rapidly stirring HEPES buffer (10 mM, pH 7.4), and continue stirring for 6 h to remove the organic solvent.

[0036] The particle size distribution and surface potential of DSPE-G2 nanoparticles were measured with a laser particle size analyzer. The results showed that the particle size of DSPE-G2 nanoparticles was 296.3±19.6nm, the dispersion coefficient was 0.269, and the surface potential was 31.5±6.1mV.

Embodiment 2

[0037] Example 2: Preparation of binary complexes

[0038] The pDNA was dissolved in HEPES buffer (10 mM, pH 7.4) to adjust the concentration to 0.2 mg / mL. The concentration of DSPE-G2 solution was adjusted to 0.1, 0.2, 0.4, 0.5, 0.6, 1, 1.5, 2 mg / mL with HEPES buffer (10 mM, pH 7.4). Mix different concentrations of DSPE-G2 solution and pDNA solution in equal volumes, corresponding to N / P ratios of 1, 2, 4, 5, 6, 10, 15, and 20, vortex for 1 min, and incubate at room temperature for 20 min.

[0039] Gel retardation experiments were performed on binary complexes to test the ability of DSPE-G2 to compress pDNA. Take 10 μL of the binary complex solution and mix it with the loading buffer, add it to the loading hole of 1% agarose gel containing Goldview nucleic acid dye, immerse in the TAE electrophoresis buffer, and electrophoresis at a constant voltage of 90V for 30 min. Gel retardation bands were observed under UV light. The results show that when the N / P ratio is ≥ 4, DSPE-...

Embodiment 3

[0041] Example 3: Preparation method of a dendritic cell-targeted pH-responsive DNA vaccine delivery system

[0042] Mix 50 μL of 0.2 mg / mL pDNA solution with 50 μL of 2 mg / mL DSPE-G2 solution, vortex for 1 min, and incubate at room temperature for 20 min. Take 100 μL of 1 mg / mL PAH-Cit solution and mix it with the binary complex solution, and incubate at room temperature for 30 min. Add 200 μL of ternary complex solution dropwise to 300 μL of 0.5 mg / mL MCS solution under stirring, and keep stirring for 30 min.

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Abstract

The invention belongs to the fields of pharmaceutical preparations and drug delivery of gene vaccine carriers and particularly relates to a dendritic cell-targeted pH-response type DNA vaccine delivery system and a preparation method thereof. According to the delivery system, second-generation lysine substituted distearoyl Phosphoethanolamine (DSPE-G2) is taken as a DNA carrier material, citraconoyl polyacrylamide (PAH-Cit) is taken as a pH-sensitive charge reversion material, mannosylated chitosan (MCS) is taken as a dendritic cell-targeted material, and the materials are organically combined in a layer-by-layer self-assembling manner to form a quaternary compound as the DNA vaccine delivery system, so that the procedural transfection of the DNA vaccine to dendritic cells is realized, and the transfection efficiency is improved. According to the dendritic cell-targeted pH-response type DNA vaccine delivery system, stable composite nanoparticles can be formed to specifically target a specific targete mannose acceptors on the surfaces of the dendritic cells so as to generate charge reversion under a weak acid environment in an endosome-lysosome system, so that the structures of the composite nanoparticles are broken, the release of DNA is promoted, and finally the gene transfection efficiency is remarkably improved. The dendritic cell-targeted pH-response type DNA vaccine delivery system has great utilization potentiality.

Description

technical field [0001] The invention belongs to the field of pharmaceutical preparations and the field of gene vaccine carrier administration, and in particular relates to a dendritic cell-targeted pH-responsive DNA vaccine delivery system and a preparation method. Background technique [0002] Vaccines are an effective way to prevent and treat diseases such as cancer, hepatitis C, AIDS, influenza, and more. Compared with traditional vaccines, DNA vaccines have the advantages of stability, easy modification and preparation, safety, and the ability to form long-term humoral immunity and cellular immunity, and have become a new direction for vaccine development. However, the delivery process of DNA vaccines has risks in terms of enzymatic hydrolysis, low absorption, low transfection efficiency, and insufficient antigen presentation. Therefore, selection of an appropriate DNA vaccine delivery vehicle is crucial for development. [0003] First, targeted delivery of DNA vaccine...

Claims

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Application Information

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IPC IPC(8): A61K39/00A61K47/54A61K47/58A61K47/61
Inventor 姜雷宗莉徐梦露
Owner CHINA PHARM UNIV
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