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Macrophage targeting carrier system and preparation method thereof

A carrier system and macrophage technology, applied in the field of macrophage targeting carrier system and its preparation, can solve the problems of lack of cell specificity of protamine carrier, increase macrophage uptake, etc., achieve good nuclear localization performance, good Targeted, mild and controllable reaction conditions

Inactive Publication Date: 2015-07-15
CHINA PHARM UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

The mannose ligand in the molecular structure of mannosaccharified protamine can bind to the mannose receptor on the surface of macrophages, and through the sugar recognition mechanism, with the help of the receptor-mediated active targeting of macrophages, macrophages can increase the ability of macrophages to Uptake of nanoparticles to improve the transfection rate of genetic material to solve the problem of lack of cell specificity in existing protamine carriers and improve the immunogenicity of nucleic acid vaccines

Method used

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  • Macrophage targeting carrier system and preparation method thereof
  • Macrophage targeting carrier system and preparation method thereof
  • Macrophage targeting carrier system and preparation method thereof

Examples

Experimental program
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Embodiment 1

[0038] Example 1: Synthesis and Characterization of Mannosylated Protamine

[0039] Add 1-α-formylmethyl-mannopyranoside (50 mg, 0.23 mM) and protamine sulfate (PS, 113.6 mg, 0.02 mM) into 15 mL of distilled water, and stir at room temperature until completely dissolved. Sodium triacetoxyborohydride (474.75 mg, 2.3 mM) was added to the reaction solution, and stirred at room temperature for 48 h (TLC monitored the reaction progress, methanol: triethylamine: water = 3:2:1, v / v / v) . After the reaction was complete, the solution was dialyzed in distilled water at 4° C. for 48 h, and then freeze-dried to remove the solvent to obtain 69 mg of white final product mannosaccharated protamine (MPS), with a yield of 65%.

[0040] The synthesized MPS is carried out to proton nuclear magnetic resonance spectrum analysis, the result shows that the hydroxyl proton peak signal on the mannose ligand appears at 3.6 and 4.1ppm, and the degree of substitution of the calculated mannose is 6.4% (a...

Embodiment 2

[0042] Embodiment 2: Preparation of MPS / pDNA nanoparticles

[0043] The pDNA aqueous solution (100 μg / mL) was added dropwise to an equal volume of the mannosaccharified protamine aqueous solution (56 μg / mL) of Example 1 under stirring, and allowed to stand at room temperature for 30 minutes to form a nanoparticle prepared with an N / P ratio of 0.8. particle.

Embodiment 3

[0044] Embodiment 3: Preparation of MPS / pDNA nanoparticles

[0045] The pDNA aqueous solution (100 μg / mL) was added dropwise to an equal volume of the mannosaccharified protamine aqueous solution (70 μg / mL) of Example 1 under stirring, and allowed to stand at room temperature for 30 minutes to form a nanoparticle prepared with an N / P ratio of 1. particle.

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Abstract

The invention belongs to the field of pharmacy, and discloses a synthesis method of a macrophage targeting carrier system and a preparation method of a gene delivery system. The macrophage targeting carrier system is characterized in that a macrophage targeting carrier is mannosylation protamine; electropositive mannosylation protamine loads electronegative nucleic acid to form a positively charged nano particle; and protamine modified by carubinose is prepared from formyl methyl mannopyranoside and protamine sulfate through a reductive amination reaction. Compared with non-viral gene carrier protamine, mannosylation protamine has a nuclear localization function and a macrophage targeting property, and can improve the gene transfection mediation efficiency of protamine in macrophage. The preparation method is simple, and mature in technology, and has good application prospects.

Description

technical field [0001] The invention belongs to the field of pharmacy, and in particular relates to a macrophage targeting carrier system and its preparation. Background technique [0002] With the development of gene therapy technology, nucleic acid vaccines have received more and more attention because they can induce specific cellular and humoral immune responses against antigens. As we all know, in the process of gene therapy, genes must enter the nucleus to play a role. However, the nuclear membrane is the last barrier with higher specificity and selectivity than the cell membrane. Therefore, it is necessary to utilize a suitable delivery vehicle that protects the gene from nuclease degradation and is able to cross multiple intracellular barriers to reach the nucleus. In recent years, many cationic polymers such as chitosan, polylysine, etc. have been used for gene delivery. However, the transfection efficiency of these cationic polymer-mediated genes still needs to b...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K48/00A61K39/00A61K47/42C12N15/87
Inventor 宗莉曾昭雁戴爽
Owner CHINA PHARM UNIV
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