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Application of crosslinked polyethylenimine as oncoprotein antigen vaccine vector

A polyethyleneimine, tumor protein technology, applied in the direction of antitumor drugs, antibody medical ingredients, medical preparations with non-active ingredients, etc., can solve problems such as application limitations, PEI cannot be degraded, etc.

Active Publication Date: 2015-11-04
SHANGHAI JIAO TONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, hydrophobic PEI cannot be degraded in vivo, which limits its further application.

Method used

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  • Application of crosslinked polyethylenimine as oncoprotein antigen vaccine vector
  • Application of crosslinked polyethylenimine as oncoprotein antigen vaccine vector
  • Application of crosslinked polyethylenimine as oncoprotein antigen vaccine vector

Examples

Experimental program
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Effect test

Embodiment 1

[0038] In this example, the degradable PEI containing imine bonds synthesized by small molecular weight PEI and DTBP was used as the antigen carrier material. For the synthesis method, refer to the literature (M.A.Gosselin, W.Guo, and R.J.Lee, "Efficient gene transfer using reversibly cross- linked low molecular weight polyethyleneimine," Bioconjugate Chemistry, vol. 12, no. 6, pp. 989–994, 2001.).

[0039] The specific steps for the preparation of the composite nanoparticles of the present embodiment are as follows:

[0040] 1. Cross-link PEI and DTBP to form a degradable PEI polymer with a molecular weight of 1000Da containing disulfide bonds (hereinafter referred to as PEI-DS) dissolved in ddH 2 In O, make a 0.2mg / mL stock solution; make OVA a 5mg / mL stock solution with 10mM HEPES buffer at pH 7.4;

[0041] 2. Add the same volume of 5 mg / mL OVA solution to 0.2 mg / mL PEI-DS solution under vortex conditions, and mix them evenly to obtain composite nanoparticles.

[0042] Th...

Embodiment 2

[0058] The synthesis method used in this example is the same as in Example 1, except that the weight average molecular weight of the small molecular weight PEI used in this example is 1000Da, and the weight average molecular weight of the synthesized polymer polymer is 50000Da.

[0059] PEI-DS / OVA composite nanoparticles were prepared according to the same method as in Example 1, and the measured particle diameter was 280nm, and the potential was -16mv. Cytotoxicity assays such as Figure 7 As shown, the antigen cross-presentation experiment was performed as Figure 8 shown. It can be seen from the figure that the antigen cross-presentation effect of the prepared complex nanoparticles is 3 times that of PEI25KDa, and the proportion of living cells is increased by 51%.

[0060] The antitumor effect of composite nanoparticles such as Figure 9 with Figure 10 As shown, the smaller the tumor after 7 days, the better the effect, and the higher the mouse survival rate, the bett...

Embodiment 3

[0062] The synthesis method used in this example is the same as Example 1, except that the weight average molecular weight of the small molecular weight PEI used in this example is 2000Da, and the weight average molecular weight of the synthesized polymer polymer is 100000Da.

[0063] PEI-DS / OVA composite nanoparticles were prepared according to the same method as in Example 1, and the measured particle diameter was 280nm, and the potential was -16mv. Cytotoxicity assays such as Figure 11 As shown, the antigen cross-presentation experiment was performed as Figure 12 shown. It can be seen from the figure that the antigen cross-presentation effect of the prepared complex nanoparticles is 3.7 times higher than that of PEI25KDa, and the corresponding ratio of living cells is increased by 54%.

[0064] The antitumor effect of composite nanoparticles such as Figure 13 with Figure 14 As shown, the smaller the tumor after 7 days, the better the effect, and the higher the mouse...

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Abstract

The present invention provides application of crosslinked polyethylenimine as an oncoprotein antigen vaccine vector, and overcomes the problem that protein antigen after penetration into the body can be degraded easily by enzyme, leading to weak immunogenicity of the vaccine. In an aqueous solution, the polymer material is capable of forming a complex antigen nanoparticle with antigen, in order to achieve loading of tumor protein antigen. The present invention uses RF33.70 as a target cell model, OVA as a model antigen, and C57BL / 6 mice bone marrow-derived dendritic cells as antigen cross presenting cells, and detects the antigen cross-presentation, cytotoxicity and in vivo anti-tumor effect of the complex nanoparticles formed by biodegradable PEI as the antigen vector and the OVA.

Description

technical field [0001] The invention relates to the direction of tumor immunotherapy in the technical field of molecular pharmacy, in particular to the use of a cross-linked polyethyleneimine as a protein antigen vaccine carrier. Background technique [0002] Tumor vaccine is an important means of tumor immunotherapy. Tumor vaccines activate the immune system, enhance the immune system's ability to recognize tumor antigens, improve the immune microenvironment, and induce the body's specific cellular and humoral immune responses, thereby preventing the growth, spread, and recurrence of tumors, and ultimately eradicating or controlling them. tumor. Research and development of vaccines and their carriers and adjuvants that are safe and can effectively stimulate the body's cellular and humoral immunity are the key issues in the development of most vaccines. [0003] The mechanism of action of tumor antigen vaccine is to use tumor antigens to activate the immune system, generat...

Claims

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Application Information

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IPC IPC(8): A61K47/34A61K39/00A61P35/00
Inventor 闫婉颖陈剑徐宇虹曾垂宇商鼎王辉邱瑶
Owner SHANGHAI JIAO TONG UNIV
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