Antigen and adjuvant co-delivery nanometer vaccine applied to liver cancer immunization therapy

An immunotherapy and nano-vaccine technology, applied in the field of biomedicine, can solve the problems of high cost of tumor vaccine raw materials, unfavorable large-scale processing and production, low immunotherapy effect, etc., and achieves easy repeatability of the preparation method, good application prospects, and enhanced immunotherapy. effect of effect

Active Publication Date: 2019-12-20
INST OF BIOMEDICAL ENG CHINESE ACAD OF MEDICAL SCI
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Problems solved by technology

[0008] The present invention designs an antigen and adjuvant co-delivery nano-vaccine for immunotherapy of liver cancer and its preparation method. The technical problem it solves is that the existing tumor vacc

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  • Antigen and adjuvant co-delivery nanometer vaccine applied to liver cancer immunization therapy
  • Antigen and adjuvant co-delivery nanometer vaccine applied to liver cancer immunization therapy
  • Antigen and adjuvant co-delivery nanometer vaccine applied to liver cancer immunization therapy

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preparation example Construction

[0034] The preparation method of antigen and adjuvant co-delivery nano-vaccine applied to the immunotherapy of liver cancer comprises the following steps:

[0035] Step 1, the GPC3 127-136 Aqueous solution and polyethylenimine aqueous solution are mixed to obtain PC3 127-136 Mix solution with polyethyleneimine, then mix sodium alginate aqueous solution and CpG aqueous solution to obtain sodium alginate and CpG mixed solution, wherein GPC3 127-136 The concentration of aqueous solution is 0.5 mg / ml~4 mg / ml; the concentration of polyethyleneimine aqueous solution is 0.5 mg / ml~3 mg / ml; the concentration of CpG aqueous solution is 0.5 mg / ml~4 mg / ml; sodium alginate The concentration of the aqueous solution is 0.5 mg / ml~2 mg / ml; GPC3 127-136 The mass ratio between the aqueous solution and the polyethyleneimine aqueous solution is 1:0.05-0.5; the mass ratio between the sodium alginate aqueous solution and the CpG aqueous solution is 1:0.8-1.5.

[0036] Step 2: Add the mixed soluti...

Embodiment 1

[0038] Preparation of Nanovaccine:

[0039] GPC3 127-136Aqueous solution (1mg / ml, 0.5ml) was mixed with PEI aqueous solution (0.5mg / ml, 0.2ml), then ALG aqueous solution (0.5mg / ml, 0.5ml) was mixed with CpG aqueous solution (0.5mg / ml, 0.5ml), Under the condition of stirring, add the mixed solution of ALG and CpG to GPC3 dropwise 127-136 and PEI mixed solution, stirred for 5 minutes to obtain antigen and adjuvant co-delivery nano-vaccine.

[0040] Such as figure 1 As shown, the particle size distribution of the nano-vaccine obtained in this embodiment is relatively uniform, and the average particle size is 111.9nm.

[0041] Evaluation of nano-vaccine promoting DC maturation:

[0042] Collect the DC of immature on the sixth day, press 10 6 cells per well, spread DC on 24-well plate, and add nano-vaccine, GPC3 after 2 hours of attachment 127-136 The concentration was 10 micrograms per milliliter, and the culture was continued for 48 hours. The cells were collected in a cen...

Embodiment 2

[0045] Preparation of Nanovaccine:

[0046] GPC3 127-136 Mix aqueous solution (2mg / ml, 1ml) and PEI aqueous solution (2mg / ml, 0.3ml), then mix ALG aqueous solution (2mg / ml, 0.1ml) and CpG aqueous solution (2mg / ml, 0.1ml), under stirring conditions, Add ALG and CpG mixed solution dropwise to GPC3 127-136 and PEI mixed solution, stirred for 2 minutes to obtain antigen and adjuvant co-delivery nano-vaccine.

[0047] Such as image 3 As shown, the particle size distribution of the nano-vaccine obtained in this example is relatively uniform, with an average particle size of 173.7nm.

[0048] Evaluation of nano-vaccine promoting DC cytokine secretion:

[0049] Collect the DC of immature on the sixth day, press 10 6 cells per well, spread DC on 24-well plate, and add nano-vaccine, GPC3 after 2 hours of attachment 127-136 The concentration was 10 micrograms per milliliter, and the culture was continued for 48 hours. The cells were collected in a centrifuge tube by gentle blowin...

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Abstract

The invention relates to an antigen and adjuvant co-delivery nanometer vaccine applied to liver cancer immunization therapy. Sodium alginate and polymine are used as carrier materials, liver cancer specific polypeptide antigen phosphatidylinositol proteoglycan 3127-136 peptides(GPC3127-136, AMFKNNYPSL) are used as immunizing antigens, CpG oligodeoxynucleotide is used as an adjuvant, and through static electricity interaction, the antigen and adjuvant co-delivery nanometer vaccine is prepared, wherein the mass ratio of the carrier materials to the immunizing antigens to the adjuvant is 1: (1-10): (0.3-0.8). The nanometer vaccine disclosed by the invention can increase the endocytosis quantity of dendritic cells (dendritic cell, DC) to the antigen and the adjuvant, and through raising costimulatory molecules on the surfaces of the DCs and promoting the secretion of cytokine of TNF-a, IL-6 and the like, arousing organisms to produce effective immune response is facilitated; and besides, the raw materials are cheap and easy to obtain, the preparation method is easy and easily repeated, large-scale processing and producing are easy, the liver cancer immunization therapy effects can be strengthened, and the antigen and adjuvant co-delivery nanometer vaccine has favorable application prospects.

Description

technical field [0001] The invention relates to the field of biomedicine, in particular to an antigen and adjuvant co-delivery nano-vaccine for liver cancer immunotherapy and a preparation method thereof. Background technique [0002] The incidence and mortality of liver cancer are increasing year by year, seriously endangering human health and safety. Immunotherapy for liver cancer is a treatment method to control and eliminate tumors by reactivating the body's normal anti-tumor immune response. [0003] Tumor vaccines play an important role in tumor immunotherapy. Tumor vaccines are processed and processed by antigen-presenting cells in vivo, and present antigen information to T cells, thereby stimulating the body's immune response. Dendritic cells (DC) are the most powerful professional antigen-presenting cells known in vivo, and are the central link in the initiation, regulation and maintenance of immune responses. [0004] Finding effective antigens is crucial for the...

Claims

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Application Information

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IPC IPC(8): A61K39/00A61K39/39A61K47/69A61K47/32A61K47/34A61P35/00
CPCA61K39/001174A61K39/39A61K47/6933A61K47/6939A61P35/00A61K2039/55561A61K2039/622A61K2039/844Y02A50/30
Inventor 张闯年王晓莉孔德领孙洪范裴萌月徐蓉
Owner INST OF BIOMEDICAL ENG CHINESE ACAD OF MEDICAL SCI
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