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Preparation method and application of hybrid nano-tumor vaccine

A tumor vaccine and nanotechnology, applied in the field of preparation of hybrid nano-tumor vaccine, can solve the problems of low activation efficiency of immune response, increase anti-tumor immune effect, low immunogenicity, etc., achieve no immune response and inflammatory response, and facilitate the Large-scale production, good anti-tumor effect

Pending Publication Date: 2019-06-11
斯潘思生命科技(武汉)有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] Aiming at the technical gap in the prior art, the purpose of the present invention is to provide a preparation method and application of a hybrid nano tumor vaccine, which can increase the cell uptake and cytoplasmic localization of the tumor cell vaccine, and activate the body's immune system to a greater extent. Innate immunity and adaptive immunity, increase anti-tumor immune effect, solve the problems of easy degradation of common tumor antigen vaccines, low immunogenicity, low activation efficiency of immune response, etc.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Embodiment 1: the preparation of hybrid nano tumor vaccine

[0032] The specific preparation method is as follows:

[0033] (1) Dissolve 20 mg of sterile Melittin-RADA32 powder in 1 ml of sterile deionized water to obtain a Melittin-RADA32 solution with a concentration of 20 mg / ml for use.

[0034] (2) Mix 8 mg / ml CpG-ODN and 40 μg / ml lysis aqueous solution in equal proportions.

[0035] (3) The Melittin-RADA32 solution in the step (1) is fully mixed with the solution in the step (2), and an aqueous sodium chloride solution (final concentration is 0.9wt%) is added to the mixed solution to obtain the Melittin-RADA32 / CpG / lysis nano vaccine, wherein the final concentrations of Melittin-RADA32, CpG-ODN, and lysis are 10mg / ml, 2mg / ml and 10μg / ml respectively. Place the mixture at 4°C for 2 hours to form a gel, as figure 1 As shown, the prepared nano-vaccine is a milky white gel under the naked eye.

[0036] The physical and chemical properties of the MLC self-assembled ...

Embodiment 2

[0037] Example 2: Assessing the security of MLC

[0038] Take B16-luc10 cells in the logarithmic growth phase, adjust the cell concentration to 1x10*7 / ml, and inoculate them subcutaneously into the right hind limb of C57 mice, with an inoculation volume of 50 μl, until the tumor grows to 40-50mm 3, the mice were randomly divided into PBS group, Melittin-RADA32 group (MR group), CpG+Lysis group (CL group), MCL group (Example 1MLC self-assembled hybrid nanovaccine), with 5-6 mice in each group. The method of subcutaneous injection at the base of the tail + foot pad was used for injection, and the dose of each injection was 50 μl. Dosing once every other day, a total of 3 administrations. When the tumor volume of the PBS control group reached 1000mm 3 Afterwards, the hearts, livers, spleens, lungs, and kidneys of the mice were taken for HE staining, and the blood of the mice in each group was taken for biochemical detection. image 3 As shown, AST, ALT, BUN, Cr, and UA were al...

Embodiment 3

[0039] Example 3: Evaluation of MCL's ability to activate DC cells

[0040] Bone marrow stem cells were obtained from the femur and tibia of 6-8-week-old male C57 mice, cultured in 1640 medium containing M-CSF for 5 days to obtain immature DC cells, and then planted in 24-well plates, divided into PBS group, Melittin- RADA32 group (MR group), CpG+Lysis group (CL group), MCL group (Example 1MLC self-assembled hybrid nanovaccine) and lipopolysaccharide LPS (positive control), each group takes 40 μ l of the corresponding solution and immature DC cells Co-incubated to stimulate the maturation of DC cells, and after 24 hours, use flow cytometry to detect the activation of CD11c, CD80, and CD86 cells (such as Figure 4 shown), the experimental results showed that the novel hybrid nanovaccine MCL with stable loading could promote the maturation of DCs.

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Abstract

The invention relates to the technical field of biological nano-medicines, and specifically relates to a preparation method and application of a hybrid nano-tumor vaccine. The preparation method comprises the following steps: fully mixing a tumor cell lysis solution, a polypeptide hydrogel aqueous solution and a CpG-ODN aqueous solution, and dropwise adding a sodium chloride aqueous solution intothe mixed solution until the concentration of sodium chloride in the mixed solution is 0.9 wt%; and allowing the mixed solution stand to obtain the hybrid nano-tumor vaccine stably loaded with a tumorkilling polypeptide / CpG / lysis. According to the method disclosed by the invention, a polypeptide hydrogel nano vector is used as a bridge for loading a tumor cell antigen and an immunoactivator; theprepared tumor vaccine has a proper particle size, and can increase cell uptake and cytoplasm positioning of the tumor cell vaccine, activate inherent immunity and adaptive immunity of an organism toa greater extent, enhance an anti-tumor immune effect, and solve the problems that a common tumor antigen vaccine is easy to degrade, low in immunogenicity, low in immune response activation efficiency and the like. The hybrid gel vaccine can be applied to preparation of tumor vaccine drugs.

Description

technical field [0001] The invention relates to the technical field of bio-nano medicine, in particular to a preparation method and application of a hybrid nano-tumor vaccine. Background technique [0002] Tumor immunotherapy is a treatment method that uses immunological methods to stimulate and induce the immune response in the body to produce specific factors that kill tumor cells to kill tumor cells, thereby inhibiting tumor growth. Because malignant melanoma is highly immunogenic and sensitive to immunotherapy, among all malignant tumor treatments, immunotherapy is most widely used in metastatic melanoma. In immunotherapy, tumor cell vaccine is one of the research hotspots in recent years. Tumor cell vaccines are prepared by treating tumor cells with physical, chemical or biological methods (heating, radiation, etc.). This kind of vaccine retains the original immunogenicity and has no tumorigenic effect. Anti-tumor effect dominated by cytotoxic T lymphocyte response. ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/00A61K39/39A61K47/42A61P35/00
Inventor 金红林黄浩周洁黄壁旺洪磊聂晓芬廖婷潘雄华
Owner 斯潘思生命科技(武汉)有限公司
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