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Tumor vaccine in mice and preparation method thereof

A tumor vaccine and mouse technology, applied in the field of mouse tumor vaccine and its preparation, to achieve the effect of inhibiting tumor growth

Pending Publication Date: 2020-03-27
天津贝罗尼生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] In view of this, the present invention provides a mouse tumor vaccine and its preparation method, which overcomes the inability to activate CD4 in combination with MHC class I molecules and short peptides in the prior art. + The technical defect of T cells, for the first time through the introduction of specific tumor adjuvants, the activation of CD4 + The purpose of T cells is to make mouse tumor vaccines have the function of inhibiting tumor growth; at the same time, it provides new ideas and basis for the development of human tumor vaccines.

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  • Tumor vaccine in mice and preparation method thereof
  • Tumor vaccine in mice and preparation method thereof
  • Tumor vaccine in mice and preparation method thereof

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Embodiment 1

[0043] The embodiment of the present invention discloses a tumor vaccine for mice, wherein the beer yeast shell glucan GP, ​​the adjuvant polyinosinic acid Poly(I:C) and the polypeptide M30 are respectively connected according to the mass ratio of 1:20:10 , to get GP-Poly(I:C)-M30 mouse tumor vaccine.

[0044] Wherein, the preparation method of wine yeast shell glucan GP is as follows:

[0045] (1) Take 5 bags (15g / bag) of Angel high-efficiency active dry yeast and place them in a 1L glass beaker, add 1L deionized water to resuspend the yeast.

[0046] (2) Place the resuspended yeast solution in a centrifuge bottle and balance, centrifuge at 2000rpm for 5min, and discard the supernatant. Repeat the washing twice by adding deionized water again (to remove the additives in the yeast).

[0047] (3) Collect the yeast cells into a beaker, add 1L NaOH (1M / L) and heat and stir at 90°C for 1h on a magnetic stirrer.

[0048] (4) Centrifuge at 5000rpm for 5min to collect the precipit...

Embodiment 2

[0061] The embodiment of the present invention discloses a mouse tumor vaccine, wherein the beer yeast shell glucan GP, ​​the adjuvant polyinosinic acid Poly(I:C) and the polypeptide M30 are respectively connected according to the mass ratio of 1:25:11 , to get GP-Poly(I:C)-M30 mouse tumor vaccine. Among them, the structure of beer yeast shell dextran GP under the electron microscope is as follows: figure 1 shown.

[0062] The preparation method is as follows: (1) Weigh 5mg of GP into a 2ml EP tube, add 2ml of oil phase solution (cyclohexane / Igepal = volume ratio 85:15) into the tube and place it in an ultrasonic cleaner for ultrasonic dispersion for 50min.

[0063] (2) After putting magnets into the tube, add 12.5 μl of 10 mg / ml fluorescently labeled polypeptide M30 solution and 5.5 μl of 10 mg / ml Poly(I:C) solution, and stir overnight on a magnetic stirrer.

[0064] (3) Take 10 μl of 0.5% chitosan solution, add it into 250 μl oil phase solution and mix evenly, then mix it ...

Embodiment 3

[0069] The embodiment of the present invention discloses a mouse tumor vaccine, wherein the beer yeast shell glucan GP, ​​the adjuvant polyinosinic acid Poly(I:C) and the polypeptide M30 are connected according to the mass ratio of 1:22:12 respectively , to get GP-Poly(I:C)-M30 mouse tumor vaccine. Among them, the structure of beer yeast shell dextran GP under the electron microscope is as follows: figure 1 shown.

[0070] The preparation method is as follows: (1) Weigh 5 mg of GP into a 2 ml EP tube, add 5 ml of oil phase solution (cyclohexane / Igepal = volume ratio 85:15) into the tube and place it in an ultrasonic cleaner for ultrasonic dispersion for 50 min.

[0071] (2) After putting magnets into the tube, add 11 μl of 10 mg / ml fluorescently labeled polypeptide M30 solution and 6 μl of 10 mg / ml Poly(I:C) solution, and stir overnight on a magnetic stirrer.

[0072] (3) Take 7 μl of 0.5% chitosan solution, add it to 220 μl oil phase solution and mix evenly, then mix it eve...

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Abstract

The invention discloses a tumor vaccine in mice and a preparation method thereof. The vaccine contains saccharomyces cerevisiae shell glucan GP, adjuvant polyinosinic-polycytidylic acid Poly(I:C) andpolypeptide M30, which are bonded in a mass ratio of 1:(20-25):(10-12) respectively. The tumor vaccine in mice and the preparation method thereof can overcome the technical defect that the combinationof an MHC class I molecule and short peptides cannot activate CD4+T cells in the prior art, can achieve the aim of activating the CD4+T cells by introducing the specific tumor adjuvant for the firsttime, so that the tumor vaccine in mice has a function of inhibiting tumor growth; and a new thought and foundation are provided to researches and development of vaccines for treating human tumors.

Description

technical field [0001] The invention relates to the technical field of vaccine development, in particular to a mouse tumor vaccine and a preparation method thereof. Background technique [0002] At present, research on tumor vaccines is in an intensified state. In 2017, PD-1 therapy covers multiple indications; CAR-T therapy has become another milestone in immunotherapy after being recommended by the FDA committee, but PD-1 The mechanism of action of / PD-L1 drugs is very complex, the beneficiary group is relatively limited, and some patients may also have the risk of drug resistance and accelerated progression; while CAR-T therapy involves genetic engineering, it may also trigger a cytokine storm, and the target Too single, and the effect is not good in solid tumors. For this reason, scholars are trying hard to find new treatment methods to make up for the deficiencies of the above two immunotherapies. The study of tumor neoantigens and the development of personalized tumo...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/00A61K39/39A61P35/00
CPCA61K39/0011A61K39/39A61P35/00A61K2039/55511
Inventor 张伯清贾正虎
Owner 天津贝罗尼生物科技有限公司
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