MRNA (messenger ribonucleic acid) tumor vaccine for improving DCs (dendritic cells) disability in tumor immune microenvironment as well as preparation method and application of mRNA tumor vaccine

A tumor vaccine and microenvironment technology, applied in the field of mRNA tumor vaccine preparation for improving the incapacitation of tumor immune microenvironment DCs, can solve the problems of complex production process, weak immunogenicity, etc., and achieve improved inhibitory effect, strong application value, The effect of activated CTL activation and proliferation

Pending Publication Date: 2022-04-08
NINGXIA MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0007] The purpose of the present invention is to construct an mRNA tumor vaccine that can improve the inability of DCs in the tumor immune microenvironment. The vaccine can improve or even reverse the immunosuppressive state of DCs in the tumor immu...

Method used

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  • MRNA (messenger ribonucleic acid) tumor vaccine for improving DCs (dendritic cells) disability in tumor immune microenvironment as well as preparation method and application of mRNA tumor vaccine
  • MRNA (messenger ribonucleic acid) tumor vaccine for improving DCs (dendritic cells) disability in tumor immune microenvironment as well as preparation method and application of mRNA tumor vaccine
  • MRNA (messenger ribonucleic acid) tumor vaccine for improving DCs (dendritic cells) disability in tumor immune microenvironment as well as preparation method and application of mRNA tumor vaccine

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0049] Example 1: A thin-film dispersion method was used to prepare a cationic complex mRNA tumor vaccine that can improve the inability of DCs in the tumor immune microenvironment.

[0050] Dissolve DDAB, Chol, DSPE-PEG2000, DSPE-PEG-mannose, and α-Galcer in 3ml chloroform at a molar ratio of 2:2:0.1:0.05:0.3, add to a 100ml round bottom flask, and place on a vortex mixer After vortex mixing for 1 min, 37°C water bath rotary evaporation for 15 min, and drying under reduced pressure to obtain a cationic liposome film. Nitrogen gas was passed into the round bottom flask where the cationic liposome film was formed, and the organic solvent was evaporated to dryness. Add pH 7.4 Tris buffer and hydrate in a water bath at 50°C for 20 minutes to obtain a blank cationic liposome solution (Lip) with a nm size of 80-600 nm. Incubate at room temperature for 10 min at a protamine:mRNA mass ratio of 1:1 to obtain a protamine-HER2 mRNA condensate. Then it was added to Lip and incubated fo...

Embodiment 2

[0052] Example 2: A thin-film dispersion method was used to prepare a cationic complex mRNA tumor vaccine that can improve the dysfunction of DCs in the tumor immune microenvironment.

[0053] Dissolve DC-Chol, Chol, DSPE-PEG2000, DSPE-PEG-mannose, and α-Galcer in 3ml of chloroform at a molar ratio of 2:3:0.3:0.1:0.5, add to a 100ml round bottom flask, and vortex to mix After vortex mixing on a device for 1 min, 37°C water bath rotary evaporation for 15 min, and drying under reduced pressure to obtain a cationic liposome film. Nitrogen gas was passed into the round bottom flask where the cationic liposome film was formed, and the organic solvent was evaporated to dryness. Add pH 7.4 HEPES buffer solution, and hydrate in a water bath at 45°C for 20 minutes to obtain a blank cationic liposome solution (Lip) of 80-600 nm. Incubate at room temperature for 10 min at a protamine:mRNA mass ratio of 1:1 to obtain a protamine-HER2 mRNA condensate. Then it was added to Lip and incubat...

Embodiment 3

[0055] Example 3: A thin-film dispersion method was used to prepare a cationic complex mRNA tumor vaccine that can improve the inability of DCs in the tumor immune microenvironment.

[0056]Dissolve DOTAP, Chol, DSPE-PEG2000, DSPE-PEG-mannose, and α-Galcer in 3ml chloroform at a molar ratio of 2:2:0.3:0.05:0.5, add to a 100ml round bottom flask, and place on a vortex mixer After vortex mixing for 1 min, 37°C water bath rotary evaporation for 20 min, and drying under reduced pressure to obtain a cationic liposome film. Nitrogen gas was passed into the round bottom flask where the cationic liposome film was formed, and the organic solvent was evaporated to dryness. Add pH 7.4 Tris buffer and hydrate in a water bath at 50°C for 20 minutes to obtain a blank cationic liposome solution (Lip) with a nm size of 80-600 nm. Incubate at room temperature for 10 min at a protamine:mRNA mass ratio of 1:1 to obtain a protamine-HER2 mRNA condensate. Then it was added to Lip and incubated fo...

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Abstract

The invention relates to an mRNA (messenger Ribonucleic Acid) tumor vaccine for improving DCs (Dendritic Cancer) disability in a tumor immune microenvironment as well as a preparation method and application thereof. The mRNA tumor vaccine is constructed from a cation compound loaded with alpha-galactosylceramide (alpha-Galcer), protamine and mRNA, the alpha-Galcer is adsorbed to a phospholipid layer of the compound, and a protamine-HER2 mRNA condensation compound is wrapped in an inner core of the compound. The mRNA tumor vaccine constructed by the invention can be used for immunotherapy through subcutaneous injection or intramuscular injection or nasal mucosa, and can stimulate DCs cells to be mature and secrete cell factors. After mice are immunized, the novel mRNA tumor vaccine can effectively promote T cell proliferation and activation, exert congenital and adaptive immune response, improve the inhibition effect of a tumor immune microenvironment and effectively enhance anti-tumor immune response.

Description

technical field [0001] The invention belongs to the technical field of nanomaterials and the field of nucleic acid vaccines, and in particular relates to an mRNA tumor vaccine for improving the incapacity of DCs in the tumor immune microenvironment, a preparation method and an application thereof. Background technique [0002] Malignant tumors seriously threaten human health. Tumor immunotherapy kills tumor cells by activating the body's own immune system, with little or no damage to normal cells. It has great potential in tumor treatment. As an important strategy for tumor immunotherapy, tumor vaccines deliver tumor-associated antigens into antigen-presenting cells (APCs), thereby activating cellular immune responses and inhibiting tumor growth. The mRNA vaccine is to deliver the mRNA small gene fragment encoding the target protein gene to the cell, translate it into protein in the cytoplasm, and then activate APCs to induce the activation and proliferation of antigen-speci...

Claims

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Application Information

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IPC IPC(8): A61K39/00A61K47/42A61K47/18A61K39/39A61P35/00
Inventor 杨建宏马世杰郭珏铄买亚萍乔芳霞李治芳
Owner NINGXIA MEDICAL UNIV
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