Whole-cell vaccine, and preparation method and application thereof

A whole-cell vaccine and single-cell technology, which can be applied to pharmaceutical formulations, medical preparations containing active ingredients, antibody medical ingredients, etc., can solve the problems of inability to perform specific identification and insufficient whole-cell vaccine antigens, and achieve enhanced application effects And practicality, improve the effect of wide application

Pending Publication Date: 2021-02-09
SHENZHEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The purpose of this application is to provide a whole cell vaccine and its preparation method and application, aiming to solve the problem of insufficient whole cell vaccine antigens and inability to perform specific recognition in the prior art tumor immunotherapy

Method used

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  • Whole-cell vaccine, and preparation method and application thereof
  • Whole-cell vaccine, and preparation method and application thereof
  • Whole-cell vaccine, and preparation method and application thereof

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preparation example Construction

[0047] The second aspect of the embodiment of the present application provides a method for preparing a whole cell vaccine, comprising the following steps:

[0048] S01. According to the whole cell vaccine, tumor cells, TLR7 small molecule agonist molecules and conjugated chain molecules are provided;

[0049] S02. Use the activated ester method to esterify the TLR7 small molecule agonist molecule and the coupled chain molecule to obtain the TLR7 small molecule agonist-coupled chain activated ester molecule or use the TLR7 small molecule agonist with NCS functional groups; tumor cells Perform digestion to obtain tumor single cells;

[0050] S03. Mix TLR7 small molecule agonist-conjugated chain activated ester molecule or TLR7 small molecule agonist with NCS functional group and tumor single cells to obtain coupled tumor single cells;

[0051] S04. Inactivate the coupled tumor single cells to obtain a whole cell vaccine.

[0052] The preparation method of the whole cell vacci...

Embodiment 1

[0070] A kind of whole cell vaccine and preparation method thereof

[0071] The whole cell vaccine includes: tumor cells, TLR7 small molecule agonist molecule compound 5, the structural formula is as follows: And the coupled chain molecule 1-ethyl-(3-dimethylaminopropyl) carbodiimide hydrochloride.

[0072] The preparation method comprises the following steps:

[0073] (1) Provide tumor cells, TLR7 small molecule agonist molecules and conjugated chain molecules;

[0074] (2) Esterifying the TLR7 small-molecule agonist molecule and the conjugated chain molecule by an activated ester method to obtain the TLR7 small-molecule agonist-conjugated chain molecule;

[0075] Specifically: Weigh 0.97mg TLR7 small molecule agonist compound 5, 0.25mg N-hydroxysuccinimide (NHS) and 0.38mg 1-ethyl-(3-dimethylaminopropyl) carbodiimide hydrochloride Salt (EDCI) was dissolved in 166 μL of dimethyl sulfoxide (DMSO), and the reaction was shaken at room temperature. The reaction formula is sho...

Embodiment 2

[0083] A kind of whole cell vaccine and preparation method thereof

[0084] The whole cell vaccine includes: tumor cells, TLR7 small molecule agonist molecule compound 2, the structural formula is as follows: The preparation method comprises the following steps:

[0085] (1) Provide tumor cells, TLR7 small molecule agonist molecule 2;

[0086] (2) The TLR7 small molecule agonist molecule 2 and tumor cells are directly subjected to thiourea reaction by using the isothiocyanate method;

[0087] (3) providing mouse breast cancer 4T1 cells as tumor cells, digesting 4T1 cells with trypsin, washing with PBS twice, and counting to obtain 4T1 tumor single cells;

[0088] (4) Mixing TLR7 small molecule agonist molecule 2 and tumor single cells to obtain coupled tumor single cells;

[0089] Specifically: at a cell concentration of 3 x 10 7 A DMSO solution (10 mM) of TLR7 small-molecule agonist Compound 2 (10 mM) was added to 4T1 tumor single cells per ml to a final concentration of...

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Abstract

The invention relates to the technical field of crossing of immunochemistry and cytobiology, and provides a whole-cell vaccine, and a preparation method and application thereof. The whole-cell vaccinecomprises tumor cells, TLR7 small-molecule agonist molecules and coupling chain molecules, the whole-cell vaccine obtained by coupling the TLR7 small-molecule agonist molecules and the tumor cells through the coupling chain molecules can recognize all antigens on the surfaces of the tumor cells, the vaccine also can be specifically recognize a three-dimensional spatial configuration of the wholetumor cells, the vaccine can be effectively enhance the application effect and practicability in immunotherapy for preventing tumorigenesis, and the wide application of a tumorigenesis immunotherapy method is improved.

Description

technical field [0001] The application belongs to the interdisciplinary technical field of immunochemistry and cell biology, and in particular relates to a whole cell vaccine and its preparation method and application. Background technique [0002] Tumor refers to the neogrowth formed by the proliferation of local tissue cells under the action of various tumorigenic factors, because this neogrowth is mostly in the form of space-occupying blocky protrusions, also known as neogrowth. Studies have found that tumor cells undergo metabolic changes different from normal cells, and tumor cells themselves can adapt to changes in the metabolic environment through the conversion between glycolysis and oxidative phosphorylation (OXPHOS). Due to the three characteristics of unlimited proliferation, transformation and easy transfer of tumor cells, the treatment of tumors has always been a major problem that plagues human health. [0003] There are many treatment methods for tumors, main...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/00A61K39/385A61P35/00
CPCA61K39/0011A61K39/385A61P35/00A61K2039/627A61K2039/60A61K2039/5152
Inventor 靳广毅阮双琛唐黎秦勉王竹林周继
Owner SHENZHEN UNIV
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