CN-121987774-A - Composite milk adjuvant, preparation method and application thereof in vaccine

Abstract

The application relates to a compound milk adjuvant, a preparation method and application thereof in vaccines, and belongs to the technical field of medicines. The technical problems to be solved are weak cellular immunity, poor immunogenicity and weak immune regulation. The main point of the technical scheme is that squalene, polyoxyethylene castor oil 35 and poloxamer 407 are used as raw materials to prepare a dairy adjuvant basic formula, a composite dairy adjuvant is prepared on the basis by adding an immunostimulant, physical and chemical properties and safety of the composite dairy adjuvant are characterized, and the synergy of the composite dairy adjuvant on humoral immunity and cellular immunity is further evaluated by matching a recombinant tuberculosis antigen Ag85B immunized mouse, so that data support is provided for the application of the dairy adjuvant in the development of novel vaccines such as tuberculosis vaccines. The composite emulsion adjuvant prepared by the application not only maintains good humoral immunity and induces high-level IgG1 and IgG2a, but also remarkably improves the response level of cellular immunity.

Inventors

• JIAO LEI
• ZHANG GUANGLEI
• WANG YUCHONG
• ZHANG TINGTING
• ZHANG XIAOXIN

Assignees

• 兰州生物制品研究所有限责任公司

Dates

Publication Date

20260508

Application Date

20260210

Claims (13)

1. 1. The composite milk adjuvant is characterized by comprising JC1 milk adjuvant and CpG oligonucleotide, wherein the content of the CpG oligonucleotide is 150-500 mug/mL, and the volume ratio of the JC1 milk adjuvant to the CpG oligonucleotide is 2-4:1-3.
2. 2. The compound milk adjuvant according to claim 1, wherein the CpG oligonucleotide is present in an amount of 300 μg/mL.
3. 3. The compound milk adjuvant according to claim 1, wherein the volume ratio of JC1 milk adjuvant to CpG oligonucleotide is 2:1.
4. 4. The compound milk adjuvant according to claim 1, wherein the JC1 milk adjuvant comprises the following components in percentage by weight: 。
5. 5. The compound milk adjuvant according to claim 4, wherein the JC1 milk adjuvant comprises the following components in percentage by weight: 。
6. 6. the compound milk adjuvant according to claim 4, wherein the mass ratio of squalene, poloxamer 407 and polyoxyethylated castor oil 35 is 8-32:2-4:1.
7. 7. The compound milk adjuvant according to claim 6, wherein the mass ratio of squalene, poloxamer 407 and polyoxyethylated castor oil 35 is 8:2:1.
8. 8. The compound milk adjuvant according to claim 1, wherein the JC1 milk adjuvant is prepared by the following steps: (1) Preparing water phase, namely dissolving poloxamer 407 in buffer solution, heating and stirring to obtain water phase; (2) The preparation of oil phase comprises dissolving polyoxyethylene castor oil 35 in squalene, heating and stirring to obtain oil phase; (3) Pre-emulsifying, namely adding the prepared oil phase into an aqueous phase to perform pre-emulsification to obtain a material 1; (4) Homogenizing and emulsifying under high pressure, namely homogenizing the material 1 for multiple times by using a high-pressure homogenizer.
9. 9. The preparation method is characterized in that CpG oligonucleotide is dissolved in buffer solution, mixed with JC1 emulsion adjuvant in proportion by vortex oscillation, and further emulsified by a dispersing instrument.
10. 10. Use of a compound milk adjuvant according to any one of claims 1 to 8 in the manufacture of a vaccine adjuvant, said use being in the manufacture of a vaccine for eliciting a Th1 and Th2 type immune response.
11. 11. An immunogenic composition comprising a complex milk adjuvant according to any one of claims 1 to 8 and an antigen capable of eliciting an immune response.
12. 12. The immunogenic composition of claim 11, wherein the antigen comprises a mycobacterium tuberculosis antigen.
13. 13. The immunogenic composition of claim 12, wherein the mycobacterium tuberculosis antigen is Ag85B.

Description

Composite milk adjuvant, preparation method and application thereof in vaccine Technical Field The invention belongs to the technical field of medicines, and particularly relates to a composite dairy adjuvant, a preparation method and application thereof in vaccines. Background Vaccines are a special commodity for preventing and treating diseases, and vaccination is currently the most economical and effective measure for controlling bacterial and viral infectious diseases for humans. Along with the development of scientific technology, various novel vaccines are continuously emerging, and compared with traditional vaccines (attenuated live vaccines and inactivated vaccines), the novel vaccines (protein vaccines, nucleic acid vaccines and polypeptide vaccines) have obviously improved safety, but have poorer immunogenicity, and can induce strong humoral immunity and cellular immunity only by being compatible with good vaccine adjuvants, so that higher-level immune protection is achieved. Modern vaccines are increasingly unadjusted to the action of adjuvants, and in particular antigens characterised by genetically engineered products must be supplemented with adjuvants in order to achieve good immune effects. It can be said that a vaccine is a generic term for an antigen and an adjuvant. The immune adjuvant is injected with antigen simultaneously or in advance, can activate the natural immune system of the organism, strengthen the immune response of the organism to the antigen, prolong the vaccine protection time, reduce the dosage of the antigen, promote the presentation capability of mononuclear phagocytes to the antigen and stimulate the proliferation and differentiation of lymphocytes, thereby strengthening the immune response of the organism to the antigen or changing the immune response type. The key role of the novel adjuvant is represented in three layers, namely first, initiation and enhancement of innate immunity. The target dendritic cells and other antigen presenting cells are used for identifying the receptor in a mode, activating a downstream signal path, promoting the secretion of costimulatory molecules and key cytokines, and laying a foundation for activating adaptive immunity. Second, the immune response type is changed, and the immune synergism is precisely exerted. Third, the breadth and persistence of the immune response is extended. In the face of rapidly mutating viruses, some adjuvants can guide the immune system to focus on more conserved epitopes, providing the possibility of developing universal vaccines. The novel adjuvant is thus a key technological fulcrum for achieving vaccines from "effective" to "efficient". While the pathways by which adjuvants activate the immune response can be generalized to intracellular and extracellular receptors, such as TLR, CLR, NLR, RLR, and signal receptors, such as NLRP3, STING, MARVs, and RANKs, as well as downstream signaling pathways and amplification circuits to which these substances are linked. The adjuvant can increase antigen surface area after adsorbing antigen, and make it easy to be phagocytized by Antigen Presenting Cell (APC), and the APC can take in and deliver antigen, and through stimulation of antigen, initial T cell (Tn) can be developed in thymus The maturation migrates into peripheral lymphoid tissues and further differentiates into Th1 cells and Th2 cells. Th1 cells are involved in cellular immunity and delayed hypersensitivity inflammatory reactions. Th2 cells can assist B cell differentiation to produce antibodies, involved in humoral immune responses. Wherein the APC can secrete interleukin-12 (IL-12) to promote differentiation of Tn into Th1 cells. Th1 cells produce natural killer cells (NK), while Th1 cells secrete gamma-interferon (IFN-gamma) and IL-2, respectively, to stimulate continuous activation of macrophages and proliferation of NK cells and killer T cells. The antigen peptide-Major Histocompatibility Complex (MHC) is recognized by T cell receptors to activate T cells, and CD 8+ T cells can differentiate into killer T cells with the ability to specifically kill target cells after activation. MHCII initiates maturation of CD 4+ T cells followed by outward Zhou Qianyi, which in turn cooperates with Dendritic Cells (DCs) to activate CD 8+ T cells. The T cells derived from Tn can secrete IL-4 to trigger B cells to generate immune response to pathogen infection. CD 4+ T cells can also activate CD 8+ T cells by a variety of mechanisms after activation to differentiate into killer T cells that assist B cells in producing antibodies for immune responses. Currently, single-function adjuvants have failed to meet the needs of new vaccine development. The development of future vaccine adjuvants will focus on improving the immunogenicity and protective effects of the vaccine, with the adoption of personalized designs and the use of novel adjuvant technologies to better address the threat of infectious diseases. Milk adjuvants (