Application of tauroursodeoxycholic acid or analogue thereof in preparation of immunopotentiator of vaccine

Abstract

The invention provides an application of tauroursodeoxycholic acid or an analogue thereof in preparation of an immunopotentiator of a vaccine, and the immunopotentiator. The application finds that tauroursodeoxycholic acid is an effective active component capable of activating immunity, enhancing antibody titer and shortening antibody generation reaction time, and can be widely applied to various vaccine immune processes. The bile acid metabolite and the derivative of the bile acid metabolite, which are analogues of tauroursodeoxycholic acid, also have immunological enhancement activity according to structural similarity.

Description

Technical Field

[0001] The present application belongs to the field of immunotechnology, and specifically, relates to the use of tauroursodeoxycholic acid or its analogs in the preparation of an immunopotentiator for a vaccine, and an immunopotentiator for a vaccine. Background Art

[0002] Tauroursodeoxycholic acid (TUDCA) is a secondary bile acid formed by the metabolism of primary bile acid to ursodeoxycholic acid (UDCA) and then combined with taurine. TUDCA cannot be directly synthesized in the human body. Endogenous TUDCA is produced by the metabolism of specific microorganisms colonizing the intestine. TUDCA is a key regulator of intestinal barrier integrity. At the same time, as an endogenous signaling molecule, it is crucial for lipid metabolism and immune regulation. Summary of the invention

[0003] The purpose of the present application is to provide the use of tauroursodeoxycholic acid or its analogs in the preparation of an immunopotentiator for a vaccine, and an immunopotentiator for a vaccine.

[0004] Specifically, this application involves the following aspects:

[0005] 1. Use of tauroursodeoxycholic acid or its analogs in preparing an immunopotentiator for a vaccine.

[0006] 2. The use according to item 1, wherein the analog of tauroursodeoxycholic acid is selected from the following:

[0007] Glycochenodeoxycholic acid, glycohyodeoxycholic acid, glycocholic acid, tallow-α-mucosaic acid, glycohyodeoxycholic acid, 7-ketocholic acid, 3-sulfate taurochenodeoxycholic acid, 3β-hydroxy-δ5-cholenic acid, allolithocholic acid, lithocholic acid, deoxycholic acid, chenodeoxycholic acid, hyodeoxycholic acid, ursodeoxycholic acid, isoursodeoxycholic acid, cholic acid, hyodeoxycholic acid, β-mucosaic acid, α-mucosaic acid, 3β-cholic acid, ursocholic acid, glycolithocholic acid, taurolithocholic acid, glycodeoxycholic acid, glycoursodeoxycholic acid, taurodeoxycholic acid, taurochenodeoxycholic acid, taurocholic acid, tallow-β-mucosaic acid, orthocholic acid, isolithocholic acid, dehydrolithocholic acid Acid, 12-ketodeoxycholic acid, omega-muricolic acid, murodeoxycholic acid, isodeoxycholic acid, taurohyocholic acid, 3-sulfatedeoxycholic acid, 3-sulfateglycochenodeoxycholic acid, 3-sulfateglycocholic acid, 3-sulfateglycodeoxylithocholic acid, 3-sulfateglycolithocholic acid, 3-sulfatelithocholic acid, 3-sulfatetaurodeoxycholic acid, 3-sulfatetaurolithocholic acid, 3-sulfatetauroursodeoxycholic acid, orthodeoxycholic acid, 24-norursodeoxycholic acid, 7-ketochenodeoxycholic acid, dehydrocholic acid, 12-ketocholic acid, 3-oxodeoxycholic acid, isochenodeoxycholic acid, isodeoxycholic acid, 3-sulfatechenodeoxycholic acid, 3-oxochenodeoxycholic acid.

[0008] 3. The use according to item 1 or 2, wherein tauroursodeoxycholic acid or an analog thereof enhances the titer of antibodies after vaccination and / or increases the duration of vaccine response.

[0009] 4. The use according to claim 3, wherein administration of tauroursodeoxycholic acid or an analog thereof increases follicular helper T cells, plasma cells, or memory cells in the vaccinated subject.

[0010] 5. The use according to item 4, wherein the subject is a mammal.

[0011] 6. The use according to any one of items 1 to 5, wherein tauroursodeoxycholic acid or an analog thereof is administered prior to vaccination.

[0012] 7. The use according to any one of items 1-6, wherein the vaccine is a rabies vaccine, an influenza vaccine, a hepatitis B vaccine, a hepatitis A vaccine, a hepatitis C vaccine, a hand, foot and mouth disease vaccine, or a HPV vaccine.

[0013] 8. The use according to any one of items 1 to 7, wherein the vaccine is an inactivated vaccine, a live attenuated vaccine, a protein vaccine, a DNA vaccine or a polypeptide vaccine.

[0014] 9. An immunopotentiator for vaccines, comprising tauroursodeoxycholic acid or its analogs.

[0015] 10. The immunopotentiator according to item 9, wherein the analog of tauroursodeoxycholic acid is selected from the following:

[0016] Glycochenodeoxycholic acid, glycohyodeoxycholic acid, glycocholic acid, tallow-α-mucosaic acid, glycohyodeoxycholic acid, 7-ketocholic acid, 3-sulfate taurochenodeoxycholic acid, 3β-hydroxy-δ5-cholenic acid, allolithocholic acid, lithocholic acid, deoxycholic acid, chenodeoxycholic acid, hyodeoxycholic acid, ursodeoxycholic acid, isoursodeoxycholic acid, cholic acid, hyodeoxycholic acid, β-mucosaic acid, α-mucosaic acid, 3β-cholic acid, ursocholic acid, glycolithocholic acid, taurolithocholic acid, glycodeoxycholic acid, glycoursodeoxycholic acid, taurodeoxycholic acid, taurochenodeoxycholic acid, taurocholic acid, tallow-β-mucosaic acid, orthocholic acid, isolithocholic acid, dehydrolithocholic acid Acid, 12-ketodeoxycholic acid, omega-muricolic acid, murodeoxycholic acid, isodeoxycholic acid, taurohyocholic acid, 3-sulfatedeoxycholic acid, 3-sulfateglycochenodeoxycholic acid, 3-sulfateglycocholic acid, 3-sulfateglycodeoxylithocholic acid, 3-sulfateglycolithocholic acid, 3-sulfatelithocholic acid, 3-sulfatetaurodeoxycholic acid, 3-sulfatetaurolithocholic acid, 3-sulfatetauroursodeoxycholic acid, orthodeoxycholic acid, 24-norursodeoxycholic acid, 7-ketochenodeoxycholic acid, dehydrocholic acid, 12-ketocholic acid, 3-oxodeoxycholic acid, isochenodeoxycholic acid, isodeoxycholic acid, 3-sulfatechenodeoxycholic acid, 3-oxochenodeoxycholic acid.

[0017] 11. The immunopotentiator according to item 9 or 10, wherein the immunopotentiator enhances the titer of antibodies after vaccination and / or increases the vaccine response time.

[0018] 12. The immunopotentiator according to claim 11, wherein administration of the immunopotentiator increases follicular helper T cells, plasma cells, or memory cells in the vaccinated subject.

[0019] 13. The immunopotentiator according to claim 12, wherein the subject is a mammal.

[0020] 14. The immunopotentiator according to any one of items 9 to 13, wherein the immunopotentiator is administered before vaccination.

[0021] 15. An immunopotentiator according to any one of items 9 to 14, wherein the vaccine is a rabies vaccine, an influenza vaccine, a hepatitis B vaccine, a hepatitis A vaccine, a hepatitis C vaccine, a hand, foot and mouth disease vaccine, or an HPV vaccine.

[0022] 16. An immunopotentiator according to any one of items 9 to 15, wherein the vaccine is an inactivated vaccine, a live attenuated vaccine, a protein vaccine, a DNA vaccine or a polypeptide vaccine.

[0023] 17. The immunopotentiator according to any one of items 9 to 16, wherein the immunopotentiator further comprises a pharmaceutically acceptable carrier.

[0024] Effects of the Invention

[0025] The present application found that TUDCA is an effective active ingredient that can activate immunity, enhance antibody titer, and shorten the reaction time of antibody generation, and can be widely used in a variety of vaccine immunization processes. In addition, TUDCA analogs - bile acid metabolites and their derivatives also have immune enhancement activity based on structural similarity. BRIEF DESCRIPTION OF THE DRAWINGS

[0026] Figure 1 Shows untargeted metabolomics identification strategy;

[0027] Figure 2 Display the analysis results of PLS-DA;

[0028] Figure 3 Significantly differential metabolites showing up / down regulation;

[0029] Figure 4 Display the weight values ​​of main metabolite variables;

[0030] Figure 5 Display the animal experiment design plan;

[0031] Figure 6 Display the relative antibody titer test results of rabies vaccine;

[0032] Figure 7 Flow cytometric analysis results showing mouse spleen cell populations;

[0033] Figure 8 Displays the results of OVA relative antibody titer determination;

[0034] Fig. 9 Predictive model showing TUDCA binding to PPARγ;

[0035] Fig.10 The results of the TUDCA-PPAR-γ binding assay are shown;

[0036] Fig.11 The results of the assay showed the binding capacity of TUDCA-like bile acid metabolites to PPAR-γ;

[0037] Fig.12 Representative candidate molecular structures are shown;

[0038] Fig.13 Prediction models showing representative candidate molecules binding to PPARγ. DETAILED DESCRIPTION

[0039] The present application is further described below in conjunction with examples. It should be understood that the examples are only used to further describe and illustrate the present application and are not used to limit the present application.

[0040] Unless otherwise defined, the technical and scientific terms in this specification have the same meaning as those generally understood by those skilled in the art. Although similar or identical methods and materials as described herein may be used in experiments or practical applications, materials and methods are described herein below. In the event of a conflict, the present specification including the definitions therein shall prevail. In addition, materials, methods and examples are provided for illustration only and are not restrictive. The present application is further described below in conjunction with specific embodiments, but is not intended to limit the scope of the present application.

[0041] As used herein, the term "vaccine" refers to any preparation of antigenic or immunogenic substance suitable for stimulating active immunity in animals or humans.

[0042] As used herein, the term "antigen" refers to any substance that, when introduced into an immunocompetent human or animal, stimulates a humoral and / or cell-mediated immune response. The antigen may be a pure substance, a mixture of substances, or particulate matter (including cells, cell fragments, or cell-derived fragments) or a live (usually attenuated) organism or virus. Examples of suitable antigens include, but are not limited to, proteins, glycoproteins, lipoproteins, peptides, carbohydrates / polysaccharides, lipopolysaccharides, toxins, viruses, bacteria, fungi, and parasites. Antigens may be natural (naturally expressed or produced), synthetic, or derived by recombinant DNA methodology familiar to those skilled in the art.

[0043] As used herein, the term "immunoenhancer" refers to a substance that can improve the immune function of an animal body. It is mainly used to enhance the body's anti-tumor effect, anti-infection ability, and correct immune deficiency. It can activate one or more immune active cells, enhance the body's specific and nonspecific immune function, and restore the low immune function to normal, or have an adjuvant effect, enhance the immunogenicity of the antigen used with it, and accelerate the induction of immune response; or replace the lack of immune active components in the body to produce an immune replacement effect; or produce a two-way regulatory effect on the body's immune function, so that excessive or low immune function tends to normal.

[0044] As used herein, the term "subject" refers to a mammal, which can be a human (i.e., a male or female of any age, e.g., a pediatric subject (e.g., an infant, child, or adolescent) or an adult subject (e.g., a young, middle-aged, or elderly person)) or a non-human animal. In some embodiments, the subject is a human.

[0045] As used herein, the term "administering" refers to implanting, absorbing, ingesting, injecting, inhaling or otherwise introducing a drug or agent into or onto a subject.

[0046] As used herein, the term "pharmaceutically acceptable" refers to compounds, materials, compositions and / or dosage forms that are suitable for contact with human or animal tissues without excessive toxicity, irritation, allergic reactions, other problems or complications, and have a reasonable benefit / risk ratio. In some embodiments, pharmaceutically acceptable compounds, materials, compositions and / or dosage forms refer to those compounds, materials, compositions and / or dosage forms approved by regulatory agencies (such as the U.S. Food and Drug Administration, China National Drug Administration, European Medicines Agency) or listed in recognized pharmacopoeias (such as the U.S. Pharmacopoeia, Chinese Pharmacopoeia, European Pharmacopoeia) for animals, especially humans.

[0047] As used herein, the term "pharmaceutically acceptable carrier" refers to a pharmaceutically acceptable material, composition or vehicle involved in carrying or transporting the tauroursodeoxycholic acid or its analogs of the present application from one location, body fluid, tissue, organ (internal or external) or body part to another location, body fluid, tissue, organ or body part, such as a liquid or solid filler, diluent, excipient, solvent or encapsulating material. A pharmaceutically acceptable carrier may be a carrier, diluent, excipient or other material that can be used to contact animal tissue without excessive toxicity or adverse reactions. Exemplary pharmaceutically acceptable carriers include, but are not limited to, sugars, starches, cellulose, malt, tragacanth gum, gelatin, Ringer's solution, alginic acid, isotonic saline, buffers, and the like.

[0048] In one aspect, the present application provides the use of tauroursodeoxycholic acid or its analogs in the preparation of an immunopotentiator for a vaccine.

[0049] Among them, the analogs of sulfoursodeoxycholic acid can be various types of bile acid metabolites, which have similar immune enhancement functions as tauroursodeoxycholic acid.

[0050] In a specific embodiment, the analog of tauroursodeoxycholic acid is selected from the following: glycochenodeoxycholic acid, glycohyodeoxycholic acid, glycocholic acid, tallow-α-murocolic acid, glycohyocholic acid, 7-ketocholic acid, 3-sulfate taurochenodeoxycholic acid, 3β-hydroxy-δ5-cholenic acid, allolithocholic acid, lithocholic acid, deoxycholic acid, chenodeoxycholic acid, hyodeoxycholic acid, ursodeoxycholic acid, isoursodeoxycholic acid, cholic acid, hyocholic acid, β-murocolic acid, α-murocolic acid, 3β-cholic acid, ursolic acid, glycolithocholic acid, taurolithocholic acid, glycodeoxycholic acid, glycoursodeoxycholic acid, taurodeoxycholic acid, taurochenodeoxycholic acid, taurocholic acid, tallow-β-murocolic acid Acid, orthocholic acid, isolithocholic acid, dehydrolithocholic acid, 12-ketodeoxycholic acid, omega-muric acid, murodeoxycholic acid, isodeoxycholic acid, taurohyocholic acid, 3-sulfatedeoxycholic acid, 3-sulfateglycochenodeoxycholic acid, 3-sulfateglycocholic acid, 3-sulfateglycodeoxylithocholic acid, 3-sulfateglycolithocholic acid, 3-sulfatelithocholic acid, 3-sulfatetaurodeoxycholic acid, 3-sulfatetaurolithocholic acid, 3-sulfatetauroursodeoxycholic acid, orthodeoxycholic acid, 24-norursodeoxycholic acid, 7-ketochenodeoxycholic acid, dehydrocholic acid, 12-ketocholic acid, 3-oxodeoxycholic acid, isochenodeoxycholic acid, isodeoxycholic acid, 3-sulfatechenodeoxycholic acid, 3-oxochenodeoxycholic acid.

[0051] In the present application, tauroursodeoxycholic acid or its analogs have an immunoenhancing function for vaccines, for example, tauroursodeoxycholic acid or its analogs can enhance the titer of antibodies after vaccination and / or increase the vaccine response time. In a specific embodiment, the administration of tauroursodeoxycholic acid or its analogs increases the follicular helper T cells involved in humoral immunity, plasma cells secreting antibodies, or memory cells of the vaccinated subject.

[0052] In the present application, there is no restriction on the order of tauroursodeoxycholic acid or its analogs and vaccination, for example, tauroursodeoxycholic acid or its analogs can be used before vaccination, administered simultaneously with the vaccine, or administered after vaccination. In a specific embodiment, tauroursodeoxycholic acid or its analogs are administered before vaccination.

[0053] In the present application, the vaccine may be any vaccine known in the art, such as rabies vaccine, influenza vaccine, hepatitis B vaccine, hepatitis A vaccine, hepatitis C vaccine, hand, foot and mouth disease vaccine, HPV vaccine, etc.

[0054] The type of vaccine is also not limited, for example, it can be an inactivated vaccine, a live attenuated vaccine, a protein vaccine, a DNA vaccine or a polypeptide vaccine, etc. In a specific embodiment, the vaccine is an inactivated vaccine.

[0055] Tauroursodeoxycholic acid or its analogs can be delivered to the subject by appropriate routes, including but not limited to, oral routes, injection routes (such as intravenous injection, intramuscular injection, subcutaneous injection, intradermal injection, intracardiac injection, intrathecal injection, intrapleural injection, intraperitoneal injection, etc.), mucosal routes (such as intranasal administration, oral administration, etc.), sublingual routes, rectal routes, transdermal routes, intraocular routes, and pulmonary routes.

[0056] Depending on the desired route of administration, tauroursodeoxycholic acid or its analogs can be formulated as tablets, capsules, pills, dragees, powders, granules, cachets, lozenges, suppositories, suspensions, emulsions, syrups, aerosols (as a solid or in a liquid medium), sprays, ointments, pastes, patches, creams, lotions, gels, inhalants, and the like.

[0057] It will be appreciated by those skilled in the art that the dosage and frequency of administration of tauroursodeoxycholic acid or its analogs may vary depending on the subject's age, weight, or individual response to the vaccine and the specific administration selected.

[0058] In a specific embodiment, tauroursodeoxycholic acid or an analog thereof is administered orally at a dose of 90 mg / kg subject / day to 550 mg / kg subject / day for 14-17 days.

[0059] On the other hand, the present application also provides an immunopotentiator for a vaccine, comprising tauroursodeoxycholic acid or an analog thereof.

[0060] In a specific embodiment, the analog of tauroursodeoxycholic acid is selected from the following: glycochenodeoxycholic acid, glycohyodeoxycholic acid, glycocholic acid, tallow-α-murocolic acid, glycohyocholic acid, 7-ketocholic acid, 3-sulfate taurochenodeoxycholic acid, 3β-hydroxy-δ5-cholenic acid, allolithocholic acid, lithocholic acid, deoxycholic acid, chenodeoxycholic acid, hyodeoxycholic acid, ursodeoxycholic acid, isoursodeoxycholic acid, cholic acid, hyocholic acid, β-murocolic acid, α-murocolic acid, 3β-cholic acid, ursolic acid, glycolithocholic acid, taurolithocholic acid, glycodeoxycholic acid, glycoursodeoxycholic acid, taurodeoxycholic acid, taurochenodeoxycholic acid, taurocholic acid, tallow-β-murocolic acid Acid, orthocholic acid, isolithocholic acid, dehydrolithocholic acid, 12-ketodeoxycholic acid, omega-muric acid, murodeoxycholic acid, isodeoxycholic acid, taurohyocholic acid, 3-sulfatedeoxycholic acid, 3-sulfateglycochenodeoxycholic acid, 3-sulfateglycocholic acid, 3-sulfateglycodeoxylithocholic acid, 3-sulfateglycolithocholic acid, 3-sulfatelithocholic acid, 3-sulfatetaurodeoxycholic acid, 3-sulfatetaurolithocholic acid, 3-sulfatetauroursodeoxycholic acid, orthodeoxycholic acid, 24-norursodeoxycholic acid, 7-ketochenodeoxycholic acid, dehydrocholic acid, 12-ketocholic acid, 3-oxodeoxycholic acid, isochenodeoxycholic acid, isodeoxycholic acid, 3-sulfatechenodeoxycholic acid, 3-oxochenodeoxycholic acid.

[0061] In the present application, the immunopotentiator can enhance the titer of antibodies after vaccination and / or increase the vaccine response time. In a specific embodiment, the immunopotentiator is administered to increase the follicular helper T cells, antibody-secreting plasma cells, or memory cells involved in humoral immunity of the vaccinated subject.

[0062] In the present application, there is no restriction on the order of immunopotentiator and vaccination, for example, immunopotentiator can be used before vaccination, administered simultaneously with the vaccine, or administered after vaccination. In a specific embodiment, the immunopotentiator is administered before vaccination.

[0063] The immunopotentiator can be delivered to the subject by an appropriate route, including but not limited to, oral route, injection route (such as intravenous injection, intramuscular injection, subcutaneous injection, intradermal injection, intracardiac injection, intrathecal injection, intrapleural injection, intraperitoneal injection, etc.), mucosal route (such as intranasal administration, oral administration, etc.), sublingual route, rectal route, transdermal route, intraocular route, pulmonary route.

[0064] Depending on the desired route of administration, the immunopotentiator can be formulated as tablets, capsules, pills, dragees, powders, granules, cachets, lozenges, suppositories, suspensions, emulsions, syrups, aerosols (as solid or in a liquid medium), sprays, ointments, pastes, patches, creams, lotions, gels, inhalants, and the like.

[0065] In a specific embodiment, the immunopotentiator further comprises a pharmaceutically acceptable carrier. Those skilled in the art can select the required carrier according to the route of use, dosage form, etc. of the immunopotentiator.

[0066] Example

[0067] Example 1

[0068] Preliminary studies of this application have found that the use of a specific nutritional supplement, arachidonic acid (ARA), for 14-17 days of intervention significantly enhanced the titer of specific antibodies after vaccination. Peripheral blood samples were collected from subjects, and non-targeted metabolomics identification of serum samples was performed using ultra-high performance liquid chromatography-high resolution mass spectrometry (UPLC-HRMS). Three complementary analytical detection methods were used to collect data in positive and negative ion modes for the same sample. The overall identification route, such as Figure 1 shown.

[0069] After partial least-squares discrimination analysis (PLS-DA) was used to process the metabolomics data, it was shown that there were significant differences in the metabolic profiles between the intervention group and the control group ( Figure 2 ). This suggests that some metabolites after nutritional intervention are closely related to the enhancement of antibody titer. Univariate statistical analysis combined with multivariate statistical analysis and fold change values ​​were used to further screen the differential metabolites. The results showed that the metabolite TUDCA in the intervention group increased significantly ( Figure 3-4 ), which suggests that TUDCA may be a key factor in producing immune-enhancing effects.

[0070] Example 2

[0071] In order to verify that TUDCA can be used as an active ingredient to effectively enhance antibody titer and has considerable safety, this study conducted animal experiments. The animal experiment protocol is as follows Figure 5 shown.

[0072] Specifically, 6-week-old C57BL / 6 mice were randomly divided into 2 groups, and were given pure water / TUDCA (100 mg / kg / day) by gavage from day 3 to day 19. The mice were given the first rabies vaccine on day 6 and the second rabies vaccine on day 20. On days 13, 19, and 26, 4 to 6 mice were randomly selected from each group to collect biological samples, including serum and spleen. After the serum was obtained, specific antibody detection was performed immediately. The results are as follows: Figure 6 As shown. * indicates p < 0.05, which is statistically significant.

[0073] The results showed that 7 days after immunization, the mice receiving TUDCA intervention had shown a trend of increased antibody titers; on the 19th and 26th days, the antibody titers of mice receiving TUDCA intervention increased significantly, which was statistically significant.

[0074] The spleen was further taken to isolate lymphocytes, and flow cytometry analysis was performed after immunostaining. The results were as follows Figure 7 The results showed that on the 19th day, the follicular helper T cell subsets involved in humoral immunity increased significantly in mice treated with TUDCA; on the 26th day, the plasma cells that secrete antibodies increased significantly in mice treated with TUDCA; at the same time, the memory B cell subsets also increased significantly, p < 0.05, which was statistically significant.

[0075] Example 3

[0076] In order to verify the universality of TUDCA intervention in enhancing the antibody titer after immunization, this example used the model antigen chicken ovalbumin (OVA) as an immunostimulant for the experiment. Six-week-old C57BL / 6 mice (N=16) were randomly divided into a control group and an intervention group. They were given pure water or TUDCA (100 mg / kg / day) by gavage from the 3rd to the 19th day, and the first OVA immunization was performed on the 6th day, and the second immunization was performed on the 20th day. On the 13th day, the 19th day, and the 26th day, tail tip blood samples were collected from the control group and the intervention group mice, and after the serum was obtained, specific antibody detection was performed immediately. The results are as follows Figure 8 As shown, where * represents p < 0.05.

[0077] The test results showed that 7 days after immunization, the antibody titers of mice receiving TUDCA intervention showed a trend of increasing; on the 19th and 26th days, the antibody titers of mice receiving TUDCA intervention were significantly increased compared with the control group, and the increase was statistically significant.

[0078] Example 4

[0079] Regarding the molecular mechanism of TUDCA's antibody enhancement effect, this example predicts potential targets, and the predicted target is peroxisome proliferator activated receptor gamma (PPAR-γ), and its predicted binding energy is -8.9 kcal / mol. Fig. 9 The predicted binding model is shown. Subsequently, the present embodiment expressed and purified PPAR-γ protein in vitro, and the binding force was measured. The results of isothermal titration calorimetry are shown in FIG. Fig.10 shown.

[0080] The results showed that TUDCA could bind to PPAR-γ, K D The value was 34 μM. This suggests that PPAR-γ can serve as a potential target of TUDCA to conduct signal transduction and activate downstream pathways.

[0081] This example conducted a structure-based high-throughput virtual screening of bile acid metabolites similar to TUDCA. After pre-processing the three-dimensional structure of the metabolite molecules and the target protein PPAR-γ, the AutoDock Vina molecular docking platform was used to screen 56 bile acid metabolite molecule candidates, all of which had antibody enhancement functions similar to TUDCA, including various primary and secondary bile acids. The results are shown in Table 1:

[0082] Table 1

[0083]

[0084]

[0085]

[0086] The binding energy values ​​in the table represent the predicted affinity of the molecule to PPAR-γ. The lower the binding energy, the stronger the interaction and better binding ability between the molecule and the target protein. The lower the binding energy, the higher the stability of the complex system after binding.

[0087] Based on the above virtual screening, this example selects representative structural candidate molecules with the highest (3-sulfate taurochenodeoxycholic acid) and lowest (lithocholic acid) binding energy, as well as isochenodeoxycholic acid and allolithocholic acid, and uses tauroursodeoxycholic acid (TUDCA) as a positive control to verify the interaction between the candidate small molecules and the protein receptor PPAR-γ using surface plasmon resonance technology (SPR). The results are as follows: Fig.11 shown.

[0088] The results showed that the affinity of tauroursodeoxycholic acid (TUDCA) to PPAR-γ was 4 μM, and the affinity of 3-sulfated taurochenodeoxycholic acid, lithocholic acid, isochenodeoxycholic acid, and allolithocholic acid to PPAR-γ were 11 μM, 51 μM, 1.67 μM, and 3.61 μM, respectively. This indicates that these candidate molecules can interact with PPAR-γ intermolecularly and have similar antibody enhancement potential as TUDCA.

[0089] Some representative candidate molecular structures and prediction models for binding to PPAR-γ are as follows: Fig.12 , Fig.13 shown. Fig.13 The metabolites shown are enclosed in a hydrophobic pocket inside PPAR-γ and interact with the side chain groups of PPAR-γ amino acids.

[0090] In summary, this application verifies that TUDCA is an effective active ingredient that can activate immunity and enhance antibody titer, and can be widely used in a variety of vaccine immunization processes. In addition, TUDCA analogs - bile acid metabolites and their derivatives also have immune enhancement activity based on structural similarity.

Claims

1. Use of tauroursodeoxycholic acid or its analogs in preparing an immunopotentiator for a vaccine.

2. The use according to claim 1, wherein the analog of tauroursodeoxycholic acid is selected from the following: Glycochenodeoxycholic acid, glycohyodeoxycholic acid, glycocholic acid, tallow-α-mucosaic acid, glycohyodeoxycholic acid, 7-ketocholic acid, 3-sulfate taurochenodeoxycholic acid, 3β-hydroxy-δ5-cholenic acid, allolithocholic acid, lithocholic acid, deoxycholic acid, chenodeoxycholic acid, hyodeoxycholic acid, ursodeoxycholic acid, isoursodeoxycholic acid, cholic acid, hyodeoxycholic acid, β-mucosaic acid, α-mucosaic acid, 3β-cholic acid, ursocholic acid, glycolithocholic acid, taurolithocholic acid, glycodeoxycholic acid, glycoursodeoxycholic acid, taurodeoxycholic acid, taurochenodeoxycholic acid, taurocholic acid, tallow-β-mucosaic acid, orthocholic acid, isolithocholic acid, dehydrolithocholic acid Acid, 12-ketodeoxycholic acid, omega-muricolic acid, murodeoxycholic acid, isodeoxycholic acid, taurohyocholic acid, 3-sulfatedeoxycholic acid, 3-sulfateglycochenodeoxycholic acid, 3-sulfateglycocholic acid, 3-sulfateglycodeoxylithocholic acid, 3-sulfateglycolithocholic acid, 3-sulfatelithocholic acid, 3-sulfatetaurodeoxycholic acid, 3-sulfatetaurolithocholic acid, 3-sulfatetauroursodeoxycholic acid, orthodeoxycholic acid, 24-norursodeoxycholic acid, 7-ketochenodeoxycholic acid, dehydrocholic acid, 12-ketocholic acid, 3-oxodeoxycholic acid, isochenodeoxycholic acid, isodeoxycholic acid, 3-sulfatechenodeoxycholic acid, 3-oxochenodeoxycholic acid.

3. The use according to claim 1 or 2, wherein tauroursodeoxycholic acid or its analogues enhance the titer of antibodies after vaccination and / or increase the vaccine response time.

4. The use according to claim 3, wherein administration of tauroursodeoxycholic acid or an analog thereof increases follicular helper T cells, plasma cells, or memory cells in the vaccinated subject.

5. The use according to claim 4, wherein the subject is a mammal.

6. The use according to any one of claims 1 to 5, wherein tauroursodeoxycholic acid or an analog thereof is administered prior to vaccination.

7. The use according to any one of claims 1-6, wherein the vaccine is a rabies vaccine, an influenza vaccine, a hepatitis B vaccine, a hepatitis A vaccine, a hepatitis C vaccine, a hand, foot and mouth disease vaccine, or a HPV vaccine.

8. The use according to any one of claims 1 to 7, wherein the vaccine is an inactivated vaccine, a live attenuated vaccine, a protein vaccine, a DNA vaccine or a polypeptide vaccine.

9. An immunopotentiator for vaccines, comprising tauroursodeoxycholic acid or its analogs.

10. The immunopotentiator according to claim 9, wherein the analog of tauroursodeoxycholic acid is selected from the following: Glycochenodeoxycholic acid, glycohyodeoxycholic acid, glycocholic acid, tallow-α-mucosaic acid, glycohyodeoxycholic acid, 7-ketocholic acid, 3-sulfate taurochenodeoxycholic acid, 3β-hydroxy-δ5-cholenic acid, allolithocholic acid, lithocholic acid, deoxycholic acid, chenodeoxycholic acid, hyodeoxycholic acid, ursodeoxycholic acid, isoursodeoxycholic acid, cholic acid, hyodeoxycholic acid, β-mucosaic acid, α-mucosaic acid, 3β-cholic acid, ursocholic acid, glycolithocholic acid, taurolithocholic acid, glycodeoxycholic acid, glycoursodeoxycholic acid, taurodeoxycholic acid, taurochenodeoxycholic acid, taurocholic acid, tallow-β-mucosaic acid, orthocholic acid, isolithocholic acid, dehydrolithocholic acid Acid, 12-ketodeoxycholic acid, omega-muricolic acid, murodeoxycholic acid, isodeoxycholic acid, taurohyocholic acid, 3-sulfatedeoxycholic acid, 3-sulfateglycochenodeoxycholic acid, 3-sulfateglycocholic acid, 3-sulfateglycodeoxylithocholic acid, 3-sulfateglycolithocholic acid, 3-sulfatelithocholic acid, 3-sulfatetaurodeoxycholic acid, 3-sulfatetaurolithocholic acid, 3-sulfatetauroursodeoxycholic acid, orthodeoxycholic acid, 24-norursodeoxycholic acid, 7-ketochenodeoxycholic acid, dehydrocholic acid, 12-ketocholic acid, 3-oxodeoxycholic acid, isochenodeoxycholic acid, isodeoxycholic acid, 3-sulfatechenodeoxycholic acid, 3-oxochenodeoxycholic acid.

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