WO2024040979A1 - Adjuvant de vaccin à base de polysaccharide acide de ginseng, composition de vaccin et son utilisation - Google Patents

Adjuvant de vaccin à base de polysaccharide acide de ginseng, composition de vaccin et son utilisation Download PDF

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WO2024040979A1
WO2024040979A1 PCT/CN2023/087301 CN2023087301W WO2024040979A1 WO 2024040979 A1 WO2024040979 A1 WO 2024040979A1 CN 2023087301 W CN2023087301 W CN 2023087301W WO 2024040979 A1 WO2024040979 A1 WO 2024040979A1
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vaccine
adjuvant
gaps
ginseng
hepatitis
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PCT/CN2023/087301
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English (en)
Chinese (zh)
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翟健秀
殷军
周荔葆
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沈阳药科大学
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K39/39—Medicinal preparations containing antigens or antibodies characterised by the immunostimulating additives, e.g. chemical adjuvants
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • C—CHEMISTRY; METALLURGY
    • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
    • C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
    • C08B37/0003—General processes for their isolation or fractionation, e.g. purification or extraction from biomass
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K2039/51—Medicinal preparations containing antigens or antibodies comprising whole cells, viruses or DNA/RNA
    • A61K2039/53—DNA (RNA) vaccination
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K2039/555—Medicinal preparations containing antigens or antibodies characterised by a specific combination antigen/adjuvant
    • A61K2039/55511—Organic adjuvants
    • A61K2039/55583—Polysaccharides
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
    • Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
    • Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change

Definitions

  • This application relates to the field of medical technology, specifically to a ginseng acidic polysaccharide (GAPS) vaccine adjuvant, vaccine composition and application thereof.
  • GAPS ginseng acidic polysaccharide
  • Vaccines can activate humoral immune responses in organisms to produce antibodies, or activate cytotoxic T cells and other lymphocytes through cellular immune responses to resist invading foreign pathogens and prevent disease (Cavallo Fet al., Vaccination for treatment and prevention of cancer animal models.Adv Immunol.2006.90:175-213.Review). Although the vaccine has the effect of activating the immune system, in clinical use it is often found that it cannot exert its due effect on certain groups whose immune systems are too weak, such as the elderly and children. Therefore, the addition of appropriate amounts of vaccine adjuvants is necessary. .
  • Vaccine adjuvants refer to substances that can non-specifically change or enhance the body's specific immune response to antigens. They are required to be non-toxic, high in purity, and have certain adsorption capacity and stable properties.
  • the main mechanism of action of vaccine adjuvants is to increase the surface area of the antigen and improve immunogenicity; to slow-release the antigen and prolong the residence time of the antigen in tissues; to promote inflammatory response and stimulate active immune response.
  • adjuvants can usually be divided into two categories. The first is to adsorb antigens and assist the antigens to be engulfed by cells, such as aluminum salts and M59 emulsifiers (O'Hagan D T, Wack A, Podda A. MF59 is a safe and potent vaccine adjuvant for flu vaccines in humans: what did we learn during its development? Clin Pharmacol Ther. 2007 Dec; 82(6):740-4; 4. Clapp T, Siebert P, Chen D, Jones Braun L. Vaccines with aluminum-containing adjuvants: optimizing vaccine efficacy and thermal stability. J Pharm Sci.
  • CFA-mycobacteria mycobacteria
  • Hoft DF Blazevic A, Abate G, Hanekom WA, KaplanG, Soler JH, Weichold F, Geiter L, Sadoff JC, Horwitz MA.
  • a new recombinantbacille Calmette-Guérin vaccine safely induces significantly enhanced tuberculosis-specific immunity in human volunteers. J Infect Dis.2008Nov15;198(10):1491-501).
  • This application provides a new vaccine adjuvant, ginseng acidic polysaccharide (GAPS), which has good activity and can improve the body's response to rabies vaccine, influenza vaccine, hepatitis B vaccine, hepatitis A vaccine, hepatitis C vaccine, hand, foot and mouth vaccine, HPV vaccine, new Coronavirus vaccine-specific antibody (or neutralizing antibody) titer levels.
  • GAPS ginseng acidic polysaccharide
  • a vaccine adjuvant comprising ginseng acidic polysaccharide (GAPS).
  • the vaccine adjuvant according to item 1 also includes physiological saline or water for vaccine injection or pharmaceutical excipients.
  • the vaccine adjuvant is composed of ginseng acidic polysaccharide (GAPS) and physiological saline or vaccine injection water or pharmaceutical excipients.
  • the vaccine adjuvant is composed of ginseng acidic polysaccharide (GAPS) and physiological saline, such as,
  • the vaccine adjuvant is composed of ginseng acidic polysaccharide (GAPS) and vaccine water for injection.
  • the vaccine adjuvant is composed of ginseng acidic polysaccharide (GAPS) and pharmaceutical excipients.
  • the vaccine adjuvant according to item 3 includes,
  • ginseng acidic polysaccharide was obtained.
  • the column of the column chromatography is a DEAE cellulose column or a macroporous resin column, preferably a DEAE cellulose column, and further preferably the column chromatography is performed at 2 to 8 mL/min. Loading.
  • the eluent is water and an alkali solution, a borax solution or a salt solution
  • the salt solution is preferably a NaCl solution
  • the concentration of the eluent is 0.3 ⁇ 0.7mol/L
  • the elution flow rate is 0.5 ⁇ 2mL/min.
  • a vaccine composition comprising the vaccine adjuvant described in any one of items 1-7 and an antigen or DNA encoding the antigen.
  • the dosage ratio of the GAPS to the antigen is: Preferably, the dosage ratio is: Preferably, the dosage ratio is: Preferably, the dosage ratio is: Preferably, the dosage ratio is: Preferably, the dosage ratio is: Preferably, the dosage ratio is: Preferably, the dosage ratio is: Preferably, the dosage ratio is: Preferably, the dosage ratio is: Preferably, the dosage ratio is: Preferably, the dosage ratio is: Preferably, the dosage ratio is:
  • the vaccine composition according to any one of items 8-10 which is a rabies vaccine, an influenza vaccine, a hepatitis B vaccine, a hepatitis A vaccine, a hepatitis C vaccine, a hand, foot and mouth vaccine, an HPV vaccine or a new coronavirus vaccine .
  • a method for preventing and/or treating diseases comprising administering the vaccine composition described in any one of items 8 to 12 to a subject.
  • the disease is selected from rabies, influenza, hepatitis B, hepatitis A, hepatitis C, hand, foot and mouth disease, HPV disease or novel coronavirus disease.
  • the active ingredient ginseng acidic polysaccharide GAPS disclosed in this application is used as a vaccine adjuvant and has high adjuvant activity. Due to its high content of sulfate radicals and carboxylate radicals, acidic polysaccharides have a high negative charge density and may participate in a variety of enzymatic reactions and immune activities in the body (for example, heparin, which is widely found in animals, is a small molecule acidic polysaccharide. ), acidic polysaccharides are widely derived from animals, plants and microorganisms. Their monosaccharide composition, the type and number of acidic groups on the sugar chain vary with the source species.
  • acidic polysaccharides derived from plants have a macromolecular structure and are mostly used as water-soluble dietary fiber. Research on adjuvant activity is very limited. This application has undergone a large number of screening tests. During the activity screening process, it was found that ginseng acidic polysaccharides have high adjuvant activity and can effectively enhance the immune response activity of various vaccines. In particular, ginseng acidic polysaccharides come from natural plants with clear sources and resources. Rich, and the ingredients have good safety and stability.
  • the ginseng acidic polysaccharide GAPS of the present application can significantly increase the specific antibody (or neutralizing antibody) titer after antigen immunization, and can effectively enhance rabies vaccine, influenza vaccine, hepatitis B vaccine, hepatitis A vaccine, hepatitis C vaccine, hand, foot and mouth vaccine Immune response activity of vaccines, HPV vaccines, and novel coronavirus vaccines. And its activity is significantly higher than that of currently commonly used vaccine adjuvants.
  • the ginseng acidic polysaccharide GAPS vaccine adjuvant of the present application has the advantages of good immune effect and easy use, and provides a new adjuvant option for vaccines.
  • Figure 1 is the elution-absorbance curve of ginseng polysaccharide
  • Figure 2 is the absorbance-glucose content standard curve
  • Figure 3 is the absorbance-protein content standard curve
  • Figure 4 shows the effects of GAPS and GPS on IgG antibody levels in mice immunized with rabies vaccine
  • Figure 5 shows the effects of GAPS and GPS on neutralizing antibody titers in mice immunized with rabies vaccine
  • Figure 6 shows the effects of GAPS and GPS on neutralizing antibody levels in mice immunized with influenza vaccine
  • FIG. 7 shows the effects of different doses of GAPS on neutralizing antibody levels in mice immunized with influenza vaccine
  • Figure 8 shows the effects of GAPS and GPS on IgG antibody titers in mice immunized with hepatitis B vaccine
  • Figure 9 shows the effects of GAPS and GPS on neutralizing antibody levels in mice immunized with hepatitis A vaccine
  • Figure 10 shows the effects of different doses of GAPS on neutralizing antibody levels in mice immunized with hepatitis A vaccine
  • Figure 11 shows the effects of GAPS and GPS on IgG antibody levels in mice immunized with hepatitis C vaccine
  • Figure 12 shows the effects of GAPS and GPS on neutralizing antibody levels in mice immunized with hand, foot and mouth vaccine
  • Figure 13 shows the effects of GAPS and GPS on IgG antibody levels in mice immunized with hand, foot and mouth vaccine
  • Figure 14 shows the effects of different doses of GAPS on neutralizing antibody levels in mice immunized with hand, foot and mouth vaccine
  • Figure 15 shows the effects of GAPS and GPS on IgG antibody titers in mice immunized with HPV vaccine
  • Figure 16 shows the effect of different doses of GAPS on IgG antibody titers in mice immunized with HPV vaccine
  • FIG. 17 shows the effects of GAPS and GPS on IgG antibody levels in mice immunized with the new coronavirus vaccine
  • FIG. 18 Effects of different doses of GAPS on IgG antibody levels in mice immunized with the new coronavirus vaccine.
  • the present application discloses a vaccine adjuvant, including ginseng acidic polysaccharide (GAPS).
  • GAPS ginseng acidic polysaccharide
  • the vaccine adjuvant further includes physiological saline or water for vaccine injection or pharmaceutical excipients.
  • vaccine refers to any preparation of an antigen or immunogenic substance suitable for stimulating active immunity in an animal or human.
  • adjuvant refers to any substance or mixture of substances that enhances, increases, up-regulates, alters, or otherwise promotes an immune response (eg, a humoral or cellular immune response) to an antigen in an animal.
  • 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.
  • suitable antigens include, but are not limited to, proteins, glycoproteins, lipoproteins, peptides, carbohydrates/polysaccharides, lipopolysaccharides, toxins, viruses, bacteria, fungi, and parasites.
  • suitable antigens include minimal components of the antigen, such as, but not limited to, epitopes, epitopes or peptides. Still other suitable antigens include those described in U.S. Patent No. 5,855,894. Antigens may be natural (naturally expressed or produced), synthetic, or derived by recombinant DNA methodologies familiar to those skilled in the art.
  • pharmaceutical excipients refers to the collective name for all pharmaceutical materials other than the main drug that are added to the prescription to improve the formability, effectiveness, stability, and safety of the preparation during the production of drugs and preparation of prescriptions. Substances that have been reasonably evaluated for their safety and are included in pharmaceutical preparations. In addition to excipients, serving as carriers, and improving stability, pharmaceutical excipients also have important functions such as solubilization, dissolution, and sustained and controlled release. They are important ingredients that may affect the quality, safety, and effectiveness of drugs.
  • the pharmaceutical excipients described in this application can be appropriate carriers or excipients, emulsifiers, wetting agents, preservatives, stabilizers, antioxidants, adjuvants (such as aluminum hydroxide adjuvant, oil adjuvant, Freund's Complete adjuvant and Freund's incomplete adjuvant), etc.
  • adjuvants such as aluminum hydroxide adjuvant, oil adjuvant, Freund's Complete adjuvant and Freund's incomplete adjuvant
  • ginseng acidic polysaccharide refers to acidic polysaccharides extracted from the Araliaceae plant ginseng.
  • the extraction from Araliaceae plant ginseng can be extracted from original ginseng, or it can be extracted from processed ginseng, such as from secondary developed products of ginseng, such as ginseng total polysaccharides extracted from ginseng.
  • ginseng Extraction from ginseng, such as extraction from other parts remaining after the total ginseng polysaccharide is extracted from ginseng, such as extraction from ginseng extraction waste, such as extraction from ginseng-containing medicinal residues, etc., as long as it contains ginseng acidic polysaccharide, any ginseng product
  • the ginseng acidic polysaccharide extracted from can be used as an adjuvant for the vaccine of this application.
  • the ginseng acidic polysaccharide is purchased. In a preferred embodiment, the ginseng acidic polysaccharide is extracted from ginseng. In a preferred embodiment, the ginseng acidic polysaccharide is Acidic polysaccharides are extracted from ginseng polysaccharides. In a preferred embodiment, the ginseng acidic polysaccharides are extracted in the following manner:
  • ginseng acidic polysaccharide was obtained.
  • ginseng polysaccharide refers to the total polysaccharides extracted from the Araliaceae plant ginseng.
  • ginseng total polysaccharide has the same meaning and can be used interchangeably.
  • the method for extracting ginseng total polysaccharide from ginseng can be any method in the art that can extract ginseng total polysaccharide from ginseng. It can also be extracted through the following methods:
  • the protein-removed retentate was extracted with ethanol to obtain total ginseng polysaccharides.
  • the step of obtaining the first extract includes:
  • the protease culture temperature is 30°C to 40°C, and the culture time is 1 to 3 hours.
  • the step of obtaining the second extract includes:
  • Neutral protease was added to the first extract, incubated at 40°C for 3 hours, and then inactivated at 100°C for 30 minutes. The mixed solution was centrifuged at 10,000 rpm to obtain the supernatant, which was evaporated to dryness to obtain the second extract.
  • the steps of obtaining ginseng crude polysaccharide include:
  • the second extract was soaked in 10 times the volume of 95% ethanol at low temperature for 2.0 h. Filter, take the filter residue, add 2 times the volume of distilled water to dissolve, extract with boiling water reflux 3 times, 1.0h each time, filter with 120 mesh nylon cloth, combine the extracts, concentrate to 500mL, centrifuge for 20 minutes (5000rpm), discard the precipitate. Add 95% ethanol to the supernatant until the final concentration of ethanol is 90%, and let it stand for 1 hour.
  • the molecular weight cutoff during dialysis is 800-2500kDa, for example, it can be 900kDa, 1000kDa, 1100kDa, 1200kDa, 1300kDa, 1400kDa, 1500kDa, 1600kDa, 1700kDa, 1800kDa, 1900kDa, 2000kDa, 2100kDa, 220 0kDa , 2300kDa, 2400kDa, preferably 1000-1500kDa.
  • Sevag reagent is used to remove proteins in the retentate.
  • the steps of dissolving crude ginseng polysaccharide and then dialyzing it, and removing the protein in the retentate include:
  • the protein-removed retentate is extracted with ethanol, and the steps of obtaining total ginseng polysaccharides include:
  • the column when performing column chromatography and then using eluent to elute, can be any column in the field that can realize column chromatography, which is a preferred embodiment.
  • the column of the column chromatography is a DEAE cellulose column or a macroporous resin column, preferably a DEAE cellulose column. It is further preferred that the column chromatography is loaded at 2 to 8 mL/min, for example, it can be 3 mL/min or 4 mL/min. ,5mL/min,6mL/min,7mL/min.
  • the eluent can be any one or more than two solutions in the art that can achieve the purpose of elution.
  • the eluent is water and an alkali solution, or water and borax. solution, or water and salt solution.
  • the eluent is water and a salt solution.
  • the eluent is water and NaCl solution.
  • the concentration of the eluent can be any concentration in the art that can achieve the purpose of elution.
  • the concentration of the eluent is 0.3-0.7 mol/L, for example, it can be 0.4 mol/L, 0.5mol/L, 0.6mol/L.
  • the elution flow rate can be any flow rate in the art that can achieve the purpose of elution.
  • the elution flow rate is 0.5-2mL/min, for example, it can be 0.6mL/min, 0.7mL /min, 0.8mL/min, 0.9mL/min, 1.0mL/min, 1.1mL/min, 1.2mL/min, 1.3mL/min, 1.4mL/min, 1.5mL/min, 1.6mL/min, 1.7mL /min, 1.8mL/min, 1.9mL/min.
  • the method before dialyzing the eluate with distilled water, the method further includes detecting A 490 of the eluate using the phenol sulfuric acid method, collecting absorption peaks, and then performing dialysis.
  • the dialysis time is 24 to 72 hours. , for example, it can be 25 hours, 26 hours, 27 hours, 28 hours, 29 hours, 30 hours, 35 hours, 40 hours, 45 hours, 50 hours, 55 hours, 60 hours, 65 hours, 70 hours.
  • the present application further provides a vaccine composition, including a vaccine adjuvant containing any one of the ginseng acidic polysaccharides in the present application and an antigen or DNA encoding the antigen.
  • the dosage of the vaccine adjuvant in the vaccine composition is an effective dosage to achieve a therapeutic effect, and the dosage is An effective amount is an amount that enhances, increases, upregulates, alters, or otherwise promotes the immune response to an antigen.
  • a therapeutically effective amount is an amount that induces immunity in an animal susceptible to a disease caused by a pathogen, cancer cell, or allergen.
  • therapeutically effective amounts will vary and be determined on a case-by-case basis.
  • the doses of vaccine adjuvants and vaccine antigens are not particularly limited and will vary depending on the method of administration, subject The patient, age of the subject, dosage form, administration route, etc. should be selected appropriately.
  • the dosage ratio of the ginseng acidic polysaccharide to the vaccine antigen is:
  • the dosage ratio can be 0.005, 0.05, 0.5, 1, 1.25, 2, 3, 4, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65 ,70,75,80,85,90,95,100,105,110,115,120,125,130,135,140,145,150,160,166.7,170,180,200,500,1000,10000 ( ⁇ g/IU or ⁇ g: ⁇ g), preferably, the dosage ratio is:
  • the dosage ratio is 1.25-125 ( ⁇ g/IU or ⁇ g: ⁇ g); preferably, the dosage ratio is 1.25-50 ( ⁇ g/IU or ⁇ g: ⁇ g); preferably, the dosage ratio is 1.25 ⁇ 25 ( ⁇ g/IU or ⁇ g: ⁇ g);
  • the dosage ratio is 1.25 ⁇ 12.5 ( ⁇ g/IU or ⁇ g: ⁇ g);
  • the dosage ratio is 1.25 ⁇ ⁇
  • the dosage ratio of the ginseng acidic polysaccharide to the vaccine antigen is: For example, it can be 17 ( ⁇ g: ⁇ g), 20 ( ⁇ g: ⁇ g), 25 ( ⁇ g: ⁇ g), 30 ( ⁇ g: ⁇ g), 35 ( ⁇ g: ⁇ g), 40 ( ⁇ g: ⁇ g), 45 ( ⁇ g: ⁇ g ), 50( ⁇ g: ⁇ g), 55( ⁇ g: ⁇ g), 60( ⁇ g: ⁇ g), 65( ⁇ g: ⁇ g), 70( ⁇ g: ⁇ g), 75( ⁇ g: ⁇ g), 80( ⁇ g: ⁇ g) , 90( ⁇ g: ⁇ g), 100( ⁇ g: ⁇ g), 110( ⁇ g: ⁇ g), 120( ⁇ g: ⁇ g), 130( ⁇ g: ⁇ g), 140( ⁇ g: ⁇ g), 150( ⁇ g: ⁇ g), 160( ⁇ g: ⁇ g), with better effect.
  • the dosage ratio of the ginseng acidic polysaccharide to the vaccine antigen is: For example, it can be 6 ( ⁇ g:IU), 7 ( ⁇ g:IU), 8 ( ⁇ g:IU), 9 ( ⁇ g:IU), 10 ( ⁇ g:IU), 15 ( ⁇ g:IU), 20 ( ⁇ g:IU ), 25( ⁇ g:IU), 30( ⁇ g:IU), 35( ⁇ g:IU), 40( ⁇ g:IU), 45( ⁇ g:IU), 49( ⁇ g:IU), have better effects.
  • the dosage ratio of the ginseng acidic polysaccharide to the vaccine antigen is: For example, it can be 1.5( ⁇ g:IU), 2( ⁇ g:IU), 2.5( ⁇ g:IU), 3( ⁇ g:IU), 4( ⁇ g:IU), 5( ⁇ g:IU), 6( ⁇ g:IU ), 7( ⁇ g:IU), 8( ⁇ g:IU), 9( ⁇ g:IU), 10( ⁇ g:IU), 15( ⁇ g:IU), 16( ⁇ g:IU), 17( ⁇ g:IU) , 18( ⁇ g:IU), 19( ⁇ g:IU), 20( ⁇ g:IU), 21( ⁇ g:IU), 22( ⁇ g:IU), 23( ⁇ g:IU), 24( ⁇ g:IU), Have better results.
  • the dosage ratio of the ginseng acidic polysaccharide to the vaccine antigen is: For example, it can be 13( ⁇ g: ⁇ g), 14( ⁇ g: ⁇ g), 15( ⁇ g: ⁇ g), 16( ⁇ g: ⁇ g), 17( ⁇ g: ⁇ g), 18( ⁇ g: ⁇ g), 19( ⁇ g: ⁇ g ), 20( ⁇ g: ⁇ g), 25( ⁇ g: ⁇ g), 30( ⁇ g: ⁇ g), 35( ⁇ g: ⁇ g), 40( ⁇ g: ⁇ g), 50( ⁇ g: ⁇ g), 60( ⁇ g: ⁇ g) , 70( ⁇ g: ⁇ g), 80( ⁇ g: ⁇ g), 90( ⁇ g: ⁇ g), 100( ⁇ g: ⁇ g), 110( ⁇ g: ⁇ g), 120( ⁇ g: ⁇ g), 121( ⁇ g: ⁇ g), 124( ⁇ g: ⁇ g), with better effect.
  • the vaccine composition provided in this application is used in a new coronavirus vaccine, and the dosage ratio of the ginseng acidic polysaccharide to the vaccine antigen is:
  • the dosage ratio of the ginseng acidic polysaccharide to the vaccine antigen is:
  • it can be 11( ⁇ g: ⁇ g), 12( ⁇ g: ⁇ g), 13( ⁇ g: ⁇ g), 14( ⁇ g: ⁇ g), 15( ⁇ g: ⁇ g), 16( ⁇ g: ⁇ g), 17( ⁇ g: ⁇ g ), 18( ⁇ g: ⁇ g), 19( ⁇ g: ⁇ g), 20( ⁇ g: ⁇ g), 25( ⁇ g: ⁇ g), 30( ⁇ g: ⁇ g), 35( ⁇ g: ⁇ g), 40( ⁇ g: ⁇ g) , 45( ⁇ g: ⁇ g), 46( ⁇ g: ⁇ g), 47( ⁇ g: ⁇ g), 48( ⁇ g: ⁇ g), 49( ⁇ g: ⁇ g), have better effects.
  • dosage and length of treatment may vary depending on the type, weight, and condition of the patient to be treated, their individual response to the vaccine composition, and the particular route of administration chosen. In some cases, dosage levels below the lower end of the foregoing ranges may be therapeutically effective, while in other cases, still larger dosages may be employed without causing any deleterious side effects, provided such larger dosages are First divide into several small doses to be administered throughout the day. Whenever secondary stress or exposure is likely to occur, a challenge dose is considered ideal.
  • the vaccine adjuvant and the antigen or the DNA encoding the antigen in the vaccine combination of the present application can be included together in one composition and can be formulated in separate compositions.
  • the vaccine adjuvant and The route of administration of the antigen or the DNA encoding the antigen may be the same or different.
  • the vaccine adjuvant and the antigen or DNA encoding the antigen may be administered simultaneously or with a temporal difference, i.e., the vaccine adjuvant and the antigen or DNA encoding the antigen may be administered simultaneously or separately (e.g., vaccine The adjuvant is administered before or after the vaccine antigen is administered).
  • Vaccine adjuvants and antigens or DNA encoding said antigens may be provided as kits containing them. However, from the perspective of reducing patient burden, it is preferable that the vaccine adjuvant and the antigen or DNA encoding the antigen are included in one composition so that they can be administered simultaneously. Whether co-administered or simultaneously administered, the mode of administration of the vaccine composition can be any suitable route that delivers the vaccine composition to Host.
  • the vaccine adjuvant or vaccine composition of the present application also includes pharmaceutical auxiliary materials and a second vaccine adjuvant.
  • the pharmaceutical excipients are as described above.
  • the vaccine adjuvants of the present application can be administered as part of a vaccine formulation, optionally containing an additional second vaccine adjuvant.
  • the second vaccine adjuvant is other adjuvant different from the ginseng acidic polysaccharide adjuvant of the present application, and may be one or more than two. Examples of appropriate second vaccine adjuvants include those known in the art. adjuvants,
  • the vaccine adjuvant or vaccine composition of the present application may further comprise one or more antioxidants selected from the group consisting of: sodium bisulfite, sodium sulfite, metabisulfite Sodium, sodium thiosulfate, sodium methoxysulfite, L-ascorbic acid, erythorbic acid, acetylcysteine, cysteine, monothioglycerol, mercaptoacetic acid, thiolactic acid, sulfide, dithiol Thiothreitol, dithioerythritol, glutathione, ascorbyl palmitolic acid, butylated hydroxytoluene, butylated hydroxytoluene, nordihydroguaiaretic acid, propyl gallate Tocopherol, a-tocopherol, and mixtures thereof.
  • antioxidants selected from the group consisting of: sodium bisulfite, sodium sulfite, metabisulfite Sodium, sodium
  • the vaccine adjuvant or vaccine composition of the present application may further comprise one or more preservatives.
  • suitable preservatives include (but are not limited to): benzalkonium chloride, ethyl alcohol, benzoic acid, ethyl alcohol, p- Formaben, ethyl p-hydroxybenzoate, propyl p-hydroxybenzoate, butyl p-hydroxybenzoate, sodium benzoate, phenol, and mixtures thereof.
  • suitable preservatives include (but are not limited to): benzalkonium chloride, ethyl alcohol, benzoic acid, ethyl alcohol, p- Formaben, ethyl p-hydroxybenzoate, propyl p-hydroxybenzoate, butyl p-hydroxybenzoate, sodium benzoate, phenol, and mixtures thereof.
  • the presence or absence of preservatives will depend on the antigen. For example, if the antigen is a live bacterial anti
  • the vaccine adjuvant or vaccine composition of the present application can be used to prevent or treat diseases caused by pathogens, cancer cells or allergens in humans or animals by administering a therapeutically effective amount of the adjuvant to humans or animals susceptible to the disease. composition or vaccine.
  • the pathogen can be any pathogen, including but not limited to: bacteria, protozoa, helminths, viruses and fungi. Diseases in animals caused by such pathogens include, but are not limited to: bovine respiratory disease, porcine respiratory disease, pneumonia, pasteurellosis, coccidiosis, anaplasmosis, and infectious keratitis.
  • the cancer cells can be any type of cancer cells in the art.
  • the allergen may be any allergen known in the art.
  • the vaccine composition of the present application can be a rabies vaccine, an influenza vaccine, a hepatitis B vaccine, a hepatitis A vaccine, a hepatitis C vaccine, a hand, foot and mouth vaccine, an HPV vaccine or a new coronavirus vaccine, etc.
  • the vaccine type of the vaccine composition of the present application can be an inactivated virus vaccine, an attenuated vaccine, an inactivated vaccine, a protein vaccine, a DNA vaccine or a polypeptide vaccine, etc.
  • the vaccine adjuvant or vaccine composition of the present application can be used to protect or treat humans and non-human animals such as livestock and livestock animals, including (but not limited to) cattle, horses, sheep, pigs, goats, rabbits, cats, dogs and other animals in need. Other mammals treated.
  • the vaccine adjuvant or vaccine composition of the present application is used to protect or treat humans.
  • the vaccine adjuvant or vaccine composition of the present application to be administered can be selected based on the patient to be protected or treated.
  • compositions of the present application can be prepared by general methods, in which one or more pharmaceutically acceptable diluents or carriers are added, for example, in an oral pharmaceutical form, such as tablets, capsules, granules, powders, and lozenges. , syrups, emulsions, suspensions, etc., or parenteral drugs, such as topical drugs, suppositories, injections, eye drops, intranasal agents, transpulmonary agents, etc.
  • parenteral drugs such as topical drugs, suppositories, injections, eye drops, intranasal agents, transpulmonary agents, etc.
  • formulations include injectable or intranasal solutions, or lyophilized formulations prepared by lyophilizing said solutions.
  • injectable solutions include emulsions and liposomes containing aqueous solutions and oleaginous compositions, such as aqueous solutions in which a vaccine adjuvant and an antigen or DNA encoding the antigen are dissolved or dispersed in water.
  • aqueous solutions in which a vaccine adjuvant and an antigen or DNA encoding the antigen are dissolved or dispersed in water.
  • a solution formulation or an aqueous suspension formulation, or an oily solution formulation or an oily suspension formulation in which the vaccine adjuvant and the antigen or DNA encoding the antigen are dissolved or dispersed in oil.
  • aqueous solutions, aqueous solution formulations or aqueous suspension formulations include aqueous solutions containing distilled water for injection and optionally containing buffers, pH adjusters, stabilizers, isotonic agents and/or emulsifiers, or Aqueous suspension, etc.
  • the vaccine adjuvant or vaccine composition of the present application can be administered by oral, intramuscular, intravenous, subcutaneous, intraocular, parenteral, topical, intravaginal or rectal routes.
  • the adjuvant composition or vaccine adjuvant may be administered orally in feed or as a gavage composition.
  • the vaccine adjuvant or vaccine composition of the present application is injected intramuscularly, intravenously or subcutaneously.
  • the ginseng acidic polysaccharide vaccine adjuvant provided by this application can significantly increase the specific antibody (or neutralizing antibody) titer after antigen immunization, and can be used for rabies vaccine, influenza vaccine, hepatitis B vaccine, hepatitis A vaccine, hepatitis C vaccine, and hand, foot and mouth vaccine. , HPV vaccine, new coronavirus vaccine and other vaccines, can effectively enhance immune response activity.
  • ginseng acidic polysaccharide adjuvant can significantly increase the level of neutralizing antibodies in the blood of vaccinated mice (P ⁇ 0.01); for some vaccines, such as hepatitis B vaccine, the antibody levels in experimental mice are more significantly improved (P ⁇ 0.001).
  • this application has gone through a large number of tests to explore the impact of using different doses of ginseng polysaccharide vaccine adjuvants on the adjuvant activity of the vaccine. It was found that different doses have a certain impact on the adjuvant activity, and also have effects on different types of vaccines. Different effects. After many attempts, the dosage of ginseng polysaccharide vaccine adjuvant with better effect has been optimized.
  • this application has further optimized the dosage of vaccine adjuvant with specific and better effects for different vaccines, such as
  • the results of the examples in this application show that for influenza vaccines, the dosage ratio of the ginseng acidic polysaccharide to the vaccine antigen is: has a better effect; for hepatitis A vaccine, the dosage ratio of the ginseng acidic polysaccharide to the vaccine antigen is: When used for hand, foot and mouth vaccine, the dosage ratio of the ginseng acidic polysaccharide to the vaccine antigen is: When used for HPV vaccine, the dosage ratio of the ginseng acidic polysaccharide to the vaccine antigen is: has better results; For use in the new coronavirus vaccine, the dosage ratio of the ginseng acidic polysaccharide to the vaccine antigen is: has better results.
  • Aluminum salt adjuvant produced by Croda Company
  • Centrifuge for 20 minutes (5000 rpm), collect the precipitate, add 400 mL of distilled water to the precipitate, then add 95% ethanol to the supernatant to a final concentration of 80%, let stand for 1 hour, and centrifuge for 20 minutes (5000 rpm) , let stand at room temperature overnight, and collect the precipitate.
  • the precipitate was washed twice with absolute ethanol and 95% ethanol.
  • the ginseng crude polysaccharide was freeze-dried and prepared into a 30% aqueous solution, added to a 1000kDa dialysis bag for dialysis, and left to stand overnight. Take the liquid in the dialysis bag, add 1/4 volume of Sevag reagent to it, let it stand and centrifuge to remove the gel-like precipitate. Repeat the above operation 5 times, combine the supernatants, concentrate under reduced pressure to remove organic reagents, add 95% ethanol to a final concentration of 80%, let stand at 4°C overnight, centrifuge, and remove the supernatant. The precipitate was washed twice with 95% ethanol and absolute ethanol. Freeze-dried ginseng polysaccharide.
  • Pretreatment Add distilled water to prevent bubbles into the column tube, add DEAE packing to the column tube, and let it stand for 24 hours; rinse 1 column volume with 0.5M hydrochloric acid; then balance 4 column volumes with ultrapure water;
  • ginseng polysaccharides Take total ginseng polysaccharides, dissolve them in distilled water, load the sample at a speed of 4 mL/min, and elute with water and 0.3 mol/L NaCl solution respectively. The flow rate is set to 0.5 mL/min.
  • the eluate was tested for A 490 using the phenol sulfuric acid method, the absorption peaks were collected, dialyzed against distilled water for 72 hours, and freeze-dried. Ginseng acidic polysaccharide was obtained, and the elution curve is shown in Figure 1.
  • the phenol-sulfuric acid method was used to determine the content of ginseng acidic polysaccharides. Using different concentrations of glucose as the abscissa and the absorbance value at OD490nm as the ordinate, draw a standard curve, as shown in Figure 2. Calculate the polysaccharide content based on the glucose standard curve and sample absorbance. The content of ginseng acidic polysaccharides is 97.3%.
  • the protein content in ginseng acidic polysaccharide was determined using the Bradford method. Taking bovine serum albumin as the abscissa and OD595nm as the ordinate, draw a standard curve, as shown in Figure 3. Calculate the protein content based on the bovine serum albumin standard curve and sample absorbance. The protein content of ginseng acidic polysaccharide is 0.3%.
  • Example 1 Immune adjuvant activity of ginseng acidic polysaccharide (GAPS) to rabies vaccine
  • Diploid inactivated rabies vaccine produced by Liaoning Chengda Biological Co., Ltd.
  • mice Female C57BL/6 mice, 6-8 weeks old, were purchased from Beijing Huafukang Biotechnology Co., Ltd.
  • High-dose GPS adjuvant vaccine composition Measure appropriate amounts of ginseng polysaccharide (GPS) and diploid inactivated rabies vaccine respectively, dissolve them in physiological saline so that each milliliter of solution contains 2500 ⁇ g of GPS and 2.5 IU of rabies vaccine, and add 0.22 Filter with ⁇ m microporous membrane and pack aseptically.
  • GPS ginseng polysaccharide
  • diploid inactivated rabies vaccine composition Measure appropriate amounts of ginseng polysaccharide (GPS) and diploid inactivated rabies vaccine respectively, dissolve them in physiological saline so that each milliliter of solution contains 2500 ⁇ g of GPS and 2.5 IU of rabies vaccine, and add 0.22 Filter with ⁇ m microporous membrane and pack aseptically.
  • High-dose GAPS adjuvant vaccine composition Measure appropriate amounts of ginseng acidic polysaccharide (GAPS) and diploid inactivated rabies vaccine respectively, and dissolve them in physiological saline so that each milliliter of solution contains 2500 ⁇ g of GAPS and 2.5 IU of rabies vaccine. Filter through 0.22 ⁇ m microporous membrane and pack aseptically.
  • GAPS ginseng acidic polysaccharide
  • diploid inactivated rabies vaccine composition Measure appropriate amounts of ginseng acidic polysaccharide (GAPS) and diploid inactivated rabies vaccine respectively, and dissolve them in physiological saline so that each milliliter of solution contains 2500 ⁇ g of GAPS and 2.5 IU of rabies vaccine. Filter through 0.22 ⁇ m microporous membrane and pack aseptically.
  • Low-dose GPS adjuvant vaccine composition Measure appropriate amounts of GPS and diploid inactivated rabies vaccine respectively, dissolve them in physiological saline so that each milliliter of solution contains 250 ⁇ g of GPS and 2.5 IU of rabies vaccine, filter with 0.22 ⁇ m micropores Membrane filtration, sterile packaging.
  • Low-dose GAPS adjuvant vaccine composition Measure appropriate amounts of GAPS and diploid inactivated rabies vaccine respectively, dissolve them in physiological saline so that each milliliter of the solution contains 250 ⁇ g of GAPS and 2.5 IU of rabies vaccine, filter with 0.22 ⁇ m micropores Membrane filtration, sterile packaging.
  • Positive control group - aluminum salt adjuvant vaccine composition measure appropriate amounts of aluminum salt and diploid inactivated rabies vaccine respectively, dissolve them in physiological saline so that each milliliter of solution contains 250 ⁇ g of aluminum salt and 2.5 IU of rabies vaccine. Filter through 0.22 ⁇ m microporous membrane and pack aseptically.
  • Negative control group - vaccine without adjuvant Dissolve an appropriate amount of diploid inactivated rabies vaccine in physiological saline so that each milliliter of the solution contains 2.5IU of rabies vaccine, filter it with a 0.22 ⁇ m microporous filter, and sterilize Pack.
  • mice were randomly divided into 6 groups, with 10 mice in each group. Inject 0.1 ml/animal of the vaccine composition prepared above intramuscularly. One week after the first immunization, the second immunization is performed. Sera from mice in each group were collected 14 days after the second immunization, and the neutralizing antibodies and IgG-specific antibody titers in each serum were detected by RFFIT and ELISA.
  • GPS high-dose adjuvant group 0.25IU rabies vaccine + 250 ⁇ g GPS/animal;
  • GAPS high-dose adjuvant group 0.25IU rabies vaccine + 250 ⁇ g GAPS/animal;
  • GPS low-dose adjuvant group 0.25IU rabies vaccine + 25 ⁇ g GPS/animal;
  • GAPS low-dose adjuvant group 0.25IU rabies vaccine + 25 ⁇ g GAPS/animal;
  • Positive control group 0.25IU rabies vaccine + 25 ⁇ g aluminum salt adjuvant/animal;
  • Negative control group 0.25IU rabies vaccine/animal.
  • Example 2 Immune adjuvant activity of ginseng acidic polysaccharide (GAPS) for influenza vaccine
  • Influenza vaccine produced by Liaoning Chengda Biological Co., Ltd.
  • mice Female C57BL/6 mice, 6-8 weeks old, were purchased from Beijing Huafukang Biotechnology Co., Ltd.
  • High-dose GPS adjuvant vaccine composition Measure appropriate amounts of ginseng polysaccharide (GPS) and influenza vaccine respectively, dissolve them in physiological saline so that each milliliter of the solution contains 2500 ⁇ g of GPS and 15 ⁇ g of influenza vaccine, and filter with a 0.22 ⁇ m microporous membrane. Passed, sterile aliquots.
  • GPS ginseng polysaccharide
  • influenza vaccine composition Measure appropriate amounts of ginseng polysaccharide (GPS) and influenza vaccine respectively, dissolve them in physiological saline so that each milliliter of the solution contains 2500 ⁇ g of GPS and 15 ⁇ g of influenza vaccine, and filter with a 0.22 ⁇ m microporous membrane. Passed, sterile aliquots.
  • High-dose GAPS adjuvant vaccine composition Measure appropriate amounts of ginseng acidic polysaccharide (GAPS) and influenza vaccine respectively, dissolve them in physiological saline so that each milliliter of solution contains 2500 ⁇ g of GPS and 15 ⁇ g of influenza vaccine, and filter with a 0.22 ⁇ m microporous membrane Filter and aliquot aseptically.
  • GAPS ginseng acidic polysaccharide
  • influenza vaccine composition dissolve them in physiological saline so that each milliliter of solution contains 2500 ⁇ g of GPS and 15 ⁇ g of influenza vaccine, and filter with a 0.22 ⁇ m microporous membrane Filter and aliquot aseptically.
  • GPS adjuvant vaccine composition Measure appropriate amounts of ginseng polysaccharide (GPS) and influenza vaccine respectively, dissolve them in physiological saline so that each milliliter of solution contains 250 ⁇ g of GPS and 15 ⁇ g of influenza vaccine, and filter with a 0.22 ⁇ m microporous membrane. Passed, sterile aliquots.
  • GAPS adjuvant vaccine composition Measure appropriate amounts of ginseng acidic polysaccharide (GAPS) and influenza vaccine respectively, dissolve them in physiological saline so that each milliliter of solution contains 250 ⁇ g of GPS and 15 ⁇ g of influenza vaccine, and filter with a 0.22 ⁇ m microporous membrane Filter and aliquot aseptically.
  • GAPS ginseng acidic polysaccharide
  • influenza vaccine composition dissolve them in physiological saline so that each milliliter of solution contains 250 ⁇ g of GPS and 15 ⁇ g of influenza vaccine, and filter with a 0.22 ⁇ m microporous membrane Filter and aliquot aseptically.
  • Positive control group-aluminum salt adjuvant vaccine composition measure appropriate amounts of aluminum salt and influenza vaccine respectively, dissolve them in physiological saline so that each milliliter of solution contains 250 ⁇ g of aluminum salt and 15 ⁇ g of influenza vaccine, and filter with a 0.22 ⁇ m microporous membrane Filter and aliquot aseptically.
  • Negative control group - vaccine without adjuvant Dissolve an appropriate amount of influenza vaccine in physiological saline so that each milliliter of the solution contains 15 ⁇ g of influenza vaccine, filter it with a 0.22 ⁇ m microporous filter, and pack aseptically.
  • mice were randomly divided into 6 groups, with 10 mice in each group. Inject 0.1ml/animal of the above composition intramuscularly, and perform the second immunization one week after the first immunization. On the 14th day after the second immunization, the blood of each mouse was collected, and the neutralizing antibody level of the mouse serum influenza vaccine was detected by hemagglutination inhibition test.
  • GPS high-dose adjuvant group 1.5 ⁇ g influenza vaccine + 250 ⁇ g GPS/bird;
  • GAPS high-dose adjuvant group 1.5 ⁇ g influenza vaccine + 250 ⁇ g GAPS/bird;
  • GPS low-dose adjuvant group 1.5 ⁇ g influenza vaccine + 25 ⁇ g GPS/bird;
  • GAPS low-dose adjuvant group 1.5 ⁇ g influenza vaccine + 25 ⁇ g GAPS/bird;
  • Positive control group 1.5 ⁇ g influenza vaccine + 25 ⁇ g aluminum salt adjuvant/animal;
  • Negative control group 1.5 ⁇ g influenza vaccine/animal.
  • both low and high doses of GAPS can significantly increase the antibody levels in influenza vaccinated mice (P ⁇ 0.01); compared with the positive control group (aluminum salt adjuvant) , both low and high doses of GAPS can significantly increase the level of neutralizing antibodies in the blood of influenza vaccinated mice (P ⁇ 0.01).
  • Example 3 Immune adjuvant activity of different doses of ginseng acidic polysaccharide (GAPS) for influenza vaccine 1.
  • GAPS ginseng acidic polysaccharide
  • Preparation of vaccine composition Measure appropriate amounts of GAPS and influenza vaccine respectively, dissolve them with physiological saline, and prepare five composition solutions respectively.
  • GAPS and 15 ⁇ g of influenza vaccine are filtered through a 0.22 ⁇ m microporous membrane and packed aseptically.
  • the ratios of GAPS adjuvant to influenza vaccine in each group of vaccine compositions were 3.3, 16.7, 83.3, 166.7, and 333.3 ( ⁇ g: ⁇ g) respectively.
  • Positive control group-aluminum salt adjuvant vaccine composition measure appropriate amounts of aluminum salt and influenza vaccine respectively, dissolve them in physiological saline so that each milliliter of solution contains 250 ⁇ g of aluminum salt and 15 ⁇ g of influenza vaccine, and filter with a 0.22 ⁇ m microporous membrane Filter and aliquot aseptically.
  • Negative control group - vaccine without adjuvant Dissolve an appropriate amount of influenza vaccine in physiological saline so that each milliliter of the solution contains 15 ⁇ g of influenza vaccine, filter it with a 0.22 ⁇ m microporous filter, and pack aseptically.
  • mice were randomly divided into 7 groups, with 10 mice in each group. Inject 0.1ml/animal of the above composition intramuscularly, and perform the second immunization one week after the first immunization. On the 14th day after the second immunization, the blood of each mouse was collected, and the neutralizing antibody level of the mouse serum influenza vaccine was detected by hemagglutination inhibition test.
  • GAPS adjuvant group 1 1.5 ⁇ g influenza vaccine + 5 ⁇ g GAPS/bird;
  • GAPS adjuvant group 2 1.5 ⁇ g influenza vaccine + 25 ⁇ g GAPS/bird;
  • GAPS adjuvant group 3 1.5 ⁇ g influenza vaccine + 125 ⁇ g GAPS/bird;
  • GAPS adjuvant group 4 1.5 ⁇ g influenza vaccine + 250 ⁇ g GAPS/bird;
  • GAPS adjuvant group 5 1.5 ⁇ g influenza vaccine + 500 ⁇ g GAPS/bird;
  • Positive control group 1.5 ⁇ g influenza vaccine + 25 ⁇ g aluminum salt adjuvant/animal;
  • Negative control group 1.5 ⁇ g influenza vaccine/animal.
  • Example 4 Immune adjuvant activity of ginseng acidic polysaccharide (GAPS) for hepatitis B vaccine
  • Hepatitis B vaccine produced by Liaoning Chengda Biological Co., Ltd.
  • mice Female C57BL/6 mice, 6-8 weeks old, were purchased from Beijing Huafukang Biotechnology Co., Ltd.
  • High-dose GPS adjuvant vaccine composition Measure appropriate amounts of ginseng polysaccharide (GPS) and hepatitis B vaccine respectively, dissolve them in physiological saline so that each milliliter of the solution contains 2500 ⁇ g of GPS and 25 ⁇ g of hepatitis B vaccine, and filter with a 0.22 ⁇ m microporous membrane. Passed, sterile aliquots.
  • GPS ginseng polysaccharide
  • High-dose GAPS adjuvant vaccine composition Measure appropriate amounts of ginseng acidic polysaccharide (GAPS) and hepatitis B vaccine respectively, dissolve them in physiological saline so that each milliliter of solution contains 2500 ⁇ g of GPS and 25 g of hepatitis B vaccine, and filter with a 0.22 ⁇ m microporous membrane Filter and aliquot aseptically.
  • GAPS ginseng acidic polysaccharide
  • hepatitis B vaccine composition dissolve them in physiological saline so that each milliliter of solution contains 2500 ⁇ g of GPS and 25 g of hepatitis B vaccine, and filter with a 0.22 ⁇ m microporous membrane Filter and aliquot aseptically.
  • Low-dose GPS adjuvant vaccine composition Measure appropriate amounts of ginseng polysaccharide (GPS) and hepatitis B vaccine respectively, dissolve them in physiological saline so that each milliliter of the solution contains 250 ⁇ g of GPS and 25 ⁇ g of hepatitis B vaccine, and filter with a 0.22 ⁇ m microporous membrane. Passed, sterile aliquots.
  • GPS ginseng polysaccharide
  • GAPS adjuvant vaccine composition Measure appropriate amounts of ginseng acidic polysaccharide (GAPS) and hepatitis B vaccine respectively, dissolve them in physiological saline so that each milliliter of solution contains 250 ⁇ g of GPS and 25 ⁇ g of hepatitis B vaccine, and filter with a 0.22 ⁇ m microporous membrane Filter and aliquot aseptically.
  • GAPS ginseng acidic polysaccharide
  • hepatitis B vaccine composition dissolve them in physiological saline so that each milliliter of solution contains 250 ⁇ g of GPS and 25 ⁇ g of hepatitis B vaccine, and filter with a 0.22 ⁇ m microporous membrane Filter and aliquot aseptically.
  • Positive control group - aluminum salt adjuvant vaccine composition measure appropriate amounts of aluminum salt and hepatitis B vaccine respectively, dissolve them in physiological saline so that each milliliter of solution contains 250 ⁇ g of aluminum salt and 25 ⁇ g of hepatitis B vaccine, and filter with a 0.22 ⁇ m microporous membrane Filter and aliquot aseptically.
  • Negative control group - vaccine without adjuvant Dissolve an appropriate amount of hepatitis B vaccine in physiological saline so that each milliliter of solution contains 25 ⁇ g of hepatitis B vaccine, filter it with a 0.22 ⁇ m microporous filter, and pack aseptically.
  • mice were randomly divided into 6 groups, with 10 mice in each group. Inject 0.1ml/animal of the above composition intramuscularly, and perform the second immunization 2 weeks after the first immunization. On the 14th day after the second immunization, the blood of the mice was collected, and the serum hepatitis B antibody levels of the mice were detected by ELISA experiment.
  • GPS high-dose adjuvant group 2.5 ⁇ g hepatitis B vaccine + 250 ⁇ g GPS/animal;
  • GAPS high-dose adjuvant group 2.5 ⁇ g hepatitis B vaccine + 250 ⁇ g GAPS/bird;
  • GPS low-dose adjuvant group 2.5 ⁇ g hepatitis B vaccine + 25 ⁇ g GPS/animal;
  • GAPS low-dose adjuvant group 2.5 ⁇ g hepatitis B vaccine + 25 ⁇ g GAPS/bird;
  • Positive control group 2.5 ⁇ g hepatitis B vaccine + 25 ⁇ g aluminum salt adjuvant/animal;
  • Negative control group 2.5 ⁇ g hepatitis B vaccine/animal.
  • Example 5 Adjuvant activity of ginseng acidic polysaccharide (GAPS) for hepatitis A vaccine
  • Diploid hepatitis A vaccine produced by Liaoning Chengda Biological Co., Ltd.
  • mice Female C57BL/6 mice, 6-8 weeks old, were purchased from Beijing Huafukang Biotechnology Co., Ltd.
  • High-dose GPS adjuvant vaccine composition Measure appropriate amounts of ginseng polysaccharide (GPS) and hepatitis A vaccine respectively, dissolve them in physiological saline so that each milliliter of solution contains 2500 ⁇ g of GPS and 50 IU of hepatitis A vaccine, and filter with a 0.22 ⁇ m microporous membrane. Passed, sterile aliquots.
  • GPS ginseng polysaccharide
  • hepatitis A vaccine hepatitis A vaccine
  • High-dose GAPS adjuvant vaccine composition Measure appropriate amounts of ginseng acidic polysaccharide (GAPS) and hepatitis A vaccine respectively, dissolve them in physiological saline so that each milliliter of solution contains 2500 ⁇ g of GPS and 50 IU of hepatitis A vaccine, and filter with a 0.22 ⁇ m microporous membrane Filter and aliquot aseptically.
  • GAPS ginseng acidic polysaccharide
  • hepatitis A vaccine composition dissolve them in physiological saline so that each milliliter of solution contains 2500 ⁇ g of GPS and 50 IU of hepatitis A vaccine, and filter with a 0.22 ⁇ m microporous membrane Filter and aliquot aseptically.
  • Low-dose GPS adjuvant vaccine composition Measure appropriate amounts of ginseng polysaccharide (GPS) and hepatitis A vaccine respectively, dissolve them in physiological saline so that each milliliter of the solution contains 250 ⁇ g of GPS and 50 IU of hepatitis A vaccine, and filter with a 0.22 ⁇ m microporous membrane. Passed, sterile aliquots.
  • GPS ginseng polysaccharide
  • hepatitis A vaccine hepatitis A vaccine
  • GAPS adjuvant vaccine composition Measure appropriate amounts of ginseng acidic polysaccharide (GAPS) and hepatitis A vaccine respectively, dissolve them in physiological saline so that each milliliter of solution contains 250 ⁇ g of GPS and 50 IU of hepatitis A vaccine, and filter with a 0.22 ⁇ m microporous membrane Filter and aliquot aseptically.
  • GAPS ginseng acidic polysaccharide
  • hepatitis A vaccine hepatitis A vaccine
  • Positive control group - aluminum salt adjuvant vaccine composition measure appropriate amounts of aluminum salt and hepatitis A vaccine respectively, dissolve them in physiological saline so that each milliliter of solution contains 250 ⁇ g of aluminum salt and 50 IU of hepatitis A vaccine, and filter with a 0.22 ⁇ m microporous membrane Filter and aliquot aseptically.
  • Negative control group - vaccine without adjuvant Dissolve an appropriate amount of influenza vaccine in physiological saline so that each milliliter of the solution contains 50IU of hepatitis A vaccine, filter it with a 0.22 ⁇ m microporous filter, and pack aseptically.
  • mice were randomly divided into 4 groups, with 10 mice in each group. 0.1 ml of the above composition was injected intramuscularly per animal. Two weeks after the first immunization, the second immunization was performed. On the 14th day after the second immunization, the blood of each mouse was collected, and the neutralizing antibody level in the serum was detected.
  • GPS high-dose adjuvant group 5IU hepatitis A vaccine + 250 ⁇ g GPS/bird;
  • GAPS high-dose adjuvant group 5IU hepatitis A vaccine + 250 ⁇ g GAPS/bird;
  • GPS low-dose adjuvant group 5IU hepatitis A vaccine + 25 ⁇ g GPS/animal;
  • GAPS low-dose adjuvant group 5IU hepatitis A vaccine + 25 ⁇ g GAPS/bird;
  • Positive control group 5IU hepatitis A vaccine + 25 ⁇ g aluminum salt adjuvant/animal;
  • Negative control group 5IU hepatitis A vaccine/animal.
  • Example 6 Adjuvant activity of different doses of ginseng acidic polysaccharide (GAPS) for hepatitis A vaccine
  • GAPS adjuvant group 1 GAPS adjuvant group 2
  • GAPS adjuvant group 3 GAPS adjuvant group 4
  • GAPS adjuvant group 5 make each group contain 125, 250, 1250, 1250, 2500 and 5000 ⁇ g of GAPS and 50 IU of hepatitis A vaccine are filtered through a 0.22 ⁇ m microporous membrane and packed aseptically.
  • the ratios of GAPS adjuvant and hepatitis A vaccine in each group of vaccine compositions were 2.5, 5, 25, 50, and 100 ( ⁇ g:IU) respectively.
  • Positive control group - aluminum salt adjuvant vaccine composition measure appropriate amounts of aluminum salt and hepatitis A vaccine respectively, dissolve them in physiological saline so that each milliliter of solution contains 250 ⁇ g of aluminum salt and 50 IU of hepatitis A vaccine, and filter with a 0.22 ⁇ m microporous membrane Filter and aliquot aseptically.
  • Negative control group - vaccine without adjuvant Dissolve an appropriate amount of influenza vaccine in physiological saline so that each milliliter of the solution contains 50IU of hepatitis A vaccine, filter it with a 0.22 ⁇ m microporous filter, and pack aseptically.
  • mice were randomly divided into 7 groups, with 10 mice in each group. Inject 0.1ml/animal of the above composition intramuscularly, and perform the second immunization one week after the first immunization. On the 14th day after the second immunization, the blood of each mouse was collected, and the neutralizing antibody level of the mouse serum influenza vaccine was detected by hemagglutination inhibition test.
  • GAPS adjuvant group 5IU hepatitis A vaccine + 12.5 ⁇ g GAPS/bird;
  • GAPS adjuvant group 2 5IU hepatitis A vaccine + 25 ⁇ g GAPS/bird;
  • GAPS adjuvant group 3 5IU hepatitis A vaccine + 125 ⁇ g GAPS/bird;
  • GAPS adjuvant group 4 5IU hepatitis A vaccine + 250 ⁇ g GAPS/bird;
  • GAPS adjuvant group 5 5IU hepatitis A vaccine + 500 ⁇ g GAPS/bird;
  • Positive control group 5IU hepatitis A vaccine + 25 ⁇ g aluminum salt adjuvant/animal;
  • Negative control group 5IU hepatitis A vaccine/animal.
  • Example 7 Adjuvant activity of ginseng acidic polysaccharide (GAPS) for hepatitis C vaccine
  • Hepatitis C vaccine produced by Liaoning Chengda Biological Co., Ltd.
  • mice Female C57BL/6 mice, 6-8 weeks old, were purchased from Beijing Huafukang Biotechnology Co., Ltd.
  • High-dose GPS adjuvant vaccine composition measure appropriate amounts of ginseng polysaccharide (GPS) and hepatitis C vaccine respectively, Dissolve in physiological saline so that each milliliter of the solution contains 2500 ⁇ g of GPS and 50 ⁇ g of hepatitis C vaccine. Filter with a 0.22 ⁇ m microporous membrane and pack aseptically.
  • GPS ginseng polysaccharide
  • hepatitis C vaccine Dissolve in physiological saline so that each milliliter of the solution contains 2500 ⁇ g of GPS and 50 ⁇ g of hepatitis C vaccine. Filter with a 0.22 ⁇ m microporous membrane and pack aseptically.
  • High-dose GAPS adjuvant vaccine composition Measure appropriate amounts of ginseng acidic polysaccharide (GAPS) and hepatitis C vaccine respectively, dissolve them in physiological saline so that each milliliter of solution contains 2500 ⁇ g of GPS and 50 ⁇ g of hepatitis C vaccine, and filter with a 0.22 ⁇ m microporous membrane Filter and aliquot aseptically.
  • GAPS ginseng acidic polysaccharide
  • hepatitis C vaccine composition dissolve them in physiological saline so that each milliliter of solution contains 2500 ⁇ g of GPS and 50 ⁇ g of hepatitis C vaccine, and filter with a 0.22 ⁇ m microporous membrane Filter and aliquot aseptically.
  • Low-dose GPS adjuvant vaccine composition Measure appropriate amounts of ginseng polysaccharide (GPS) and hepatitis C vaccine respectively, dissolve them in physiological saline so that each milliliter of solution contains 250 ⁇ g of GPS and 50 ⁇ g of hepatitis C vaccine, and filter with a 0.22 ⁇ m microporous membrane. Passed, sterile aliquots.
  • GPS ginseng polysaccharide
  • hepatitis C vaccine hepatitis C vaccine
  • GAPS adjuvant vaccine composition Measure appropriate amounts of ginseng acidic polysaccharide (GAPS) and hepatitis C vaccine respectively, dissolve them in physiological saline so that each milliliter of solution contains 250 ⁇ g of GPS and 50 ⁇ g of hepatitis C vaccine, and filter with a 0.22 ⁇ m microporous membrane Filter and aliquot aseptically.
  • GAPS ginseng acidic polysaccharide
  • hepatitis C vaccine composition dissolve them in physiological saline so that each milliliter of solution contains 250 ⁇ g of GPS and 50 ⁇ g of hepatitis C vaccine, and filter with a 0.22 ⁇ m microporous membrane Filter and aliquot aseptically.
  • Positive control group - aluminum salt adjuvant vaccine composition measure appropriate amounts of aluminum salt and hepatitis C vaccine respectively, dissolve them in physiological saline so that each milliliter of solution contains 250 ⁇ g of aluminum salt and 50 ⁇ g of hepatitis C vaccine, and filter with a 0.22 ⁇ m microporous membrane Filter and aliquot aseptically.
  • Negative control group - vaccine without adjuvant Dissolve an appropriate amount of hepatitis C vaccine in physiological saline so that each milliliter of solution contains 50 ⁇ g of hepatitis C vaccine, filter it with a 0.22 ⁇ m microporous filter, and pack aseptically.
  • mice were randomly divided into 6 groups, with 10 mice in each group. 0.1 ml of the above composition was injected intramuscularly per animal. Two weeks after the first immunization, the second immunization was performed. On the 14th day after the second immunization, the blood of each mouse was collected, and the level of specific antibody IgG in the serum was detected.
  • GPS high-dose adjuvant group 5 ⁇ g hepatitis C vaccine + 500 ⁇ g GPS/animal;
  • GAPS high-dose adjuvant group 5 ⁇ g hepatitis C vaccine + 500 ⁇ g GAPS/bird;
  • GPS low-dose adjuvant group 5 ⁇ g hepatitis C vaccine + 50 ⁇ g GPS/animal;
  • GAPS low-dose adjuvant group 5 ⁇ g hepatitis C vaccine + 50 ⁇ g GAPS/bird;
  • Positive control group 5 ⁇ g hepatitis C vaccine + 50 ⁇ g aluminum salt adjuvant/animal;
  • Negative control group 5 ⁇ g hepatitis C vaccine/animal.
  • Example 8 Adjuvant activity of ginseng acidic polysaccharide (GAPS) for hand, foot and mouth vaccine
  • Hand, foot and mouth vaccine produced by Liaoning Chengda Biological Co., Ltd.
  • mice Female C57BL/6 mice, 6-8 weeks old, were purchased from Beijing Huafukang Biotechnology Co., Ltd.
  • High-dose GPS adjuvant vaccine composition Measure appropriate amounts of ginseng polysaccharide (GPS) and hand, foot and mouth vaccine respectively, dissolve them in physiological saline so that each milliliter of solution contains 2500 ⁇ g of GPS and 200 IU of hand, foot and mouth vaccine, filter with 0.22 ⁇ m micropores Membrane filtration, sterile packaging.
  • GPS ginseng polysaccharide
  • hand, foot and mouth vaccine dissolve them in physiological saline so that each milliliter of solution contains 2500 ⁇ g of GPS and 200 IU of hand, foot and mouth vaccine, filter with 0.22 ⁇ m micropores Membrane filtration, sterile packaging.
  • High-dose GAPS adjuvant vaccine composition Measure appropriate amounts of ginseng acidic polysaccharide (GAPS) and hand, foot and mouth vaccine respectively, dissolve them in physiological saline, so that each milliliter of solution contains 2500 ⁇ g of GPS and 200 IU of hand, foot and mouth vaccine, and use 0.22 ⁇ m micropores Filter through membrane and aliquot aseptically.
  • GAPS ginseng acidic polysaccharide
  • Low-dose GPS adjuvant vaccine composition Measure appropriate amounts of ginseng polysaccharide (GPS) and hand, foot and mouth vaccine respectively, dissolve them in physiological saline so that each milliliter of solution contains 250 ⁇ g of GPS and 200 IU of hand, foot and mouth vaccine, filter with 0.22 ⁇ m micropores Membrane filtration, sterile packaging.
  • GPS ginseng polysaccharide
  • hand, foot and mouth vaccine dissolve them in physiological saline so that each milliliter of solution contains 250 ⁇ g of GPS and 200 IU of hand, foot and mouth vaccine, filter with 0.22 ⁇ m micropores Membrane filtration, sterile packaging.
  • GAPS adjuvant vaccine composition Measure appropriate amounts of ginseng acidic polysaccharide (GAPS) and hand, foot and mouth vaccine respectively, dissolve them in physiological saline so that each milliliter of solution contains 250 ⁇ g of GPS and 200 IU of hand, foot and mouth vaccine, and use 0.22 ⁇ m micropores Filter through membrane and aliquot aseptically.
  • GAPS ginseng acidic polysaccharide
  • Positive control group - aluminum salt adjuvant vaccine composition measure appropriate amounts of aluminum salt and hand, foot and mouth vaccine respectively, dissolve them in physiological saline so that each milliliter of solution contains 250 ⁇ g of aluminum salt and 200 IU of hand, foot and mouth vaccine, and use 0.22 ⁇ m micropores Filter through membrane and aliquot aseptically.
  • Negative control group - vaccine without adjuvant Dissolve an appropriate amount of hand, foot and mouth vaccine in physiological saline so that each milliliter of solution contains 200IU of hand, foot and mouth vaccine, filter it with a 0.22 ⁇ m microporous filter, and pack aseptically.
  • Immunization scheme Mice were randomly divided into 6 groups, 10 mice in each group. Inject 0.1ml/mouse of the above composition intramuscularly. 2 weeks after the first immunization, perform the second immunization. On the 14th day after the second immunization, collect the blood of each mouse and detect serum neutralizing antibodies and specific antibody IgG. level.
  • GPS high-dose adjuvant group 20IU hand, foot and mouth vaccine + 250 ⁇ g GPS/bird;
  • GAPS high-dose adjuvant group 20IU hand, foot and mouth vaccine + 250 ⁇ g GAPS/bird;
  • GPS low-dose adjuvant group 20IU hand, foot and mouth vaccine + 25 ⁇ g GPS/bird;
  • GAPS low-dose adjuvant group 20IU hand, foot and mouth vaccine + 25 ⁇ g GAPS/bird;
  • Positive control group 20IU hand, foot and mouth vaccine + 25 ⁇ g aluminum salt adjuvant/animal;
  • Negative control group 20IU hand, foot and mouth vaccine/animal.
  • Example 9 Adjuvant activity of different doses of ginseng acidic polysaccharide (GAPS) for hand, foot and mouth vaccine
  • Positive control group - aluminum salt adjuvant vaccine composition measure appropriate amounts of aluminum salt and hand, foot and mouth vaccine respectively, dissolve them in physiological saline so that each milliliter of solution contains 250 ⁇ g of aluminum salt and 200 IU of hand, foot and mouth vaccine, and use 0.22 ⁇ m micropores Filter through membrane and aliquot aseptically.
  • Negative control group - vaccine without adjuvant Dissolve an appropriate amount of hand, foot and mouth vaccine in physiological saline so that each milliliter of solution contains 200IU of hand, foot and mouth vaccine, filter it with a 0.22 ⁇ m microporous filter, and pack aseptically.
  • mice were randomly divided into 7 groups, 10 mice in each group. 0.1 ml of the above composition was injected intramuscularly per animal. Two weeks after the first immunization, the second immunization was performed. On the 14th day after the second immunization, the blood of each mouse was collected, and the serum neutralizing antibody level was detected.
  • GAPS adjuvant group 1 20IU hand, foot and mouth vaccine + 12.5 ⁇ g GPS/bird;
  • GAPS adjuvant group 2 20IU hand, foot and mouth vaccine + 25 ⁇ g GPS/bird;
  • GAPS adjuvant group 3 20IU hand, foot and mouth vaccine + 125 ⁇ g GPS/bird;
  • GAPS adjuvant group 4 20IU hand, foot and mouth vaccine + 250 ⁇ g GPS/animal;
  • GAPS adjuvant group 5 20IU hand, foot and mouth vaccine + 500 ⁇ g GPS/bird;
  • Positive control group 20IU hand, foot and mouth vaccine + 25 ⁇ g aluminum salt adjuvant/animal;
  • Negative control group 20IU hand, foot and mouth vaccine/animal.
  • Example 10 Adjuvant activity of ginseng acidic polysaccharide (GAPS) to HPV vaccine
  • HPV vaccine produced by Liaoning Chengda Biological Co., Ltd.
  • mice Female C57BL/6 mice, 6-8 weeks old, were purchased from Beijing Huafukang Biotechnology Co., Ltd.
  • High-dose GPS adjuvant vaccine composition Measure appropriate amounts of ginseng polysaccharide (GPS) and HPV vaccine respectively, dissolve them in physiological saline so that each milliliter of solution contains 2500 ⁇ g of GPS and 20 ⁇ g of HPV vaccine, and filter with a 0.22 ⁇ m microporous membrane. Passed, sterile aliquots.
  • GPS ginseng polysaccharide
  • HPV vaccine HPV vaccine
  • High-dose GAPS adjuvant vaccine composition Measure appropriate amounts of ginseng acidic polysaccharide (GAPS) and HPV vaccine respectively, dissolve them in physiological saline so that each milliliter of solution contains 2500 ⁇ g of GPS and 20 ⁇ g of HPV vaccine, and filter with a 0.22 ⁇ m microporous membrane. Filter and aliquot aseptically.
  • Low-dose GPS adjuvant vaccine composition Measure appropriate amounts of ginseng polysaccharide (GPS) and HPV vaccine respectively, dissolve them in physiological saline so that each milliliter of solution contains 250 ⁇ g of GPS and 20 ⁇ g of HPV vaccine, and filter with a 0.22 ⁇ m microporous membrane. Passed, sterile aliquots.
  • GPS ginseng polysaccharide
  • HPV vaccine HPV vaccine
  • GAPS adjuvant vaccine composition Measure appropriate amounts of ginseng acidic polysaccharide (GAPS) and HPV vaccine respectively, dissolve them in physiological saline so that each milliliter of solution contains 250 ⁇ g of GPS and 20 ⁇ g of HPV vaccine, and filter with a 0.22 ⁇ m microporous membrane. Filter and aliquot aseptically.
  • GAPS ginseng acidic polysaccharide
  • HPV vaccine HPV vaccine
  • Positive control group-aluminum salt adjuvant vaccine composition measure appropriate amounts of aluminum salt and HPV vaccine respectively, dissolve them in physiological saline so that each milliliter of solution contains 250 ⁇ g of aluminum salt and 20 ⁇ g of HPV vaccine, and filter with a 0.22 ⁇ m microporous membrane Filter and aliquot aseptically.
  • Negative control group - vaccine without adjuvant Dissolve an appropriate amount of HPV vaccine in physiological saline so that each milliliter of solution contains 20 ⁇ g of HPV vaccine, filter it with a 0.22 ⁇ m microporous filter, and pack aseptically.
  • Immunization scheme Mice were randomly divided into 6 groups, 10 mice in each group. 0.1 ml of the above composition was injected intramuscularly per animal. Two weeks after the first immunization, the second immunization was performed. On the 14th day after the second immunization, the blood of each mouse was collected, and the level of specific antibody IgG in the serum was detected.
  • GPS high-dose adjuvant group 2 ⁇ g HPV vaccine + 250 ⁇ g GPS/bird;
  • GAPS high-dose adjuvant group 2 ⁇ g HPV vaccine + 250 ⁇ g GAPS/bird;
  • GPS low-dose adjuvant group 2 ⁇ g HPV vaccine + 25 ⁇ g GPS/bird;
  • GAPS low-dose adjuvant group 2 ⁇ g HPV vaccine + 25 ⁇ g GAPS/bird;
  • Positive control group 2 ⁇ g HPV vaccine + 25 ⁇ g aluminum salt adjuvant/animal;
  • Negative control group 2 ⁇ g HPV vaccine/animal.
  • Example 11 Adjuvant activity of different doses of ginseng acidic polysaccharide (GAPS) for HPV vaccine
  • Positive control group-aluminum salt adjuvant vaccine composition measure appropriate amounts of aluminum salt and HPV vaccine respectively, dissolve them in physiological saline so that each milliliter of solution contains 250 ⁇ g of aluminum salt and 20 ⁇ g of HPV vaccine, and filter with a 0.22 ⁇ m microporous membrane Filter and aliquot aseptically.
  • Negative control group - vaccine without adjuvant Dissolve an appropriate amount of HPV vaccine in physiological saline so that each milliliter of solution contains 20 ⁇ g of HPV vaccine, filter it with a 0.22 ⁇ m microporous filter, and pack aseptically.
  • mice were randomly divided into 7 groups, 10 mice in each group. Inject 0.1 of the above composition intramuscularly ml/mouse, 2 weeks after the first immunization, the second immunization was performed. On the 14th day after the second immunization, the blood of each mouse was collected, and the level of specific antibody IgG in the serum was detected.
  • GAPS adjuvant group 1 2 ⁇ g HPV vaccine + 12.5 ⁇ g GAPS/bird;
  • GAPS adjuvant group 2 2 ⁇ g HPV vaccine + 25 ⁇ g GAPS/bird;
  • GAPS adjuvant group 3 2 ⁇ g HPV vaccine + 125 ⁇ g GAPS/bird;
  • GAPS adjuvant group 4 2 ⁇ g HPV vaccine + 250 ⁇ g GAPS/bird;
  • GAPS adjuvant group 5 2 ⁇ g HPV vaccine + 500 ⁇ g GAPS/bird;
  • Positive control group 2 ⁇ g HPV vaccine + 25 ⁇ g aluminum salt adjuvant/animal;
  • Negative control group 2 ⁇ g HPV vaccine/animal.
  • mice in the GAPS adjuvant groups 2, 3, and 4 were significantly higher than the negative control group and the positive control group.
  • the antibody levels in mice in GAPS adjuvant groups 1 and 5 increased to a certain extent, they were significantly lower than those in adjuvant groups 2 to 4, and were not significantly different from the positive control group.
  • Example 12 Adjuvant activity of ginseng acidic polysaccharide (GAPS) on novel coronavirus vaccine (COVID-19 vaccine)
  • COVID-19 vaccine produced by Liaoning Chengda Biological Co., Ltd.
  • mice Female C57BL/6 mice, 6-8 weeks old, were purchased from Beijing Huafukang Biotechnology Co., Ltd.
  • High-dose GPS adjuvant vaccine composition Measure appropriate amounts of ginseng polysaccharide (GPS) and new coronavirus vaccine respectively, dissolve them in physiological saline so that each milliliter of solution contains 5000 ⁇ g of GPS and 50 ⁇ g of new coronavirus vaccine, and filter with 0.22 ⁇ m micropores. Membrane filtration, sterile packaging.
  • GPS ginseng polysaccharide
  • new coronavirus vaccine sterile packaging.
  • High-dose GAPS adjuvant vaccine composition Measure appropriate amounts of ginseng acidic polysaccharide (GAPS) and new coronavirus vaccine respectively, dissolve them in physiological saline, so that each milliliter of solution contains 5000 ⁇ g of GPS and 50 ⁇ g of new coronavirus vaccine, and use 0.22 ⁇ m micropores Filter through membrane and aliquot aseptically.
  • GAPS ginseng acidic polysaccharide
  • new coronavirus vaccine composition Measure appropriate amounts of ginseng acidic polysaccharide (GAPS) and new coronavirus vaccine respectively, dissolve them in physiological saline, so that each milliliter of solution contains 5000 ⁇ g of GPS and 50 ⁇ g of new coronavirus vaccine, and use 0.22 ⁇ m micropores Filter through membrane and aliquot aseptically.
  • Low-dose GPS adjuvant vaccine composition Measure appropriate amounts of ginseng polysaccharide (GPS) and new coronavirus vaccine respectively, dissolve them in physiological saline so that each milliliter of solution contains 500 ⁇ g of GPS and 50 ⁇ g of new coronavirus vaccine, and filter with 0.22 ⁇ m micropores. Membrane filtration, sterile packaging.
  • GPS ginseng polysaccharide
  • new coronavirus vaccine sterile packaging.
  • GAPS adjuvant vaccine composition measure ginseng acidic polysaccharide (GAPS) and COVID-19 Dissolve an appropriate amount of coronavirus vaccine in physiological saline so that each milliliter of the solution contains 500 ⁇ g of GPS and 50 ⁇ g of COVID-19 vaccine. Filter it with a 0.22 ⁇ m microporous filter and pack aseptically.
  • Aluminum salt adjuvant vaccine composition Measure appropriate amounts of aluminum salt and new coronavirus vaccine respectively, dissolve them in physiological saline, so that each milliliter of solution contains 500 ⁇ g of aluminum salt and 50 ⁇ g of new coronavirus vaccine, and use 0.22 ⁇ m micropores Filter through membrane and aliquot aseptically.
  • Negative control group - vaccine without adjuvant Dissolve an appropriate amount of the new coronavirus vaccine in physiological saline so that each milliliter of the solution contains 50 ⁇ g of the new coronavirus vaccine, filter it with a 0.22 ⁇ m microporous filter, and pack aseptically.
  • Immunization scheme Mice were randomly divided into 6 groups, 10 mice in each group. 0.1 ml of the above composition was injected intramuscularly per animal. Two weeks after the first immunization, the second immunization was performed. On the 14th day after the second immunization, the blood of each mouse was collected, and the level of specific antibody IgG in the serum was detected.
  • GPS high-dose group 5 ⁇ g COVID-19 vaccine + 500 ⁇ g GPS/animal;
  • GAPS high-dose group 5 ⁇ g COVID-19 vaccine + 500 ⁇ g GAPS/bird;
  • GPS low-dose group 5 ⁇ g COVID-19 vaccine + 50 ⁇ g GPS/animal;
  • GAPS low-dose group 5 ⁇ g COVID-19 vaccine + 50 ⁇ g GAPS/bird;
  • Positive control group 5 ⁇ g COVID-19 vaccine + 50 ⁇ g aluminum salt adjuvant/animal;
  • Negative control group 5 ⁇ g new coronavirus vaccine/animal.
  • Example 13 Adjuvant activity of different doses of ginseng acidic polysaccharide (GAPS) on novel coronavirus vaccine (COVID-19 vaccine)
  • Aluminum salt adjuvant vaccine composition Measure appropriate amounts of aluminum salt and COVID-19 vaccine respectively, dissolve them in physiological saline so that each milliliter of solution contains 250 ⁇ g of aluminum salt and 50 ⁇ g of COVID-19 vaccine, filter with a 0.22 ⁇ m microporous membrane Filter and aliquot aseptically.
  • Negative control group - vaccine without adjuvant Dissolve an appropriate amount of the new coronavirus vaccine in physiological saline so that each milliliter of the solution contains 50 ⁇ g of the new coronavirus vaccine, filter it with a 0.22 ⁇ m microporous filter, and pack aseptically.
  • Immunization scheme Mice were randomly divided into 7 groups, 10 mice in each group. 0.1 ml of the above composition was injected intramuscularly per animal. Two weeks after the first immunization, the second immunization was performed. On the 14th day after the second immunization, the blood of each mouse was collected, and the level of specific antibody IgG in the serum was detected.
  • GAPS adjuvant group 1 5 ⁇ g COVID-19 vaccine + 25 ⁇ g GPS/bird;
  • GAPS adjuvant group 2 5 ⁇ g COVID-19 vaccine + 50 ⁇ g GPS/bird;
  • GAPS adjuvant group 3 5 ⁇ g COVID-19 vaccine + 250 ⁇ g GPS/bird;
  • GAPS adjuvant group 4 5 ⁇ g COVID-19 vaccine + 500 ⁇ g GPS/bird;
  • GAPS adjuvant group 5 5 ⁇ g COVID-19 vaccine + 1000 ⁇ g GPS/bird;
  • Positive control group 5 ⁇ g COVID-19 vaccine + 25 ⁇ g aluminum salt adjuvant/animal;
  • Negative control group 5 ⁇ g new coronavirus vaccine/animal.

Abstract

L'invention concerne un adjuvant de vaccin comprenant un polysaccharide acide de ginseng (GAPS), qui peut améliorer significativement le titre d'un anticorps spécifique (ou d'un anticorps neutralisant) après immunisation par antigène et peut améliorer efficacement l'activité de réponse immunitaire d'un vaccin contre la rage, d'un vaccin contre la grippe, d'un vaccin contre l'hépatite B, d'un vaccin contre l'hépatite A, d'un vaccin contre l'hépatite C, d'un vaccin contre la maladie pieds-mains-bouche, d'un vaccin contre les HPV et d'un vaccin contre le nouveau coronavirus.
PCT/CN2023/087301 2022-08-23 2023-04-10 Adjuvant de vaccin à base de polysaccharide acide de ginseng, composition de vaccin et son utilisation WO2024040979A1 (fr)

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