CN107281483A - The preparation method of immune activator and composition and its application and composition - Google Patents

The preparation method of immune activator and composition and its application and composition Download PDF

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CN107281483A
CN107281483A CN201710361460.0A CN201710361460A CN107281483A CN 107281483 A CN107281483 A CN 107281483A CN 201710361460 A CN201710361460 A CN 201710361460A CN 107281483 A CN107281483 A CN 107281483A
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hepatitis
immune
hbsag
composition
formula
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靳广毅
王竹林
胡云龙
唐黎
刘冰
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SHENZHEN KANGJUZHENG MEDICAL TECHNOLOGY Co Ltd
Shenzhen University
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SHENZHEN KANGJUZHENG MEDICAL TECHNOLOGY Co Ltd
Shenzhen University
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Priority to PCT/CN2018/084771 priority Critical patent/WO2018214699A1/en
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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
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
    • A61K31/505—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim
    • A61K31/519—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim ortho- or peri-condensed with heterocyclic rings
    • A61K31/52—Purines, e.g. adenine
    • 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
    • A61K39/12—Viral antigens
    • 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
    • A61K39/385—Haptens or antigens, bound to carriers
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D473/00—Heterocyclic compounds containing purine ring systems
    • C07D473/02—Heterocyclic compounds containing purine ring systems with oxygen, sulphur, or nitrogen atoms directly attached in positions 2 and 6
    • C07D473/18—Heterocyclic compounds containing purine ring systems with oxygen, sulphur, or nitrogen atoms directly attached in positions 2 and 6 one oxygen and one nitrogen atom, e.g. guanine
    • 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
    • A61K2039/55505—Inorganic 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
    • A61K2039/57—Medicinal preparations containing antigens or antibodies characterised by the type of response, e.g. Th1, Th2
    • A61K2039/575—Medicinal preparations containing antigens or antibodies characterised by the type of response, e.g. Th1, Th2 humoral response
    • 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
    • A61K2039/60—Medicinal preparations containing antigens or antibodies characteristics by the carrier linked to the antigen

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Abstract

The present invention relates to a kind of preparation method of immune activator and composition and its application and composition, it is in animal and human body on the basis of the hepatitis B specific antibody of original preventative vaccine is kept, the cell for infecting hepatitis B can be inactivated and removed, the double action with prevention and treatment；It has stronger immune activation effect than original HBsAg antigens, show that the immune cell factor induction of reinforcement, higher antibody tell on, particularly the immune generation of high level mark Th1 types of interferon and IgG2a antibody, has significant innovation and practicality for removing hepatitis B.

Description

The preparation method of immune activator and composition and its application and composition
Technical field
The present invention relates to the technical field of hepatitis B vaccine prevention and treatment, more specifically to immune activator and group Compound and its application and the preparation method of composition.
Background technology
Hepatitis B vaccine is the medicine for preventing hepatitis B.After vaccine inoculation, can stimulating immune system produce protection antibody, So that human body is provided with the immunity of prevention hepatitis B, reaches and prevent hepatitis B infected purpose.But China's infection hepatitis B (HBV) crowd has 90,000,000 people, and wherein Chronic Hepatitis B has 20,000,000 people.For chronic hepatitis B infections person and patient, in advance Anti- property hepatitis B vaccine is substantially invalid.Therefore it is highly desirable to a kind of " treatment that can prevent and hepatitis B chronic infection can be removed Property hepatitis B vaccine ".
The content of the invention
It is an object of the invention to provide immune activator and composition and its application and the preparation method of composition, solution Having determined hepatitis B vaccine of the prior art can not be while the problem of preventing and removes hepatitis B chronic infection.
The technical proposal for solving the technical problem of the invention is：A kind of immune activator, its formula is as follows：
Wherein, R is OH or SH.
In the immune activator of the present invention, the immune activator is to be selected from following structural formula I, formula II, structure The compound of formula III or structural formula IV：
Application of the above-mentioned immune activator in therapeutic hepatitis B vaccine is prepared of the present invention and preparing people and dynamic Application in thing immunoregulation medicament.
The present invention also provides a kind of composition for being used to prepare prevention and treatment hepatitis B vaccine, includes above-mentioned immune excitement Agent, recombination hepatitis B surface antigen and aluminium adjuvant, the ratio of components of the recombination hepatitis B surface antigen, aluminium adjuvant and immune activator Example is X：Y：Z, wherein, X, Y, Z are the numeral between 1-120 respectively and represent molal quantity, volume number or mass number.
In the present compositions, the proportion of composing of the recombination hepatitis B surface antigen, aluminium adjuvant and immune activator For X：Y：Z=1：20.36：20~31.8.
In the present compositions, the recombination hepatitis B surface antigen is HBsAg, and HBsAg amino acid sequence includes 200-400 amino acid, HBsAg molecular weight is 20000Da-44000Da.
In the present compositions, HBsAg amino acid sequence includes 200-250 amino acid；HBsAg molecular weight For 20000Da-25000Da.
In the present compositions, the aluminium adjuvant is gel aluminum hydroxide, aluminum phosphate, aluminum sulfate, ammonia-alum or potassium Alum.
Present invention also offers the preparation method of combinations of the above thing, it is characterised in that including：Will be with DMSO or physiology salt Water is that the HBsAg mother liquors of solvent and the aluminium adjuvant solution of 9 times of volumes are mixed to get mixed liquor, then by the mixed liquor with excitement is immunized Agent presses 9：1 volume ratio is mixed to get the composition for preparing therapeutic hepatitis B vaccine.
The combinations of the above thing of the present invention is for improving antibody titer and effect T for hepatitis B surface antigen HBsAg Applied in cell and the application in the vaccine and medicine for preparing prevention and treatment hepatitis B.
Implement the immune activator of the present invention and the preparation method of composition and its application and composition, have with following Beneficial effect：The present invention provides a kind of immune activator and for preparing prevention and treatment hepatitis B vaccine for chronic hepatitis B infections person Composition, its in animal and human body on the basis of the hepatitis B specific antibody of original preventative vaccine is kept, can The cell for infecting hepatitis B is inactivated and removed, the double action with prevention and treatment；It is than original HBsAg antigens With stronger immune activation effect, show that the immune cell factor induction of reinforcement, higher antibody tell on, particularly The immune generation of high level mark Th1 types of gamma interferon and IgG2a antibody, has significant innovation for removing hepatitis B Property and practicality.
Brief description of the drawings
Fig. 1 is the immune activator SZU-103 of formula I synthesis technique figure；
Fig. 2 schemes for the immune activator SZU-103 of formula I ESI-MS；
Fig. 3 is the immune activator SZU-117 of formula II synthesis technique figure；
Fig. 4 schemes for the immune activator SZU-117 of formula II ESI-MS；
Fig. 5 is the immune activator SZU-114 of formula III synthesis technique figure；
Fig. 6 schemes for the immune activator SZU-114 of formula III ESI-MS；
Fig. 7 is the synthesis technique figure for the immune activator SZU-122 that structural formula IV is represented；
The ESI-MS figures that Fig. 8 is the immune activator SZU-122 that structural formula IV is represented；
Fig. 9 A are the priming effect figure of immune activator that formula I is representative to immune cell factor IL-6；
Fig. 9 B are the priming effect figure of immune activator that formula I is representative to immune cell factor IFN γ；
Fig. 9 C are the priming effect figure of immune activator that formula I is representative to immune cell factor IL-12；
Figure 10 A are that the immune activator and control group that formula I is representative induce HBsAg antigentic specificity IgG1 antibody respectively Immune response comparison diagram；
Figure 10 B are that the immune activator and control group that formula I is representative induce HBsAg antigentic specificities IgG2a to resist respectively Body immune response comparison diagram；
Figure 11 A are that the immune activator that formula I is representative reacts as hepatitis B vaccine adjuvant inducing antigen-specific T cell Photo；
Figure 11 B are that the immune activator that formula I is representative is special as hepatitis B vaccine adjuvant and control group difference inducing antigen The comparison diagram of specific T cell reaction；
Figure 12 A are immune activator that formula I is representative as hepatitis B vaccine adjuvant inducing antigen-specific CD4+T cell The fluorogram of the effect of reaction；
Figure 12 B are that the immune activator that formula I is representative is special as hepatitis B vaccine adjuvant and control group difference inducing antigen Different in nature CD4+The comparison diagram of the effect of t cell responses；
Figure 12 C are immune activator that formula I is representative as hepatitis B vaccine adjuvant inducing antigen-specific CD8+T cell The fluorogram of the effect of reaction；
Figure 12 D are that the immune activator that formula I is representative is special as hepatitis B vaccine adjuvant and control group difference inducing antigen Different in nature CD8+The comparison diagram of the effect of t cell responses.
Embodiment
With reference to the accompanying drawings and examples, to the present invention immune activator and composition and its application and composition Preparation method is described further：
Advantage using small molecule immune activator as vaccine adjuvant is its Stability Analysis of Structures, it is easy to artificial synthesized purifying and Manufacture, but the shortcoming of small molecule immune adjuvant is body that to absorb fast, metabolism fast, therefore can not well be cooperateed with antigen protein Distribution and metabolism.The structural formula for the small molecule immune activator applied in the present invention carries α, beta-unsaturated carbonyl group, to antigen Albumen (the particularly albumen containing sulfhydryl residue) has Covalent attachment effect；To aluminium atom and aluminum ions empty Atomic Orbits stage property There is electron adsorption effect.These advantages of the immune activator of the present invention substantially increase the immune synergism of itself and antigen, Therefore also have from mechanism significant innovative.
The immune activator, its formula is as follows：
SM360320 (1,2) is TLR7 activators (Proc Natl Acad Sci U S A.2006Feb 7 in formula I； 103(6):1828-33；Blood.2011May 26；117(21):5683-91).In addition, the oh group (- OH) in formula I It can also be the sulfydryl (- SH) of homologue.
Wherein, the SM360320 in formula I has such as the dynamic isomer of formula 1 and the same compound of formula 2：
Specifically, immune activator is the chemical combination selected from following structural formula I, formula II, formula II I or structural formula IV Thing：
Structure above is characterized as comprising (TLR7) the activator SM360320 of Toll-like receptor 7 with having α, β-unsaturation carbonyl The chemical combination objects system of base group.It should be noted that structural formula I, formula II, formula II I or structural formula IV compound Can also be 8 hydroxyls and hydroxy tautomer.
The present invention also provides a kind of composition for being used to prepare prevention and treatment hepatitis B vaccine, includes above-mentioned immune excitement Agent and recombination hepatitis B surface antigen and aluminium adjuvant, the proportion of composing of recombination hepatitis B surface antigen, aluminium adjuvant and immune activator For X：Y：Z, wherein, X, Y, Z are the numeral between 1-120 respectively and represent molal quantity, volume number or mass number.Preferably, The mass ratio of recombination hepatitis B surface antigen, aluminium adjuvant and immune activator is X：Y：Z=1：20.36：20~31.8.
Wherein, recombination hepatitis B surface antigen is HBsAg, and HBsAg amino acid sequence includes 200-400 amino acid, HBsAg molecular weight is 20000Da-44000Da.Preferably, HBsAg amino acid sequence includes 200-250 amino acid； HBsAg molecular weight is 20000Da-25000Da.HBsAg amino acid Typical Representative sequence is following (containing 226 amino acid) (《Chinese biological engineering magazine》1995,15 (2)：43-45)：
Met Glu Asn Ile Thr Ser Gly Phe Leu Gly Pro Leu Leu Val Leu Gln Ala Gly Phe Phe Leu Leu Thr Arg Ile Leu Thr Ile Pro Gln Ser Leu Asp Ser Trp Trp Thr Ser Leu Asn Phe Leu Gly Gly Ser Pro Val Cys Leu Gly Gln Asn Ser Gln Ser Pro Thr Ser Asn His Ser Pro Thr Ser Cys Pro Pro Ile Cys Pro Gly Tyr Arg Trp Met Gys Leu Arg Arg Phe Ile Ile Phe Leu Phe Ile Leu Leu Leu Cys Leu Ile Phe Leu Leu Val Leu Leu Asp Tyr Gln Gly Met Leu Pro Val Cys Pro Leu Ile Pro Gly Ser Thr Thr Thr Ser Thr GlyPro Cys Lys Thr Cys Thr Thr Pro Ala Gln Gly Asn Ser Met Phe Pro Ser Cys Cys Cys Thr Lys Pro ThrAsp GlyAsn Cys Thr Cys Ile ProIle Pro Ser SerTrpAla Phe Ala Lys Tyr Leu TrpGlu Trp Ala Ser Val Arg Phe Ser Trp Leu Ser Leu Leu Val Pro Phe Val Gln Trp Phe Val Gly Leu Ser Pro Thr Val Trp Leu Ser Ala Ile Trp Met Met Trp Tyr Trp Gly Pro Ser Leu Tyr Ser Ile Val Ser Pro Phe Ile Pro Leu Leu Pro Ile Phe Phe Cys Leu Trp Val Tyr Ile
Above-mentioned aluminium adjuvant can be gel aluminum hydroxide, aluminum phosphate, aluminum sulfate, ammonia-alum or potassium alum.
The present invention provides a kind of immune activator and for preparing prevention and treatment hepatitis B epidemic disease for chronic hepatitis B infections person The composition of seedling, its in animal and human body on the basis of the hepatitis B specific antibody of original preventative vaccine is kept, energy Enough cells to infecting hepatitis B are inactivated and removed, the double action with prevention and treatment；It is more anti-than original HBsAg Original has stronger immune activation effect, shows that the immune cell factor induction of reinforcement, higher antibody tell on, especially It is the immune generation of high level mark Th1 types of gamma interferon and IgG2a antibody, there is significant wound for removing hepatitis B New property and practicality.
Above-mentioned immune activator can be used for preparing hepatitis vaccine, preventative vaccine and therapeutic epidemic disease for hepatitis B Seedling.
It is described in detail below by specific embodiment.
Embodiment 1：The synthesis of the immune activator (SZU-103) of formula I
As shown in figure 1, weighing 344mg (1mmol) SZU-008T and 341mg (1.125mmol) ethacrynic acid is dissolved in 8mL DMF In, the DMAP of HBTU (427mg, 1.125mmol), triethylamine (416 μ L, 3mmol) and catalytic amount is added, stirs anti-at room temperature Should, overnight.Reaction solution is poured into 100mL water, suction filtration, filter residue is through washing, dry crude product.Again through column chromatography for separation (DCM: MeOH=20:1) white solid 475mg, yield 75.5%, are obtained.As shown in Fig. 2 ESI-MS:M/z=629.1 [M+H]+。
Embodiment 2：The synthesis of the immune activator (SZU-117) of formula II
As shown in figure 3, in SZU-101 (1g, 0.002mol) anhydrous DMF solution, addition EDCI (0.0025mol), Half an hour is stirred at room temperature after HOBT (0.0025mol).DIEA (0.004mol) is added dropwise at 0 DEG C and adds N-Boc- piperazines afterwards (0.0025mol), room temperature reaction is stayed overnight.Reaction solution is poured into water, suction filtration, added after washing and drying to terminating by TLC monitoring reactions Enter 1:4TFA/DCM mixed solvents are stirred overnight.After the completion of reaction, decompression removes solvent, is dissolved again with a small amount of ethanol, adds 2N hydrochloric acid is into salt.After being spin-dried for, triturated under ether is added, filtering obtains SZU-130, yield 56%.
After SZU-130 (100mg), acryloyl chloride and triethylamine are mixed at 0 DEG C, room temperature reaction 2h is transferred to, by reaction solution It is poured into water, suction filtration, filter residue is through washing, dry crude product.Again through column chromatography for separation, white solid 80mg, yield 72% are obtained.Such as Shown in Fig. 4, ESI-MS:M/z=567.2 [M+H]+。
Embodiment 3：The synthesis of the immune activator (SZU-114) of formula III
As shown in figure 5, SZU-008T (350mg), EDCI (172mg), HOBT (172mg) are dissolved in dry DMF, 0 DEG C Lower dropwise addition DIEA (0.35mL).After drop finishes, it is transferred to room temperature and continues to react after 30min, add the trans -4- dimethyl amidos of 117mg Cronate hydrochlorate, room temperature reaction is stayed overnight.LCMS is monitored, and after completion of the reaction, reaction solution is poured into water, suction filtration, filter residue is through water Wash, dry crude product.Again through preparing liquid phase separation, white solid 130mg, yield 33% are obtained.As shown in fig. 6, ESI-MS:m/z =456.2 [M+H]+。
Embodiment 4：The synthesis for the immune activator (SZU-122) that structural formula IV is represented is as shown in fig. 7, SZU-008T After (300mg), acryloyl chloride and triethylamine are mixed at 0 DEG C, room temperature reaction 2h is transferred to, reaction solution is poured into water, suction filtration, is filtered Slag is through washing, dry crude product.Again through column chromatography for separation, white solid 280mg, yield 80% are obtained.As shown in figure 8, ESI-MS: M/z=399.1 [M+H]+。
Embodiment 5：Preparation method of composition for preparing prevention and treatment hepatitis B vaccine
By HBsAg mother liquor (Shenzhen Kangtai Biological Product Co., Ltd. provide) and 9 times bodies of the concentration for 220 μ g/mL Long-pending aluminium adjuvant solution ((OH) containing Al3Concentration is 1.44mg/mL) mixed liquor that concentration containing HBsAg is 22 μ g/mL is mixed to get, The mixed liquor and 10mM immune activator are pressed 9:1 volume mixture (the volume of vaccine solution 9：The immune volume of Agonist solutions 1) Obtain the hepatitis B vaccine of final composition.Weight ratio (HBsAg)：(Al)：(M)=X：Y：Z=0.22mg：4.48mg：7mg=1: 20.36:31.8.Wherein M is immune activator SZU-103.
With SZU-114 replacements SZU-103 preparation method ibid, weight ratio (HBsAg) is prepared：(Al)：(M)=X：Y：Z =0.22mg：4.48mg：5mg=1:20.36:22.7.
With SZU-117 replacements SZU-103 preparation method ibid, weight ratio (HBsAg) is prepared：(Al)：(M)=X：Y：Z =0.22mg：4.48mg：6.3mg=1:20.36:28.6.
With SZU-122 replacements SZU-103 preparation method ibid, weight ratio (HBsAg) is prepared：(Al)：(M)=X：Y：Z =0.22mg：4.48mg：4.4mg=1:20.36:20.
It is DMSO or physiological saline that activator mother liquor configuration solvent, which is immunized, in wherein 10mM；Other aluminium adjuvants replace having with aluminium Effect content is defined (initial aluminum assist agent solution contains aluminium 0.5mg/mL).HBsAg form can be monomer or many condensates, its mother liquor Concentration range is calculated as between 220 μ g/mL-250 μ g/mL using HBsAg.
Biological activity test is specifically described below.
Embodiment 6：Cell in vitro factor excitation
The spleen lymphocyte originated using Balb/c mouse is used as test system.1 × 106/mL spleen lymphocyte It is small for 0.03-40 μM of SZU-101, SZU-103, SZU-114, SZU-117 and SZU-122 co-incubation 24 with concentration range When, collect and IL6 and IFN γ concentration in Elisa methods detection supernatant are used after supernatant, R848 is no added as positive control It is used as negative control.As a result such as Fig. 9 A, 9B and 9C, immune activator swashs to immune cell factor IL-6, IL-12 and IFN-γ Send out effect notable.
Embodiment 7：Formula I is used as hepatitis B vaccine adjuvant inducing antigen-specific humoral immunity for the immune activator of representative The effect of reaction
Balb/c mouse (every group 5) are 0, carry out within 14 and 28 days it is immune (2 μ g HBsAg antigen mixing 100nmol's TLR7 activators, 1.12mg/mL Al (OH) 3 are used as another addition adjuvant).Mice serum is obtained in separation in the 35th day, is used Elisa methods detection HBsAg in serum specific IgG 1 and IgG2a antibody titers.R848, commercial hepatitis B vaccine and PBS injections are made For control group.As a result see Figure 10 A and 10B, activator is immunized anti-as hepatitis B vaccine adjuvant inducing antigen-specific humoral immunity The effect answered is obvious.
Embodiment 8：Formula I is anti-as hepatitis B vaccine adjuvant inducing antigen-specific T cell for the immune activator of representative The effect answered
Balb/c mouse (every group 5) are 0, carry out within 14 and 28 days it is immune (2 μ g HBsAg antigen mixing 100nmol's TLR7 activators, 1.12mg/mL Al (OH) 3 are used as another addition adjuvant).It is thin that mouse spleen lymph is obtained in separation in the 35th day Born of the same parents, HBsAg specific T-cells levels in spleen lymphocyte are detected with Elispot methods.R848, commercial hepatitis B vaccine and PBS Injection is as a control group.See Figure 11 A and Figure 11 B, as a result show immune activators of the formula I for representative as hepatitis B vaccine adjuvant The effect of inducing antigen-specific T cell reaction is obvious.
Embodiment 9：Formula I for representative immune activator as hepatitis B vaccine adjuvant inducing antigen-specific CD4+ and The effect of CD8+T cell effects
Balb/c mouse (every group 5) are 0, carry out within 14 and 28 days it is immune (2 μ g HBsAg antigen mixing 100nmol's TLR7 activators, 1.12mg/mL Al (OH) 3 are used as another addition adjuvant).It is thin that mouse spleen lymph is obtained in separation in the 35th day Born of the same parents, HBsAg specific C D4+ and CD8+T cell proportions are identified with the dyeing of intracellular IFN γ and Flow Cytometry.R848 and business With hepatitis B vaccine as a control group.See Figure 12 A, 12B, 12C and 12D, as a result show the immune activator conduct that formula I is representative Hepatitis B vaccine adjuvant inducing antigen-specific CD4+And CD8+The effect of t cell responses is obvious.
It should be appreciated that for those of ordinary skills, can according to the above description be improved or converted, All these improvement or conversion should all belong within the protection domain of appended claims of the present invention.

Claims (10)

1. a kind of immune activator, its formula is as follows：
Wherein, R is OH or SH.
2. immune activator according to claim 1, it is characterised in that the immune activator is to be selected from following structural formula I, formula II, formula II I or structural formula IV compound：
3. application of the immune activator described in claim 1 or 2 in therapeutic hepatitis B vaccine is prepared and prepare people and Application in animal immune regulating drug.
4. a kind of composition for being used to prepare prevention and treatment hepatitis B vaccine, it is characterised in that comprising described in claim 1 or 2 Immune activator, recombination hepatitis B surface antigen and aluminium adjuvant, the recombination hepatitis B surface antigen, aluminium adjuvant and immune excitement The proportion of composing of agent is X：Y：Z, wherein, X, Y, Z are the numeral between 1-120 respectively and represent molal quantity, volume number or matter Measure number.
5. composition according to claim 4, it is characterised in that the recombination hepatitis B surface antigen, aluminium adjuvant and immune The proportion of composing of activator is X：Y：Z=1：20.36：20~31.8.
6. composition according to claim 4, it is characterised in that the recombination hepatitis B surface antigen is HBsAg, HBsAg Amino acid sequence include 200-400 amino acid, HBsAg molecular weight is 20000Da-44000Da.
7. composition according to claim 6, it is characterised in that HBsAg amino acid sequence includes 200-250 amino Acid；HBsAg molecular weight is 20000Da-25000Da.
8. composition according to claim 4, it is characterised in that the aluminium adjuvant is gel aluminum hydroxide, aluminum phosphate, sulphur Sour aluminium, ammonia-alum or potassium alum.
9. the preparation method of any described composition in claim 4-8, it is characterised in that including：Will be with DMSO or physiology Salt solution is that the HBsAg mother liquors of solvent and the aluminium adjuvant solution of 9 times of volumes are mixed to get mixed liquor, then the mixed liquor is swashed with immune Dynamic agent presses 9：1 volume ratio is mixed to get the composition for preparing therapeutic hepatitis B vaccine.
10. any described composition is for improving the antibody titer for hepatitis B surface antigen HBsAg in claim 4-8 With the application applied in effector T cell and in the vaccine and medicine for preparing prevention and treatment hepatitis B.
CN201710361460.0A 2017-04-25 2017-05-22 The preparation method of immune activator and composition and its application and composition Pending CN107281483A (en)

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Cited By (4)

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WO2018214699A1 (en) * 2017-04-25 2018-11-29 深圳大学 Immune agonist and composition and use thereof, and preparation method for composition
WO2019192454A1 (en) * 2018-04-03 2019-10-10 深圳大学 Novel small molecule immune agonists and immune targeting compounds and application thereof
CN112336852A (en) * 2020-10-14 2021-02-09 深圳大学 Whole cell vaccine and preparation method and application thereof
CN113663067A (en) * 2021-02-22 2021-11-19 深圳市康居正医药科技有限公司 Immune activation type antibody and application thereof

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