CN118001383A - HPV vaccine composition, preparation method and application thereof - Google Patents
HPV vaccine composition, preparation method and application thereof Download PDFInfo
- Publication number
- CN118001383A CN118001383A CN202311488835.1A CN202311488835A CN118001383A CN 118001383 A CN118001383 A CN 118001383A CN 202311488835 A CN202311488835 A CN 202311488835A CN 118001383 A CN118001383 A CN 118001383A
- Authority
- CN
- China
- Prior art keywords
- component
- hpv
- adjuvant
- vaccine composition
- oil
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 239000000203 mixture Substances 0.000 title claims abstract description 85
- 229960002566 papillomavirus vaccine Drugs 0.000 title claims description 19
- 238000002360 preparation method Methods 0.000 title abstract description 23
- 239000002671 adjuvant Substances 0.000 claims abstract description 150
- 239000000427 antigen Substances 0.000 claims abstract description 85
- 102000036639 antigens Human genes 0.000 claims abstract description 85
- 108091007433 antigens Proteins 0.000 claims abstract description 85
- 229960005486 vaccine Drugs 0.000 claims abstract description 71
- 229910052782 aluminium Inorganic materials 0.000 claims abstract description 55
- XAGFODPZIPBFFR-UHFFFAOYSA-N aluminium Chemical compound [Al] XAGFODPZIPBFFR-UHFFFAOYSA-N 0.000 claims abstract description 52
- GVJHHUAWPYXKBD-IEOSBIPESA-N α-tocopherol Chemical compound OC1=C(C)C(C)=C2O[C@@](CCC[C@H](C)CCC[C@H](C)CCCC(C)C)(C)CCC2=C1C GVJHHUAWPYXKBD-IEOSBIPESA-N 0.000 claims abstract description 42
- 239000007764 o/w emulsion Substances 0.000 claims abstract description 32
- YYGNTYWPHWGJRM-UHFFFAOYSA-N (6E,10E,14E,18E)-2,6,10,15,19,23-hexamethyltetracosa-2,6,10,14,18,22-hexaene Chemical compound CC(C)=CCCC(C)=CCCC(C)=CCCC=C(C)CCC=C(C)CCC=C(C)C YYGNTYWPHWGJRM-UHFFFAOYSA-N 0.000 claims abstract description 23
- BHEOSNUKNHRBNM-UHFFFAOYSA-N Tetramethylsqualene Natural products CC(=C)C(C)CCC(=C)C(C)CCC(C)=CCCC=C(C)CCC(C)C(=C)CCC(C)C(C)=C BHEOSNUKNHRBNM-UHFFFAOYSA-N 0.000 claims abstract description 23
- PRAKJMSDJKAYCZ-UHFFFAOYSA-N dodecahydrosqualene Natural products CC(C)CCCC(C)CCCC(C)CCCCC(C)CCCC(C)CCCC(C)C PRAKJMSDJKAYCZ-UHFFFAOYSA-N 0.000 claims abstract description 23
- 229940031439 squalene Drugs 0.000 claims abstract description 23
- TUHBEKDERLKLEC-UHFFFAOYSA-N squalene Natural products CC(=CCCC(=CCCC(=CCCC=C(/C)CCC=C(/C)CC=C(C)C)C)C)C TUHBEKDERLKLEC-UHFFFAOYSA-N 0.000 claims abstract description 23
- 229940087168 alpha tocopherol Drugs 0.000 claims abstract description 21
- 235000010482 polyoxyethylene sorbitan monooleate Nutrition 0.000 claims abstract description 21
- 229920000053 polysorbate 80 Polymers 0.000 claims abstract description 21
- 229960000984 tocofersolan Drugs 0.000 claims abstract description 21
- 239000002076 α-tocopherol Substances 0.000 claims abstract description 21
- 235000004835 α-tocopherol Nutrition 0.000 claims abstract description 21
- 239000004411 aluminium Substances 0.000 claims abstract description 16
- 108090000623 proteins and genes Proteins 0.000 claims description 35
- 102000004169 proteins and genes Human genes 0.000 claims description 33
- WNROFYMDJYEPJX-UHFFFAOYSA-K aluminium hydroxide Chemical compound [OH-].[OH-].[OH-].[Al+3] WNROFYMDJYEPJX-UHFFFAOYSA-K 0.000 claims description 22
- 238000000034 method Methods 0.000 claims description 22
- ILRRQNADMUWWFW-UHFFFAOYSA-K aluminium phosphate Chemical compound O1[Al]2OP1(=O)O2 ILRRQNADMUWWFW-UHFFFAOYSA-K 0.000 claims description 8
- 239000000243 solution Substances 0.000 claims description 7
- DIZPMCHEQGEION-UHFFFAOYSA-H aluminium sulfate (anhydrous) Chemical compound [Al+3].[Al+3].[O-]S([O-])(=O)=O.[O-]S([O-])(=O)=O.[O-]S([O-])(=O)=O DIZPMCHEQGEION-UHFFFAOYSA-H 0.000 claims description 6
- 238000009472 formulation Methods 0.000 claims description 5
- 239000000725 suspension Substances 0.000 claims description 4
- 229940037003 alum Drugs 0.000 claims description 3
- 230000002265 prevention Effects 0.000 claims description 3
- 108091036414 Polyinosinic:polycytidylic acid Proteins 0.000 claims description 2
- 201000010099 disease Diseases 0.000 claims description 2
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 claims description 2
- 239000003814 drug Substances 0.000 claims description 2
- 239000012931 lyophilized formulation Substances 0.000 claims description 2
- 238000004806 packaging method and process Methods 0.000 claims description 2
- 229930182490 saponin Natural products 0.000 claims description 2
- 150000007949 saponins Chemical class 0.000 claims description 2
- 238000004519 manufacturing process Methods 0.000 claims 1
- 235000017709 saponins Nutrition 0.000 claims 1
- 239000000839 emulsion Substances 0.000 abstract description 62
- 230000000694 effects Effects 0.000 abstract description 8
- 241000701806 Human papillomavirus Species 0.000 description 106
- 230000000052 comparative effect Effects 0.000 description 20
- 239000012071 phase Substances 0.000 description 17
- 230000003472 neutralizing effect Effects 0.000 description 16
- 238000002156 mixing Methods 0.000 description 15
- 239000002245 particle Substances 0.000 description 15
- 230000003053 immunization Effects 0.000 description 14
- 238000002649 immunization Methods 0.000 description 14
- 241000699670 Mus sp. Species 0.000 description 13
- 238000000265 homogenisation Methods 0.000 description 12
- 239000008346 aqueous phase Substances 0.000 description 11
- 230000001900 immune effect Effects 0.000 description 11
- 125000003275 alpha amino acid group Chemical group 0.000 description 10
- 229910017119 AlPO Inorganic materials 0.000 description 9
- 241001112090 Pseudovirus Species 0.000 description 9
- 230000000890 antigenic effect Effects 0.000 description 9
- 230000028993 immune response Effects 0.000 description 8
- 238000002347 injection Methods 0.000 description 8
- 239000007924 injection Substances 0.000 description 8
- 239000008363 phosphate buffer Substances 0.000 description 8
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 8
- 239000000126 substance Substances 0.000 description 7
- 238000001514 detection method Methods 0.000 description 6
- 210000002966 serum Anatomy 0.000 description 6
- 230000001939 inductive effect Effects 0.000 description 5
- 239000008055 phosphate buffer solution Substances 0.000 description 5
- 239000004094 surface-active agent Substances 0.000 description 5
- 150000001413 amino acids Chemical class 0.000 description 4
- 229940124866 human papillomavirus vaccine Drugs 0.000 description 4
- 230000005847 immunogenicity Effects 0.000 description 4
- 238000001179 sorption measurement Methods 0.000 description 4
- 238000011725 BALB/c mouse Methods 0.000 description 3
- 230000000240 adjuvant effect Effects 0.000 description 3
- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid Chemical compound OC(=O)CC(O)(C(O)=O)CC(O)=O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 description 3
- 238000001914 filtration Methods 0.000 description 3
- 210000003205 muscle Anatomy 0.000 description 3
- 238000006386 neutralization reaction Methods 0.000 description 3
- 239000007787 solid Substances 0.000 description 3
- 238000003756 stirring Methods 0.000 description 3
- 239000011550 stock solution Substances 0.000 description 3
- 241000341655 Human papillomavirus type 16 Species 0.000 description 2
- 101000641175 Human papillomavirus type 18 Major capsid protein L1 Proteins 0.000 description 2
- 101710135729 Major capsid protein L1 Proteins 0.000 description 2
- 241000699666 Mus <mouse, genus> Species 0.000 description 2
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 2
- 235000014680 Saccharomyces cerevisiae Nutrition 0.000 description 2
- PRXRUNOAOLTIEF-ADSICKODSA-N Sorbitan trioleate Chemical compound CCCCCCCC\C=C/CCCCCCCC(=O)OC[C@@H](OC(=O)CCCCCCC\C=C/CCCCCCCC)[C@H]1OC[C@H](O)[C@H]1OC(=O)CCCCCCC\C=C/CCCCCCCC PRXRUNOAOLTIEF-ADSICKODSA-N 0.000 description 2
- 238000003917 TEM image Methods 0.000 description 2
- 241000700605 Viruses Species 0.000 description 2
- AZDRQVAHHNSJOQ-UHFFFAOYSA-N alumane Chemical class [AlH3] AZDRQVAHHNSJOQ-UHFFFAOYSA-N 0.000 description 2
- 230000015572 biosynthetic process Effects 0.000 description 2
- 239000000872 buffer Substances 0.000 description 2
- 230000032798 delamination Effects 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 108700042300 human papillomavirus type 52 L1 Proteins 0.000 description 2
- 230000002209 hydrophobic effect Effects 0.000 description 2
- 230000003902 lesion Effects 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 210000004400 mucous membrane Anatomy 0.000 description 2
- 239000000244 polyoxyethylene sorbitan monooleate Substances 0.000 description 2
- 229940068968 polysorbate 80 Drugs 0.000 description 2
- 230000006798 recombination Effects 0.000 description 2
- 238000005215 recombination Methods 0.000 description 2
- 239000007762 w/o emulsion Substances 0.000 description 2
- QKNYBSVHEMOAJP-UHFFFAOYSA-N 2-amino-2-(hydroxymethyl)propane-1,3-diol;hydron;chloride Chemical compound Cl.OCC(N)(CO)CO QKNYBSVHEMOAJP-UHFFFAOYSA-N 0.000 description 1
- GHCZTIFQWKKGSB-UHFFFAOYSA-N 2-hydroxypropane-1,2,3-tricarboxylic acid;phosphoric acid Chemical compound OP(O)(O)=O.OC(=O)CC(O)(C(O)=O)CC(O)=O GHCZTIFQWKKGSB-UHFFFAOYSA-N 0.000 description 1
- 229940124957 Cervarix Drugs 0.000 description 1
- 206010008342 Cervix carcinoma Diseases 0.000 description 1
- 108020004414 DNA Proteins 0.000 description 1
- 102000053602 DNA Human genes 0.000 description 1
- 241000588724 Escherichia coli Species 0.000 description 1
- 229940124897 Gardasil Drugs 0.000 description 1
- 101100209954 Human papillomavirus type 16 L1 gene Proteins 0.000 description 1
- 206010061218 Inflammation Diseases 0.000 description 1
- 101710163801 Minor capsid protein L2 Proteins 0.000 description 1
- 206010028980 Neoplasm Diseases 0.000 description 1
- 208000009608 Papillomavirus Infections Diseases 0.000 description 1
- 241000235648 Pichia Species 0.000 description 1
- 241001631648 Polyomaviridae Species 0.000 description 1
- 102000007056 Recombinant Fusion Proteins Human genes 0.000 description 1
- 108010008281 Recombinant Fusion Proteins Proteins 0.000 description 1
- 208000007313 Reproductive Tract Infections Diseases 0.000 description 1
- 230000024932 T cell mediated immunity Effects 0.000 description 1
- 208000006105 Uterine Cervical Neoplasms Diseases 0.000 description 1
- 239000008351 acetate buffer Substances 0.000 description 1
- 229910021502 aluminium hydroxide Inorganic materials 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 230000030741 antigen processing and presentation Effects 0.000 description 1
- 238000013459 approach Methods 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 239000007853 buffer solution Substances 0.000 description 1
- 210000000234 capsid Anatomy 0.000 description 1
- 230000036755 cellular response Effects 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 201000010881 cervical cancer Diseases 0.000 description 1
- 239000007979 citrate buffer Substances 0.000 description 1
- 210000003022 colostrum Anatomy 0.000 description 1
- 235000021277 colostrum Nutrition 0.000 description 1
- 239000002131 composite material Substances 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 235000013365 dairy product Nutrition 0.000 description 1
- 230000018044 dehydration Effects 0.000 description 1
- 238000006297 dehydration reaction Methods 0.000 description 1
- 230000003111 delayed effect Effects 0.000 description 1
- 238000013461 design Methods 0.000 description 1
- 230000001066 destructive effect Effects 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 238000004945 emulsification Methods 0.000 description 1
- 239000003995 emulsifying agent Substances 0.000 description 1
- 230000001804 emulsifying effect Effects 0.000 description 1
- 230000002708 enhancing effect Effects 0.000 description 1
- 229940102767 gardasil 9 Drugs 0.000 description 1
- 230000016784 immunoglobulin production Effects 0.000 description 1
- 230000003308 immunostimulating effect Effects 0.000 description 1
- 238000009169 immunotherapy Methods 0.000 description 1
- 230000004054 inflammatory process Effects 0.000 description 1
- 230000003993 interaction Effects 0.000 description 1
- 239000010410 layer Substances 0.000 description 1
- 230000007774 longterm Effects 0.000 description 1
- 230000003211 malignant effect Effects 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 239000002105 nanoparticle Substances 0.000 description 1
- 229920001184 polypeptide Polymers 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 102000004196 processed proteins & peptides Human genes 0.000 description 1
- 108090000765 processed proteins & peptides Proteins 0.000 description 1
- 230000002062 proliferating effect Effects 0.000 description 1
- 229940032313 prophylactic HPV vaccine Drugs 0.000 description 1
- 230000000069 prophylactic effect Effects 0.000 description 1
- 239000001397 quillaja saponaria molina bark Substances 0.000 description 1
- 238000010008 shearing Methods 0.000 description 1
- 238000007086 side reaction Methods 0.000 description 1
- 239000002356 single layer Substances 0.000 description 1
- 230000000087 stabilizing effect Effects 0.000 description 1
- 230000004936 stimulating effect Effects 0.000 description 1
- 230000002195 synergetic effect Effects 0.000 description 1
- 210000001519 tissue Anatomy 0.000 description 1
- 230000010415 tropism Effects 0.000 description 1
- 238000002255 vaccination Methods 0.000 description 1
Landscapes
- Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
Abstract
The present invention relates to a vaccine composition comprising a first component comprising an HPV antigen and an aluminium adjuvant and a second component comprising an oil-in-water emulsion adjuvant comprising squalene, alpha-tocopherol and tween 80, packaged separately. The vaccine composition does not need to prepare two components into a finished product preparation during preparation, and the two components can be independently packaged, so that the mutual influence between an antigen or/and an aluminum adjuvant and an emulsion adjuvant can be avoided, and the technical effects of higher efficiency, safer and more stable simultaneously are achieved.
Description
Technical Field
The invention belongs to the field of biomedical engineering, and in particular relates to an HPV vaccine composition, a preparation method thereof and application thereof in the field of immunotherapy and prevention.
Background
Human papillomavirus (human papillomavirus, HPV) is a non-enveloped, closed-loop, double-stranded DNA virus belonging to the subfamily polyomaviridae of papovaviridae, which mainly invades the epithelial mucosal tissue of the human body, thereby inducing various benign and malignant proliferative lesions. Over 200 types of HPV have been identified at present, HPV infection has obvious tissue specificity, different types of HPV have different tropisms to skin and mucous membrane, different papillary lesions can be induced, about 30 types of HPV are associated with genital tract infection, and about 20 types of HPV are associated with tumors.
Vaccination with prophylactic HPV vaccines is an effective and viable prophylactic means for the prevention of cervical cancer, and vaccines that have been marketed at present are vaccines based on Virus-like particles (VLPs) formed from HPV major capsid protein L1 as antigens, and the L1 protein expressed by gene recombination can form VLPs resembling viruses under certain conditions. Adjuvants which are important components of vaccines are mainly aluminum salt adjuvants, which exert an effect of enhancing immune responses through an antigen-depot effect and an immune stimulating effect of inducing inflammation and the like. However, aluminum salt adjuvants also have a number of limitations, such as inability to induce cellular immune responses, undesirable antigen presentation effects, delayed immune response generation time, susceptibility to side reactions, and the like.
Emulsion-type adjuvants (including oil-in-water emulsions, water-in-oil emulsions, etc.) are an important branch of new adjuvants, and emulsions can be used in combination with a variety of weak antigens (recombinant proteins, polypeptides, etc.) and elicit high titers of antigen-specific antibodies. Milk-type adjuvants generally comprise an oil phase component, an aqueous phase component and an emulsifier. The oil-in-water emulsion mainly comprises water phase components, has high tolerance to human body and has good compatibility with most vaccine antigens.
Although both dairy-based adjuvants and aluminum adjuvants have been widely used, there remains a need to date for new immune formulations and methods of immunization that provide vaccine formulations that are safer and more effective, easier to formulate, and less costly, and that are capable of rapidly inducing an immune response that is more efficient.
Disclosure of Invention
The invention aims to provide an HPV vaccine composition which has good safety and immunostimulation activity, is convenient to prepare, has low cost and can quickly induce high-efficiency immune response. Meanwhile, the invention provides a simple preparation method and application with wide prospect.
To achieve the above object, the present invention provides in one aspect an HPV vaccine composition comprising a first component comprising HPV antigen and an aluminium adjuvant and a second component comprising an oil-in-water emulsion adjuvant comprising squalene, alpha-tocopherol and tween 80, packaged separately.
The capsid of HPV is composed of a major capsid protein L1 and a minor capsid protein L2. The existing vaccines are all vaccines based on HPV L1 Virus-LIKE PARTICLE (VLP) as antigens, and the L1 protein expressed by gene recombination can form Virus-like particles under certain conditions, so that the vaccine has better immunogenicity.
In the present invention, the antigen derived from Human Papillomavirus (HPV) is the L1 protein and/or L2 protein of each type of HPV. In embodiments of the invention, HPV may be a low-risk HPV (e.g., HPV6, 11, 40, 42, 43, 44, 54, 61, 70, 72, 81, 89), a medium-risk HPV (e.g., HPV26, 53, 66, 73, 82), or a high-risk HPV (e.g., HPV16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59, 68). In preferred embodiments, the antigen comprises L1 protein and/or L2 protein of one or more of HPV types 6, 11, 16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58 and 59.
In the NCBI database, there are a number of existing HPV-associated L1 protein (HPV 6L1, 11L1, 16L1, 18L1, 31L1, 33L1, 45L1, 52L1 and 58L 1) sequences available for selection by those skilled in the art, which can be used as the basis for the desired selection of antigenic proteins. In some embodiments of the invention, sequences with a high degree of conservation are employed, as follows: the amino acid sequence of HPV 6L1 was recorded in NCBI database in 1995 under accession No. AAA74218.1; the amino acid sequence of HPV 11L1 was recorded in NCBI database in 1994 under accession No. AAA46935.1; the amino acid sequence of HPV 16L1 was recorded in NCBI database in 1998 under accession No. AAC09292.1; the amino acid sequence of HPV 18L1 protein was recorded in NCBI database in 2003 under accession number AAQ92369.1; the amino acid sequence of HPV 31L1 protein has been recorded in NCBI database in 1994 under accession No. AAA46956.1; the amino acid sequence of HPV 33L1 protein has been recorded in the NCBI database in 2009 under accession No. ACL12333.1; the amino acid sequence of HPV 45L1 protein was recorded in the NCBI database in 2009 under accession number ABP99831.1 (N-terminal 26 amino acids were truncated, the 26 amino acids being hydrophobic regions that could affect the formation of the L1 protein into VLPs, thus truncated); the amino acid sequence of HPV 52L1 protein was recorded in NCBI database 2005 under accession number CAA52590.1 (truncated by 27 amino acids at the N-terminus, the 27 amino acids being hydrophobic regions that could affect the formation of VLP from the L1 protein, thus truncated); the amino acid sequence of HPV 58L1 protein was recorded in the NCBI database in 2009 under accession number CAX48979.1. The antigen of the present invention is not limited to the L1 protein having the above amino acid sequence, and may be other L1 proteins having immunogenicity, including L1 proteins used in commercially available vaccines.
According to some embodiments of the invention, the content of antigenic HPV types L1 and/or L2 proteins in the first component is between 1-100 μg, more preferably between 5-50 μg, such as 10 μg, 20 μg, 30 μg or 40 μg.
In some embodiments, the aluminum adjuvant is selected from at least one of aluminum hydroxide, aluminum phosphate, aluminum sulfate, and alum.
In some embodiments, the amount of aluminum adjuvant in the first component is between about 10-1000 μg, preferably between 50-800 μg, more preferably between 100-600 μg, for example 200 μg, 300 μg, 400 μg, or 500 μg.
In some embodiments, the first component is a lyophilized formulation, solution, or suspension.
In some embodiments, the solution or suspension of the first component has a volume of 0.2 to 1ml, preferably 0.5ml.
In some embodiments, the first component is a HPV vaccine finished formulation, e.g., a divalent, tetravalent, nine or higher valent HPV vaccine, commercially available.
In some embodiments, the oil-in-water emulsion adjuvant comprises 5-15mg squalene, 5-15mg alpha-tocopherol and 2-10mg tween 80, preferably the oil-in-water emulsion adjuvant comprises 10.69mg squalene, 11.86mg alpha-tocopherol and 4.86mg tween 80.
In some embodiments, the volume of the oil-in-water emulsion adjuvant is 0.2-1ml, preferably 0.5ml.
In some embodiments, the oil-in-water emulsion adjuvant further comprises one or more of MPL, saponin, poly I: C, and CpG.
The invention also provides a preparation method of the HPV vaccine composition, which comprises the following steps:
(i) Preparing a first component comprising HPV antigen and an aluminium adjuvant;
(ii) Preparing as a second component an oil-in-water emulsion adjuvant comprising squalene, alpha-tocopherol and tween 80;
(iii) And packaging the first component and the second component independently to obtain the HPV vaccine composition.
Wherein steps (i) and (ii) may be performed in any order, including sequentially, simultaneously or in a split site.
For specific preparation of the first component according to the invention, reference is made, for example, to VACCINE DESIGN: the Subunit and Adjuvant Approach (ISBN: 0-306-44867-X). In some embodiments, the invention provides methods of preparation by adsorbing VLPs of HPV-type L1 proteins to an aluminum adjuvant to maintain VLP stability and enhance their immunogenicity.
According to an embodiment of the present invention, a method of preparing a second component includes: firstly, mixing squalene and alpha-tocopherol, then adding phosphate buffer solution containing tween 80, emulsifying by stirring, and finally homogenizing to obtain the oil-in-water emulsion adjuvant.
The invention also provides application of the vaccine composition in preparing medicines for treating or preventing diseases.
In another aspect, the invention provides a kit comprising the vaccine composition comprising a first component comprising HPV antigen and an aluminium adjuvant and a second component comprising an oil-in-water emulsion adjuvant comprising squalene, alpha-tocopherol and tween 80, packaged separately.
In another aspect, the invention provides a multi-compartment syringe comprising the vaccine composition comprising a first compartment and a second compartment, the first compartment containing a first component comprising HPV antigen and an aluminium adjuvant; the second compartment contains a second component comprising an oil-in-water emulsion adjuvant comprising squalene, alpha tocopherol and tween 80.
In another aspect, the invention provides a method of inducing an immune response using the vaccine composition, which separately administers the first component and the second component to a subject. The method comprises the following steps:
(i) Administering the first component to the subject;
(ii) Administering the second component to the subject,
Wherein the steps may be performed simultaneously in any order or at specific times intervals.
In some embodiments, the first component and the second component are administered to the same site in the subject. In other embodiments, the first component and the second component are administered to different sites of the subject separately.
The invention also provides another method of inducing an immune response using the vaccine composition, which simultaneously administers the first component and the second component to a subject. The method comprises the following steps:
(i) Mixing the first component and the second component;
(ii) Administering the mixture obtained in step (i) to the subject.
In another aspect, the invention provides a method of improving a CD 4T-cell immune response and/or improving a B-memory cell response using the vaccine composition, wherein the first component and the second component are administered to a subject simultaneously or not simultaneously, respectively, or wherein the first component and the second component are administered to a subject after mixing.
Compared with the prior art, the invention has the following beneficial effects:
the invention further adds emulsion adjuvant on the basis of immune component containing antigenic substance and aluminium adjuvant, and the two components can be separately applied or co-applied and can basically achieve synergistic adjuvant effect consistent with or even better than traditional preparation form, can effectively improve immune response and quickly stimulate antibody production.
On the other hand, the vaccine composition of the invention does not need to prepare the immune component containing the antigenic substance and the aluminum adjuvant and the emulsion adjuvant into finished preparations when being prepared, the emulsion adjuvant and the immune component containing the antigenic substance and the aluminum adjuvant can be independently packaged, so that the interaction between the antigenic substance or/and the aluminum adjuvant and the emulsion adjuvant can be avoided, the aluminum adjuvant can absorb the antigenic substance to play a role of protecting the antigen, meanwhile, the aluminum adjuvant and the antigenic substance form a whole, and when being mixed with the emulsion, the antigenic substance can be absorbed by the aluminum adjuvant or carried by the emulsion, and the double adjuvant effect of the aluminum adjuvant and the emulsion can be simultaneously exerted, so that the vaccine composition of the invention can achieve more efficient, safer and more stable technical effects.
Drawings
The following drawings are only for purposes of illustration and explanation of the present invention and are not intended to limit the scope of the invention. Wherein:
FIG. 1 is a transmission electron micrograph of an aluminum hydroxide adjuvant according to example 3 of the present invention;
FIG. 2 is a schematic representation of the adsorption of antigen to aluminum adjuvants in example 3 of the present invention;
FIG. 3 is a transmission electron micrograph of the emulsion adjuvant of example 3 of the present invention;
FIG. 4 is a graph showing layering of the adjuvant system according to comparative example 2 of the present invention;
FIG. 5 is a graph showing layering of the adjuvant system according to comparative example 3 of the present invention;
FIG. 6 is a schematic representation of a vaccine complex of comparative example 4 of the present invention consisting of an aluminum adjuvant, an oil-in-water emulsion and an antigen;
FIG. 7 shows the neutralizing antibody titres against HPV type 45 pseudoviruses after immunization of mice with HPV type 45 antigen in combination with various adjuvants in section 4.1 of example 4 of the present invention;
FIG. 8 is a graph showing the neutralizing antibody titres against HPV type 45 pseudoviruses after immunization of mice with HPV type 45 antigen in combination with various adjuvants in section 4.2 of example 4 of the present invention;
FIG. 9 shows the neutralizing antibody titres against HPV type 31 pseudoviruses after immunization of mice with nine HPV antigens in example 5 in combination with different adjuvants according to the present invention;
FIG. 10 shows the titers of neutralizing antibodies against HPV type 6, type 45 and type 58 pseudoviruses after immunization of mice with nine HPV antigens in example 5 of the invention in combination with different adjuvants;
FIG. 11 shows the neutralizing antibody titres of HPV vaccine and emulsion adjuvant according to example 6 of the present invention after immunization of mice at different times and different sites,
Wherein, injection 1 represents that an antigen adsorbed to an aluminum hydroxide adjuvant is injected on a horse after the emulsion adjuvant is injected at the same place;
Injection 2 means that the antigen adsorbed to the aluminum hydroxide adjuvant is injected 2 hours after the emulsion adjuvant is injected at the same site;
Injection 3 represents injection of antigen adsorbed to aluminium hydroxide adjuvant on horses after injection of emulsion adjuvant at different sites.
Detailed Description
The present invention will be further described in detail below with reference to specific embodiments and with reference to the accompanying drawings, in order to make the objects, technical solutions and advantages of the present invention more apparent.
The HPV vaccine composition of the present invention comprises a first component comprising HPV antigen and an aluminum adjuvant and a second component comprising an oil-in-water emulsion adjuvant, packaged separately.
In embodiments of the invention, the antigen derived from Human Papillomavirus (HPV) may be the L1 protein of each type of HPV. In embodiments of the invention, HPV may be a low-risk HPV (e.g., HPV6, 11, 40, 42, 43, 44, 54, 61, 70, 72, 81, 89), a medium-risk HPV (e.g., HPV26, 53, 66, 73, 82), or a high-risk HPV (e.g., HPV16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59, 68).
In a preferred embodiment, the antigen derived from Human Papillomavirus (HPV) comprises HPV virus-like particles assembled from L1 proteins and/or L2 proteins of one or more of HPV types 6, 11, 16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58 and 59.
In a preferred embodiment, the antigen derived from Human Papillomavirus (HPV) comprises HPV virus-like particles assembled from L1 and/or L2 proteins of HPV types 6 and 11.
In a preferred embodiment, the antigen derived from Human Papillomavirus (HPV) comprises HPV virus-like particles assembled from L1 and/or L2 proteins of HPV types 16 and 18.
In a preferred embodiment, the antigen derived from Human Papillomavirus (HPV) comprises HPV virus-like particles assembled from L1 proteins and/or L2 proteins of HPV types 6, 11, 16 and 18.
In a preferred embodiment, the antigen derived from Human Papillomavirus (HPV) comprises HPV virus-like particles assembled from L1 proteins and/or L2 proteins of HPV types 6, 11, 16, 18, 31, 33, 45, 52 and 58.
In a preferred embodiment, the antigen derived from Human Papillomavirus (HPV) comprises HPV virus-like particles assembled from L1 proteins and/or L2 proteins of one or more of HPV types 6, 11, 16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58 and 59.
HPV antigens in the present invention may be prepared using any of the existing techniques. The HPV antigens prepared may be further mixed with an aluminium adjuvant to produce the first component of the present invention. Wherein the aluminum adjuvant may be selected from aluminum hydroxide, aluminum phosphate, aluminum sulfate and alum. The aluminum adjuvant may be prepared using any of the existing techniques, and commercially available aluminum adjuvants such as CRODA's aluminum hydroxide adjuvant ALHYDROGEL may also be used.
In a preferred embodiment, the first component of the present invention may also be a commercially available HPV vaccine finished formulation, e.g., tetravalent human papilloma virus vaccine (saccharomyces cerevisiae) -jiadau (GARDASIL) and nine-valent human papilloma virus vaccine (saccharomyces cerevisiae) -jiadau 9 (GARDASIL 9) of moxadong corporation, bivalent human papilloma virus adsorbing vaccine of GSK-greedy (CERVARIX), bivalent human papilloma virus vaccine (escherichia coli) -xinkening (Cecolin) of the mansion vantagetes biotechnology limited, bivalent human papilloma virus vaccine (pichia) -Wo Zehui of the yuxi zerun biotechnology limited, and the like.
In the present invention, the oil-in-water emulsion adjuvant as the second component comprises an aqueous phase, an oil phase, and a surfactant. Wherein the aqueous phase may be a buffer solution selected from phosphate buffer, citrate buffer, tris-HCl buffer, acetate buffer or citric acid-phosphate buffer. The oil phase comprises a metabolisable oil, preferably squalene, and preferably further comprises alpha-tocopherol. For example, the oil-in-water emulsion adjuvant may comprise squalene, span 85 and tween 80, more preferably the oil-in-water emulsion adjuvant comprises squalene, alpha-tocopherol and tween 80.
Example 1 preparation of a two-component vaccine composition comprising an emulsion adjuvant and an HPV antigen adsorbed to an aluminium adjuvant
Preparation of the first component: the antigen stock solution of each type of L1 VLP of HPV was mixed with an aluminum hydroxide adjuvant (ALHYDROGEL, CRODA, the same applies below) or an aluminum phosphate adjuvant, and the mixture was shaken at 15℃and 100rpm for 10 hours to adsorb VLP to the aluminum hydroxide or aluminum phosphate adjuvant, thereby obtaining HPV antigen adsorbed to the aluminum adjuvant as a first component.
Preparation of the second component: firstly, mixing 2.14g of squalene and 2.37g of alpha-tocopherol, then adding 45ml of phosphate buffer solution containing 5wt.% of Tween 80, mixing, stirring at 8000rpm for 15min at 15 ℃, carrying out slit homogenization at 80MPa pressure at 15 ℃, carrying out micro-jet homogenization at 100MPa pressure, reducing the particle size to 160.5nm after 2 cycles of homogenization, stopping homogenization, and filtering by a 0.22 mu m filter to obtain the oil-in-water emulsion.
Example 2 preparation of a two-component vaccine composition comprising an emulsion adjuvant and an HPV antigen adsorbed to an aluminium adjuvant
Preparation of the first component: the antigen stock solution of each type L1 VLP of HPV is mixed with an aluminum hydroxide adjuvant or an aluminum phosphate adjuvant, and the mixture is vibrated for 10 hours at 15 ℃ and 100rpm, so that the VLP is adsorbed on the aluminum hydroxide or the aluminum phosphate adjuvant, and the HPV antigen adsorbed on the aluminum adjuvant is obtained as a first component.
Preparation of the second component: 2.47g of Tween 80 was weighed and dissolved in a citric acid buffer, the pH was adjusted to 6.5, and the volume was set to 500ml as the aqueous phase. 3.90g of squalene and 0.47g of span 85 are weighed and mixed uniformly to obtain an oil phase. At room temperature, adding water phase to 4.37g of oil phase to 100ml, mixing uniformly, and shearing at 10000rpm for 15min to obtain colostrum.
Slit-type homogenization is carried out at 20 ℃ under the pressure of 50MPa, then micro-jet homogenization is carried out under the pressure of 120MPa, the particle size can be reduced to 165.3nm after 2 cycles of homogenization, PDI is 0.062, homogenization is stopped, and a filter with the size of 0.22 mu m is used for filtering, thus obtaining the oil-in-water emulsion.
Comparative example 1 preparation of vaccine compositions comprising a Complex adjuvant and an antigen
4.29G squalene, 4.74g alpha-tocopherol, 1.94g polysorbate 80 were weighed out and mixed as an oil phase. 10ml of aluminum hydroxide adjuvant was added to 80ml of phosphate buffer to give a white crushed flocculent, milky liquid. Dispersing at 20 ℃ at 8000rpm for 10min, homogenizing by a 1bar pre-stage valve and a 120MPa micro-jet valve, and circulating for 8 times to obtain the oily emulsion. 27. Mu.l of HPV antigen was mixed with 873. Mu.l of phosphate buffer as an aqueous phase, and the oily emulsion and the aqueous phase were mixed in equal volumes to obtain a vaccine composition.
Comparative example 2 preparation of vaccine compositions comprising a Complex adjuvant and an antigen
1.28G squalene, 1.43g alpha-tocopherol, 0.58g polysorbate 80 were weighed and mixed as an oil phase. 360. Mu.l of aluminum hydroxide adjuvant, 108. Mu.l of HPV antigen was added to 132. Mu.l of phosphate buffer and mixed as an aqueous phase. Mu.l of the aqueous phase was added to about 2.5ml of the oil phase and phacoemulsified (300W, 4min,5s/5 s) to give a water-in-oil emulsion, which was finally diluted with the same volume of phosphate buffer.
Comparative example 3 preparation of vaccine compositions comprising a Complex adjuvant and an antigen
1.29G squalene and 1.43g alpha-tocopherol were weighed out and mixed as an oil phase. 400. Mu.l of aluminum hydroxide nanoparticles were added to 3080. Mu.l of phosphate buffer, 120. Mu.l of HPV antigen stock solution was added and mixed as an aqueous phase. Mixing 300 μl of oil phase with 2.7ml of water phase, performing ultrasonic emulsification (300W, 4min,5s/5 s) to obtain Pickering emulsion, mixing Pickering emulsion with phosphate buffer solution, obtaining vaccine composition, and centrifuging to obtain sample which is obviously layered.
Comparative example 4 preparation of vaccine composition comprising aluminium adjuvant, emulsion adjuvant and antigen
Firstly, mixing 2.14g of squalene and 2.37g of alpha-tocopherol, then adding 45ml of phosphate buffer solution containing 5wt.% of Tween 80, mixing, stirring at 8000rpm for 15min at 15 ℃, carrying out slit homogenization at 80MPa pressure at 15 ℃, carrying out micro-jet homogenization at 100MPa pressure, reducing the particle size to 160.5nm after 2 cycles of homogenization, stopping homogenization, and filtering by a 0.22 mu m filter to obtain the oil-in-water emulsion.
400 Μl of aluminum hydroxide or aluminum phosphate adjuvant with aluminum content of 10mg/ml and the above oil-in-water emulsion of 4ml were added into a centrifuge tube, and the mixture was stirred for 1min by vortex shaker, and mixed well. Mu.l of HPV antigen was added to 1740. Mu.l of phosphate buffer and mixed as antigen solution. To 1.1ml of the adjuvant mixture was added 900. Mu.l of the antigen solution, followed by shaking at 15℃and 100rpm for 10 hours, to prepare a vaccine composition.
EXAMPLE 3 analysis of vaccine composition properties
The aluminum hydroxide adjuvant is actually an incomplete dehydration product of Al (OH) 3, namely aluminum metahydroxide AlO (OH) in fibrous crystalline form, as shown in fig. 1. In examples 1 and 2, HPV VLPs were adsorbed onto aluminum hydroxide adjuvant when mixed with HPV VLPs as antigens, as shown in fig. 2.
The oil phase of the oil-in-water emulsions of examples 1 and 2 and the droplets formed by the surfactant were uniformly distributed in water as shown in fig. 3. The stability of both HPV antigens adsorbed to aluminum adjuvants and oil-in-water emulsions in examples 1 and 2 was higher, and the stability of the prepared two-component vaccine composition was also higher since the two components were independent of each other and did not affect each other.
In comparative example 1, since the formed complex adjuvant was an oily emulsion, delamination was found after mixing it with an aqueous phase containing an antigen and standing for a long period of time. Similarly, the compound adjuvant of comparative example 2 was a water-in-oil emulsion, and white flocculent, precipitated and layered phenomena were found after dilution with phosphate buffer solution and standing, as shown in fig. 4. The Pickering emulsion prepared in comparative example 3 is an emulsion system with solid particles instead of surfactant. The emulsion stabilizing mechanism is mainly that solid particles are adsorbed on an oil-water interface to form a solid particle single-layer or multi-layer structure, so that the emulsion is stabilized. However, this technique has high requirements on the preparation process conditions and poor emulsion stability, and the inventors found that delamination phenomenon occurs after long-term standing or centrifugation, as shown in fig. 5.
In comparative example 4, three components of an aluminum adjuvant, an oil-in-water emulsion, and an antigen were sequentially mixed, and droplets formed of an oil phase and a surfactant, and the antigen were sequentially adsorbed onto the aluminum adjuvant, as shown in fig. 6. The adsorption between the liquid drops formed by the oil phase and the surfactant and the aluminum adjuvant can compete with the adsorption between the antigen and the aluminum adjuvant, the aluminum adjuvant is difficult to effectively adsorb and protect the antigen, on the other hand, the aluminum adjuvant can destroy the balance of the water-oil interface of the emulsion, thereby destroying the form of the emulsion, and the advantages of the emulsion per se can not be exerted after the emulsion is mixed with the antigen.
Example 4 monovalent HPV vaccine mouse immunization experiments
4.1 Comparison of two-component vaccine with vaccine composition comprising Complex adjuvant and antigen
BALB/c mice were immunized with the vaccine compositions of example 1 and comparative examples 1-3, with an Al (OH) 3 control group, 10 per group, were muscle immunized, HPV antigens contained 5 μg of HPV 45L 1 VLP, and adjuvant amounts are shown in Table 1. Mice were immunized on days 0 and 14, respectively, mice were killed and serum was isolated 7 days after the second immunization, and serum neutralization antibody detection was performed by HPV pseudovirus-based method.
TABLE 1
The detection results are shown in FIG. 7. It can be seen that the immune compositions prepared in example 1 and comparative example 1 each induced a higher titer of neutralizing antibodies than the Al (OH) 3 control group, but no significant difference was found between the two groups. The immune compositions prepared in comparative examples 2 and 3 induced a lower neutralizing antibody titer than the Al (OH) 3 control, which may be due to the poor stability of the immune compositions in the two comparative examples on the one hand, and the emulsion of the antigen and the aluminum adjuvant as an aqueous phase together with an oil phase on the other hand, the process of forming the emulsion resulted in a great destructive effect on the antigen, possibly affecting the immune effect of the antigen.
In the comprehensive view, the independently packaged bi-component vaccine composition is adopted, so that on one hand, the stability problem caused by mixing the aluminum adjuvant and the emulsion adjuvant can be avoided, and on the other hand, the immune effect of the antigen can be obviously improved.
4.2 Comparison of two-component vaccine with vaccine composition comprising adjuvant mixture and antigen
Vaccine compositions were prepared according to the methods of example 1 and comparative example 4, respectively. Wherein, in preparing the vaccine composition according to the method of comparative example 4, the adjuvant mixture of aluminum hydroxide and oil-in-water emulsion was formulated 14 days, 7 days, 3 days and the day before immunization, respectively, the adjuvant mixture was mixed with HPV 45L 1 antigen on the day of immunization, the corresponding vaccine compositions were designated as H-14, H-7, H-3 and H-0, respectively, and the two-component vaccine in example 1 was mixed on the day of immunization.
BALB/c mice were immunized with the vaccine compositions prepared in example 1 and comparative example 4, 9 mice per group were immunized by muscle, HPV antigen contained HPV 45L 1 VLP 2. Mu.g, emulsion adjuvants were used in an amount of 25. Mu.l, and Al (OH) 3 adjuvants were used in an amount of 50. Mu.g. Mice were immunized on days 0 and 21, respectively, mice were killed and serum was isolated 14 days after the second immunization, and serum neutralization antibody detection was performed by HPV pseudovirus-based method.
As can be seen from fig. 8, when preparing the vaccine composition according to the method of comparative example 4, the time of the placement of the adjuvant mixture may have an influence on the immune effect, i.e., the immune effect of the prepared vaccine composition is gradually enhanced after the placement of the adjuvant mixture for at least 3 days. This may be an initial stage after mixing the aluminum hydroxide and the oil-in-water emulsion, the adjuvant mixture is unstable due to adsorption of the aluminum hydroxide, and the immune effect of the antigen is not sufficiently improved, and after a period of time, the whole system becomes more stable and uniform, thereby being able to sufficiently exert the adjuvant effect. The two-component vaccine composition of example 1 was immunized after mixing, and a better immune effect was obtained, probably because the antigen in the first component was adsorbed to aluminum hydroxide, and the oil-in-water emulsion was inoculated immediately after mixing with it, and the stability of the whole system was not significantly affected.
EXAMPLE 5 nine-valent HPV vaccine mouse immunization experiment
BALB/c mice were immunized with the prepared vaccine composition, 8 mice per group, were immunized with muscle, HPV antigens contained HPV6 L1 3μg、HPV11 L1 4μg、HPV16 L1 6μg、HPV18 L1 4μg、HPV31 L1 2μg、HPV33L1 2μg、HPV45 L1 2μg、HPV52 L1 2μg and HPV 58L 12. Mu.g, and adjuvant amounts are shown in Table 2. Mice were immunized on days 0 and 14, respectively, mice were killed and serum was isolated 7 days after the second immunization, and serum neutralization antibody detection was performed by HPV pseudovirus-based method.
TABLE 2
Groups 1 and 3 are vaccine compositions comprising Al (OH) 3/emulsion or AlPO 4/ emulsion composite adjuvant and HPV antigen prepared according to the method of comparative example 1, groups 2 and 4 are two-component vaccine compositions prepared according to the method of example 1 or 2 (HPV antigen adsorbed on Al (OH) 3 or AlPO 4 as a first component, emulsion as a second component), the two components are mixed prior to administration, group 5 is a mixture of emulsion and HPV antigen as a vaccine composition, no aluminum adjuvant is included, and groups 6 and 7 are HPV antigen adsorbed on Al (OH) 3 or AlPO 4 as a vaccine composition, no emulsion adjuvant is included.
The results of the detection of neutralizing antibodies against HPV type 31 pseudoviruses are shown in FIG. 9. It can be seen that the highest neutralizing antibody titer was induced when using the emulsion adjuvant and the lowest neutralizing antibody titer was induced when using the AlPO 4 adjuvant. While the neutralizing antibody titer induced by group 2 (labeled "emulsion+al (OH) 3") was significantly higher when using both the emulsion and Al (OH) 3 or AlPO 4 adjuvants, indicating that a vaccine comprising Al (OH) 3 adjuvant in combination with an emulsion adjuvant may significantly enhance the immune effect, the neutralizing antibody titer induced by groups 4 (labeled "emulsion+alpo 4") and groups 1 and 3 (labeled "emulsion/Al (OH) 3" and "emulsion/AlPO 4") was slightly lower when using the emulsion adjuvant alone but significantly higher when using Al (OH) 3 or AlPO 4 alone.
The detection results of neutralizing antibodies against pseudoviruses of HPV types 6, 45 and 58 are shown in FIG. 10. The results also show that group 2 (labeled "emulsion+al (OH) 3") or group 4 (labeled "emulsion+alpo 4") induced a significantly higher neutralizing antibody titer than that induced by Al (OH) 3 or AlPO 4 adjuvants alone, and greater than that induced by group 1 (labeled "emulsion/Al (OH) 3"), demonstrating that a vaccine combination emulsion adjuvant comprising Al (OH) 3 or AlPO 4 adjuvants can significantly enhance immune efficacy and is superior to vaccine compositions "antigen+emulsion/Al (OH) 3 complex adjuvants" that are similar in composition but distinct in preparation.
The results show that the emulsion adjuvant can obviously enhance the immunogenicity of HPV antigen, and can obtain better immune effect by simply adding the emulsion adjuvant on the basis of the existing HPV vaccine containing aluminum adjuvant.
Example 6 Effect of emulsion adjuvant and HPV antigen adsorbed to aluminium adjuvant injection site and administration sequence on the immune Effect
The vaccine composition of example 1 of the present invention comprises two independent components, and the present invention investigated the effect of injection sites and administration sequences of the two components on the immune effect, and explored various combinations of injection sites and administration sequences in order to screen the administration modes that can obtain the optimal immune effect.
In the case of simultaneous administration, the antigen adsorbed to the aluminum adjuvant and the two components of the emulsion adjuvant may be mixed and administered, or the two components may be administered separately using two syringes. The application interval of the two components is short (e.g. within 1min, preferably within 30 s) and still considered to be simultaneous.
The two components of the antigen and emulsion adjuvant adsorbed to the aluminium adjuvant should be administered at least 1h apart when not administered simultaneously, in this example at intervals of 2h apart. The grouping situation is shown in table 3. The dose of each group of antigens was the same as in example 5.
TABLE 3 Table 3
/>
As shown in fig. 11, both components were administered sequentially or at different sites, the vaccine composition induced a lower neutralizing antibody titer than when the two components were mixed and administered at the same site, with the neutralizing antibody titer at the different sites being the lowest. Thus, the results of the study found that the optimal way was to mix the two components and apply them, followed by the two components in the same place.
The foregoing description of the embodiments has been provided for the purpose of illustrating the general principles of the invention, and is not meant to limit the invention thereto, but to limit the invention thereto, and any modifications, equivalents, improvements and equivalents thereof may be made without departing from the spirit and principles of the invention.
Claims (15)
1. A vaccine composition comprising a first component and a second component in separate packages, the first component comprising HPV antigen and an aluminium adjuvant, the second component comprising an oil-in-water emulsion adjuvant comprising squalene, alpha tocopherol and tween 80.
2. The vaccine composition of claim 1, wherein the HPV antigen comprises L1 protein and/or L2 protein of one or more of HPV types 6, 11, 16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58 and 59.
3. Vaccine composition according to claim 2, characterized in that the content of HPV each type L1 and/or L2 protein in the first component is between 1 and 100 μg, more preferably between 5 and 50 μg.
4. The vaccine composition of claim 1, wherein the aluminum adjuvant is selected from at least one of aluminum hydroxide, aluminum phosphate, aluminum sulfate, and alum.
5. Vaccine composition according to claim 1, characterized in that the content of aluminium adjuvant in the first component is between 10-1000 μg, preferably between 50-800 μg.
6. The vaccine composition of claim 1, wherein the first component is a lyophilized formulation, solution or suspension.
7. Vaccine composition according to claim 6, characterized in that the volume of the solution or suspension of the first component is 0.2-1ml, preferably 0.5ml.
8. The vaccine composition according to any one of claims 1-7, wherein the first component is a final HPV vaccine formulation, such as a commercially available bivalent, tetravalent, nine-valent or higher order HPV vaccine.
9. The vaccine composition according to claim 1, wherein the oil-in-water emulsion adjuvant comprises 5-15mg squalene, 5-15mg alpha-tocopherol and 2-10mg tween 80, preferably the oil-in-water emulsion adjuvant comprises 10.69mg squalene, 11.86mg alpha-tocopherol and 4.86mg tween 80.
10. Vaccine composition according to claim 1 or 9, characterized in that the volume of the oil-in-water emulsion adjuvant is 0.2-1ml, preferably 0.5ml.
11. The vaccine composition of claim 1, wherein the oil-in-water emulsion adjuvant further comprises MPL, saponins, poly I: c and CpG.
12. A method of preparing the vaccine composition of any one of claims 1-11, the method comprising the steps of:
(i) Preparing a first component comprising HPV antigen and an aluminium adjuvant;
(ii) Preparing as a second component an oil-in-water emulsion adjuvant comprising squalene, alpha-tocopherol and tween 80;
(iii) Packaging the first component and the second component separately to obtain HPV vaccine composition,
Wherein steps (i) and (ii) may be performed in either order, including sequentially, simultaneously or in separate sites.
13. Use of an HPV vaccine composition of any one of claims 1-11 in the manufacture of a medicament for the treatment or prevention of a disease.
14. A kit comprising the vaccine composition of any one of claims 1-11, characterized in that it comprises a first component comprising HPV antigen and an aluminium adjuvant and a second component comprising an oil-in-water emulsion adjuvant comprising squalene, alpha-tocopherol and tween 80, packaged separately.
15. A multi-chamber syringe comprising the vaccine composition of any one of claims 1-11, characterized by comprising a first chamber and a second chamber, said first chamber containing a first component comprising HPV antigens and an aluminum adjuvant; the second compartment contains a second component comprising an oil-in-water emulsion adjuvant comprising squalene, alpha tocopherol and tween 80.
Applications Claiming Priority (4)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2022113884433 | 2022-11-08 | ||
| CN202211388443 | 2022-11-08 | ||
| CN202310881092 | 2023-07-18 | ||
| CN2023108810928 | 2023-07-18 |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN118001383A true CN118001383A (en) | 2024-05-10 |
Family
ID=90949245
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202311488835.1A Pending CN118001383A (en) | 2022-11-08 | 2023-11-08 | HPV vaccine composition, preparation method and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN118001383A (en) |
-
2023
- 2023-11-08 CN CN202311488835.1A patent/CN118001383A/en active Pending
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| JP5956974B2 (en) | Papillomavirus vaccine composition | |
| CN104013955B (en) | A kind of not containing O/w emulsion and uses thereof of surfactant | |
| EP2293813A2 (en) | Nanoemulsion vaccines | |
| JP2018065821A (en) | Particulate vaccine formulations | |
| CN107188932B (en) | Truncated human papilloma virus 16 type L1 protein and application thereof | |
| CN104338126B (en) | It is a kind of that there is the vaccine combination for treating or preventing HPV viruse and its application | |
| Wang et al. | Immune responses to varicella-zoster virus glycoprotein E formulated with poly (lactic-co-glycolic acid) nanoparticles and nucleic acid adjuvants in mice | |
| CN109701010A (en) | Vaccine composite adjuvant system and its application in antigen | |
| CN101146551A (en) | Adjuvant composition comprising aluminium phosphate and 3D-MPL | |
| CN103784953A (en) | Oil-in-water submicron emulsion serving as vaccine adjuvant and preparation method thereof | |
| Taleghani et al. | Immunogenicity of HPV and HBV vaccines: adjuvanticity of synthetic analogs of monophosphoryl lipid A combined with aluminum hydroxide | |
| CN104043119B (en) | A kind of new vaccine adjuvant and preparation method thereof | |
| CN118001383A (en) | HPV vaccine composition, preparation method and application thereof | |
| US20080019989A1 (en) | Immunity Adjuvant Containing a Complexed Metal Cation and Vaccine Containing Same | |
| CN115192703A (en) | Herpes zoster vaccine and application thereof | |
| TWI811248B (en) | Nasal hepatitis b vaccine composition and preparation process thereof | |
| CN113967252A (en) | Immunopotentiator for poultry, vaccine composition containing immunopotentiator and application of vaccine composition | |
| CN109876140A (en) | A kind of vaccine and its preparation method and application for treating chronic hepatitis B | |
| CN112545992B (en) | Veterinary vaccine nanoemulsion and preparation method thereof | |
| CN105031645B (en) | A kind of Antirabic Vaccine and preparation method thereof | |
| CN117582491A (en) | Influenza vaccine composition, preparation method and application thereof | |
| CN117462667A (en) | Composite adjuvant and application thereof in vaccine preparation | |
| CN117281899A (en) | Adjuvant system and preparation method and application thereof | |
| TWI756948B (en) | Manufacturing method and use of peptide nanotube and immunogenic composition including the same | |
| US20150150967A1 (en) | Composition comprising a colloidal synthetic bioresorbable vector and a viral vector |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination |