Recombinant poliovirus-like particle composition, preparation method therefor, and use thereof

Abstract

Provided are a recombinant poliovirus-like particle immune composition, a preparation method therefor, and use thereof. The recombinant poliovirus-like particle immune composition comprises type I, type II, and / or type III poliovirus-like particles, wherein the type I, type II, and / or type III poliovirus-like particles have a suitable particle content. The prepared recombinant poliovirus-like particle immune composition and vaccine have a simple constituent and a weak side effect, and have significant advantages in the aspects of safety, immune efficacy, stability, and adaptability relative to existing inactivated vaccines and attenuated vaccines.

Description

A recombinant poliovirus-like particle composition and its preparation method and application Technical Field

[0001] The present invention belongs to the technical field of vaccines, and in particular relates to a recombinant poliovirus-like particle composition, a preparation method and an application thereof. Background Art

[0002] Poliomyelitis, also known as poliomyelitis, is an acute intestinal infectious disease caused by the poliovirus, primarily causing limb paralysis. It primarily affects children under five years old. Currently, there is no effective treatment, and the disease can only be prevented through vaccination.

[0003] Currently, there is no specific treatment for polio, and the primary preventive measure is vaccination. The vaccines commonly used to prevent polio both domestically and internationally include the inactivated poliovirus vaccine (IPV) and the oral attenuated poliovirus vaccine (OPV). OPV was successfully developed and marketed in the United States in 1958 and later globally. The advantages of OPV include simple oral administration, low cost, stable intestinal mucosal immunity, and effective blocking of poliovirus transmission. However, as the global goal of polio eradication progresses, the role of biosafety in curbing the spread of poliovirus is becoming increasingly important. Cases of vaccine-associated paralytic poliomyelitis (VAPP) following oral immunization with oral vaccine (OPV), as well as vaccine-derived polioviruses (VDPVs) arising from reversion to attenuated vaccine strains, including circulating vaccine-derived polioviruses (cVDPVs) and immunodeficiency-related vaccine-derived polioviruses (iVDPVs), which can cause further polio outbreaks, have become the primary cause of polio cases following the eradication of wild-type polioviruses and are therefore receiving increasing attention. IPV, a vaccine composed of a mixture of three poliovirus types, is prepared through cell culture, supernatant harvesting, purification, and formaldehyde inactivation. It exhibits excellent immunogenicity and safety, effectively preventing polio outbreaks without the risk of causing VAPP or cVDPVs. Its herd immunity is superior to that of OPV. IPV vaccine has better immune safety than OPV, but IPV vaccination cannot form obvious local intestinal immunity and its cost is significantly higher than OPV.

[0004] Poliovirus (PV) belongs to the genus Enterovirus in the family Picornaviridae. Its genome consists of a single-stranded positive-sense RNA approximately 7.5 kb long. The genome is divided into three parts: the 5' noncoding region, the polyprotein coding region, and the 3' noncoding region. Upon infection, the polyprotein coding region is directly translated to produce a polyprotein precursor, consisting of three regions: P1, P2, and P3. The P1 region is cleaved by its own 3CD protease into three structural subunits: VP0, VP1, and VP3. These three structural subunits form the viral protomer. Five protomers assemble into pentamers, and 12 pentamers further assemble into a symmetrical icosahedron. The primary antigenic epitopes of poliovirus are located on the viral structural proteins. Therefore, virus-like particles (VLPs) can also stimulate an effective protective immune response, making them promising vaccine candidates.

[0005] In the existing technology, inactivated polio vaccines are mainly divided into two categories: Salk IPV, a trivalent inactivated vaccine composed of wild strains Mahoney, MEF-1, and Saukett, developed by Salk; and Sabin IPV, a trivalent inactivated vaccine composed of attenuated strains Sabin1, Sabin2, and Sabin3 developed by Sabin. Both Salk IPV and Sabin IPV have certain problems in terms of safety and immune efficacy. Salk IPV uses inactivated pathogens, and generally requires multiple vaccinations to produce sufficient quantities to achieve the expected immune efficacy. In addition, the excipients in existing inactivated vaccines are mainly M199 culture medium, which has many and complex components and does not contain adjuvants. The storage condition is low temperature storage at 2-8°C. However, the inactivated vaccine itself has a large vaccination dose, a short immunity period, and a single immunization route. In addition, there is a risk of virus leakage during the production process of inactivated polio vaccines, which poses safety issues.

[0006] VLP (virus-like particle) vaccines have obvious advantages in safety, immune efficacy, stability and adaptability compared to the above-mentioned inactivated vaccines and attenuated vaccines. Based on the problems existing in the existing technology, there is an urgent need to develop a new vaccine against polio virus. Summary of the Invention

[0007] The primary object of the present invention is to provide a composition containing poliovirus-like particles.

[0008] The technical solution adopted by the present invention is:

[0009] A composition comprising type I, type II and / or type III poliovirus-like particles;

[0010] Specifically, the content of type I polio virus-like particles in the composition is 6-60 DU / dose.

[0011] Specifically, the content of type II polio virus-like particles in the composition is 4-32 DU / dose.

[0012] Specifically, the content of type III polio virus-like particles in the composition is 5-45 DU / dose.

[0013] In some specific embodiments, the content of type I poliovirus-like particles in the composition of the present invention can be 6DU / dose, 7DU / dose, 8DU / dose, 9DU / dose, 10DU / dose, 11DU / dose, 12DU / dose, 13DU / dose, 14DU / dose, 15DU / dose, 16DU / dose, 17DU / dose, 18DU / dose, 19DU / dose, 20DU / dose, 21DU / dose, 22DU / dose, 23DU / dose, 24DU / dose, 25DU / dose, 26DU / dose, 27DU / dose, 28DU / dose, 29DU / dose, 30DU / dose, 31DU / dose, e. 32DU / dose, 33DU / dose, 34DU / dose, 35DU / dose, 36DU / dose, 37DU / dose, 38DU / dose, 39DU / dose, 40DU / dose, 41DU / dose, 42DU / dose, 43DU / dose, 44DU / dose, 45DU / dose, 46DU / dose, 47DU / dose, 48DU / dose, 49DU / dose, 50DU / dose, 51DU / dose, 52DU / dose, 53DU / dose, 54DU / dose, 55DU / dose, 56DU / dose, 57DU / dose, 58DU / dose, 59DU / dose or 60DU / dose.

[0014] In some specific embodiments, the content of type II poliovirus-like particles in the composition of the present invention can be 4DU / dose, 5DU / dose, 6DU / dose, 7DU / dose, 8DU / dose, 9DU / dose, 10DU / dose, 11DU / dose, 12DU / dose, 13DU / dose, 14DU / dose, 15DU / dose, 16DU / dose, 17DU / dose, 18DU / dose, 19DU / dose, 20DU / dose, 21DU / dose, 22DU / dose, 23DU / dose, 24DU / dose, 25DU / dose, 26DU / dose, 27DU / dose, 28DU / dose, 29DU / dose, 30DU / dose, 31DU / dose or 32DU / dose.

[0015] In some specific embodiments, the content of type III poliovirus-like particles in the composition of the present invention can be 5DU / dose, 6DU / dose, 7DU / dose, 8DU / dose, 9DU / dose, 10DU / dose, 11DU / dose, 12DU / dose, 13DU / dose, 14DU / dose, 15DU / dose, 16DU / dose, 17DU / dose, 18DU / dose, 19DU / dose, 20DU / dose, 21DU / dose, 22DU / dose, 23DU / dose e. 24DU / dose, 25DU / dose, 26DU / dose, 27DU / dose, 28DU / dose, 29DU / dose, 30DU / dose, 31DU / dose, 32DU / dose, 33DU / dose, 34DU / dose , 35DU / dose, 36DU / dose, 37DU / dose, 38DU / dose, 39DU / dose, 40DU / dose, 41DU / dose, 42DU / dose, 43DU / dose, 44DU / dose or 45DU / dose.

[0016] Specifically, the type I, II, and III poliovirus-like particles are derived from wild-type or attenuated poliovirus strains.

[0017] Specifically, the type I poliovirus-like particles are derived from the Mahoney strain or Sabin 1, the type II poliovirus-like particles are derived from the MEF-1 strain or Sabin 2, and the type IIII poliovirus-like particles are derived from the Saukett strain or Sabin 3.

[0018] Specifically, at least one of the type I, type II and type III virus-like particles is derived from an attenuated poliovirus strain.

[0019] Specifically, the type I and / or type III poliovirus particles are attenuated strains

[0020] Specifically, the type I is the sabin1 strain, the type II is the MEF-1 strain, and the type III is the sabin3 strain.

[0021] Specifically, the composition further comprises an adjuvant. The composition of the present invention may optionally comprise any adjuvant. The adjuvant of the present invention may be any one or more of an oil adjuvant, an oil-in-water adjuvant, an oil-in-water adjuvant, an oil-in-water adjuvant, an aluminum salt, and a polysaccharide adjuvant.

[0022] Specifically, the aluminum adjuvant is selected from any one or more of the following four aluminum adjuvants: aluminum hydroxide, aluminum hydroxide unsaturated with phosphoric acid, aluminum hydroxide saturated with phosphoric acid, or aluminum phosphate.

[0023] Specifically, the aluminum adjuvant is any one or more of aluminum phosphate (AP), aluminum hydroxide treated with different concentrations of phosphate (PTAH-unsa), and aluminum hydroxide saturated with phosphate (PTAH-sa). More preferably, the concentration of the phosphate is any one of saturated and unsaturated.

[0024] Specifically, the adjuvant is prepared using a buffer system prepared with aluminum products, and the buffer system prepared with aluminum products includes a phosphate buffer system, a histidine buffer system, a citric acid buffer system, or a succinic acid buffer system, preferably a histidine buffer.

[0025] In some embodiments, an aluminum phosphate adjuvant may be added to the composition preparation of the present invention. Specifically, 0.05-1 mg of aluminum phosphate adjuvant may be added to 0.5 ml of the preparation of the present invention.

[0026] In some specific embodiments, 0.05 mg, 0.10 mg, 0.15 mg, 0.20 mg, 0.25 mg, 0.30 mg, 0.35 mg, 0.40 mg, 0.45 mg, 0.50 mg, 0.55 mg, 0.60 mg, 0.65 mg, 0.70 mg, 0.75 mg, 0.80 mg, 0.85 mg, 0.90 mg, 0.95 mg or 1.00 mg of aluminum phosphate adjuvant can be added per 0.5 ml of the composition preparation of the present invention.

[0027] Specifically, the adjuvant can also be selected from any one or more of calcium, iron or zinc.

[0028] Specifically, the adjuvant can also be any one or more of a cationically or anionically derived carbohydrate, polyphosphazene, biodegradable microspheres, monophosphoryl lipid A (MPL), lipid A derivatives, 3-O-deacylated MPL, quil A, Saponin, QS21, tocol (EP 0382271), Freund's incomplete adjuvant (Difco Laboratories, Detroit, MI), Merck Adjuvant 65 (Merck and Company, Inc., Rahway, NJ), AS-2 (Smith-Kline Beecham, Philadelphia, PA), CpG oligonucleotides, bioadhesives and mucoadhesives, microparticles, liposomes, polyoxyethylene ether preparations, polyoxyethylene ester preparations, muramyl peptides or imidazoquinolone compounds (e.g., imiquimod and its homologues).

[0029] Specifically, the adjuvant can also be a human immunomodulator including cytokines, such as interleukins (such as IL-1, IL-2, IL-4, IL-5, IL-6, IL-7, IL-12, etc.), macrophage colony stimulating factor (M-CSF), tumor necrosis factor (TNF), granulocyte macrophage colony stimulating factor (GM-CSF), any one or more of them.

[0030] Specifically, the adjuvant can induce a TH1 immune response and high levels of TH1 cytokines, such as any one or more of IFN-γ, TNFα, JL-2, and IL-12.

[0031] Further, suitable adjuvant systems that promote a predominantly TH1 response include derivatives of lipid A (e.g., with reduced toxicity), monophosphoryl lipid A (MPL) or its derivatives, particularly 3-de-O-acylated monophosphoryl lipid A (3D-MPL), and combinations of monophosphoryl lipid A, optionally 3-de-O-acylated monophosphoryl lipid A, and aluminum salts.

[0032] Specifically, the vaccine further includes excipients, which include osmotic pressure regulators, surfactants, and excipients.

[0033] Specifically, the osmotic pressure regulator is preferably sodium chloride, and the sodium chloride content is 50-200mM; 50-150mM; 50-100mM; 50-75mM; 75-200mM; 75-150mM; 75-100mM.

[0034] Specifically, the surfactant is preferably Tween-80; the content of Tween-80 is preferably less than 0.01%; less than 0.005%, less than 0.0025%; less than 0.0015%; less than 0.001%.

[0035] Specifically, the pH range of the vaccine is 5-8, preferably, the pH is 5.0-7.5, 5.0-7.0, 5.0-6.5, 5.0-6.0, 5.0-5.5.

[0036] Specifically, the dosage form of the composition is any one or more of a liquid preparation, a lyophilized preparation, a powder and an inhalation preparation; more preferably, the composition is a subcutaneous injection, an intramuscular injection or a microneedle preparation.

[0037] The attenuated poliovirus strain described in the present invention refers to a safe and effective strain obtained by reducing the pathogenicity of the virus through a series of methods and techniques, wherein the attenuation methods and techniques include but are not limited to: continuous passage, temperature-sensitive screening, genetic engineering, antisense mutation and / or recombinant virus.

[0038] The wild-type poliovirus strains described herein refer to strains isolated from naturally infected animals. Currently discovered wild-type poliovirus strains include, but are not limited to, type I strains such as Mahoney, Brunenders, and Lansing; type II strains such as MEF-1, MEF-2, and MEF-3; and type III strains such as Leon, Saukett, and P3 / Leon.

[0039] The DU mentioned in the present invention is an effective antigen content unit, and the method for measuring DU is well known in the art.

[0040] When testing the antigen content of the same product using different antibodies, the measured antigen content results may be different. The different results do not mean that the antigen content has changed. They are only due to the different antibodies used.

[0041] For the poliovirus-like particles described in this application, CanSino Biologics Inc.'s self-produced antibodies were used to detect the content of type I, type II, and type III poliovirus-like particles, respectively, which were 15-135 DU / dose, 4-16 DU / dose, and 20-50 DU / dose. Using standard antibodies certified by the World Health Organization (WHO) (international standard cIPV standard (12 / 104), international standard detection antibodies Type 1 (20 / 250), Type 2 (20 / 252), and Type 3 (20 / 254)), the poliovirus-like particles described in this application were detected, and the content of type I, type II, and type III was 4-32 DU / dose, 6-60 DU / dose, and 5-45 DU / dose, respectively.

[0042] The dose of the present invention is generally the amount of the vaccine administered once, i.e., generally the amount of a single injection. A typical human dose is 0.5 mL. Of course, various doses can be used in the vaccine administration protocol.

[0043] The present invention further provides use of the above composition in preparing a medicament for preventing and / or treating poliovirus infection.

[0044] The present invention further provides a method for preparing a poliovirus-like particle composition, which comprises mixing purified PV1-VLPs, PV2-VLPs and PV3-VLPs, adding an adjuvant, mixing thoroughly, and storing.

[0045] Furthermore, the preparation method includes preparing an adjuvant, and when preparing the adjuvant, the pH and / or osmotic pressure of the adjuvant stock solution is first adjusted with a buffer solution.

[0046] The present invention provides a combined immune composition comprising the composition described above and optionally: respiratory syncytial virus vaccine, herpes zoster vaccine, Haemophilus influenzae type B (Hib) vaccine, rotavirus vaccine, SARS vaccine, tuberculosis vaccine, hepatitis A vaccine, hepatitis B vaccine, measles vaccine, polio vaccine, diphtheria and pertussis vaccine, Japanese encephalitis vaccine, pertussis vaccine, diphtheria vaccine, tetanus vaccine, rabies vaccine, influenza vaccine, pneumonia vaccine, varicella vaccine, hand, foot and mouth disease vaccine, meningitis vaccine, epidemic encephalitis Japanese vaccine, new coronavirus vaccine, HPV vaccine, mumps vaccine, rubella vaccine, tumor vaccine or one or more vaccines of the above disease mutants.

[0047] Specifically, it comprises a Haemophilus influenzae type b antigen (Hib) carrier protein conjugate, wherein the capsular saccharide of Haemophilus influenzae type B and the carrier protein are covalently bound; preferably, Hib is conjugated to the carrier protein by reductive amination.

[0048] Specifically, the carrier is selected from the group consisting of: tetanus toxoid (TT), diphtheria toxoid (DT), CRM197, recombinant diphtheria toxin, OMPC of Neisseria meningitidis, pneumolysin (PLY) of Streptococcus pneumoniae, pertussis toxoid, PsaA, cholera toxoid and Haemophilus influenzae protein D (PD) or a combination thereof, an inactivated bacterial toxin carrier protein; preferably, tetanus toxoid (TT).

[0049] Specifically, it includes a pneumococcal capsular polysaccharide protein conjugate, which includes a pneumococcal capsular polysaccharide selected from the group consisting of serotypes 1, 2, 3, 4, 5, 6A, 6B, 6C, 6D, 7F, 8, 9N, 9V, 10A, 11A, 12F, 14, 15A, 15B, 15C, 16F, 17F, 18C, 19F, 19A, 20A, 20B, 22F, 23A, 23B, 23F, 24B, 24F, 31, 33F, 34, 35B, 35F, 38, 39 and 45; preferably, the capsule The membrane polysaccharide is selected from 1, 2, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 9N, 10A, 11A, 12F, 14, 15B, 17F, 18C, 19A, 19F, 20, 22F, 23F or 33F; preferably, the capsular polysaccharide is selected from 1, 2, 3, 4, 5, 6A, 6B, 7F, 9V, 14, 18C, 19A, 19F, 23F or 33F, or the capsular polysaccharide is selected from 1, 3, 4, 5, 6A, 6B, 7F, 9V, 14, 18C, 19A, 19F or 23F.

[0050] Specifically, the preparation combination includes at least one first carrier protein conjugated with capsular polysaccharides from different pneumococcal serotypes and at least one second carrier protein conjugated with capsular polysaccharides from different pneumococcal serotypes, the first carrier protein is CRM197, and the second carrier protein is TT, DT or HID, wherein capsular polysaccharides of the same serotype are conjugated to only one carrier protein.

[0051] Specifically, the pneumococcal conjugate vaccine is selected from a 7-valent pneumococcal conjugate vaccine, a 10-valent pneumococcal conjugate vaccine, a 13-valent pneumococcal conjugate vaccine, a 15-valent pneumococcal conjugate vaccine or a 24-valent pneumococcal conjugate vaccine; further, the serotypes included in the 7-valent pneumococcal conjugate vaccine are 4, 6B, 9V, 14, 18C, 19F or 23F, and the serotypes included in the 10-valent pneumococcal conjugate vaccine are 1, 4, 5, 6B, 7F, 9V, 14, 18C, 19F or 23F, the serotype included in the 13-valent pneumococcal conjugate vaccine is 1, 3, 4, 5, 6A, 6B, 7F, 9V, 14, 18C, 19A, 19F or 23F, and the serotype included in the 15-valent pneumococcal conjugate vaccine is 1, 2, 3, 4, 5, 6A, 6B, 7F, 9V, 14, 18C, 19A, 19F, 23F or 33F.

[0052] In some embodiments, the combined immune composition of the present invention includes any of the above-mentioned compositions and other immune compositions, and optionally includes a pharmaceutically acceptable excipient. The other immune composition is derived from one or more of Bordetella pertussis, Bordetella tetani, Corynebacterium diphtheriae, Hepatitis B virus, Haemophilus influenzae, Meningococcus A, Meningococcus C, Meningococcus W, Meningococcus Y, Meningococcus B, Pneumococcus, Salmonella typhi, and Hepatitis A virus;

[0053] The polysaccharide is present in the combination vaccine as any single type, or different combinations of two types, or different combinations of three types, or different combinations of four types from serogroups A, C, W135, and Y. The outer membrane protein complex is present in the combination vaccine as any single type, or different combinations of two types, or different combinations of three types, or four types, or different combinations of multiple types from serogroup B.

[0054] In another embodiment of the present invention, a multivalent vaccine is provided, which includes the composition of the present invention as described above and other optional inactivated whole-cell Bordetella pertussis (Pw), tetanus toxoid (TT), diphtheria toxoid (DT), a conjugate of a carrier protein and a capsular saccharide of Haemophilus influenzae type B (Hib-optionally conjugated to TT, DT or CRM197), and the immunogenicity of the conjugate is equal to or improved over such compositions including a larger amount of conjugates. Optionally, hepatitis B surface antigen may also be included. Optionally, capsular polysaccharides derived from group A, C, W135, and Y meningococci and outer membrane protein complexes derived from group B meningococci may also be included. Optionally, the other immune compositions are also derived from capsular polysaccharides in group A, C, W135, and Y meningococci and outer membrane protein complexes derived from group B meningococci.

[0055] The polysaccharide is present in the combination vaccine in an independent state or in a coupled state. The polysaccharide can be present in the combination vaccine as any single component from groups A, C, W135, or Y, or in different combinations of two components, three components, or four components. The group B meningococcal outer membrane protein complex is a monovalent, bivalent, or multivalent outer membrane protein antigen derived from different strains of group B meningococci.

[0056] Alternatively, a pneumococcal capsular polysaccharide protein conjugate comprises a pneumococcal capsular polysaccharide selected from the group consisting of serotypes 1, 2, 3, 4, 5, 6A, 6B, 6C, 6D, 7F, 8, 9N, 9V, 10A, 11A, 12F, 14, 15A, 15B, 15C, 16F, 17F, 18C, 19F, 19A, 20A, 20B, 22F, 23A, 23B, 23F, 24B, 24F, 31, 33F, 34, 35B, 35F, 38, 39 and 45; preferably, the capsule The membrane polysaccharide is selected from 1, 2, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 9N, 10A, 11A, 12F, 14, 15B, 17F, 18C, 19A, 19F, 20, 22F, 23F or 33F; preferably, the capsular polysaccharide is selected from 1, 2, 3, 4, 5, 6A, 6B, 7F, 9V, 14, 18C, 19A, 19F, 23F or 33F, or the capsular polysaccharide is selected from 1, 3, 4, 5, 6A, 6B, 7F, 9V, 14, 18C, 19A, 19F or 23F.

[0057] In another embodiment of the present invention, there is provided the use of the immune composition and / or vaccine of the present invention in the preparation of a medicament for the treatment or prevention of a disease caused by infection with poliovirus and optionally Bordetella pertussis, Bordetella tetani, Corynebacterium diphtheria, Hepatitis B virus, Haemophilus influenzae, Meningococcus A, Meningococcus C, Meningococcus W, Meningococcus Y, Meningococcus B, Pneumococcus, Salmonella typhi, Hepatitis A or Hepatitis B.

[0058] In addition, a method for immunizing a human host against a disease caused by poliovirus and optionally Bordetella pertussis, Bordetella tetani, Corynebacterium diphtheria, hepatitis B virus, Haemophilus influenzae, meningococcus A, meningococcus C, meningococcus W, meningococcus Y, meningococcus B, pneumococcus, Salmonella typhi or hepatitis A is provided, the method comprising administering to the host an immunoprotective dose of the vaccine of the present invention. BRIEF DESCRIPTION OF THE DRAWINGS

[0059] Figure 1 Type I neutralizing antibody titer in monovalent antigen immunogenicity experiment

[0060] Figure 2 Type II neutralizing antibody titer in monovalent antigen immunogenicity experiment

[0061] Figure 3 Type III neutralizing antibody titer in monovalent antigen immunogenicity experiment

[0062] Figure 4 Type I neutralizing antibody titer in trivalent antigen immunogenicity experiment

[0063] Figure 5 Type II neutralizing antibody titer in trivalent antigen immunogenicity experiment

[0064] Figure 6 Type III neutralizing antibody titer in trivalent antigen immunogenicity experiment DETAILED DESCRIPTION

[0065] The present invention will now be described in more detail with reference to the following examples, which are provided for the sole purpose of illustrating the present invention and are not to be construed as limiting the scope and substance of the present invention.

[0066] Unless otherwise specified, the terms used in the present invention generally have the meanings commonly understood by those skilled in the art.

[0067] Example 1 Screening of recombinant poliovirus-like particle antigen doses

[0068] Experimental group: The structural gene (VP0, VP1 and VP3) sequences of type I: sabin1, type II: MEF-1, and type III: sabin3 were used to prepare recombinant monovalent poliovirus-like particle preparations to investigate the immunogenicity of the monovalent products.

[0069] Control group: Prepare monovalent sIPV1, sIPV2, and sIPV3 inactivated preparations composed of attenuated strains salk1, salk2, and salk3, with dosages of salk1: 40DU, salk2: 8DU, and salk3: 32DU / dose / 0.5ml, as the experimental group to investigate the monovalent immunogenicity.

[0070] Table 1. Antigen dose screening of type I monovalent poliovirus-like particle preparations

[0071] Table 2. Antigen dose screening of type II monovalent poliovirus-like particle preparations

[0072] Table 3. Antigen dose screening of type III monovalent poliovirus-like particle preparations

[0073] Table 4. Control group trivalent inactivated poliomyelitis vaccine

[0074] Experimental Procedure: Female Wister rats weighing approximately 160-200g were selected for SPF vaccination and received two intramuscular immunizations on days 0 and 21, respectively. Blood was collected from the retroorbital venous plexus 21 days after the first and second immunizations. Serum was separated after centrifugation at 8000 rpm for 6 minutes at 4°C. The serum was then heated at 56°C for 30 minutes, and the levels of neutralizing antibodies against poliovirus types I, II, and III in each serum were determined using a pseudovirus assay.

[0075] The results showed that all rats were 100% healthy and no adverse reactions were observed. Neutralizing antibody titers are shown in Figures 1-3.

[0076] Example 2 Recombinant trivalent poliovirus-like particles

[0077] Experimental group: Based on the dosage of the recombinant trivalent polio vaccine preparation in Example 1, the immune effects of different ratios of the trivalent polio vaccine preparation were evaluated.

[0078] Positive control (Group 16): sIPV: trivalent inactivated vaccine sIPV composed of attenuated strains sa1k1, sa1k2, and sa1k3, salk1:salk2:salk3 = 40 DU:8 DU:32 DU / dose / 0.5 ml.

[0079] Experimental Procedure: SPF-qualified female Wister rats weighing approximately 160-200g were immunized twice intramuscularly on days 0 and 21. Twenty-one days after the second immunization, blood was collected from the retroorbital venous plexus. Serum was separated after centrifugation at 8000 rpm for 6 minutes at 4°C. The serum was then heated at 56°C for 30 minutes, and the level of neutralizing antibodies against poliovirus types I, II, and III in each serum was determined using a pseudovirus assay.

[0080] Table 5. Trivalent poliovirus-like particle antigen dose screening

[0081] The results showed that all rats were 100% healthy and no adverse reactions were observed. Neutralizing antibody titers are shown in Figures 4-6.

Claims

1. A composition, characterized in that Contains type I, type II and / or type III poliovirus-like particles; the content of type II poliovirus-like particles in the composition is 4-32 DU / dose.

2. The composition according to claim 1, characterized in that The content of type I poliovirus-like particles is 6-60DU / dose.

3. The composition according to any one of claims 1 to 2, characterized in that The content of type III polio virus-like particles in the composition is 5-45 DU / dose.

4. The composition according to any one of claims 1 to 3, characterized in that The type I, II, and III poliovirus-like particles are derived from wild-type or attenuated poliovirus strains.

5. The composition according to claim 4, characterized in that At least one of the type I, type II and type III virus-like particles is derived from an attenuated poliovirus strain.

6. The composition according to claim 4 or 5, characterized in that The type I and / or type III poliovirus particles are attenuated strains.

7. The composition according to any one of claims 4 to 6, characterized in that The type I polio virus-like particles are derived from the Mahoney strain or Sabin 1, the type II polio virus-like particles are derived from the MEF-1 strain or Sabin 2, and the type IIII polio virus-like particles are derived from the Saukett strain or sabin 3; preferably, the type I is the sabin 1 strain, the type II is the MEF-1 strain, and the type III is the sabin 3 strain.

8. The composition according to any one of claims 1 to 7, characterized in that The composition further includes an adjuvant, which is any one or more of an oil adjuvant, an oil-in-water adjuvant, an oil-in-water adjuvant, a water-in-oil adjuvant, an aluminum salt and a polysaccharide adjuvant; the aluminum adjuvant is selected from any one or more of the four aluminum adjuvants: aluminum hydroxide, aluminum hydroxide unsaturated with phosphate, aluminum hydroxide saturated with phosphate or aluminum phosphate; the aluminum adjuvant is any one or more of aluminum phosphate (AP), aluminum hydroxide treated with different concentrations of phosphate (PTAH-unsa) and aluminum hydroxide saturated with phosphate (PTAH-sa); more preferably, the concentration of the phosphate is any one of saturated or unsaturated concentrations.

9. The composition according to claim 8, characterized in that The adjuvant is prepared with a buffer system prepared with aluminum finished products, and the buffer system prepared with aluminum finished products includes a phosphate buffer system, a histidine buffer system, a citric acid buffer system or a succinic acid buffer system; preferably, a histidine buffer.

10. The composition according to any one of claims 1 to 9, characterized in that The vaccine also includes excipients, which include osmotic pressure regulators, surfactants, and excipients; preferably, the osmotic pressure regulator is preferably sodium chloride, and the sodium chloride content is 50-200mM; 50-150mM; 50-100mM; 50-75mM; 75-200mM; 75-150mM; 75-100mM; preferably, the surfactant is preferably Tween-80; the content of Tween-80 is preferably less than 0.01%; less than 0.005%, less than 0.0025%; less than 0.0015%; less than 0.001%.

11. The composition according to any one of claims 1 to 10, characterized in that The pH range of the composition is 5-8, preferably, the pH is 5.0-7.5, 5.0-7.0, 5.0-6.5, 5.0-6.0, 5.0-5.

5.

12. The composition according to any one of claims 1 to 11, characterized in that The dosage form of the composition is any one or more of a liquid preparation, a lyophilized preparation, a powder and an inhalation preparation; more preferably, the composition is a subcutaneous injection, an intramuscular injection or a microneedle preparation.

13. Use of the composition according to any one of claims 1 to 12 in the preparation of a medicament for preventing and / or treating poliovirus infection.

14. A method for preparing the composition according to any one of claims 1 to 13, comprising mixing purified PV1-VLPs, PV2-VLPs and PV3-VLPs, adding an adjuvant, mixing thoroughly, and storing.

15. The preparation method according to claim 14, characterized in that The method comprises preparing the adjuvant. When preparing the adjuvant, the pH and / or osmotic pressure of the adjuvant stock solution is first adjusted with a buffer.

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