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CN-122005771-A - Composition of 31-valent pneumococcal capsular polysaccharide and application thereof

CN122005771ACN 122005771 ACN122005771 ACN 122005771ACN-122005771-A

Abstract

The invention provides a composition of 31-valent pneumococcal capsular polysaccharide and application thereof, wherein the composition comprises 31 pneumococci Capsular polysaccharides of the serotype, which form conjugates with carrier proteins. The safety and the immune protection effect of the vaccine are obviously improved by integrating a multi-carrier compatibility strategy, an adjuvant-free preparation technology and serotype specific antigen dosage optimization. The vaccine effectively overcomes carrier inhibition in multivalent vaccine, adopts an adjuvant-free formula to thoroughly avoid local and systemic adverse reactions related to aluminum adjuvant, improves vaccination compliance, implements antigen dose enhancement on low-immunogenicity serotypes, and synergistically enhances the breadth and intensity of overall immune response. The product has a stable process, is suitable for large-scale production, and can provide safer and broad-spectrum protection for sensitive people such as infants, the elderly and the like.

Inventors

  • ZHAO ZHIQIANG
  • SUN ZUYONG
  • ZHANG YINCHUAN
  • LI LINQIANG
  • SUN YINGYING
  • WU JINJUN
  • MA GUODI
  • YANG JUN
  • YANG WEI
  • WEI LIQIN
  • ZHANG XIAOYAN

Assignees

  • 江苏坤力生物制药有限责任公司
  • 山东坤力生物制药有限责任公司

Dates

Publication Date
20260512
Application Date
20251114
Priority Date
20250523

Claims (12)

  1. 1. A composition of a 31-valent pneumococcal capsular polysaccharide, the composition comprising 31 pneumococci Capsular polysaccharides of serotypes 1,2,3,4, 5, 6A, 6B, 7F, 8, 9N, 9V, 10A, 11A, 12F, 14, 15A, 15B, 15C, 16F, 17F, 18C, 19A, 19F, 20, 22F, 23A, 23B, 23F, 31, 33F and 35B, wherein the individual capsular polysaccharides of types 1,3, 6B, 19A and 19F are present in the composition in an amount greater than the other types.
  2. 2. The composition of claim 1, wherein the composition is divided into the following three groups according to the content of individual capsular polysaccharides: 1) Group 1, 3, 6B, 19A and 19F capsular polysaccharides; 2) Group 15A, 15C, 16F, 23A, 23B, 31 and 35B capsular polysaccharides, and, 3) 2,4, 5, 6A, 7F, 8, 9N, 9V, 10A, 11A, 12F, 14, 15B, 17F, 18C, 20, 22F, 23F and 33F capsular polysaccharides, wherein the content of single type capsular polysaccharides of different serotypes in the group is the same, and the content of single type capsular polysaccharides in the group is different; Preferably, the capsular polysaccharide content of the composition for a single type is 1) 2-6 times, e.g. 4-5 times, the capsular polysaccharide content of group 2), and/or 3) 1-3 times, e.g. 2 times, the capsular polysaccharide content of group 2).
  3. 3. The composition of claim 2, wherein the composition meets one or more conditions selected from the group consisting of: a. 1) The content of the single capsular polysaccharide in the group is 4-10 mug/0.5 mL, for example, 10 mug/0.5 mL, 8 mug/0.5 mL, 6 mug/0.5 mL or 4 mug/0.5 mL; b. 2) The content of the single capsular polysaccharide in the group is 1.5-2.5 mug/0.5 mL, and, C. 3) The content of single capsular polysaccharide in the group is 3.5-4.5 mug/0.5 mL; Preferably, the composition comprises 8-10 mug/0.5 mL of the single type capsular polysaccharide in group 1), 2 mug/0.5 mL of the single type capsular polysaccharide in group 2), and 4 mug/0.5 mL of the single type capsular polysaccharide in group 3).
  4. 4. The composition of any one of claims 1-3, wherein the composition further comprises a carrier protein, wherein the capsular polysaccharide forms a conjugate with the carrier protein, and/or wherein the composition does not contain an adjuvant; Preferably, the carrier protein is selected from one or more of pneumolysin, tetanus toxoid, diphtheria toxin nontoxic mutant and recombinant streptococcus pneumoniae histidine triple protein, preferably tetanus toxoid is a tetanus toxin C fragment; More preferably, the carrier protein comprises pneumolysin and/or a tetanus toxin C fragment, and further comprises one or more of a tetanus toxoid, a diphtheria toxin non-toxic mutant and a recombinant pneumococcal histidine triple protein, preferably the carrier protein comprises pneumolysin, a tetanus toxin C fragment and a diphtheria toxin non-toxic mutant; Even more preferably, in the composition: 1) One or more capsular polysaccharides from the 31 serotypes are conjugated to pneumolysin, respectively; 2) One or more capsular polysaccharides from 31 serotypes are conjugated to non-toxic mutants of diphtheria toxin, respectively, or, 3) One or more capsular polysaccharides from 31 serotypes are conjugated to the tetanus toxin C fragment separately.
  5. 5. The composition of claim 4, wherein the composition is divided into three groups depending on the carrier protein to which the capsular polysaccharide is bound: i) A group comprising one or more of the capsular polysaccharides of serotypes 1,3, 4, 6A, 6B, 11A, 14, 17F, 19A, 19F, 20, 22F, 23B, 23F, 31 and 35B, preferably 9-13, for example 10, 11 or 13; ii) one or more, preferably 10-13, such as 11 or 12, capsular polysaccharides comprising serotypes 1,2, 5, 7F, 8, 9V, 9N, 10A, 11A, 14, 15A, 15B, 15C, 16F, 18C, 20, 31 and 33F, and, Iii) A group comprising one or more of the capsular polysaccharides of serotypes 3,5, 7F, 8, 9N, 9V, 10A, 12F, 15B, 18C, 19A, 22F, 23A and 23B, preferably 5-11, e.g., 6, 9 or 10, wherein the carrier proteins bound by individual capsular polysaccharides of different serotypes within the group are identical, the carrier proteins bound by individual capsular polysaccharides of the group are different from one another, and the individual capsular polysaccharides occur only 1 time in three groups; Preferably, the composition satisfies one or more of the following conditions: i) The capsular polysaccharides in the group were conjugated to pneumolysin, respectively; ii) conjugated to diphtheria toxin nontoxic mutants, respectively, and, Iii) The capsular polysaccharides in the group were conjugated to tetanus toxin C fragment, respectively; more preferably, the composition comprises: 1) Conjugates of capsular polysaccharides from serotypes 1, 3, 4, 6A, 6B, 14, 17F, 19A, 19F, 20, 23B, 23F and 35B with pneumolysin, respectively, conjugates of capsular polysaccharides from serotypes 2, 9V, 9N, 10A, 11A, 15B, 15C, 16F, 18C, 31 and 33F with non-toxic mutants of diphtheria toxin, conjugates of capsular polysaccharides from serotypes 5, 7F, 8, 12F, 22F and 23A with tetanus toxin C fragments, respectively; 2) Conjugates of capsular polysaccharides from serotypes 1, 3, 6B, 14, 17F, 19F, 20, 23B, 23F, 33F and 35B with pneumolysin, respectively, conjugates of capsular polysaccharides from serotypes 2, 4, 6A, 9V, 9N, 10A, 11A, 12F, 15A, 15C, 16F, 18C, 19A, 20, 23A and 31 with non-toxic mutants of diphtheria toxin, conjugates of capsular polysaccharides from serotypes 5, 7F, 8 and 14 with fragments of tetanus toxin C, respectively; 3) Conjugates of capsular polysaccharides from serotypes 1, 3, 4, 6A, 6B, 14, 17F, 19A, 19F, 20, 23A, 23B and 23F with pneumolysin, respectively, conjugates of capsular polysaccharides from serotypes 2, 9V, 9N, 11A, 15B, 15C, 16F, 18C, 31, 33F and 35B with non-toxic mutants of diphtheria toxin, conjugates of capsular polysaccharides from serotypes 5, 7F, 8, 10A, 12F and 22F with tetanus toxin C fragments, respectively; 4) Conjugates of capsular polysaccharides from serotypes 1, 3, 6B, 14, 17F, 19F, 23B, 23F, 31, 33F and 35B with pneumolysin, respectively, conjugates of capsular polysaccharides from serotypes 2, 4, 6A, 9V, 9N, 11A, 15B, 15C, 16F and 18C with non-toxic mutants of diphtheria toxin, respectively, and conjugates of capsular polysaccharides from serotypes 5, 7F, 8, 10A, 12F, 15A, 19A, 20, 22F and 23A with tetanus toxin C fragment, respectively; 5) Conjugates of capsular polysaccharides from serotypes 1, 3, 6A, 6B, 14, 17F, 19F, 23B, 23F, 31 and 35B with pneumolysin, respectively, conjugates of capsular polysaccharides from serotypes 2, 4, 9V, 9N, 11A, 15B, 15C, 16F, 18C and 33F with non-toxic mutants of diphtheria toxin, respectively, and conjugates of capsular polysaccharides from serotypes 5, 7F, 8, 10A, 12F, 15A, 19A, 20, 22F and 23A with tetanus toxin C fragment, respectively; 6) Conjugates of capsular polysaccharides from serotypes 4, 6A, 6B, 11A, 17F, 19F, 22F, 23F, 31 and 35B with pneumolysin, respectively, conjugates of capsular polysaccharides from serotypes 1, 2, 5, 7F, 8, 14, 15A, 15C, 16F, 20 and 33F with non-toxic mutants of diphtheria toxin, conjugates of capsular polysaccharides from serotypes 3, 9N, 9V, 10A, 12F, 15B, 18C, 19A, 23A and 23B with fragments of tetanus toxin C, respectively, or, 7) Conjugates of capsular polysaccharides from serotypes 3, 4, 6A, 6B, 11A, 14, 17F, 19F, 23F, 31 and 35B with pneumolysin, respectively, conjugates of capsular polysaccharides from serotypes 1, 2, 9V, 10A, 15B, 15C, 16F, 18C, 20 and 33F with non-toxic mutants of diphtheria toxin, respectively, and conjugates of capsular polysaccharides from serotypes 5, 7F, 8, 9N, 12F, 19A, 22F, 23A and 23B with tetanus toxin C fragments, respectively.
  6. 6. The composition of claim 4 or 5, wherein the composition meets one or more conditions selected from the group consisting of: 1) Capsular polysaccharides of serotypes 1, 2,3, 4,5, 6A, 6B, 7F, 8, 9N, 9V, 10A, 11A, 12F, 14, 15A, 15B, 15C, 16F, 17F, 18C, 19F, 20, 22F, 23A, 23B, 23F, 31, 33F and 35B are hydrolyzed capsular polysaccharides; 2) The pneumolysin has an amino acid sequence shown as SEQ ID NO. 1; 3) The tetanus toxin C fragment has an amino acid sequence shown as SEQ ID NO. 2; 4) The diphtheria toxin nontoxic mutant has an amino acid sequence shown as SEQ ID NO. 3, and, 5) The mass ratio of capsular polysaccharide to carrier protein in the composition is (0.5-1.5): 1.
  7. 7. A vaccine comprising the composition of any one of claims 1-6 for inducing an immune response in an organism against pneumococci, said vaccine further comprising an adjuvant; Preferably, the auxiliary materials comprise one or more of sodium chloride, sodium acetate, citric acid, polysorbate 80, trehalose, sucrose, dipotassium hydrogen phosphate, sodium dihydrogen phosphate and arginine; more preferably, the vaccine satisfies one or more selected from the following conditions: 1) The content of sodium chloride in the vaccine is 3.75-4.75 mg/0.5mL, for example 4.5mg/0.5mL; 2) The content of sodium acetate in the vaccine is 1-3 mg/0.5mL, such as 1.23mg/0.5mL, and, 3) The content of polysorbate 80 in the vaccine is 80-2500 mug/0.5 mL, for example 100 mug/0.5 mL.
  8. 8. The vaccine of claim 7, wherein said vaccine does not contain an adjuvant.
  9. 9. Vaccine according to claim 7 or 8, characterized in that the vaccine has a content of individual capsular polysaccharides of type 1, 3, 6B, 19A and 19F of 8-10 μg/0.5mL, a content of individual capsular polysaccharides of type 15a, 15C, 16F, 23A, 23B, 31 and 35B of 2 μg/0.5mL, and a content of individual capsular polysaccharides of type 2, 4, 5, 6A, 7F, 8, 9N, 9V, 10A, 11A, 12F, 14, 15B, 17F, 18C, 20, 22F, 23F and 33F of 4 μg/0.5mL; Preferably, in the vaccine: 1) Conjugates of capsular polysaccharides from serotypes 1, 3, 4, 6A, 6B, 14, 17F, 19A, 19F, 20, 23B, 23F and 35B with pneumolysin, respectively, conjugates of capsular polysaccharides from serotypes 2, 9V, 9N, 10A, 11A, 15B, 15C, 16F, 18C, 31 and 33F with diphtheria toxin nontoxic mutants, respectively, conjugates of capsular polysaccharides from serotypes 5, 7F, 8, 12F, 22F and 23A with tetanus toxin C fragment, respectively; 2) Conjugates of capsular polysaccharides from serotypes 1, 3, 6B, 14, 17F, 19F, 20, 23B, 23F, 33F and 35B with pneumolysin, respectively, conjugates of capsular polysaccharides from serotypes 2, 4, 6A, 9V, 9N, 10A, 11A, 12F, 15A, 15C, 16F, 18C, 19A, 20, 23A and 31 with non-toxic mutants of diphtheria toxin, respectively, conjugates of capsular polysaccharides from serotypes 5, 7F, 8 and 14 with fragments of tetanus toxin C, respectively; 3) Conjugates of capsular polysaccharides from serotypes 1, 3, 4, 6A, 6B, 14, 17F, 19A, 19F, 20, 23A, 23B and 23F with pneumolysin, respectively, conjugates of capsular polysaccharides from serotypes 2, 9V, 9N, 11A, 15B, 15C, 16F, 18C, 31, 33F and 35B with diphtheria toxin nontoxic mutants, respectively, conjugates of capsular polysaccharides from serotypes 5, 7F, 8, 10A, 12F and 22F with tetanus toxin C fragment, respectively; 4) Conjugates of capsular polysaccharides from serotypes 1, 3, 6B, 14, 17F, 19F, 23B, 23F, 31, 33F and 35B with pneumolysin, respectively, conjugates of capsular polysaccharides from serotypes 2, 4, 6A, 9V, 9N, 11A, 15B, 15C, 16F and 18C with non-toxic mutants of diphtheria toxin, respectively, and conjugates of capsular polysaccharides from serotypes 5, 7F, 8, 10A, 12F, 15A, 19A, 20, 22F and 23A with tetanus toxin C fragment, respectively; 5) Conjugates of capsular polysaccharides from serotypes 1, 3, 6A, 6B, 14, 17F, 19F, 23B, 23F, 31 and 35B with pneumolysin, respectively, conjugates of capsular polysaccharides from serotypes 2, 4, 9V, 9N, 11A, 15B, 15C, 16F, 18C and 33F with non-toxic mutants of diphtheria toxin, respectively, and conjugates of capsular polysaccharides from serotypes 5, 7F, 8, 10A, 12F, 15A, 19A, 20, 22F and 23A with tetanus toxin C fragment, respectively; 6) The capsular polysaccharides of serotypes 4, 6A, 6B, 11A, 17F, 19F, 22F, 23F, 31 and 35B are conjugated to pneumolysin, respectively, the capsular polysaccharides of serotypes 1, 2, 5, 7F, 8, 14, 15A, 15C, 16F, 20 and 33F are conjugated to diphtheria toxin nontoxic mutants, respectively, the capsular polysaccharides of serotypes 3, 9N, 9V, 10A, 12F, 15B, 18C, 19A, 23A and 23B are conjugated to tetanus toxin C fragment, respectively, or, 7) Conjugates of capsular polysaccharides from serotypes 3, 4, 6A, 6B, 11A, 14, 17F, 19F, 23F, 31 and 35B with pneumolysin, respectively, conjugates of capsular polysaccharides from serotypes 1, 2, 9V, 10A, 15B, 15C, 16F, 18C, 20 and 33F with non-toxic mutants of diphtheria toxin, respectively, and conjugates of capsular polysaccharides from serotypes 5, 7F, 8, 9N, 12F, 19A, 22F, 23A and 23B with tetanus toxin C fragment, respectively; more preferably, the vaccine satisfies one or more selected from the following conditions: 1) Capsular polysaccharides of serotypes 1, 2,3, 4,5, 6A, 6B, 7F, 8, 9N, 9V, 10A, 11A, 12F, 14, 15A, 15B, 15C, 16F, 17F, 18C, 19F, 20, 22F, 23A, 23B, 23F, 31, 33F and 35B are hydrolyzed capsular polysaccharides; 2) The pneumolysin has an amino acid sequence shown as SEQ ID NO. 1; 3) The tetanus toxin C fragment has an amino acid sequence shown as SEQ ID NO. 2; 4) The diphtheria toxin nontoxic mutant has an amino acid sequence shown as SEQ ID NO. 3, and, 5) The mass ratio of capsular polysaccharide to carrier protein in the composition is (0.5-1.5): 1.
  10. 10. Use of a composition according to any one of claims 1 to 6 in the manufacture of a medicament for the prophylaxis of pneumococcal related diseases, preferably the medicament is a vaccine.
  11. 11. A process for preparing the composition of any one of claims 1-6, comprising the steps of: 1) Conjugation of capsular polysaccharides of each serotype with a carrier protein, respectively, to obtain conjugates, and, 2) Mixing the conjugates obtained in step 1) to obtain a composition; Preferably, the capsular polysaccharide in step 1) is conjugated to a carrier protein after activation, said activation being a chemical activation, such as cyanogen bromide, CDAP or periodic acid oxidation, and preferably further comprising a step of hydrolysis, such as acid hydrolysis, thermal hydrolysis or enzymatic hydrolysis, prior to activation of the capsular polysaccharide.
  12. 12. A method of preparing a pneumococcal vaccine, characterized in that the method comprises the step of adding adjuvants to the composition of any of claims 1-6; preferably, the adjuvant is as defined in the vaccine as claimed in any one of claims 7 to 9, and/or the method further comprises a step of sterilization, for example by filtration.

Description

Composition of 31-valent pneumococcal capsular polysaccharide and application thereof The present application claims priority from China patent application 2025106757070 with the application date 2025/5/23. The present application incorporates the entirety of the above-mentioned chinese patent application. Technical Field The invention relates to the field of biological medicine, in particular to a composition of 31-valent pneumococcal capsular polysaccharide and application thereof. Background PneumococcusIs a common bacterium belonging to the genus Streptococcus. It is one of the major pathogenic bacteria causing pneumonia, and can also cause other infections such as otitis media, sinusitis, meningitis, and the like. Pneumococcal serotypes are more than 100 according to capsular polysaccharide serotype classification, some of which are more pathogenic. Such bacteria are commonly present in the nasopharynx of humans and spread by droplets, and pneumococci may invade the lower respiratory tract when the immunity of the human body is lowered, causing pneumonia. Symptoms of pneumococcal pneumonia include high fever, chills, cough, expectoration, chest pain, etc., and in severe cases may lead to respiratory failure. Children, the elderly, immunocompromised persons, and persons suffering from chronic diseases are more susceptible to pneumococci. Currently, there are a variety of conjugate pneumonia vaccines on the market, such as 7-valent, 13-valent, 20-valent, 21-valent, etc. These vaccines are widely used worldwide, significantly reducing the incidence of pneumococcal disease. For example, conjugate 7-valent pneumococcal vaccines have been widely vaccinated and are effective against infections caused by pneumococci of specific serotypes. The conjugate vaccine for the 13-valent pneumonia further expands the protection range, covers more common pathogenic serotypes, is incorporated into immune planning, and plays an important role in public health. The pneumococcal conjugate vaccine on the market has remarkable effect in preventing the streptococcus pneumoniae infection. However, with the widespread use of these vaccines, serum replacement phenomena are increasingly emerging. So-called serum replacement, i.e. vaccination, is effective in controlling the serotype of streptococcus pneumoniae infection to which the vaccine is directed, but the relative increase in infection cases caused by non-vaccine serotype streptococcus pneumoniae, which somewhat detracts from the overall protective efficacy of the existing vaccine. And the prevalence of strains uncovered by existing vaccines is increasing. The market demand for conjugate pneumonic vaccines is moving towards higher prices. Higher order vaccines can theoretically cover more streptococcus pneumoniae serotypes, providing more extensive immunoprotection. However, the increase of the number of the price is not without risk, on one hand, the complexity of the vaccine production process is obviously increased along with the increase of the number of the price, which may lead to the reduction of the quality stability of the product, and on the other hand, the combination of a plurality of antigen components may trigger immune competition, influence the effectiveness of the vaccine, for example, antigens of different serotypes may interfere with each other in the immune response process, lead to the insufficient antibody level generated by an organism aiming at certain serotypes, and the ideal protection effect cannot be achieved. The antigen is increased to possibly bring higher adverse reaction risks, the antigen content is properly increased to improve the antibody level from the efficacy point of view, but to a certain extent, the immunization effect brought by continuously increasing the antigen content is limited, and secondly, the carrier protein content is possibly increased due to the increase of the antigen, so that the carrier is inhibited, the immunogenicity is reduced, the high-concentration stock solution is needed when the semi-finished product is prepared with high antigen content from the process point of view, the viscosity of the high-concentration stock solution is higher, the stability is poorer, and the antigen is improved from the cost point of view, so that the higher cost is meant. In view of the foregoing, there is a strong need in the art for a new generation of high-valent pneumococcal conjugate vaccines that address both the carrier inhibitory effects of multivalent pneumococcal conjugate vaccines, some of the safety issues raised by adjuvants, and the challenges of insufficient immunogenicity of the weak serotypes. Disclosure of Invention In order to solve the technical problems, the invention provides a multi-carrier compatibility strategy, an adjuvant-free formulation strategy and a serotype specific antigen dose adjustment strategy, and provides a multivalent pneumococcal conjugate vaccine. According to the scheme, the optimal carrier p