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WO2025176145A1 - Vaccin contre le papillomavirus humain nonavalent et son utilisation - Google Patents

Vaccin contre le papillomavirus humain nonavalent et son utilisation

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Publication number
WO2025176145A1
WO2025176145A1 PCT/CN2025/078034 CN2025078034W WO2025176145A1 WO 2025176145 A1 WO2025176145 A1 WO 2025176145A1 CN 2025078034 W CN2025078034 W CN 2025078034W WO 2025176145 A1 WO2025176145 A1 WO 2025176145A1
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WIPO (PCT)
Prior art keywords
particle size
vlp
protein
type
antigen
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Pending
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PCT/CN2025/078034
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English (en)
Chinese (zh)
Inventor
刘永江
银飞
于泓洋
高文双
陈晓
伍树明
张海江
姜绪林
高俊
刘玉莹
沈迩萃
张瑞霞
陈丹
王艳
冯慧彬
李玲
薛俊莲
张佳涛
王学红
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Beijing Health Guard Biotechnology Inc
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Beijing Health Guard Biotechnology Inc
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Filing date
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Priority claimed from CN202410202916.9A external-priority patent/CN118059223B/zh
Priority claimed from CN202410202934.7A external-priority patent/CN118059224B/zh
Application filed by Beijing Health Guard Biotechnology Inc filed Critical Beijing Health Guard Biotechnology Inc
Publication of WO2025176145A1 publication Critical patent/WO2025176145A1/fr
Pending legal-status Critical Current
Anticipated expiration legal-status Critical

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K39/12—Viral antigens
    • A61K39/295—Polyvalent viral antigens; Mixtures of viral and bacterial antigens
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K39/39—Medicinal preparations containing antigens or antibodies characterised by the immunostimulating additives, e.g. chemical adjuvants
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/12—Antivirals
    • A61P31/20—Antivirals for DNA viruses

Definitions

  • the present application relates to the field of medicine, specifically to a human papillomavirus vaccine, and in particular to a nine-valent human papillomavirus vaccine and its application.
  • HPV infection Human papillomavirus
  • cervical cancer The relationship between HPV infection and cervical cancer was first proposed in 1974, and it was eventually proved that HPV infection is the main cause of cervical cancer.
  • HPV infection is the most common viral infection of the reproductive tract. Human infection with HPV may not only cause cervical cancer, but also anogenital cancer and genital warts. In addition, the occurrence of oropharyngeal cancer and other head and neck cancers, colon cancer, and rectal cancer is also related to HPV infection.
  • Low-risk HPV mainly includes HPV6/11/30/42/43/44/61, etc., of which 90% of genital warts are caused by HPV6 and 11 infections; the World Health Organization (WHO) International Agency for Research on Cancer (International Agency for Research on Cancer)
  • WHO World Health Organization
  • IARC International Institute of Cancer Research
  • High-risk HPV is carcinogenic and is the cause of almost all cervical cancers. In addition, it also causes 88% of anal cancer, 78% of vaginal cancer, 15% to 48% of vulvar cancer (age-related), 51% of penile cancer and 13% to 60% of oropharyngeal cancer.
  • the beneficial effects of the present invention are as follows: the antigen components in the nine-valent HPV vaccine of the present invention are all derived from the major capsid protein L1 of HPV and are truncated.
  • the truncated L1 protein expressed by the E. coli system improves the yield of the L1 protein and reduces the production cost of the vaccine.
  • FIG. 1 Analysis of neutralizing antibodies against HPV11, HPV16, and HPV18.
  • A-4 represents that each type of antigen is placed at 4°C in prescription A
  • B-4 represents that each type of antigen is placed at 4°C in prescription B
  • C-4 represents that each type of antigen is placed at 4°C in prescription C
  • D-4 represents that each type of antigen is placed at 4°C in prescription D
  • E-4 represents that each type of antigen is placed at 4°C in prescription E
  • A-37-14d represents that each type of antigen is placed at 37°C in prescription A for 14 days
  • B-37-14d represents that each type of antigen is placed at 37°C in prescription B for 14 days
  • C-37-14d represents that each type of antigen is placed at 37°C in prescription C for 14 days
  • D-37-14d represents that each type of antigen is placed at 37°C in prescription D for 14 days
  • E-37-14d represents that each type of antigen is placed at 37°C in prescription E for 14 days.
  • Figure 4 Analysis of immunogenicity of nine-valent HPV vaccines placed at high temperature.
  • 16-4 represents type 16 antigen placed at 4°C
  • 16-37/7d represents type 16 antigen placed at 37°C for 7 days
  • 16-37/14d represents type 16 antigen placed at 37°C for 14 days
  • 18-4 represents type 18 antigen placed at 4°C
  • 18-37/7d represents type 18 antigen placed at 37°C for 7 days
  • 18-37/14d represents type 18 antigen placed at 37°C for 14 days
  • 58-4 represents type 58 antigen placed at 4°C.
  • 58-37/7d means 58 antigen is placed at 37°C for 7 days, 58-37/14d means 58 antigen is placed at 37°C for 14 days; 6-4 means 6 antigen is placed at 4°C, 6-37/7d means 6 antigen is placed at 37°C for 7 days, 6-37/14d means 6 antigen is placed at 37°C for 14 days; 11-4 means 11 antigen is placed at 4°C, 11-37/7d means 11 antigen is placed at 37 °C for 7 days, 11-37/14d means type 11 antigen is placed at 37°C for 14 days; 31-4 means type 31 antigen is placed at 4°C, 31-37/7d means type 31 antigen is placed at 37°C for 7 days, 31-37/14d means type 31 antigen is placed at 37°C for 14 days; 33-4 means type 33 antigen is placed at 4°C, 33-37/7d means type 33 antigen is placed at 37°C for 7 days, 33-37/14d means type 31 antigen is placed at 37°C for
  • the present invention studies and truncates each type of L1 protein based on the wild-type sequence to remove the positively charged portion, thereby obtaining optimized L1 proteins of each type.
  • the specific L1 proteins used in this example are as follows:
  • HPV6L1-VLP proteins with an average particle size of 45-60 nm and a PdI ⁇ 0.1 were obtained.
  • HPV11L1-VLP protein with an average particle size of 45-60 nm and a PdI ⁇ 0.1 was obtained.
  • HPV18L1-VLP protein with an average particle size of 55-75 nm and a PdI ⁇ 0.1 was obtained.
  • HPV45L1-VLP protein with an average particle size of 50-65 nm and a PdI ⁇ 0.1 was obtained.
  • HPV52L1-VLP protein with an average particle size of 50-65 nm and PdI ⁇ 0.1 was obtained.
  • HPV58L1-VLP protein with an average particle size of 45-60 nm and a PdI ⁇ 0.1 was obtained.
  • the nucleotide sequence of the artificially synthesized truncated HPV6L1 is (SEQ ID NO.2).
  • the DNA fragment of HPV6L1 was first amplified by PCR.
  • the L1 gene PCR fragment containing NdeI and Xho1 restriction sites and the recombinant vector pKL1 were double-digested with NdeI/Xho1 and the digested fragments were recovered.
  • T4 DNA ligase was used to ligate the recovered gene fragment with the vector fragment pKL1 containing the corresponding sticky ends. The ligation was carried out at 16°C for 10 to 15 hours.
  • the ligation system is as follows: 6 ⁇ l of the pKL1 vector fragment, 2 ⁇ l of the HPV6 L1 gene fragment, 1 ⁇ l of T4 DNA ligase, and 1 ⁇ l of T4 DNA Ligase buffer. After the ligation reaction, the ligation product is transformed into E. coli DH5 ⁇ for recombinant screening. The selected monoclonal colonies are expanded and the plasmid is extracted, which is then verified by sequencing to obtain the recombinant expression vector pKL1-HPV6L1.
  • the recombinant vectors identified by sequencing were transformed into Escherichia coli BL21 host cells and used as engineered bacteria to express the HPV6L1 protein.
  • the engineered bacteria culture medium was 2YT medium (10 g/L tryptone; 5 g/L yeast extract; 10 g/L NaCl).
  • a single bacterial plaque containing the recombinant plasmid was placed in 10 ml of 2YT medium (containing 100 ⁇ g/ml ampicillin) and cultured overnight at 37°C with shaking at 230 rpm.
  • Affinity chromatography of GST-tagged recombinant protein Load 5 ml of GST agarose affinity chromatography medium into the affinity column, equilibrate the column with buffer L (pH 8.0, 50 mM Tris, 200 mM NaCl, 5 mM DTT), and then load the GST-tagged protein solution. After completion, add 3C enzyme for enzymatic digestion. After enzymatic digestion, wash with Buffer L until no protein flows out, collect the eluate, and the affinity chromatography is complete.
  • buffer L pH 8.0, 50 mM Tris, 200 mM NaCl, 5 mM DTT
  • the HPV6L1 pentamer protein collected in the previous step after affinity chromatography purification is further purified.
  • the HPV6L1 pentamer protein can be collected by ion exchange chromatography first, or further molecular sieve chromatography can be performed directly using Superdex200 (manufactured by GE) molecular sieve gel filtration medium without going through the ion exchange chromatography step.
  • the molecular sieve mobile phase is pH 8.0, 10mM Tris, 100mM NaCl, and the sample corresponding to the ultraviolet absorption peak of the HPV6L1 pentamer protein is collected.
  • the HPV6L1 pentameric protein obtained above self-assembled to form VLPs.
  • the particle size and particle size distribution were measured using a Malvern Zetasizer Nano ZS dynamic light scattering particle size analyzer (the particle size distribution coefficient PdI value is an index of particle size dispersion, less than 0.05 is a highly uniform sample; 0.05-0.1 is a quasi-uniform sample, 0.1-0.3 is a sample with poor uniformity, and greater than 0.3 is an inhomogeneous sample).
  • the HPV6L1 pentameric protein was assembled to obtain VLPs with uniform particle size (PdI ⁇ 0.1).
  • HPV11 L1-VLP, HPV16 L1-VLP, HPV18 L1-VLP, HPV31 L1-VLP, HPV33 L1-VLP, HPV45 L1-VLP, HPV52 L1-VLP, and HPV58 L1-VLP were prepared according to the above method, except that the nucleotide sequence of the synthesized human papillomavirus coat protein L1 was different.
  • Preparation process of aluminum hydroxide adjuvant Aluminum chloride and sodium hydroxide are selected, and Al 3+ and OH - are reacted in a molar ratio of 1: (2-5). No sodium chloride is added to the reaction system. After the reaction at room temperature or 50°C ⁇ 70°C, the temperature is raised to 80°C to maintain the pH at about 6.0-7.0 for 50-70min. After being placed in the tank, the adjuvant is washed with 0.9% sodium chloride solution (resuspended mass ratio ⁇ 6.5:1) for more than 4 times.
  • the washed adjuvant is resuspended with injection water or 0.9% sodium chloride solution according to the feed amount to an aluminum content of 7mg/ml, and then divided into 500ml blue-capped reagent bottles (about 400ml/bottle) and sterilized by high-pressure steam at 121°C for 20min.
  • the aluminum hydroxide adjuvant obtained in this example was fully tested according to the pharmacopoeia method, and the test results were in compliance with the corresponding standard requirements.
  • Adjuvant preparation Prepare a 0.9% sodium chloride solution with water for injection and sterilize by filtration through a 0.22 ⁇ m sterile microporous filter or autoclaving. Each adjuvant was diluted with 0.9% sodium chloride solution to an aluminum content of 2mg/ml. The adjuvant was then further diluted to intermediate concentrations of 400 ⁇ g/ml, 200 ⁇ g/ml, and 100 ⁇ g/ml for the preparation of the respective immunization samples. Each batch of adjuvant dilution was mixed with 0.5ml of 20 ⁇ g/ml and 4 ⁇ g/ml HPV16 L1-VLP protein dilutions at a 1:1 ratio to prepare the final immunization samples. Each group of experimental mice was immunized intramuscularly with 100 ⁇ l per mouse. The immunization results are shown below.
  • the neutralizing antibody level induced by the aluminum hydroxide adjuvant of the present invention is higher than that of the commercially available adjuvant.
  • Neutralizing antibodies are the key to the immune protection effect of HPV vaccines. Therefore, the immune enhancement effect of the aluminum hydroxide adjuvant of the present invention is not lower than that of the commercially available aluminum hydroxide adjuvant.
  • the aluminum hydroxide adjuvant of the present invention also has a good immune enhancement effect at low antigen doses and can also induce a high level of immune response faster in the early stage of immunization.
  • the results of the neutralizing antibody levels of other types at different time points show that the neutralizing antibody level induced by the aluminum hydroxide adjuvant of the present invention is slightly higher than that of the commercially available adjuvant, and has a good immune enhancement effect.
  • the present invention studies the effects of different buffering conditions (see Table 3) on the thermal stability of HPV L1-VLP protein in different buffering conditions of different types of HPV L1-VLP protein stock solutions in acetic acid-sodium acetate buffer system and histidine-hydrochloric acid buffer system.
  • the nine-valent HPV protein stock solution samples were placed under different storage temperatures (4°C, 25°C, 37°C, -80°C (repeated freeze-thaw)) and the particle size and other stability indicators of the sample solution VLP under different conditions were detected.
  • Dynamic light scattering (DLS) was used to detect the particle size, and the detection results of HPV6L1-VLP are shown in Tables 4 and 5.
  • HPLC was used to detect the protein purity, and the detection results of HPV6L1-VLP are shown in Tables 6 and 7.
  • HPV nine-valent L1-VLP protein solutions of various types were matured using a constant temperature water bath at 37°C, and samples were taken at four time points (0h, 12h, 24h, and 36h) for purity and in vitro relative potency testing.
  • the purity of the nine-valent HPV vaccine protein was tested using HPLC, and the test results are shown in Table 12.
  • the relative potency of samples of each type, batch, and maturation time point was determined using a double antibody sandwich (monoclonal antibody) enzyme-linked immunosorbent assay, and the relative potency of each sample is shown in Figure 5.
  • HPV type 11 L1-VLP protein whose relative titer decreased in vitro after maturation, still demonstrated good immunogenicity in mice.
  • the vaccine preparation steps of the present invention are as follows:
  • VLPs The various types of HPV L1 proteins obtained according to the method of Example 1 are purified and self-assembled in vitro to form VLPs.
  • the VLPs are further subjected to chromatography column liquid exchange: the liquid exchange chromatography column is fully equilibrated with a buffer solution (10-30 mM histidine-hydrochloric acid, 400-1500 mM NaCl, 0.001%-0.3% polysorbate 80, pH 5.5-6.5) in advance, and then the assembled VLPs are loaded onto the equilibrated chromatography column. After the loading is completed, the sample is eluted with a buffer solution to collect the components containing the target protein.
  • a buffer solution 10-30 mM histidine-hydrochloric acid, 400-1500 mM NaCl, 0.001%-0.3% polysorbate 80, pH 5.5-6.5
  • HPVL1-VLP protein stock solution The collected components are sterilized and filtered through a sterile filter to obtain an HPVL1-VLP protein stock solution; the obtained HPVL1-VLP protein stock solution is stored below -65°C, or directly matured at 37°C for 12-36 hours to obtain a matured HPVL1-VLP protein stock solution;
  • Example 3 Dilute and mix the aluminum hydroxide adjuvant prepared in Example 2 in the prescribed amount, filter through a capsule filter, and set aside;
  • each type of protein dilution is mixed with an aluminum hydroxide adjuvant dilution in proportion to prepare a monovalent adsorption product, and finally the required amount of each type of monovalent adsorption product is taken and thoroughly mixed to obtain the finished vaccine preparation.
  • the finished product packaging is also included: pre-filled syringe filling machines or vial filling machines are used for packaging. After packaging, the finished vaccine preparations are stored in a cold storage at 2°C ⁇ 8°C for future use.
  • HPV vaccines were formulated with varying antigen dose ratios and aluminum hydroxide adjuvant doses and used to immunize BALB/c mice to investigate immunogenicity. Specifically, a monovalent antigen immunization group was established to investigate immune interference; Gardasil 9 was used as a control vaccine, and different aluminum hydroxide adjuvant dose groups were established to determine the adjuvant dose in the vaccine; and different antigen dose ratios were set to determine the antigen dose ratio for each type. Animal grouping information, antigen dose ratios, and aluminum hydroxide adjuvant dose settings are shown in Table 13.
  • the immunization schedule involved immunizations at weeks 0 and 4, with neutralizing antibody titers measured 4 weeks after the first immunization and 3 weeks after the second immunization. Intergroup comparisons were performed to determine the dose ratio of each antigen type in the vaccine and the dosage of aluminum adjuvant.
  • mice were immunized with the monovalent and nine-valent mixed antigens listed in Table 13 (same dose of antigens of the same type). The differences in immune response levels between the two groups were compared to investigate immune interference. The results showed that the neutralizing antibody titers of all types of antigens prepared by the present invention were significantly higher when immunized in a monovalent form than in the nine-valent HPV vaccine form, and were also greater than those of the corresponding types of the control vaccine (Gardasil 9).
  • the fixed antigen dose was 1/20 ⁇ of the human dose (for example, the human dose is 0.5 ml, and 1/20 of the human dose (0.5 ml) was used to immunize mice).
  • Four aluminum hydroxide adjuvant doses were set: 37.5 ⁇ g (1/20 ⁇ of the nine-valent HPV vaccine), 0 ⁇ g, 25 ⁇ g, and 50 ⁇ g.
  • antigens with different average particle sizes were prepared by adjusting different sodium chloride concentrations and pH values according to the preparation process, and dynamic light scattering (DLS) was used to measure the antigen particle size.
  • DLS dynamic light scattering
  • Each 0.5 ml of HPV6, 11, 16, 18, 31, 33, 45, 52, and 58 antigens contained 30 ⁇ g, 40 ⁇ g, 60 ⁇ g, 40 ⁇ g, 20 ⁇ g, 20 ⁇ g, 20 ⁇ g, 20 ⁇ g, 20 ⁇ g, 20 ⁇ g, and 0.75mg of the above-mentioned aluminum hydroxide adjuvant, and then different nine-valent HPV vaccines were prepared.
  • Gardasil 9 and aluminum hydroxide adjuvant were used as controls, and BALB/c mice were immunized at 1/50 ⁇ human dose to compare the differences in immunogenicity of each nine-valent HPV vaccine.
  • the immunization program used 0 and 4 weeks of immunization, and the neutralizing antibody titer was measured 3 weeks after the second immunization. The results are shown in Table 15. Inter-group comparisons were performed to optimize and control the particle size of each type of antigen in the vaccine.
  • the vaccine of the present invention containing various types of antigens in the selected particle size range can achieve a good immune response level, and the immune response level to types 6, 11, 16, 18, 31, 45 and 58 is higher than that of the marketed Gardasil 9.
  • the present invention finally determined that the key parameters for the preparation of the optimal nine-valent HPV vaccine are as follows:
  • Each 0.5 ml vaccine contains 30 ⁇ g of HPV6, 40 ⁇ g of HPV11, 60 ⁇ g of HPV16, 40 ⁇ g of HPV18, 20 ⁇ g of HPV31, 20 ⁇ g of HPV33, 20 ⁇ g of HPV45, 20 ⁇ g of HPV52, 20 ⁇ g of HPV58 antigens, and 0.75 mg of aluminum hydroxide adjuvant;
  • the particle size of HPV6 antigen is 45-60 nm
  • the particle size of HPV11 antigen is 4
  • the particle size of HPV16 antigen is 45-60nm
  • the particle size of HPV18 antigen is 55-75nm
  • the particle size of HPV31 antigen is 55-75nm
  • the particle size of HPV33 antigen is 45-60nm
  • the particle size of HPV45 antigen is 50-65nm
  • the particle size of HPV52 antigen is 50-65nm
  • the particle size of HPV58 antigen is 45-60nm
  • each 0.5 ml of the finished vaccine formulation contained 30 ⁇ g, 40 ⁇ g, 60 ⁇ g, 40 ⁇ g, 20 ⁇ g, 20 ⁇ g, 20 ⁇ g, 20 ⁇ g, and 20 ⁇ g of HPV6, 11, 16, 18, 31, 33, 45, 52, and 58 antigens, respectively, and 0.75 mg of aluminum hydroxide adjuvant.
  • the buffer solution consisted of 10 mM histidine, 0.01% polysorbate 80, a pH of 5.5-6.5, and a sodium chloride concentration of 154-500 mM.
  • the present invention used three dose groups for immunization: 1/10 ⁇ , 1/20 ⁇ , and 1/40 ⁇ the human dose (0.5 ml).
  • Merck's Gardasil 9 (1/40 ⁇ the human dose), which has the same antigenic composition, was used as a positive control.
  • the present invention determined the immunogenicity and dose effect of the nine-valent HPV vaccine in Wistar rats.
  • the animal grouping, immunization sample preparation, immunization schedule, and immunization route of the Wistar rats are shown in Table 16.
  • Wistar rats were immunized once with the three dose groups of the nine-valent HPV vaccine of the present invention and a positive control group (Gardasil 9).
  • the neutralizing antibody titers measured at each time point are shown in Table 17.
  • Wistar rats were immunized three times (2 weeks apart) with the three dose groups of the nine-valent HPV vaccine and a positive control group (Gardasil 9).
  • the neutralizing antibody titers measured at each time point are shown in Table 18.
  • Example 7 Evaluation of the immunogenicity of the nine-valent HPV vaccine of the present invention in 120 healthy women aged 18-26 years
  • the nine-valent HPV vaccine used in this example was prepared, wherein the nine-valent HPV vaccine contains HPV6, 11, 16, 18, 31, 33, 45, 52, 58 L1-VLP antigens and aluminum hydroxide adjuvant, the amount of each antigen in each 0.5 ml is 20-80 ⁇ g, the amount of aluminum hydroxide adjuvant is 0.75 mg; the particle size of HPV6 antigen is 45-60 nm, the particle size of HPV11 antigen is The particle size of the HPV16 antigen is 45-60 nm, the particle size of the HPV18 antigen is 55-75 nm, the particle size of the HPV31 antigen is 55-75 nm, the particle size of the HPV33 antigen is 45-60 nm, the particle size of the HPV45 antigen is 50-65 nm, the particle size of the HPV52 antigen is 50-65 nm, and the particle size of the HPV58 antigen is 45
  • the particle size of the aluminum hydroxide adjuvant is 3-10 ⁇ m.
  • the buffer comprises: 10 mM histidine, 0.01% polysorbate 80, a pH of 5.5-6.5, and a sodium chloride concentration of 154-500 mM.
  • each group had 40 subjects (including 10 positive controls). After enrollment, the subjects were randomly vaccinated with the test vaccine or the positive control vaccine at a ratio of 3:1. First, the 40 subjects in the low-dose group were sequentially enrolled in four batches of 5, 10, 10 and 15 subjects, and were randomly vaccinated with the first dose of the low-dose test vaccine or the positive control vaccine at a ratio of 3:1. The enrollment interval for each batch was no less than 3 days.
  • the vaccines administered in the low-dose group were: V_220: HPV-6/11/16/18/31/33/45/52/58 (20 ⁇ g, 40 ⁇ g, 40 ⁇ g, 20 ⁇ g, 20 ⁇ g, 20 ⁇ g, 20 ⁇ g, 20 ⁇ g, 20 ⁇ g);
  • the vaccines administered in the medium-dose group were: V_270: HPV-6/11/16/18/31/33/45/52/58 (30 ⁇ g, 40 ⁇ g, 60 ⁇ g, 40 ⁇ g, 20 ⁇ g, 20 ⁇ g, 20 ⁇ g, 20 ⁇ g);
  • the vaccine administered in the high-dose group was: V_360: HPV-6/11/16/18/31/33/45/52/58 (30 ⁇ g, 40 ⁇ g, 80 ⁇ g, 60 ⁇ g, 30 ⁇ g, 30 ⁇ g, 30 ⁇ g, 30 ⁇ g, 30 ⁇ g).
  • the immunization schedule for this trial is three doses, administered at 0, 2, and 6 months.
  • the injection site is the deltoid muscle on the outer side of the upper arm, preferably the deltoid muscle of the non-dominant hand. Vaccination in the buttocks or other areas is prohibited.
  • Immunogenicity results are usually evaluated by calculating the geometric mean titers (GMT), geometric mean intensity (GMI), and antibody positive conversion rates of neutralizing antibodies and IgG antibodies in different groups and their 95% CI.
  • the immunogenicity results in Tables 19 and 20 show that both the binding antibody and neutralizing antibody positivity rates were 100%.
  • the medium-dose HPV18 group had higher binding antibody GMT and GMI than the low-dose, high-dose, and control groups (P ⁇ 0.05)
  • the high-dose HPV58 group had higher binding antibody GMT and GMI than the control group (P ⁇ 0.05).
  • the low-dose and high-dose HPV33 groups had higher neutralizing antibody GMT and GMI than the control group (P ⁇ 0.05)
  • the high-dose HPV52 group had higher neutralizing antibody GMT and GMI than the medium-dose group (P ⁇ 0.05).
  • GMT mean antibody titer calculated for all subjects; 95% CI: 95% confidence interval; N: number of subjects in each group; LL: lower limit; UL: upper limit; RRE: before vaccination; M3: 3 months after vaccination; M7: 7 months after vaccination;
  • V_220 HPV-6/11/16/18/31/33/45/52/58 (20 ⁇ g, 40 ⁇ g, 40 ⁇ g, 20 ⁇ g, 20 ⁇ g, 20 ⁇ g, 20 ⁇ g, 20 ⁇ g, 20 ⁇ g);
  • V_ 270 HPV-6/11/16/18/31/33/45/52/58 (30 ⁇ g, 40 ⁇ g, 60 ⁇ g, 40 ⁇ g, 20 ⁇ g, 20 ⁇ g, 20 ⁇ g, 20 ⁇ g, 20 ⁇ g);
  • Gardas IL 9 Jiadaxiu, HPV-6/11/16/18/31/33/45/52/58 (30 ⁇ g, 40 ⁇ g, 60 ⁇ g, 40 ⁇ g, 20 ⁇ g, 20 ⁇ g, 20 ⁇ g, 20 ⁇ g).

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Abstract

L'invention concerne un vaccin contre le papillomavirus humain nonavalent et son utilisation. Le vaccin contre le papillomavirus humain nonavalent comprend des antigènes L1-VLP de type HPV6, HPV11, HPV16, HPV18, HPV31, HPV33, HPV45, HPV52 et HPV58. Tous les composants antigènes dans le vaccin contre le papillomavirus humain nonavalent fourni sont dérivés de L1, d'une protéine de capside majeure du papillomavirus humain, et tronqués. Le rendement de la protéine L1 est amélioré au moyen d'une protéine L1 tronquée exprimée par un système d'Escherichia coli, ce qui permet de réduire le coût de production du vaccin. L'optimisation de la taille de particule de chaque type d'antigènes VLP et de la taille de particule de l'adjuvant d'hydroxyde d'aluminium, combinée à un contrôle du rapport de chaque type d'antigènes VLP à l'adjuvant et au système tampon et similaire, permet au vaccin contre le papillomavirus humain nonavalent obtenu d'induire une réponse immunitaire de haut niveau, en particulier pour démontrer un effet clinique significatif.
PCT/CN2025/078034 2024-02-23 2025-02-19 Vaccin contre le papillomavirus humain nonavalent et son utilisation Pending WO2025176145A1 (fr)

Applications Claiming Priority (4)

Application Number Priority Date Filing Date Title
CN202410202916.9A CN118059223B (zh) 2024-02-23 2024-02-23 九价人乳头瘤病毒疫苗及其应用
CN202410202934.7 2024-02-23
CN202410202934.7A CN118059224B (zh) 2024-02-23 2024-02-23 九价人乳头瘤病毒疫苗制剂及其应用
CN202410202916.9 2024-02-23

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WO2025176145A1 true WO2025176145A1 (fr) 2025-08-28

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WO2001028585A1 (fr) * 1999-10-15 2001-04-26 Merck & Co., Inc. Obtention de particules a l'aspect viral de papillomavirus presentant des proprietes ameliorees
CN102349996A (zh) * 2011-10-17 2012-02-15 沈阳三生制药有限责任公司 人乳头瘤病毒药物组合物及其应用
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WO2022152204A1 (fr) * 2021-01-14 2022-07-21 神州细胞工程有限公司 Préparation stable de vaccin à particules de type virus du papillomavirus humain
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CN116200416A (zh) * 2023-02-15 2023-06-02 北京康乐卫士生物技术股份有限公司 一种基于Tac启动子的质粒表达载体构建及其用途
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