CN111821432A - Multivalent pneumococcal conjugate vaccine - Google Patents
Multivalent pneumococcal conjugate vaccine Download PDFInfo
- Publication number
- CN111821432A CN111821432A CN202010774340.5A CN202010774340A CN111821432A CN 111821432 A CN111821432 A CN 111821432A CN 202010774340 A CN202010774340 A CN 202010774340A CN 111821432 A CN111821432 A CN 111821432A
- Authority
- CN
- China
- Prior art keywords
- pneumococcal
- conjugate
- polysaccharide
- conjugate vaccine
- prepared
- 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.)
- Granted
Links
Classifications
-
- 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
- A61K39/02—Bacterial antigens
- A61K39/09—Lactobacillales, e.g. aerococcus, enterococcus, lactobacillus, lactococcus, streptococcus
- A61K39/092—Streptococcus
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K39/385—Haptens or antigens, bound to carriers
-
- 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
- A61P31/04—Antibacterial agents
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K2039/54—Medicinal preparations containing antigens or antibodies characterised by the route of administration
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K2039/555—Medicinal preparations containing antigens or antibodies characterised by a specific combination antigen/adjuvant
- A61K2039/55505—Inorganic adjuvants
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K2039/60—Medicinal preparations containing antigens or antibodies characteristics by the carrier linked to the antigen
- A61K2039/6031—Proteins
- A61K2039/6037—Bacterial toxins, e.g. diphteria toxoid [DT], tetanus toxoid [TT]
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K2039/70—Multivalent vaccine
Abstract
The invention provides a multivalent pneumococcal conjugate vaccine. The multivalent pneumococcal conjugate vaccine consists of a liquid injection and a freeze-dried powder, wherein the liquid injection contains 15 pneumococcal serotypes of 1, 2, 3, 4, 5, 6A, 6B, 7F, 9V, 12F, 14, 18C, 19A, 19F and 23F, and conjugates prepared by using diphtheria toxoid as carrier proteins; the freeze-dried powder contains 8 pneumococcal serotypes 8, 10A, 11A, 15B, 22F, 24F, 33F and 35B, and a conjugate prepared by using tetanus toxoid as a carrier protein.
Description
Technical Field
The invention belongs to the field of medical biology, and relates to a pneumonia vaccine which consists of 2 parts, respectively contains 15 types and 8 types of pneumococcal serotypes, is used in a combined way, and is used for preventing infection caused by the contained serotypes of pneumococcus.
Background
Pneumococcus (Streptococcus pneumoniae) is a major pathogenic bacterium causing pneumonia, meningitis, bacteremia and the like in children and old people, and has high morbidity and mortality in the world. Prophylactic formulations (vaccines) are currently available for use in different populations, including pneumococcal polysaccharide vaccines for use in people over 2 years of age and pneumococcal conjugate vaccines that can be used in infants and young children. There is only one pneumococcal polysaccharide vaccine, 23 valent pneumococcal polysaccharide vaccine, including pneumococcal serotypes of 1, 2, 3, 4, 5, 6B, 7F, 8, 9N, 9V, 10A, 11A, 12F, 14, 15B, 17F, 18C, 19F, 19A, 20, 22F, 23F and 33F. The pneumococcal conjugate vaccine comprises 10 valences and 13 valences, wherein the 10 valences pneumococcal conjugate vaccine comprises serotypes of 1, 4, 5, 6B, 7F, 9V, 14, 18C, 19F and 23F, and the 13 valences pneumococcal conjugate vaccine comprises serotypes of 1, 3, 4, 5, 6A, 6B, 7F, 9V, 14, 18C, 19A, 19F and 23F. Pneumococcal conjugate vaccines that have entered the clinical trial internationally and are not yet marketed also include 15-valent and 20-valent pneumococcal conjugate vaccines, wherein the 15-valent pneumococcal conjugate vaccine includes serotypes 1, 3, 4, 5, 6A, 6B, 7F, 9V, 14, 18C, 19A, 19F, 22F, 23F and 33F, and the 20-valent pneumococcal conjugate vaccine includes serotypes 1, 3, 4, 5, 6A, 6B, 7F, 8, 9V, 10A, 11A, 12F, 14, 15B, 18C, 19F, 19A, 22F, 23F and 33F.
According to the situation that the pneumococcus is epidemic serotypes in Asia and Africa regions, the inventor develops a 15-valent pneumococcal conjugate vaccine which comprises serotypes 1, 2, 3, 4, 5, 6A, 6B, 7F, 9V, 12F, 14, 18C, 19A, 19F and 23F, clinical research is carried out by clinical test batches (batch number: 2017L04692) of the national drug administration, and preliminary human serological results show that the proportion of specific antibodies >0.35 mu g/ml is basically more than 90% after the children of 2 and 3 months of age complete basic immunization.
With the use of the pneumonia conjugate vaccine at home and abroad, the serotypes in different regions are respectively obviously changed, the incidence of some non-vaccine serotypes is obviously increased, and the development of the pneumococcal conjugate vaccine with higher price in the world is also in the process of routing and compacting. However, as the price increases, the selection of carrier protein also needs to be considered, and several researches report the carrier inhibition phenomenon of multivalent conjugate vaccines (Zhanna et al, sugar conjugate vaccine and the research progress of immune interference thereof, China medical Innovation, 24 years in 2013, Tan Asia Jun et al, research and application of carrier protein in polysaccharide protein conjugate vaccine, China vaccine and immunization, 04 years in 2013), and the more types are selected, the more carriers need to be selected to avoid the carrier inhibition effect. Based on the original 15-valent pneumococcal serotype, the inventor selects 8 serotypes, 10A, 11A, 15B, 22F, 24F, 33F and 35B to prepare conjugates according to the current situation and trend of the prevalence of the current international and domestic pneumococcal serotypes, and adopts different carrier proteins. The combination of different carrier proteins to produce conjugates that produce effective immune responses requires validation data on animals and humans, and in addition, different conjugation methods and combinations may produce different immune effects. The inventor tests and finds that the specific antibody titer induced by the combined immunization of mice after 8-valent pneumococcal conjugate (tetanus toxoid carrier) is dissolved in the liquid 15-valent pneumococcal conjugate (diphtheria toxoid carrier) is not inferior to that induced by the liquid injection and the freeze-dried powder which are singly immunized, and the individual types are even superior to that induced by the single immunization, which indicates that the product combination prepared in the mode can induce the generation of specific antibodies aiming at each serotype contained in the combination, and the combined use does not interfere with and inhibit the induction of the antibodies.
The inventor adopts the liquid injection and the freeze-dried powder to form the whole of the combined vaccine, and each component unit adopts a carrier protein, so that the uniformity and the stability of the product are kept. Significant immune induction was observed in animal experiments with mixing prior to injection. This combination has the following distinct advantages:
1. increase product stability
The product is a conjugate of polysaccharide and protein, has a flocculent structure, and is easy to agglomerate. The higher the price of the product, the higher the concentration of the binder per unit volume, and the more easily the binder will agglomerate, increasing the risk of product safety. Partial types are freeze-dried and stored, and are dissolved and mixed before use, so that the stability of the product can be effectively improved, and the risk of the product can be controlled.
2. Easy quality control
One development difficulty of multivalent products is the detection of products, the higher the price, the more complex the internal components, the more interference factors during the detection, and the lower the reliability of the detection result. After the product is divided into 2 components, the price times of each component can be effectively controlled, and the non-relevant influence of detection is reduced to a controllable range, so that the establishment and verification of a verification method are facilitated. Meanwhile, only one carrier protein is used for each of the 2 components, the contents of diphtheria toxoid and tetanus toxoid can be measured by a conventional physicochemical formula, and the influence on detection and the trouble of establishing a certain specific detection method caused by mixing a plurality of carrier proteins in the same preparation can be avoided.
3. More flexible to use
The product adopts a combination mode of 1+1, and the liquid injection and the freeze-dried powder can be independently formed into a product, wherein the liquid injection of the inventor is a pneumonia conjugate with a price of 15, has been approved by the national drug administration to carry out clinical tests, and is used for the conventional immunity of people in the future to prevent the most common pneumococcal serotypes; lyophilized powders are additional serotypes, primarily of pneumococcal serotypes that have increased in incidence in recent years. The product combination of the inventor can prevent the infection of 23 pneumococcal serotypes, and the component 2 can also be a supplement of 10-valent and 13-valent pneumococcal conjugate vaccines in the current market and can be injected with the 10-valent or 13-valent pneumococcal conjugate vaccines at the same time so as to expand the immune protection range.
Disclosure of Invention
The invention aims to provide a multivalent pneumococcal conjugate vaccine for preventing infection caused by contained serotype pneumococcus.
The multivalent pneumococcal conjugate vaccine consists of a liquid injection and freeze-dried powder, wherein the liquid injection is mainly prepared from 15 pneumococcal conjugates, and the 15 pneumococcal conjugates are prepared from 1, 2, 3, 4, 5, 6A, 6B, 7F, 9V, 12F, 14, 18C, 19A, 19F and 23F pneumococcal serum and diphtheria toxoid serving as carrier proteins; the freeze-dried powder is mainly prepared from 8 pneumococcal conjugates, wherein the 8 pneumococcal conjugates are 8, 10A, 11A, 15B, 22F, 24F, 33F and 35B pneumococcal sera, and are prepared by using tetanus toxoid as carrier proteins.
Wherein, the liquid injection and the freeze-dried powder are mixed according to the proportion of 1:1 for use.
In the invention, conjugates of each serotype are prepared by chemical coupling of bacterial capsular polysaccharide and carrier protein, and the polysaccharide and the protein can be directly connected or connected through a spacer. Chemical coupling methods for conjugate preparation are well known in the art and include the use of chemical groups already present on the polysaccharide and carrier protein, either directly or via spacer groups; also included are the activation of polysaccharides with hydrogen bromide or 1-cyano-4-dimethylaminopyridine tetrafluoroborate (CDAP) to form cyanate esters, which are coupled to a carrier protein either directly or via a spacer group; also included are the oxidation with sodium periodate to form aldehyde groups, followed by coupling to a carrier protein, either directly or via a spacer group. The pneumococcal capsular polysaccharide for preparing the conjugate can be purified original polysaccharide, and also can be polysaccharide fragments degraded by physical and/or chemical means, wherein the physical degradation means comprises ultrasonic, microwave, reduced pressure and the like, and the chemical degradation means comprises acid hydrolysis, alkali hydrolysis, oxidation hydrolysis and the like.
The liquid injection also contains an adjuvant, wherein the adjuvant is an aluminum adjuvant, and the aluminum adjuvant is an aluminum phosphate adjuvant.
The freeze-dried powder also contains an excipient which is lactose.
In the unit dose of vaccine, the content of each type of capsular polysaccharide is 1, 2, 3, 4, 5, 6A, 6B, 7F, 9V, 12F, 14, 18C, 19A, 19F and 23F respectively at 4 μ g/ml, and 6B at 8 μ g/ml.
In the unit dose of vaccine, the content of each type of capsular polysaccharide is respectively 4 mug/ml for 8, 10A, 11A, 15B, 22F, 24F, 33F and 35B.
Another object of the present invention is to provide a method for preparing a multivalent pneumococcal conjugate vaccine.
The multivalent pneumococcal conjugate vaccine consists of two parts of liquid injection and freeze-dried powder,
the preparation method of the liquid injection comprises the following steps:
mixing 15 pneumococcal conjugate stock solutions (1, 2, 3, 4, 5, 6A, 6B, 7F, 9V, 12F, 14, 18C, 19A, 19F and 23F) according to a certain proportion, adsorbing with a proper amount of aluminum phosphate adjuvant, preparing into semi-finished products with the content of various polysaccharides of pneumococcus of 4 mu g/ml (8 mu g/ml for 6B) and the content of aluminum ions of not higher than 0.4mg/ml, and subpackaging with 0.5 ml/branch to obtain the pneumococcus conjugate.
The preparation method of the freeze-dried powder comprises the following steps:
mixing 8 pneumococcal conjugate stock solutions (8, 10A, 11A, 15B, 22F, 24F, 33F and 35B) at a certain ratio until the content of each polysaccharide is 4 μ g/ml, adding lactose to final concentration of 0.5%, packaging with 0.5 ml/piece, and lyophilizing.
Shaking the liquid injection evenly, sucking and injecting the liquid injection into freeze-dried powder, fully dissolving and shaking evenly to obtain the vaccine of the invention.
In 15 kinds of pneumococcal conjugates, amine reduction method is selected for preparing each type of conjugate, pneumococcal envelope polysaccharide is oxidized by sodium periodate to form aldehyde group, and then the aldehyde group is coupled to carrier protein directly or via spacer group, and the carrier protein is diphtheria toxoid.
Among 8 pneumococcal conjugates, each conjugate is prepared by selective cyanation reaction, wherein pneumococcal envelope polysaccharide is activated by cyanogen bromide or 1-cyano-4-dimethylaminopyridine tetrafluoroborate to form cyanate, and then coupled to a carrier protein directly or via a spacer group, wherein the carrier protein is tetanus toxoid.
In the various types of combination substances, the content of each combination substance is 1-5 mu g/agent. In the conjugate combination, each dose (1 human dose) contains 0.1 to 50 μ g of each polysaccharide, which amount may vary depending on the pneumococcal serotype, and generally, each vaccine dose contains 0.1 to 50 μ g, preferably 0.5 to 10 μ g, more preferably 1 to 5 μ g of each polysaccharide.
The multivalent pneumococcal conjugate vaccine can be inoculated by adopting the conventional routes in the vaccine field, and comprises a mucosal route and an parenteral route, wherein the mucosal route comprises oral administration, nasal spray, eye drop, anus and/or vaginal administration, the parenteral route comprises intramuscular injection, subcutaneous injection, intradermal injection and microneedle patch administration, and the vaccination routes can be used singly or in combination. The route of administration is preferably oral, nasal spray, intramuscular, subcutaneous, most preferably intramuscular. Compared with the existing vaccine, the multivalent pneumococcal conjugate vaccine of the invention better conforms to the protection spectrum required by the pneumococcus to cause diseases at present. Compared with the 23-valent pneumococcal polysaccharide vaccine which is already marketed, the 23-valent pneumococcal conjugate vaccine of the invention eliminates 3 serotypes 9N, 17F and 20 which are basically free of cases or have cross protection at present, adds 3 new serotypes of 6A, 24F and 35B which have high morbidity and serious drug resistance (Everyn B, et al. Serial distribution of Streptococcus pneumoniae) in which new serotype is in the same as PCV. A. systematic new derivatives and strain one.2017,12(5) e 0177113; Stephanie WL, et al. pneumococcal. 76associated with the serum replacement and antigenic replacement, and has wide range of PCV. 9-serotype conjugate of PCV. 9, 9-serotype polysaccharide conjugate of PCV. 9, 9-serotype, 9. 12, 9. PCV. 9, and the protective agent is more fit with the current dominant epidemic bacteria of pneumococcus, and the expected protective effect is better. Compared with the marketed 23-valent pneumococcal polysaccharide vaccine, the combined vaccine can be used for children under 2 years old, and enlarges immune population, particularly the population with highest incidence and greatest harm severity.
The following explains the english terms appearing in the specification:
ADH: adipic acid dihydrazide
EDAC: carbodiimides
CDAP: 1-cyano-4-dimethylamino-pyridine tetrafluoroboric acid
Sepharose 4 FF: agarose gel 4FF
V0: volume of external water
Detailed Description
In order that the invention may be more clearly understood, it will now be further described with reference to the following examples. The examples are for illustration only and do not limit the invention in any way.
Example 1: reduced amine method for conjugate preparation:
dissolving pneumococcal polysaccharide, hydrolyzing with acetic acid solution, adjusting pH to 6.4, adding sodium periodate for oxidation, stopping reaction with glycerol, dialyzing or ultrafiltering to remove small molecular substances to obtain oxidized polysaccharide. ADH and sodium borocyanide were added to the oxidized polysaccharide to final concentrations of 40mg/ml and 2mg/ml, and the reaction was carried out at room temperature for 3 days to prepare a derivative. And (3) mixing the derivative polysaccharide and diphtheria toxoid according to a certain mass ratio, adding EDAC to the final concentration of 0.02mol/L, maintaining the pH value to be 5.6, and reacting for more than 150 minutes to prepare the conjugate. The conjugate was purified by Sepharose 4FF chromatography, and the peak eluted at around V0 was collected to obtain a conjugate stock solution. 1. 2, 3, 4, 5, 6A, 6B, 7F, 9V, 12F, 14, 18C and 23F were prepared using the reduced amine method.
Example 2, the following: cyanation reactions to prepare conjugates
Dissolving pneumococcal polysaccharide, adjusting pH value to about 9.0, adding CDAP acetonitrile solution (concentration is 100mg/ml) according to the mass ratio of 1: 0.75, maintaining the pH value to be 9.00-10.00 for reaction, adding 2% triethylamine solution into the same volume with CDAP, the pH value is 10.00-11.00 for reaction for 5min, adjusting the pH value to about 8.60, adding tetanus toxoid according to the proper mass ratio of multiple glycoproteins, maintaining the pH value to be about 8.6 for reaction for more than 120 min, terminating the reaction by glycine, and obtaining polysaccharide conjugate after ultrafiltration or dialysis. The conjugate was purified by Sepharose 4FF chromatography, and the peak eluted at around V0 was collected to obtain a conjugate stock solution. 8. 10A, 11A, 15B, 19A, 19F, 22F, 24F, 33F and 35B were prepared using a cyanation reaction.
EXAMPLE 3 preparation of liquid injection
Mixing 15 pneumococcal conjugate stock solutions (1, 2, 3, 4, 5, 6A, 6B, 7F, 9V, 12F, 14, 18C, 19A, 19F and 23F) according to a certain proportion, adsorbing with a proper amount of aluminum phosphate adjuvant, preparing into semi-finished products with the content of various polysaccharides of pneumococcus of 4 mu g/ml (8 mu g/ml for 6B) and the content of aluminum ions of not higher than 0.4mg/ml, and subpackaging with 0.5 ml/branch to obtain the pneumococcus conjugate.
Example 4, formulation of lyophilized powder:
mixing 8 pneumococcal conjugate stock solutions (8, 10A, 11A, 15B, 22F, 24F, 33F and 35B) at a certain ratio until the content of each polysaccharide is 4 μ g/ml, adding lactose to final concentration of 0.5%, packaging with 0.5 ml/piece, and lyophilizing.
EXAMPLE 5 formulation of combination vaccine
Shaking the liquid injection in the example 3 uniformly, sucking and injecting the liquid injection into the freeze-dried powder in the example 4, fully dissolving and shaking uniformly to obtain the combined vaccine of the liquid injection and the freeze-dried powder.
Example 6 immunogenicity assessment:
the immunogenicity of the combined vaccine is evaluated by adopting the combined vaccine of 'liquid injection', 'freeze-dried powder' and 'liquid injection + freeze-dried powder' and taking a mouse as an animal model. The immunization was divided into 4 groups, and a physiological saline control group was provided in addition to the above 3 groups.
NIH mice were immunized subcutaneously with 2 needles at 2 week intervals at a dose of 0.1 ml/mouse. Blood was collected from the orbit 28 days after completion of the immunization, and the serum was isolated and the antibody titers of 23 specific sera were determined by ELISA. The results show that the IgG antibody levels of other types of mouse after the combined vaccine of the 'liquid injection and the freeze-dried powder' are not different from those of the 'liquid injection' and the 'freeze-dried powder' except 3, 5, 9V and 22F, while the antibody titers of 3, 5, 9V and 22F after the combined vaccine of the 'liquid injection and the freeze-dried powder' is obviously superior to those of the respective vaccine, which indicates that animals immunized in the form of the combined vaccine of the 'liquid injection and the freeze-dried powder' do not generate immune interference, each type of antibody can normally respond, and the individual type of antibody also shows an immune enhancement effect.
Claims (10)
1. A multivalent pneumococcal conjugate vaccine is characterized by comprising a liquid injection and freeze-dried powder, wherein the liquid injection is mainly prepared from 15 pneumococcal conjugates, the 15 pneumococcal conjugates are serotype 1, 2, 3, 4, 5, 6A, 6B, 7F, 9V, 12F, 14, 18C, 19A, 19F and 23F pneumococcal capsular polysaccharide, and are prepared by taking diphtheria toxoid as carrier proteins; the freeze-dried powder is mainly prepared from 8 pneumococcal conjugates, wherein the 8 pneumococcal conjugates are 8, 10A, 11A, 15B, 22F, 24F, 33F and 35B serotype pneumococcal capsular polysaccharides, and are prepared by taking tetanus toxoid as carrier proteins.
2. The multivalent pneumococcal conjugate vaccine of claim 1, wherein 15 pneumococcal conjugates, each type of conjugate is prepared by an amine reduction method, wherein pneumococcal capsular polysaccharide is oxidized with sodium periodate to form aldehyde groups, and then coupled to a carrier protein, which is diphtheria toxoid, either directly or via spacer groups.
3. The multivalent pneumococcal conjugate vaccine of claim 1, wherein 8 pneumococcal conjugates are prepared by a cyanation reaction selected from the group consisting of cyanogen bromide and 1-cyano-4-dimethylaminopyridine tetrafluoroborate to form a cyanate ester, which is coupled to a carrier protein, either directly or via a spacer group, wherein the carrier protein is tetanus toxoid.
4. The multivalent pneumococcal conjugate vaccine of claim 1, wherein the liquid injection further comprises an adjuvant, wherein the adjuvant is an aluminum adjuvant, and the aluminum adjuvant is an aluminum phosphate adjuvant.
5. The multivalent pneumococcal conjugate vaccine of claim 1, wherein the lyophilized powder further comprises an excipient, wherein the excipient is lactose.
6. The multivalent pneumococcal conjugate vaccine of claim 1, wherein each dose comprises 1 to 5 μ g of each polysaccharide.
7. The multivalent pneumococcal conjugate vaccine of claim 2, wherein 15 pneumococcal conjugates are prepared: dissolving pneumococcal polysaccharide, hydrolyzing by using acetic acid solution, adjusting the pH value to 6.4, adding sodium periodate for oxidation, stopping reaction by using glycerol, dialyzing or ultrafiltering to remove small molecular substances to obtain oxidized polysaccharide, adding ADH and sodium borocyanide into the oxidized polysaccharide to the final concentration of 40mg/ml and 2mg/ml, reacting at room temperature for 3 days to prepare derivatives, mixing the derivatized polysaccharide and diphtheria toxoid according to a certain mass ratio, adding EDAC to the final concentration of 0.02mol/L, maintaining the pH value to 5.6, reacting for more than 150 minutes to prepare a conjugate, purifying the conjugate by Sepharose 4FF chromatography, and collecting an elution peak near V0 to obtain a conjugate stock solution.
8. The multivalent pneumococcal conjugate vaccine of claim 3, wherein 8 pneumococcal conjugates are prepared: dissolving pneumococcal polysaccharide, adjusting the pH value to about 9.0, adding a CDAP acetonitrile solution according to the mass ratio of 1: 0.75, maintaining the pH value at 9.00-10.00 for reaction, adding a 2% triethylamine solution into the mixture with the same volume as the CDAP, the pH value at 10.00-11.00 for reaction for 5min, adjusting the pH value to about 8.60, adding tetanus toxoid according to the mass ratio of proper multiple glycoproteins, maintaining the pH value at about 8.6 for reaction for more than 120 min, stopping the reaction with glycine, obtaining a polysaccharide conjugate after ultrafiltration or dialysis, purifying the conjugate by sepharose 4FF chromatography, and collecting an elution peak near V0 to obtain a conjugate stock solution.
9. The method for preparing the multivalent pneumococcal conjugate vaccine of claim 1, wherein the lyophilized powder is dissolved with a liquid injection and then used in combination.
10. Use of a multivalent pneumococcal conjugate vaccine according to claim 1 in the manufacture of a medicament for the prevention and/or treatment of infection caused by pneumococci.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010774340.5A CN111821432B (en) | 2020-08-05 | 2020-08-05 | Multivalent pneumococcal conjugate vaccine |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010774340.5A CN111821432B (en) | 2020-08-05 | 2020-08-05 | Multivalent pneumococcal conjugate vaccine |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN111821432A true CN111821432A (en) | 2020-10-27 |
| CN111821432B CN111821432B (en) | 2022-10-18 |
Family
ID=72919443
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202010774340.5A Active CN111821432B (en) | 2020-08-05 | 2020-08-05 | Multivalent pneumococcal conjugate vaccine |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN111821432B (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US11116828B2 (en) | 2017-12-06 | 2021-09-14 | Merck Sharp & Dohme Corp. | Compositions comprising Streptococcus pneumoniae polysaccharide-protein conjugates and methods of use thereof |
| US11642406B2 (en) | 2018-12-19 | 2023-05-09 | Merck Sharp & Dohme Llc | Compositions comprising Streptococcus pneumoniae polysaccharide-protein conjugates and methods of use thereof |
| GB2614916A (en) * | 2022-01-25 | 2023-07-26 | Optivalent Ltd | Intradermal vaccine complement |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20030180316A1 (en) * | 2000-06-29 | 2003-09-25 | Dominique Boutriau | Multivalent vaccine composition |
| CN104383531A (en) * | 2014-11-18 | 2015-03-04 | 北京智飞绿竹生物制药有限公司 | Combined vaccine for preventing meningitis in children and preparation method of combined vaccine |
| WO2015144031A1 (en) * | 2014-03-26 | 2015-10-01 | 天津康希诺生物技术有限公司 | Pneumococcus polysaccharide protein conjugated vaccine and preparation method therefor |
| CN105999254A (en) * | 2016-06-27 | 2016-10-12 | 北京智飞绿竹生物制药有限公司 | 15-valence pneumococcus conjugate combined vaccine containing 2-type and 12 F serotypes |
| US20200030439A1 (en) * | 2017-06-10 | 2020-01-30 | Inventprise, Llc | Mixtures of Polysaccharide-Protein Pegylated Compounds |
-
2020
- 2020-08-05 CN CN202010774340.5A patent/CN111821432B/en active Active
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20030180316A1 (en) * | 2000-06-29 | 2003-09-25 | Dominique Boutriau | Multivalent vaccine composition |
| WO2015144031A1 (en) * | 2014-03-26 | 2015-10-01 | 天津康希诺生物技术有限公司 | Pneumococcus polysaccharide protein conjugated vaccine and preparation method therefor |
| CN104383531A (en) * | 2014-11-18 | 2015-03-04 | 北京智飞绿竹生物制药有限公司 | Combined vaccine for preventing meningitis in children and preparation method of combined vaccine |
| CN105999254A (en) * | 2016-06-27 | 2016-10-12 | 北京智飞绿竹生物制药有限公司 | 15-valence pneumococcus conjugate combined vaccine containing 2-type and 12 F serotypes |
| US20200030439A1 (en) * | 2017-06-10 | 2020-01-30 | Inventprise, Llc | Mixtures of Polysaccharide-Protein Pegylated Compounds |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US11116828B2 (en) | 2017-12-06 | 2021-09-14 | Merck Sharp & Dohme Corp. | Compositions comprising Streptococcus pneumoniae polysaccharide-protein conjugates and methods of use thereof |
| US11850278B2 (en) | 2017-12-06 | 2023-12-26 | Merck Sharp & Dohme Llc | Compositions comprising Streptococcus pneumoniae polysaccharide-protein conjugates and methods of use thereof |
| US11642406B2 (en) | 2018-12-19 | 2023-05-09 | Merck Sharp & Dohme Llc | Compositions comprising Streptococcus pneumoniae polysaccharide-protein conjugates and methods of use thereof |
| GB2614916A (en) * | 2022-01-25 | 2023-07-26 | Optivalent Ltd | Intradermal vaccine complement |
Also Published As
| Publication number | Publication date |
|---|---|
| CN111821432B (en) | 2022-10-18 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN111821432B (en) | Multivalent pneumococcal conjugate vaccine | |
| CN110859957B (en) | Immunogenic compositions comprising conjugated capsular saccharide antigens and uses thereof | |
| EP3313436B1 (en) | Multivalent pneumococcal conjugate vaccine | |
| JP2023036953A (en) | Enhancing immunogenicity of streptococcus pneumoniae polysaccharide-protein conjugates | |
| KR101538535B1 (en) | 15-valent pneumococcal polysaccharide-protein conjugate vaccine composition | |
| JP7030235B2 (en) | Polyvalent pneumococcal polysaccharide-protein complex composition | |
| EP1296715B2 (en) | Multivalent vaccine composition | |
| CN109862908A (en) | Multivalent pneumococcal polysaccharide-protein conjugate composition | |
| CZ20013379A3 (en) | Vaccine | |
| CN108079286B (en) | 13-valent pneumococcal polysaccharide-protein conjugate composition and preparation method and application thereof | |
| WO2015144031A1 (en) | Pneumococcus polysaccharide protein conjugated vaccine and preparation method therefor | |
| JP2021512870A (en) | Polyvalent pneumococcal polysaccharide-protein complex composition | |
| EA012214B1 (en) | Immunogenic composition | |
| KR20140130567A (en) | Multivalent pneumococcal polysaccharide-protein conjugate composition | |
| BRPI0620418A2 (en) | method for immunizing a human patient against a disease, uses at least two and at least seven, ten, eleven, thirteen or fourteen conjugates and vaccines, and | |
| CN109890415A (en) | Multivalent pneumococcal polysaccharide-protein conjugate composition | |
| CN110251667A (en) | A kind of immune combination preparation and its preparation method and application | |
| CN110652585B (en) | Polysaccharide-protein conjugate immune preparation and application thereof | |
| US20240139303A1 (en) | Enhancing immunogenicity of streptococcus pneumoniae polysaccharide-protein conjugates | |
| EP4285925A1 (en) | Immunogenic composition comprising multivalent streptococcus pneumoniae polysaccharide-protein conjugates | |
| CN117919397A (en) | Pneumococcal conjugate combined vaccine and preparation method thereof | |
| CN115006522A (en) | Preparation method of bivalent dysentery conjugate combined vaccine | |
| CN116898960A (en) | Bacterial polysaccharide protein conjugates and uses thereof |
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 | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |