CN112245394A - Vaccine protective agent - Google Patents

Vaccine protective agent Download PDF

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CN112245394A
CN112245394A CN202011135455.6A CN202011135455A CN112245394A CN 112245394 A CN112245394 A CN 112245394A CN 202011135455 A CN202011135455 A CN 202011135455A CN 112245394 A CN112245394 A CN 112245394A
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vaccine
virus
protective agent
rabies
parts
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CN112245394B (en
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梁智杰
黄林
崔利凯
陈坤
黄磊
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Boaovax Biotechnology Co ltd
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/14—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles
    • A61K9/19—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles lyophilised, i.e. freeze-dried, solutions or dispersions
    • 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
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
    • A61K47/16—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite containing nitrogen, e.g. nitro-, nitroso-, azo-compounds, nitriles, cyanates
    • A61K47/18—Amines; Amides; Ureas; Quaternary ammonium compounds; Amino acids; Oligopeptides having up to five amino acids
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
    • A61K47/26—Carbohydrates, e.g. sugar alcohols, amino sugars, nucleic acids, mono-, di- or oligo-saccharides; Derivatives thereof, e.g. polysorbates, sorbitan fatty acid esters or glycyrrhizin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/30—Macromolecular organic or inorganic compounds, e.g. inorganic polyphosphates
    • A61K47/36—Polysaccharides; Derivatives thereof, e.g. gums, starch, alginate, dextrin, hyaluronic acid, chitosan, inulin, agar or pectin
    • 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
    • A61P31/14—Antivirals for RNA viruses
    • 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
    • A61K2039/51—Medicinal preparations containing antigens or antibodies comprising whole cells, viruses or DNA/RNA
    • A61K2039/525—Virus
    • A61K2039/5252—Virus inactivated (killed)
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2760/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA ssRNA viruses negative-sense
    • C12N2760/00011—Details
    • C12N2760/20011—Rhabdoviridae
    • C12N2760/20111—Lyssavirus, e.g. rabies virus
    • C12N2760/20134—Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein

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  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Veterinary Medicine (AREA)
  • Public Health (AREA)
  • General Health & Medical Sciences (AREA)
  • Animal Behavior & Ethology (AREA)
  • Medicinal Chemistry (AREA)
  • Epidemiology (AREA)
  • General Chemical & Material Sciences (AREA)
  • Virology (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Engineering & Computer Science (AREA)
  • Molecular Biology (AREA)
  • Oil, Petroleum & Natural Gas (AREA)
  • Mycology (AREA)
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  • Microbiology (AREA)
  • Biochemistry (AREA)
  • Inorganic Chemistry (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Communicable Diseases (AREA)
  • Immunology (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Organic Chemistry (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
  • Medicinal Preparation (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)

Abstract

The invention discloses a vaccine protective agent, which consists of the following components: 2.5-5% of sucrose, 2.5-5% of maltose, 0.3-0.5% of urea, 1-3% of dextran 40, and the balance of phosphate buffer solution with NaCl content of 0.2-0.9%. The protective agent can effectively improve the heat stability of the vaccine, reduce the loss of freeze-drying activity, and reduce the cost without adding trehalose.

Description

Vaccine protective agent
Technical Field
The invention belongs to the field of biological products.
Background
Inactivated virus vaccines are the main type of rabies vaccines at present, and the production mainly comprises: in vitro culture and propagation of rabies virus, inactivation of virus, purification of virus, addition of vaccine protective agent, freeze-drying and the like.
The vaccine protective agent plays an indispensable role in protecting the activity of the vaccine in the storage of the virus liquid before freeze-drying, the freeze-drying process and the storage stage after freeze-drying.
The optimum formula of the freeze-drying protective agent is determined to be 4% of trehalose, 3% of dextran 40, 1% of sucrose, 0.5% of mannitol and 2% of human serum albumin by changing the contents of 3 main components of protective agents such as trehalose, dextran 40 and human serum albumin, so that the titer of a rabies vaccine semi-finished product in the freeze-drying process is averagely reduced by 7.8%, the heat stability efficacy is reduced by 15.6%, and the freeze-dried product is not greatly reduced within 12 months. And considered as follows: trehalose can reduce the harmful effect of the freeze-drying process on virus envelopes, and hydroxyl groups of trehalose and polar groups in protein form hydrogen bonds to replace water molecules around the polar groups of the protein and in phospholipid, so that a hydrated film is formed around the protein, the high-level structure of the protein is stabilized, the protein is prevented from being denatured due to freeze-drying, and the structural and functional integrity of the vaccine is still maintained under the conditions of low-temperature freezing and drying dehydration (screening of freeze-drying protective agents for rabies vaccines (Vero cells) for freeze-drying, volume 30, 3 rd of 2017, 3 th month, volume 30, 3 rd of China journal of biological products).
However, trehalose is costly and requires higher concentrations to be effective, thus increasing the cost of vaccine production.
Disclosure of Invention
The purpose of the invention is: provides a novel rabies vaccine protective agent without trehalose.
The technical scheme of the invention is as follows:
a vaccine protectant, the protectant comprising:
2.5 to 5 parts of cane sugar, 2.5 to 5 parts of maltose, 0.3 to 0.5 part of urea and 1 to 3 parts of dextran 40.
The protective agent as described above, which consists of the following components:
2.5 parts of cane sugar, 2.5 percent of maltose, 0.3 part of urea and 3 parts of dextran 40.
The application of the vaccine protective agent in preparing rabies vaccines.
As the application is described, the rabies vaccine is a freeze-dried rabies vaccine for human.
A preparation method of an inactivated vaccine freeze-dried powder preparation comprises the following steps:
(1) dissolving the vaccine protective agent in a phosphate buffer solution with the NaCl content of 0.2-0.9% (w/v), and adding the inactivated and purified virus solution to ensure that the final concentration of sucrose in the obtained mixture is 2.5-5% (m/v);
(2) and (5) freeze drying.
As in the above-mentioned preparation method, in step (1), the resulting mixture had a final sucrose concentration of 2.5% (m/v), a final maltose concentration of 2.5% (m/v), a final urea concentration of 0.3% (m/v), and a final dextran 40 concentration of 3% (m/v).
The virus is produced by using Vero cells cultured in a serum-free medium as a host, as described above.
The preparation method as described above, wherein the virus is rabies virus.
The virus is rabies virus fixed virus rPV-2061 strain.
The invention also provides the inactivated vaccine freeze-dried powder preparation prepared by the method.
The invention has the beneficial effects that:
the vaccine protective agent disclosed by the invention is simple in component, can effectively protect the titer of the vaccine without trehalose, and the titer after freeze-drying is almost unchanged compared with the titer before freeze-drying after the protective agent disclosed by the invention is added.
Obviously, many modifications, substitutions, and variations are possible in light of the above teachings of the invention, without departing from the basic technical spirit of the invention, as defined by the following claims.
The present invention will be described in further detail with reference to the following examples. This should not be understood as limiting the scope of the above-described subject matter of the present invention to the following examples. All the technologies realized based on the above contents of the present invention belong to the scope of the present invention.
Detailed Description
Note: the percentage contents are all mass to volume (1g/100 ml-1%) without specific reference. Example 1 serum-free human rabies vaccine protective agent of the invention
The formula of the vaccine protective agent is as follows: 2.5 parts of cane sugar, 2.5 parts of maltose, 0.3 part of urea and 3 parts of dextran 40.
When in use, the vaccine protective agent is dissolved in a phosphate buffer solution with the NaCl content of 0.2-0.9% (m/v), and the purified inactivated virus solution is added to ensure that the final concentration of sucrose, maltose, urea and dextran 40 in the mixture is 2.5% (m/v), 0.3% (m/v) and 3% (m/v).
Example 2 serum-free human rabies vaccine protective agent of the invention
The formula of the vaccine protective agent is as follows: 5 parts by weight of sucrose, 5 parts by weight of maltose, 0.5 part by weight of urea and 1 part by weight of dextran 40.
When in use, the vaccine protective agent is dissolved in a phosphate buffer solution with the NaCl content of 0.2-0.9% (m/v), and the purified inactivated virus solution is added to ensure that the final concentration of sucrose, maltose, urea and dextran 40 in the mixture is 5% (m/v), 0.5% (m/v) and 1% (m/v).
The present invention is further illustrated below in the form of experimental examples.
Experimental example 1 comparison of Effect of rabies vaccine protective agent
Preparation of inactivated virus liquid
1. Production cell
Production cells were Vero cells cultured in serum-free medium (serum-free Vero cells for short) and were introduced from ATCC in the USA.
2. Poison seed
The production seed virus is rabies virus fixed virus rPV-2061 strain, introduced from the U.S. CDC.
3. Stock preparation
(1) Cell culture: and (3) after the serum-free Vero cells are recovered, culturing at 37 ℃, and carrying out continuous passage amplification according to the seed separation rate of 1: 3. After expansion, the cells were made into cell suspensions at 6X 105cells/ml were inoculated into a fixed bed bioreactor (abbreviated as "bioreactor", hereinafter the same) and perfusion culture was carried out using a serum-free culture medium. Culture parameters: the temperature was 37.0 ℃, pH 7.20, dissolved oxygen 60%, and rotation speed 100 rpm. The serum-free culture solution comprises the following formula:
Figure BDA0002736276430000031
(2) and (3) virus culture: when the cell density in the bioreactor reaches 0.7X 107Inoculating rabies virus rPV-2061 with MOI of 0.005 at cells/ml. And (5) performing perfusion culture by using a virus maintenance solution 6-8 hours after virus inoculation. Culture parameters: the temperature is 34 ℃, the pH is 7.50, the dissolved oxygen is 60%, and the rotation speed is 100 rpm. The virus maintenance liquid comprises the following formula:
Figure BDA0002736276430000032
Figure BDA0002736276430000041
(3) and (3) toxin collection: virus fluid was harvested starting on day 3 after virus inoculation and continuously harvested for 15 days. The virus titer of the single virus harvest liquid can reach 6.5-7.5 lgLD50/ml。
(4) Concentration: the virus harvest liquid is roughly filtered by a filter element with the diameter of 0.65 mu m, and is concentrated by 20-30 times by adopting a 300kD membrane pack.
(5) Inactivation: adding beta-propiolactone into the concentrated virus harvest liquid according to the proportion of 1:4000, placing the mixture at the temperature of 2-8 ℃ for 24 hours to inactivate rabies viruses; the mixture was left at 37 ℃ for 2 hours to hydrolyze beta-propiolactone.
(6) And (3) purification: 140U/ml nuclease (non-restriction endonuclease) was added and the digestion was carried out at 37 ℃ for 24 hours. Sepharose 4FF was selected as the packing for molecular sieve chromatography purification with phosphate buffer pH 7.6 as the mobile phase and the loading volume was not more than 5% of the column volume.
Thus obtaining the purified inactivated virus liquid.
Secondly, adding a vaccine protective agent
Vaccine protectors were prepared according to examples 1 and 2, respectively, and a control vaccine protector consisting of 5 parts by weight of sucrose and 1 part by weight of dextran 40 was prepared.
Dissolving different vaccine protective agents in phosphate buffer solution with NaCl content of 0.2% (m/v), adding purified inactivated virus solution to obtain semi-finished vaccine, wherein the final concentration of each component is shown in table 1, and each group has 3 repeats.
TABLE 1 Final concentration of the vaccine protective agent Components in the vaccine intermediate product
Group of Sucrose Maltose Urea Dextran 40
A (example 1) 2.5％ 2.5％ 0.3％ 3％
B (example 2) 5％ 5％ 0.5％ 1％
C (control group) 5％ — — 1％
Third, titer change test before and after freeze-drying
The results of the potency changes before and after lyophilization are shown in table 2. It can be seen that the potency variation before and after lyophilization of A, B groups representing the vaccine protector of the present invention is significantly less than that of group C. The variation range is not higher than that of a vaccine protective agent prepared by trehalose formula such as Lichuyan (screening of freeze-dried rabies vaccine (Vero cells) for freeze-dried human, and 3 rd stage of volume 30 in 3 months and 3 months in 2017 in China biological products journal).
TABLE 2 comparison of potency before and after lyophilization
Figure BDA0002736276430000042
Figure BDA0002736276430000051
In conclusion, the vaccine protective agent has simple components and low cost, and has outstanding protective effect on the activity of the vaccine in the vaccine freeze-drying process.

Claims (10)

1. A vaccine protection agent characterized by: the protective agent consists of the following components:
2.5 to 5 parts of cane sugar, 2.5 to 5 parts of maltose, 0.3 to 0.5 part of urea and 1 to 3 parts of dextran 40.
2. The protective agent of claim 1, wherein: the protective agent consists of the following components:
2.5 parts of cane sugar, 2.5 percent of maltose, 0.3 part of urea and 3 parts of dextran 40.
3. Use of the vaccine protective agent according to claim 1 or 2 for the preparation of a rabies vaccine.
4. Use according to claim 3, characterized in that: the rabies vaccine is a freeze-dried rabies vaccine for human use.
5. A preparation method of an inactivated vaccine freeze-dried powder preparation is characterized by comprising the following steps:
(1) dissolving the vaccine protectant according to claim 1 or 2 in a phosphate buffer with a NaCl content of 0.2-0.9% (w/v), adding the inactivated and purified virus solution to obtain a final sucrose concentration of 2.5-5% (m/v);
(2) and (5) freeze drying.
6. The method of claim 5, wherein: in the step (1), the final concentration of sucrose, maltose, urea and dextran 40 in the mixture is 2.5% (m/v), 0.3% (m/v) and 3% (m/v).
7. The method of claim 5, wherein: the virus is produced by taking Vero cells cultured by a serum-free culture medium as a host.
8. The method of any of claims 5 to 7, wherein: the virus is rabies virus.
9. The method of claim 8, wherein: the virus is rabies virus fixed virus rPV-2061 strain.
10. The inactivated vaccine lyophilized powder preparation prepared by the method of any one of claims 5-9.
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Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20060275776A1 (en) * 2005-06-01 2006-12-07 Angelika Banzhoff Rabies vaccine
US20110081380A1 (en) * 2009-10-07 2011-04-07 Sanofi Pasteur Stabilizing Excipient for Inactivated Whole Virus Vaccine
CN103690943A (en) * 2013-12-12 2014-04-02 大连汉信生物制药有限公司 Freeze-dried rabies vaccine for human use and preparation method thereof
WO2014168510A1 (en) * 2013-04-11 2014-10-16 Федеральное Казенное Предприятие "Щелковский Биокомбинат" Method for producing rabies vaccine
WO2015150394A1 (en) * 2014-04-01 2015-10-08 Pharmacosmos A/S Cryoprotective agent, cryoprotective and cryopreserved compositions, uses thereof, and methods of cryopreservation
CN110101864A (en) * 2019-05-31 2019-08-09 辽宁茂康源生物科技有限公司 The protective agent of serum-free Antirabic Vaccine a kind of and its application

Patent Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20060275776A1 (en) * 2005-06-01 2006-12-07 Angelika Banzhoff Rabies vaccine
US20110081380A1 (en) * 2009-10-07 2011-04-07 Sanofi Pasteur Stabilizing Excipient for Inactivated Whole Virus Vaccine
WO2014168510A1 (en) * 2013-04-11 2014-10-16 Федеральное Казенное Предприятие "Щелковский Биокомбинат" Method for producing rabies vaccine
CN103690943A (en) * 2013-12-12 2014-04-02 大连汉信生物制药有限公司 Freeze-dried rabies vaccine for human use and preparation method thereof
WO2015150394A1 (en) * 2014-04-01 2015-10-08 Pharmacosmos A/S Cryoprotective agent, cryoprotective and cryopreserved compositions, uses thereof, and methods of cryopreservation
CN110101864A (en) * 2019-05-31 2019-08-09 辽宁茂康源生物科技有限公司 The protective agent of serum-free Antirabic Vaccine a kind of and its application

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
MARIA PELLICCIA等: "Additives for vaccine storage to improve thermal stability of adenoviruses from hours to months", 《NATURE COMMUNICATIONS》 *
任平: "《兽用生物制品技术》", 31 August 2007, 中国农业出版社 *
马超等: "冻干人用狂犬病疫苗(Vero细胞)稳定剂配方的筛选", 《微生物学免疫学进展》 *

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Denomination of invention: A vaccine protectant

Granted publication date: 20221011

Pledgee: Chengdu Rural Commercial Bank Co.,Ltd. Jinquan Branch

Pledgor: BOAOVAX BIOTECHNOLOGY CO.,LTD.

Registration number: Y2024510000043