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Oral hexavalent reassortment live rotavirus vaccine

A technology of rotavirus and live vaccines, which is applied in antiviral agents, virus antigen components, viruses/bacteriophages, etc., and can solve the problems of small serotype coverage and coverage

Pending Publication Date: 2021-12-03
WUHAN INST OF BIOLOGICAL PROD CO LTD
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] The diversity of rotavirus strains determines that multivalent vaccines are the most effective way to prevent rotavirus diarrhea. However, the main rotavirus vaccines currently on the market, Lanzhou Institute of Biological Products Rotway and GSK Rotarix is ​​a monovalent serotype rotavirus vaccine, and the serotype coverage is small. Although Merck’s Rotateq is a pentavalent rotavirus vaccine, its serotype does not contain G8 and G9 serotypes, and covers the main serotype circulating strains. The scope is small, and its clinical trial results in Malawi, Africa (G8 serotype is the dominant epidemic strain) show that the protection rate against severe rotavirus diarrhea is only 40%

Method used

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  • Oral hexavalent reassortment live rotavirus vaccine
  • Oral hexavalent reassortment live rotavirus vaccine
  • Oral hexavalent reassortment live rotavirus vaccine

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] Embodiment 1. Preparation of each serotype virus stock solution

[0046] Cell passage: The culture substrate of each serotype virus is Vero cells. First, resuscitate the working seeds of Vero cells. After culturing at 37°C for 3-5 days, carry out passage and expansion culture according to a certain ratio. The culture containers are T25, T75, and T175 cells respectively. Culture flasks, 10-layer cell factories, and 40-layer cell factories were inoculated with G1, G2, G3, G4, G8, and G9 serotype rotavirus after culturing 3-5 generations in the 40-layer cell factory.

[0047] Virus culture: After the cell confluence in the cell factory reaches more than 90%, the virus inoculation operation can be carried out. Before virus inoculation, select a cell factory for cell counting. According to the counting results, calculate according to MOI (multiplicity of virus infection) 0.005-0.2 to obtain the required amount of virus, and activate the virus with trypsin. The activation co...

Embodiment 2

[0050] Embodiment 2. Development of vaccine protective agent

[0051] (1) Determination of virus resistance to acid

[0052] The minimum pH value that the hexavalent rotavirus vaccine can tolerate is determined by experiment.

[0053] Mix the hexavalent serotype virus mixed vaccine (the stock solution of the hexavalent vaccine is mixed in equal volumes) and 1% sodium citrate solution in equal volumes.

[0054] (2) Prepare 7 disposable sterile 50ml centrifuge tubes, 6 of which were added with 2ml of hexavalent seedlings mixed in step (1), and the other 1 was added with 1ml of hexavalent mixed seedlings and 1ml of DMEM as a control.

[0055] (3) To adjust the pH, add 0.1M hydrochloric acid 0ml, 0.4ml, 0.6ml, 0.7ml, 0.8ml, 1.2ml to 6 centrifuge tubes respectively, and the corresponding pH values ​​are 7.84, 5.02, 4.00, 3.50, 3.05, 2.00 .

[0056] (4) Place 7 centrifuge tubes in a 37°C water bath and incubate for 2 hours.

[0057] (5) After the incubation time is over, immedia...

Embodiment 3

[0085] Example 3. Screening of different formulations

[0086] See Table 4 for the composition contents of 5 kinds of vaccine protective agent formulations

[0087] The content of each component of different protective agents in table 4

[0088]

[0089]

[0090] Each formula was mixed with the most temperature-sensitive serotype G9 virus, and an accelerated stability test was carried out. The results are shown in Table 5

[0091] The virus stability test of table 5 different protection formulations (all data are the mean value of three detection results, unit: lgFFU / ml)

[0092]

[0093] It can be seen that the protection of the above five formulas to G9 all meets the requirements, among which F5 is stored at 37°C for 7 days, the drop in G9 titer is the lowest, and the protection to G9 is the best, so F5 is the best for rotavirus vaccine Preservative formula.

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Abstract

The invention relates to an oral hexavalent reassortment live rotavirus vaccine. The vaccine contains six main epidemic strain serotypes G1, G2, G3, G4, G8 and G9, covers 99.6% of G serotype virus of group A rotavirus, and is expected to have a good prevention and protection effect on rotavirus diarrhea. The invention aims to provide an oral inoculated live vaccine consisting of serotypes of hexavalent rotavirus for preventing infantile diarrhea caused by rotavirus, and the vaccine comprises the following components: (1) G1, G2, G3, G4, G8 and G9 serotype live vaccines with a stock solution titer of each serotype being 1-5 * 10 < 6 > FFU / ml; and (2) a protective agent: 0.5-2 g / L of citric acid, 50-150 g / L of sodium citrate, 250-450 g / L of sucrose, 5-10 mM of zinc chloride and 10-20 mM of calcium chloride.

Description

technical field [0001] The invention belongs to the field of vaccine preparations, in particular to an oral hexavalent reassortment rotavirus live vaccine. Background technique [0002] Human rotavirus belongs to the genus Rotavirus of the Reoviridae family, and is a non-enveloped RNA virus with a particle diameter of about 75 nm and consists of three layers of icosahedral protein capsids. Its genome is a double-stranded RNA containing 11 segments, encoding 6 structural proteins (VP1, VP2, VP3, VP4, VP6, VP7) and 5 non-structural proteins (NSP1, NSP2, NSP3, NSP4, NSP5). The laminin is VP4 and VP7, the inner shell protein is VP6, and the nucleoprotein is VP1, VP2 and VP3. Genetic reassortment can occur between viruses of the same group. HRV can be divided into 7 groups (A~G) according to the different antigenicity of the viral inner coat protein VP6, and only three groups A, B and C cause human diarrhea. Group A is the most common cause of diarrhea in infants and young chi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/15A61K39/295A61K47/12A61K47/26A61K47/02A61P31/14C12N7/00C12N7/02
Inventor 杨晓明徐葛林李庆亮何泗涛白萱姜志军程满荣张久威马涛董犇胡蓉刘涛梁婧陈金华姜礼朋冯冬扬杨彪林楠段凯杨邦玲徐晓曾凯
Owner WUHAN INST OF BIOLOGICAL PROD CO LTD
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