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Rabies live vaccine and method for preparing rabies live vaccine for oral administration

A technology for rabies and live vaccines, applied in antiviral agents, pharmaceutical formulations, medical preparations with inactive ingredients, etc., can solve problems such as difficulty in carrying out and easily harm medical personnel, and achieve the effect of improving efficiency and quality

Active Publication Date: 2014-06-25
北京中联康生物科技股份有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The use of intramuscular injection requires the dog to be kept in place. For large dogs and strong dogs, it is particularly easy to hurt the medical staff who are immunized. In addition, stray dogs are difficult to catch, so it is sometimes difficult to perform intramuscular injection.

Method used

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  • Rabies live vaccine and method for preparing rabies live vaccine for oral administration
  • Rabies live vaccine and method for preparing rabies live vaccine for oral administration
  • Rabies live vaccine and method for preparing rabies live vaccine for oral administration

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0090] The preparation of embodiment 1 rabies virus antigen

[0091] (1) Suspension acclimatization of BHK-21 cells

[0092] Select the well-grown monolayer adherent cells of the BHK-21 cell clone, digest them with 0.01% trypsin-0.01% EDTA digestion solution, and suspend them to a final concentration of 2×10 5 / mL~5×10 5 / mL of cell fluid, the inoculum volume depends on the size of the tank and the amount of culture fluid. Suspension nutrient solution composition comprises: 88% MEM, 8% newborn bovine serum, 1% L-glutamine and antibiotic etc., then use (7.2%) NaHCO 2 Adjust the pH value to about 7.4, inoculate a 5L bioreactor, and cultivate at 37°C with 100rpm stirring. Follow this method for 60 passages, observe the cell morphology and count.

[0093] (2) Antigen preparation

[0094] Select the 13th generation of rabies ERAg3m strain adapted by BHK-21 cells, LD 50 for 10 -7.0 / ml, inoculated with 55-60 generations of suspension cells, the initial concentration of suspen...

Embodiment 2

[0095] The preparation of embodiment 2 microspheres

[0096] Dissolve 1g of chitosan in 1% acetic acid solution, and then add rabies virus antigen to the chitosan solution at a stirring speed of 200r / min, so that the final concentration of rabies virus antigen is 1×10 8.0 TCID 50 / ml, then slowly add sodium tripolyphosphate aqueous solution, mix evenly, centrifuge at 4 degrees Celsius and 15000-18000rpm and collect the precipitate, wash the precipitate with double distilled water and freeze-dry to obtain chitosan nanospheres , wherein the mass ratio of chitosan to sodium tripolyphosphate is (3:1) to (8:1).

Embodiment 3

[0097] The application of embodiment 3 rabies virus microspheres oral live vaccine

[0098] Inoculate 10 60-day-old stray dogs without rabies virus antibodies with live rabies virus microspheres orally, and set up 10 inactivated vaccination control dogs and 10 blank control dogs; 21 days after immunization, rabies virus street virus The above-mentioned dogs inoculated with rabies virus microspheres oral live vaccine and inactivated vaccine and blank control dogs were challenged with lethal dose. The specific results are shown in Table 1.

[0099] Table 1 rabies vaccine challenge protection results

[0100] group

[0101] Conclusion: 90% protection of dogs with live vaccine orally, 80% protection of dogs with inactivated vaccine, and blank control dogs all died without protection. Therefore, the challenge protection rate of the rabies virus microsphere oral live vaccine is significantly higher than that of the inactivated vaccine.

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Abstract

The invention discloses a rabies live vaccine and a method for preparing the rabies live vaccine for oral administration. The rabies live vaccine comprises a rabies virus ERAg3m. A strain of the rabies virus ERAg3m is obtained by using a reverse genetic method for transforming a rabies virus strain ERA and has high safety. Therefore, the immune efficacy of the rabies live vaccine containing the rabies ERAg3m virus can be further improved.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular, the invention relates to a rabies live vaccine and a method for preparing a rabies oral live vaccine. Background technique [0002] Rabies, also known as mad dog disease or hydrophobia, is an acute zoonotic infectious disease caused by rabies virus. Rabies mainly affects the central nervous system. Once a person develops the disease, there is no effective clinical treatment. It is the only acute infectious disease with a fatality rate as high as 100%. About 55,000 people die of rabies every year in the world. Since 2003, the annual death toll of rabies in my country has exceeded 2,000. It is the second last in the world, after India. The spread of rabies virus has seriously affected the public health security of our country and has become a very prominent social problem. [0003] The existing rabies vaccines in my country are attenuated live vaccines and inactivated vaccines produced...

Claims

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Application Information

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IPC IPC(8): A61K39/205A61K47/36A61K47/24A61P31/14
Inventor 熊炜高杨刘延亭刘俊生李耀东郑杰
Owner 北京中联康生物科技股份有限公司
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