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Aerosolizable botulinum toxin type A AHc subunit vaccine dry powder inhalant

A type A botulinum toxin and dry powder inhaler technology, which is applied in powder delivery, freeze-dry delivery, medical preparations of non-active ingredients, etc., can solve problems such as high risk, complicated process, and limited popularization and application, and achieve The effect of increasing the time of contact reaction and improving the effect of immune protection

Active Publication Date: 2019-10-15
ACADEMY OF MILITARY MEDICAL SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

It can make up for the shortcomings of BoNT toxoid vaccine (PBT) such as complex process, high risk, and limited popularization and application

Method used

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  • Aerosolizable botulinum toxin type A AHc subunit vaccine dry powder inhalant
  • Aerosolizable botulinum toxin type A AHc subunit vaccine dry powder inhalant
  • Aerosolizable botulinum toxin type A AHc subunit vaccine dry powder inhalant

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0060] Embodiment 1, the preparation of albumen

[0061] 1. Preparation of botulinum neurotoxin type A (BoNT / A)

[0062] Preparation of botulinum neurotoxin type A. It was prepared according to the method recorded in the literature ("Establishment of a Convenient and Easy-to-Use Method for Detecting the Activity of Botulinum Toxin Type A", Liu Jing). Using Clostridium botulinum typeA 62A, first enrichment and toxin production culture, then precipitation and extraction, then ammonium sulfate precipitation (60% saturation) and nucleic acid removal, and then gel Filtration and ion exchange chromatography to obtain botulinum neurotoxin type A solution. See the literature for specific steps.

[0063] Take botulinum neurotoxin type A solution, and measure the protein concentration by BCA method, the protein concentration is 0.3mg / mL.

[0064] 2. Preparation of AHc protein

[0065] The AHc protein is the nontoxic 50 kD carboxy-terminal fragment of the heavy chain of botulinum ne...

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Abstract

The invention discloses an aerosolizable botulinum toxin type A AHc subunit vaccine dry powder inhalant. The AHc subunit vaccine dry powder is successfully prepared by a spray freeze drying method. The mass median aerodynamic diameter (MMAD) is close to 2.5 [mu]m. The particles are spherical, and loose and porous in shape, and the distribution of particle size is uniform. The particle size of therange is beneficial to the deposition of the vaccine to the deep part of the lung, the contact reaction time between the antigen and the mucosa tissue is increased, and the immune protection effect isimproved. The recombinant AHc subunit vaccine dry powder prepared by the invention meets the requirements of lung delivery immunity, can protect mice against 1440 times LD<50> BoNT / A complex attackthrough lung delivery immunity, lays a foundation for the development of botulinum toxin subunit vaccine dry powder vaccine and the research of lung delivery immunity protection effect, and provides an advanced technical scheme for the development of inhalation dry powder vaccine.

Description

technical field [0001] The invention relates to an aerosolized dry powder inhalation agent of type A botulinum toxin AHc subunit vaccine. Background technique [0002] Botulinum toxin is a neurotoxin produced by Clostridium botulinum. It is currently the most toxic biological toxin known. It can be divided into eight serotypes A-H according to antigenicity. Among them, A, B, E and F that exist naturally Type A can cause human poisoning and is one of the most harmful Class A biological warfare agents. [0003] Botulinum toxin type A is a classic AB protein toxin with a molecular weight of 150kDa. It links a 50kDa light chain and a 100kDa heavy chain through a disulfide bond; the C-terminus of the heavy chain is the receptor binding region (Hc domain), It can be used to prepare a subunit vaccine. The recombinant protein Hc subunit vaccine has the advantages of safe production process and low cost, and can stimulate the body to produce a strong immune response after multiple i...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K9/72A61K9/19A61K39/08A61P31/04
CPCA61K9/0075A61K9/19A61K39/08A61K47/10A61K47/183A61K47/26A61K2039/55561A61P31/04
Inventor 杨慧盈干长姣周冬生孙岩松法云智高波邱业峰焦周光郝淮杰杨文慧胡凌飞赵月娥于学东熊小路焦俊殷喆
Owner ACADEMY OF MILITARY MEDICAL SCI
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