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Anthrax rPA dry powder inhalant and application thereof

A dry powder inhaler and anthrax technology, applied in powder delivery, freeze-drying delivery, aerosol delivery, etc., can solve the problems of low antibody titer and unsatisfactory immune protection effect, achieve good immune response and improve self-inoculation ability, resistance to lethal effects

Pending Publication Date: 2020-04-14
ACADEMY OF MILITARY MEDICAL SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, most of the anthrax rPA vaccines remain in the way of subcutaneous injection or intranasal drip as the way of immunization. The antibody titer after immunization is generally not high, and the immune protection effect is not ideal.

Method used

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  • Anthrax rPA dry powder inhalant and application thereof
  • Anthrax rPA dry powder inhalant and application thereof
  • Anthrax rPA dry powder inhalant and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0064] Embodiment 1, the preparation of anthrax rPA dry powder inhalation

[0065] 1. Preparation of samples before spraying of dry powder inhaler

[0066] 1. Samples of anthrax rPA dry powder inhaler before spraying

[0067] Anthrax rPA dry powder inhaler spray before sample (pH 7.2) is made up of solute and solvent; Described solute and its concentration in the sample before spray are: anthrax rPA protein 0.1% (mass percentage composition), CpG 0.1% (mass percentage content), mannitol 1% (mass percentage), inositol 1% (mass percentage), leucine 0.5% (mass percentage), poloxamer 0.05% (mass percentage) ; The solvent is deionized water.

[0068] 2. Samples before CpG dry powder spraying

[0069] The sample (pH 7.2) before the CpG dry powder spray is made up of solute and solvent; The solute and its concentration in the sample before the spray are: CpG 0.1% (mass percentage composition), mannitol 1% (mass percentage composition), Inositol 1% (mass percentage), leucine 0.5% ...

Embodiment 2

[0080] Embodiment 2, the characteristic analysis of anthrax rPA dry powder inhaler

[0081] 1. Biological activity detection

[0082] 1. Take the anthrax rPA dry powder inhaler prepared in Example 1, and use SDS-PAGE to detect the target protein. Anthrax rPA protein was used as a positive control.

[0083] The result is as figure 2 shown. figure 2 Among them, the dry powder is the detection result of anthrax rPA dry powder inhaler; the liquid is the detection result of anthrax rPA protein. The results showed that the protein content of anthrax rPA protein was consistent before and after being made into dry powder, and the anthrax rPA protein was not degraded.

[0084] 2. Using the ELISA method, refer to the ELISA auxiliary kit (article number: 1030011) of Shenzhen Dakowei Bioengineering Co., Ltd. for operation, and use anthrax rPA protein (rPA liquid) and anthrax rPA dry powder complex solution (PBS solution) for packaging The antibody titer was determined to verify the...

Embodiment 3

[0110] Example 3, Immunogenicity and Immunoprotective Evaluation of Anthrax rPA Dry Powder

[0111] 1. Animal Immunization Experiment

[0112] Experimental animals: BALB / c mice and DBA / 2 mice, 6-8 weeks old, Beijing Weitong Lihua Experimental Animal Co., Ltd.

[0113] The experimental animals were divided into five groups (50 in each group) as follows;

[0114]rPA dry powder inhalation group: the mice were inoculated with a hand-held dry powder aerosol lung delivery device, 0.5 mg rPA dry powder inhalation / mouse, impact gas volume 0.3 ml (using a 1 ml syringe), and two consecutive impacts. The dry powder completely entered the lungs of the mice through pulmonary delivery, and the mice did not die non-specifically. Respectively in the 0th week (first immunization), the 3rd week (second immunization), and the 6th week (third immunization).

[0115] PBS resuspension group of rPA dry powder inhalation: 50 μl / piece (prepared by dissolving 0.5 mg rPA dry powder in 50 μl physiolog...

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Abstract

The invention discloses an anthrax rPA dry powder inhalant and application thereof. The anthrax rPA vaccine dry powder inhalant prepared according to the invention achieves immunization by a lung delivery inhalation route, so that mice can generate better immune response and generate very high IgG antibody titer, thus reaching certain effect of resisting the infection and progress of bacillus anthracis, and effectively resisting the killing effect of anthrax toxin.

Description

technical field [0001] The invention relates to an anthrax rPA dry powder inhaler and application thereof. Background technique [0002] Anthrax caused by Bacillus anthracis (BA) is a severe zoonotic disease. Anthrax spores have strong resistance to a series of unfavorable environments such as physical and chemical properties, and are difficult to be eliminated, making them the preferred pathogenic microorganisms for biological warfare agents. Bacillus anthracis mainly has two pathogenic factors, capsule and anthrax toxin. Anthrax toxin mainly consists of protective antigen (Protective antigen, PA), lethal factor (Lethal factor, LF) and edema factor (Edemafactor, EF). PA is the main factor of Bacillus anthracis pathogenesis and immunity, and is one of the important protective immunogens of human vaccines. At present, most of the anthrax rPA vaccines remain in the way of subcutaneous injection or intranasal drip as the way of immunization. The antibody titer after immuniza...

Claims

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Application Information

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IPC IPC(8): A61K39/07A61K9/72A61P31/04
CPCA61K39/07A61K9/19A61K9/0073A61K47/26A61P31/04
Inventor 周冬生杨文慧李越孙岩松高波焦俊赵月峨于学东殷喆杨慧盈熊小路胡凌飞
Owner ACADEMY OF MILITARY MEDICAL SCI
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