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A kind of vaccine for preventing tuberculosis and combination drug and preparation method and application

A technology for tuberculosis and vaccines, which is applied in the combined drug and application field for preventing tuberculosis, can solve the problems of many types of adjuvants, the existence of hypersensitivity and Koch reaction risks, and poor efficacy, so as to reduce the treatment cycle, reduce the risk of Koch reaction, Effect of Absolute Usage Reduction

Active Publication Date: 2022-07-08
ANHUI ZHIFEI LONGCOM BIOPHARM CO LTD +3
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0018] Aiming at the problems that existing subunit vaccines are ineffective or poor in treating latent tuberculosis infected people, use many types of adjuvants, and have hypersensitivity and Koch reaction risks, the present invention provides a preventive vaccine containing mycobacterium-like microbial compound adjuvants. tuberculosis vaccine

Method used

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  • A kind of vaccine for preventing tuberculosis and combination drug and preparation method and application
  • A kind of vaccine for preventing tuberculosis and combination drug and preparation method and application
  • A kind of vaccine for preventing tuberculosis and combination drug and preparation method and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0073] Preparation of Tuberculosis Vaccine Containing Mycobacterial Microorganism Compound Adjuvant

[0074] 1. Preparation process of Ag85b protein

[0075] Find the gene sequence of Ag85b protein from GeneBank, and design primers as follows: see Table 1.

[0076] Table 1 Primer sequences, restriction sites and protected bases of Ag85b target gene

[0077]

[0078] Remarks: The underline is the restriction site of the endonuclease, and the italic part is the start codon.

[0079] The Ag85b gene was amplified by PCR using the whole genome of Mycobacterium tuberculosis H37Rv as a template. The PCR reaction system was 50 μL (ddH 2 O 30 μL, 10×Buffer 5 μL, dNTP 4 μL, P1 primer 4 μL, P2 primer 4 μL, DNA template 2.5 μL, DNA polymerase 0.5 μL), the amplification conditions are: 96 ℃ pre-denaturation for 3 minutes; 96 ℃ denaturation for 45 seconds, 62 Annealing at °C for 45 seconds, extension at 72 °C for 1 minute, a total of 30 cycles; extension at 72 °C for 7 minutes.

[0...

Embodiment 2

[0109] Animal Protection Test of Tuberculosis Vaccine Containing Mycobacterial Microorganism Compound Adjuvant

[0110] 1. Experimental method

[0111] Immunotherapy of Mtb-infected guinea pigs

[0112] Sixteen SPF Hartley guinea pigs (300-350 g / pig), half male and half male, were divided into two groups (vaccine group and control group), with 8 animals in each group. The guinea pigs of the vaccine group and the control group were subcutaneously challenged with 5.0×10 3 CFU Mtb, 4 weeks later, the vaccine group was injected with this vaccine for treatment, a total of 6 injections, each injection was 1 week apart, and the dose of each injection was [Ag85b(10μg)+EC(10μg)+MV(22.5μg)] / 0.5mL / Only. The guinea pigs in the control group were injected with the same amount of normal saline as a control. After 12 weeks of Mtb infection, guinea pigs were dissected. The comprehensive lesion index of liver, spleen and lung and the bacterial load in spleen and lung were analyzed. Mtb ...

Embodiment 3

[0124] Comparative test of animal protection between tuberculosis vaccine containing mycobacterial microbial compound adjuvant and subunit tuberculosis vaccine

[0125] 1. Experimental method

[0126] Immunotherapy of Mtb-infected guinea pigs

[0127] Sixteen SPF Hartley guinea pigs (300-350 g / pig), half male and half male, were divided into two groups (vaccine group and control group), with 8 animals in each group. The guinea pigs of the vaccine group and the control group were subcutaneously challenged with 5.0×10 3 CFU Mtb, 4 weeks later, the vaccine group was injected with this vaccine for treatment, a total of 6 injections, each injection was 1 week apart, and the dose of each injection was [Ag85b(10μg)+EC(10μg)+MV(22.5μg)] / 0.5mL / Only. Guinea pigs in the control group were injected with the same amount of subunit tuberculosis vaccine ([Ag85b(10μg)+EC(10μg)+BCG-CpG-DNA(75μg)+Al(OH) 3 (200 μg)] / 0.5 mL / piece) as a control. After 12 weeks of Mtb infection, guinea pigs wer...

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Abstract

The invention discloses a vaccine for preventing tuberculosis, a combined medicine, a preparation method and an application, and belongs to the field of medicines for treating latent infection of tuberculosis. Aiming at the problems that the existing subunit vaccines are ineffective or poor in treating people with latent tuberculosis infection, many types of adjuvants are used, and there are risks of hypersensitivity reactions and Koch reactions, the present invention provides a prevention method containing a mycobacterial microbial compound adjuvant Tuberculosis vaccine, which comprises Ag85b protein, ESAT6-CFP10 protein and Mycobacterium vaccae cell extract. The tuberculosis prevention vaccine containing the mycobacterial microbial compound adjuvant of the present invention is used for treating latent tuberculosis infection, and the effect is remarkable.

Description

technical field [0001] The invention belongs to the field of medicines for treating latent infection of tuberculosis, in particular to a tuberculosis prevention vaccine containing a mycobacterial microbial compound adjuvant, a preparation method and application thereof, and a combined medicine for preventing tuberculosis and application thereof. Background technique [0002] Tuberculosis is a chronic infectious disease caused by Mycobacterium tuberculosis, which is mainly transmitted by the respiratory tract. It once seriously endangered human health. Since the discovery of the pathogen, BCG has been developed for the treatment and prevention of tuberculosis and a variety of anti-tuberculosis drugs. However, due to the emergence of drug-resistant strains and the limitations of the BCG immunization effect, tuberculosis has not been effectively controlled. Since the mid-1980s, the tuberculosis epidemic has rebounded significantly worldwide, and its incidence and prevalence are...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K39/04A61K39/39A61K45/06A61P31/06
CPCA61K39/04A61K39/39A61K45/06A61P31/06A61K2039/55594
Inventor 张凯程琛舒钟再新倪兰芳吴德兰沈煜陈伟韦芬江秋虹仇晶晶朱银猛梁永培程兴徐守腾蒲江卢锦标都伟欣王国治徐苗陶立峰
Owner ANHUI ZHIFEI LONGCOM BIOPHARM CO LTD
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