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Vaccine and combination medicine for preventing tuberculosis, method for preparing vaccine and application

A tuberculosis and vaccine technology, applied in the field of combined drugs and applications for the prevention of tuberculosis, can solve the problems of hypersensitivity reaction and Koch reaction risk, many types of adjuvants, poor effect, etc., so as to reduce the risk of Koch reaction, reduce the treatment time, good immunogenicity

Active Publication Date: 2018-12-25
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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  • Vaccine and combination medicine for preventing tuberculosis, method for preparing vaccine and application
  • Vaccine and combination medicine for preventing tuberculosis, method for preparing vaccine and application
  • Vaccine and combination medicine for preventing tuberculosis, method for preparing vaccine and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0073] Preparation of Tuberculosis Vaccine Containing Mycobacterium-like Microorganism Compound Adjuvant

[0074] 1. Preparation process of Ag85b protein

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

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

[0077]

[0078] Remarks: The underlines are the cleavage sites of the endonucleases, and the parts in italics are the start codons.

[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°C pre-denaturation for 3 minutes; 96°C denaturation for 45 seconds, 62 Annealing at ℃ for 45 seconds, extending at 72℃ for 1 minute, a total of 30 cycles; extending at 72℃ for 7 m...

Embodiment 2

[0109] Animal Protection Test of Tuberculosis Vaccine Containing Mycobacterium-like Microorganism Compound Adjuvant

[0110] 1. Experimental method

[0111] Immunotherapy of guinea pigs infected with Mtb

[0112] Sixteen Hartley guinea pigs of SPF grade (300-350 g / guinea pig), half male and half male, were divided into two groups (vaccine group and control group), with 8 guinea pigs 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, with an interval of 1 week between each injection, 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. Guinea pigs were dissected 12 weeks after Mtb infection. The comprehensive lesion index of liver, spleen and lung organs and the bac...

Embodiment 3

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

[0125] 1. Experimental method

[0126] Immunotherapy of guinea pigs infected with Mtb

[0127] Sixteen Hartley guinea pigs of SPF grade (300-350 g / guinea pig), half male and half male, were divided into two groups (vaccine group and control group), with 8 guinea pigs 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, with an interval of 1 week between each injection, 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 (10 μg) + EC (10 μg) + BCG-CpG-DNA (75 μg) + Al(OH) 3 (200μg)] / 0.5mL / only) as a control. Guinea...

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Abstract

The invention discloses vaccine and a combination medicine for preventing tuberculosis, a method for preparing the vaccine and application, and belongs to the field of medicines for treating tuberculosis latent infection. By the aid of the vaccine with mycobacterium microbial compound adjuvants for preventing the tuberculosis, the problems of ineffectiveness or poor effects of existing subunit vaccine for treating tubercle bacillus latent infected population, diversified adjuvants, hypersensitivity and Koch reaction risks can be pertinently solved. The vaccine with mycobacterium microbial compound adjuvants for preventing the tuberculosis comprises Ag85b proteins, ESAT6-CFP10 proteins and mycobacterium vaccae thallus extract. The vaccine, the combination medicine, the method and the application have the advantages that the vaccine with the mycobacterium microbial compound adjuvants for preventing the tuberculosis is used for treating latent tuberculosis infection, and obvious effects can be realized; the vaccine only comprises a single type of adjuvants, and accordingly hypersensitivity and Koch reaction risks can be prevented.

Description

technical field [0001] The invention belongs to the field of drugs for treating latent infection of tuberculosis, and specifically relates to a vaccine for preventing tuberculosis containing a mycobacterium-like microorganism compound adjuvant, a preparation method and application thereof, a combined drug for preventing tuberculosis and its application. Background technique [0002] Tuberculosis is a chronic infectious disease mainly caused by respiratory transmission caused by Mycobacterium tuberculosis, which 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 immune effect of BCG, tuberculosis has not been effectively controlled. Since the mid-1980s, the epidemic of tuberculosis has rebounded sharply worldwide, and its incidence and prevalence are still ...

Claims

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Application Information

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Patent Type & Authority Applications(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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