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Application of miRNA inhibitor in preparation of drugs for resisting Mycobacterium tuberculosis

An anti-tuberculosis and inhibitor technology, applied in the field of pharmacy, can solve the problem of underestimation of the prevalence rate and achieve the effect of inhibiting infection and broad clinical application prospects

Inactive Publication Date: 2019-01-04
SHANGHAI PULMONARY HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Mycobacterium microti is also rare and occurs almost exclusively in immunocompromised populations, but its prevalence may be grossly underestimated

Method used

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  • Application of miRNA inhibitor in preparation of drugs for resisting Mycobacterium tuberculosis
  • Application of miRNA inhibitor in preparation of drugs for resisting Mycobacterium tuberculosis
  • Application of miRNA inhibitor in preparation of drugs for resisting Mycobacterium tuberculosis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] The C57BL / 6 mouse peritoneal primary macrophages were divided into 1*10 5Each cell / well was inoculated in a 48-well cell culture plate. After about 2 hours for the cells to adhere to the wall, the complete medium 1640 was removed, and then fresh complete medium was added to culture overnight. The next day, before transfection, replace with fresh 1640 containing double antibody and 10% serum, and transfect NC, miR-27a-M (mimic: mimic) and miR-27a-I (inhibitor: inhibitors). Wherein, the nucleotide sequence of miR-27a-I is shown in SEQ ID NO.2:

[0027] 5'-UGCUCACAAGCAGCUAAGCCCU-3'.

[0028] Next, the cells were cultured in an incubator for 48 hours, and then infected with Mycobacterium tuberculosis (H37Rv) at a dose of MOI=5. After 2-3 hours of infection, discard the supernatant, culture the cells with the medium containing amikacin for 2 hours, then discard the supernatant and replace with 1640 containing 10% serum without double antibody and continue at 37°C, 5% CO ...

Embodiment 2

[0031] C57BL / 6 mice were divided into two groups, miR-27a - / - Mice (i.e. miR-27a knockout mice) were divided into two groups, 6 in each group, were aerosol-infected with Mycobacterium tuberculosis (H37Rv) at a dose of 200CFU / mouse, and administered miR intraperitoneally after 2 weeks of infection -27a antagomir and NC antagomir (NC means negative control). Drug preparation method: pretreat miR-27aantagomir and NC antagomir with in vivo transfection reagent; first dissolve 50nmol miR-27aantagomir or NC antagomir in 375μL autoclaved ddH 2 O, then mixed with 375 μL sterile 10% glucose solution, as solution A; then, mixed 375 μL sterile 10% glucose solution with 375 μL Entranster TM-in vivo transfection reagent, as solution B; finally, mixed solution A and B solution (1:1) to obtain a working solution. The inventors administered 300 μl working solution (about 10 nmol miR-27a antagomir / NC antagomir) by intraperitoneal injection per mouse, and administered twice a week for 1 month...

Embodiment 3

[0034] Take one-third of the mouse lung tissue administered for 1 month after infection in Example 2, and grind it with 1 ml of PBS containing 1% triton-100, serially dilute, and take 10 -3 、10 -4 100ml of the tissue suspension was spread evenly on the MiddleBook 7H10 agar culture plate containing amphotericin B, and then placed in a 37°C incubator for 2-3 weeks for colony counting. The results were as follows: image 3 shown.

[0035] The experimental results of this example show that miR-27a antagomir significantly reduces the bacterial load of Mycobacterium tuberculosis in the lungs of mice.

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Abstract

The invention belongs to the technical field of pharmacy, particularly relates to application of an miRNA inhibitor in the preparation of drugs for resisting Mycobacterium tuberculosis, and relates toa Mycobacterium-tuberculosis-resistant drug. A series of experiments prove that miR-27a antagomir can lower the intracellular survival of Mycobacterium tuberculosis after the Mycobacterium tuberculosis infects macrophage. Related animal experiment results show that the miR-27a antagomir can effectively relieve the pathological injury of the lung and can evidently lower the content of the Mycobacterium tuberculosis in the lung. The new application of the miRNA inhibitor miR-27a antagomir in the preparation of the drugs for resisting the Mycobacterium tuberculosis is provided, and the miR-27a antagomir can evidently effectively inhibit Mycobacterium tuberculosis infection, is capable of favorably providing effective drug support for the treatment of tuberculosis and is promising in clinicalapplication prospect.

Description

technical field [0001] The invention belongs to the technical field of pharmacy, and particularly relates to the application of a miRNA inhibitor in the preparation of an anti-tuberculosis drug, and also relates to an anti-tuberculosis drug. Background technique [0002] As we all know, tuberculosis can occur in any part of the body, but most often occurs in the lungs, which is called pulmonary tuberculosis. Extrapulmonary TB (ie, TB that occurs in organs other than the lungs) may co-exist with pulmonary TB. Common clinical signs and symptoms of tuberculosis include fever, chills, night sweats, loss of appetite, weight loss, and fatigue. [0003] Open TB most commonly occurs in the lungs (about 90% of cases), known as pulmonary tuberculosis. Symptoms of tuberculosis can include chest pain and a prolonged cough with phlegm, while about 25% of people may not show any symptoms. Occasionally, patients may present with a small amount of hemoptysis, coughing, and, in extremely ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K31/7105A61P31/06
CPCA61K31/7105A61P31/06
Inventor 戈宝学陈建霞刘峰李蒿蒿刘海鹏刘忠华黄晓辰王洁
Owner SHANGHAI PULMONARY HOSPITAL
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