Micro-molecule polypeptide and application thereof

A technology of small molecule polypeptides and mycobacteria, applied in the field of biomedicine, can solve the problems of long-term infectivity, prolongation of the course of disease, increase of treatment costs, etc., and achieve the effect of breast-feeding cells safety, good application prospects, and low synthesis cost

Active Publication Date: 2017-11-21
SUZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The emergence of drug-resistant tuberculosis strains prolongs the course of the disease and increases the risk of death. It may also lead to long-term infectivity, expand the chance of drug-resistant transmission, and increase treatment costs.
With the rapid growth of drug-resistant tuberculosis strains and the lack of effective new anti-tuberculosis drugs, it is urgent to develop biological anti-tuberculosis drugs with unique bactericidal mechanisms

Method used

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  • Micro-molecule polypeptide and application thereof
  • Micro-molecule polypeptide and application thereof
  • Micro-molecule polypeptide and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Example 1 Preparation of AK15

[0030] Find the gene (GeneID: 4156925) encoding gp65 in the genome of Mycobacterium phage Che12 (Mycobacterium phage Che12) (NC_008203.1) or the gene ADZZY_66 in the genome of Mycobacterium phage Adzzy (Mycobacteriumphage Adzzy) (NC_022058.1) (GeneID : 16546204) encodes a linear polypeptide with unknown function, its precursor is composed of 47 amino acid residues (Mycobacterium phage Che12: YP_655644.1, Mycobacterium phage Adzzy: YP_008409360.1), and further use bioinformatics methods for this The peptide precursor was analyzed and predicted, and the complete sequence of AK15 was synthesized by an automatic peptide synthesizer (433A, Applied Biosystems), and desalted and purified by HPLC reverse-phase column chromatography. The purity of AK15 was identified by high performance liquid chromatography (HPLC), its molecular weight was determined by matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF), and ...

Embodiment 2

[0032] Embodiment 2 AK15 measures the minimum inhibitory concentration (MIC) of Mycobacterium tuberculosis

[0033] Mycobacterium tuberculosis H37Rv (ATCC 27294) and H37Ra strains were placed in Middlebrook containing 0.05% Tween 80 and 10% oleic acid, albumin, glucose and catalase (oleic acid, albumin, dextrose, and catalase, OADC) enrichment solution 7H9 broth culture medium (purchased from Qingdao Rishui Biotechnology Co., Ltd.) was cultivated for about 4 weeks, until the density of Mycobacterium tuberculosis reached the McFarland No.1 standard for turbidity (about 3×10 7CFU / ml), the bacterial solution was diluted to 1×10 with Middlebrook 7H9 broth medium containing 0.05% Tween 80 and 10% OADC enrichment solution 5 CFU / ml.

[0034] Add 100 μl of 7H9 liquid medium to each well of a sterile 96-well plate in advance, then add 100 μl of AK15 sample solution diluted to 800 μg / ml with 7H9 liquid medium to the first well, mix well and take 100 μl into the second well , sequentia...

Embodiment 3

[0041] Embodiment 3 AK15 is to the time-bactericidal effect of Mycobacterium tuberculosis, concentration-germicidal effect measurement

[0042] The Mycobacterium tuberculosis standard strain H37Rv (ATCC27294) was inoculated into Middlebrook 7H9 broth medium containing 0.05% Tween 80 and 10% OADC, and cultured in an incubator at 37°C for about 4 weeks to logarithmic growth phase. Dilute the bacterial solution to 1×10 with Middlebrook 7H9 broth medium 6 CFU / ml, the sample was added to the diluted bacterial solution to make the final concentration 0.5×, 1× and 2×MIC, and an equal volume of normal saline was added to the negative control (Control). The bacterial solution added to the sample was quickly placed in a 37°C incubator for culture, and 10 μl of the bacterial solution was diluted 1000 times with sterilized physiological saline on day 0, 3, 7 and 21 respectively, and 50 μl was spread on Middlebrook 7H9 agar for culture On the base, culture it upside down in a 37°C incubat...

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Abstract

The invention relates to a micro-molecule polypeptide. An amino acid sequence of a mature peptide of the micro-molecule polypeptide is as shown in SEQ ID No.1. The invention further discloses an application of the micro-molecule polypeptide to preparation of drugs for resisting mycobacterium tuberculosis. The micro-molecule polypeptide for resisting the mycobacterium tuberculosis stemmed from mycobacteriophage is small in molecular weight and low in synthetic cost and has obvious effects on resisting the mycobacterium tuberculosis H37Rv and the function of killing H37Rv resisting rifampin, other normal gram-positive bacteria, gram-negative bacteria and fungi are unaffected, and the micro-molecule polypeptide has a good application prospect in preparation of the drugs for resisting the mycobacterium tuberculosis.

Description

technical field [0001] The invention relates to the field of biomedicine, in particular to a small molecule polypeptide and its application. Background technique [0002] Tuberculosis is listed as one of the major infectious diseases in the world. Mycobacterium tuberculosis is the pathogenic bacteria that causes the disease, which can infect the whole body organs, but tuberculosis is the most common and is a serious respiratory infectious disease. Due to the ineffectiveness of conventional BCG vaccine in preventing tuberculosis, the emergence of drug-resistant strains of tuberculosis and the co-infection of tuberculosis caused by HIV infection, the incidence and death of tuberculosis are currently high and the situation is very serious. One-third of the world's population is infected with Mycobacterium tuberculosis, and about 10% of those infected show clinical symptoms. Every year, there are 9 million new cases of Mycobacterium tuberculosis infection, and 2 million people...

Claims

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Application Information

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IPC IPC(8): C07K14/01A61K38/16A61P31/06
CPCA61K38/00C07K14/005C12N2795/10022C12N2795/10033
Inventor 卫林徐薇
Owner SUZHOU UNIV
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