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A kind of antimicrobial peptide and its application

A technology of antimicrobial peptides and antibacterial agents, applied in the field of antimicrobial peptides and their applications, can solve the problems of complicated process, microscopic antimicrobial peptides, and time-consuming, etc., and achieve the effect of broad-spectrum killing activity and strong bactericidal activity

Active Publication Date: 2022-06-03
川北医学院附属医院
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The content of antimicrobial peptides in animals is very small, and the extraction of antimicrobial peptides from animals is low-yield, time-consuming, complicated and expensive, which has become the biggest obstacle restricting the practical application of antimicrobial peptides

Method used

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  • A kind of antimicrobial peptide and its application
  • A kind of antimicrobial peptide and its application
  • A kind of antimicrobial peptide and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0012] Example 1: Solid-phase synthesis of target product Core-A and reference antimicrobial peptide LfcinB6

[0013] The target antimicrobial peptide Core-A (Arg-Trp-Arg-Phe) and the control antimicrobial peptide LfcinB6 (Phe-Arg-Trp-Trp-Gln-Arg) were synthesized according to the standard Fmoc solid-phase procedure, and synthesized one by one from the C-terminus to the N-terminus It is carried out under the automatic control of the synthesizer (for the synthetic operation of the peptide, please refer to the operation guide of the pioneer peptide synthesizer). The synthesized product was purified by reversed-phase liquid chromatography (Vydac 218TP1022 column, 2.2*25 cm), eluted with acetonitrile / water system and subjected to mass spectrometry analysis to obtain the corresponding antimicrobial peptides. The prepared Core-A polypeptide sequence is: R-W-R-F-NH 2 (Arg-Trp-Arg-Phe-NH 2 ).

[0014] After the successful synthesis of the antimicrobial peptide, 100 mg of the dried ...

Embodiment 2

[0017] Example 2: Detection of bactericidal activity of antimicrobial peptides

[0018] The bactericidal activity of Core-A antimicrobial peptide and reference antimicrobial peptide LfcinB6 was detected by agar plate diffusion method to evaluate the bactericidal activity of the core sequence Core-A in the present invention. The specific method for the detection of bactericidal activity of antimicrobial peptides is as follows:

[0019] First, the bacteria (Gram-positive bacteria and Gram-negative bacteria, purchased from the China Institute of Biological Products) were recovered by conventional bacteria recovery methods, inoculated with slants, and cultured at 37 °C overnight; then the bacteria were picked in ordinary LB culture medium at 37°C overnight; then dilute the bacterial solution to make the bacterial concentration 10 5 ~10 6 cfu / ml, inoculate 100 μl of bacterial liquid per plate into a 15 ml LB solid petri dish, after solidification, spread evenly, punch 5 mm holes,...

Embodiment 3

[0031] Example 3: In vitro hemolytic activity detection

[0032] This example is used to detect whether the cationic antimicrobial peptide Core-A has hemolytic activity on animal erythrocytes, and the control antimicrobial peptide LfcinB6 is used as a control. The blood samples used were from defibrillated sheep blood. The hemolytic activity of cationic antimicrobial peptides was detected by the agar plate hole diffusion method, specifically:

[0033] Add the defibrillated sheep red blood cells into the LB medium at 40°C in a volume ratio of 1:20, pour 15ml of each plate to mix the LB solid medium, and after coagulation, punch 5mm holes, 4 cells / plate, and add 50μl respectively. The cationic antimicrobial peptide Core-A solution, LfcinB6 solution, Tween 80 and ultrapure water with a concentration of 1mg / ml (Tween 80 was used as a positive control, and ultrapure water was used as a negative control). Incubate at 37°C for 24h, observe whether there is a hemolysis circle within...

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Abstract

The invention discloses an antibacterial peptide and its application. The antibacterial peptide comprises an amino acid sequence of Arg-Trp-Arg-Phe-NH 2 The polypeptide and the peptidomimetic of the polypeptide, wherein the amino acid in the polypeptide is an L-type amino acid or a D-type amino acid, and the peptidomimetic is a modified product of the N-terminal and / or C-terminal of the polypeptide. The antimicrobial peptide has broad-spectrum killing activity against Gram-positive bacteria and Gram-negative bacteria, and has stronger bactericidal activity than natural antibacterial peptides, and has no toxic effect on animal and plant cells, and can be used to prepare a Antibacterial agents for bacterial infections in humans.

Description

technical field [0001] The invention belongs to the technical field of polypeptides, and in particular relates to an antibacterial peptide and its application. Background technique [0002] Since the invention of antibiotics, humans have made great achievements in the control and treatment of microbial infections, but with the continuous use of antibiotics, microbial resistance has become a major problem in microbial infection control, so that some microbial bacteria have been lost. A new substance that allows it to be controlled to kill. For example, in clinical drug treatment, vancomycin-resistant Staphylococcus, Enterococcus and other Gram-negative bacterial infections are currently a worldwide clinical problem. [0003] Antibacterial peptides are a class of small molecular polypeptides that widely exist in microorganisms, animals and plants. They are usually formed by more than 30 amino acid residues and have a molecular weight of 2 to 7KDa. The bactericidal mechanism ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K5/11A61K38/07A61P31/04
CPCY02A50/30
Inventor 钟晓武郭晓兰李丽华廖何斌彭元洪马强黄光成徐磊
Owner 川北医学院附属医院
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