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Targeting antibacterial peptide and preparation method and application thereof

An antibacterial peptide, targeting technology, applied in the field of targeted antibacterial peptide and its preparation, can solve the problems of microbial resistance mutation, prolonging the treatment period, destroying the microecological balance and the like

Active Publication Date: 2017-04-26
NORTHEAST AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] However, antimicrobial peptides, like traditional antibiotics, are mostly broad-spectrum antibacterial drugs, which not only exert great selective pressure on microbial resistance mutations
More importantly, broad-spectrum antimicrobials kill the beneficial normal flora while killing pathogenic bacteria, causing serious damage to the microecological balance and causing antibiotic-related complications such as secondary infections that are common in clinical practice. Leading to serious negative consequences such as further deterioration of the condition and prolongation of the treatment cycle

Method used

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  • Targeting antibacterial peptide and preparation method and application thereof
  • Targeting antibacterial peptide and preparation method and application thereof
  • Targeting antibacterial peptide and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0017] In this example, the two methods of combining pheromone labeling technology and optimizing the number of positive charges are used to achieve the optimal specificity effect on the target bacteria. Enterococcus faecalis was used as the target bacteria to design the antimicrobial peptide cCF10-C4. A series of multi-domain antimicrobial peptide molecules selective for Enterococcus faecalis were constructed in a combined manner. The molecular design of this series of antimicrobial peptides mainly includes two independent functional areas, namely the bactericidal area and the recognition area.

[0018] Design of Antimicrobial Peptides

[0019] Using the broad-spectrum antimicrobial peptide C6 as the parent peptide, the pheromone cCF10, which has a specific recognition effect on Enterococcus faecalis, was selected as the recognition region, and GGG was used as the linker to connect the two functional regions. The antimicrobial peptide cCF10-C6, on this basis, replaced the K ...

Embodiment 2

[0023] Synthesis of pheromone-labeled antimicrobial peptides by solid-phase chemical synthesis

[0024] 1. The preparation of antimicrobial peptides is carried out one by one from the C-terminal to the N-terminal, and is completed by a peptide synthesizer. First, Fmoc-X (X is the first amino acid at the C-terminal of each antimicrobial peptide) is inserted into Wang resin, and then the Fmoc group is removed to obtain X-Wang resin; then Fmoc-Y-Trt-OH (9 -Fmoxy-trimethyl-Y, Y is the second amino acid at the C-terminus of each antimicrobial peptide); according to this procedure, it is synthesized from the C-terminus to the N-terminus until the synthesis is completed, and the side of the Fmoc group is removed chain protection resin;

[0025] 2. Add a cleavage reagent to the peptide resin obtained above, react for 2 hours at 20°C in the dark, filter; wash the precipitate with TFA (trifluoroacetic acid), mix the washing liquid with the above filtrate, concentrate with a rotary evap...

Embodiment 3

[0029] Determination of Antimicrobial Peptide Activity

[0030] 1. Determination of antibacterial activity: the antimicrobial peptides are configured into a certain storage solution for use. The minimum inhibitory concentrations of several antimicrobial peptides were determined by the broth microdilution method. Using 0.01% acetic acid (containing 0.2% BSA) as the diluent, a series of gradient antimicrobial peptide solutions were sequentially prepared using the double dilution method. Take 100 μL of the above solution and place it in a 96-well cell culture plate, then add an equal volume of the bacteria solution to be tested (~10 5 per mL) in each well. Positive controls (containing bacterial fluid but not antimicrobial peptides) and negative controls (neither bacterial fluid nor peptides) were set up. Incubate at a constant temperature of 37°C for 20 hours, and the minimum inhibitory concentration is the one where no turbidity is seen at the bottom of the well with the nak...

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Abstract

The invention discloses a targeting antimicrobial peptide and a preparation method and application thereof. A sequence of the targeting antimicrobial peptide cCF10-C4 is shown in a sequence table SEQ ID No.1. The preparation method comprises the steps that enterococcus faecalis serves as a target bacterium to obtain the targeting antimicrobial peptide cCF10-C4 through design, a series of multi-structural-domain antimicrobial peptide molecules having selectivity to the enterococcus faecalis are established in a heterozygous mode. The design of a series of antimicrobial peptide molecules mainly includes two independent functional areas, namely a sterilization area and an identification area respectively. The targeting antimicrobial peptide has targeting selection capability to the target bacterium. The targeting antimicrobial peptide cCF10-C4 is conductive to reestablishment of microecological balance after treatment.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a targeted antibacterial peptide and its preparation method and application. Background technique [0002] Antimicrobial peptides are a class of small molecule polypeptides that exist in the natural immune defense system of organisms, participate in the immune defense function of the body, and are the first barrier for the body to resist the invasion of pathogenic microorganisms. Antimicrobial peptides are usually composed of 12-50 amino acid residues, with a molecular weight of less than 10kDa. Their common feature is that they contain positively charged amino acids and hydrophobic amino acids, and are amphipathic and cationic. Insects and even cancer cells have killing or inhibiting effect. [0003] However, antimicrobial peptides, like traditional antibiotics, are mostly broad-spectrum antibacterial drugs, which not only exert great selective pressure on microbial res...

Claims

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Application Information

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IPC IPC(8): C07K19/00C07K1/04C07K1/06A61K47/64A61K38/10A61P31/04
CPCA61K38/10C07K7/08C07K14/315C07K2319/00
Inventor 单安山徐林丑淑丽王家俊
Owner NORTHEAST AGRICULTURAL UNIVERSITY
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