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Antibacterial peptide derivative from forest frog and application thereof

A technology of antimicrobial peptides and derivatives, which is applied in the field of preparation of antibacterial drugs and antimicrobial peptide derivatives. It can solve the problems of low antibacterial and anticancer activity, limited popularization and application, and cytotoxicity of host cells, so as to promote skin wound healing, The effect of broad-spectrum antibacterial activity and simple and convenient preparation method

Inactive Publication Date: 2015-05-06
LIAONING NORMAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Of course, antimicrobial peptide derivatives from natural sources are immunogenic due to their large molecular weight, low antibacterial and anticancer activities, cytotoxicity to host cells, or hemolysis, which limits their use as antibacterial and anticancer drugs. Therefore, while looking for new antimicrobial peptide derivatives, scientists have also begun to work on structural modification or redesign of the original natural antimicrobial peptide derivatives, such as replacing certain amino acid residues or directly designing antibacterial peptides as needed. The primary structure of amino acids of peptide derivatives, in order to obtain antimicrobial peptide derivatives with higher activity, more targeted and non-toxic effects on host cells

Method used

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  • Antibacterial peptide derivative from forest frog and application thereof
  • Antibacterial peptide derivative from forest frog and application thereof
  • Antibacterial peptide derivative from forest frog and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Preparation of Antimicrobial Peptide Derivatives

[0023] 1. Chemical synthesis method of antimicrobial peptide derivatives: According to the amino acid sequence described in the summary of the invention, an automatic peptide synthesizer is used to synthesize according to the standard Fmoc solid-phase peptide synthesis procedure. The synthetic peptide was purified by reverse HPLC (Vydac218TP1022 column); 2. The molecular weight was determined by matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF);

[0024] 3. The purity of the purified antimicrobial peptide derivatives is identified by high performance liquid chromatography (HPLC), and the purity used in the test is 95%. Determine the amino acid sequence structure with an automatic amino acid sequencer;

[0025] 4. Antimicrobial peptide derivatives contain a total of 18 amino acid residues, with a molecular weight of 2542.9 Da and an isoelectric point of 12.6. The full amino acid se...

Embodiment 2

[0028] Antibacterial experiments of antimicrobial peptide derivatives against Gram-positive bacteria:

[0029] The pathogenic bacteria in the following examples all come from the General Microbiology Center (CGMCC) of China Microbiological Culture Collection Management Committee and the China Industrial Microbiological Culture Collection Management Center (CICC), wherein: Staphylococcus aureus (AS 1.72), Bacillus cereus Bacillus (AS 1.126), Streptococcus lactis (AS 1.1690), Enterococcus faecalis (CGMCC 1.595), Enterococcus faecium (CGMCC 1.2334), Staphylococcus epidermidis (CICC 23664).

[0030] Minimum inhibitory concentration (minimal inhibitory concentration, MIC): the lowest concentration of the sample at which no bacterial growth can be detected. The two-fold dilution method was used. The specific method is as follows:

[0031] Bacteria were inoculated on LB solid medium and cultured upside down in a 37°C incubator. After the colonies grow out, use an inoculation loop ...

Embodiment 3

[0037] Antibacterial experiments of antimicrobial peptide derivatives against Gram-negative bacteria:

[0038] The pathogenic bacteria in the following examples all come from the General Microbiology Center (CGMCC) of China Microbiological Culture Collection Management Committee and the China Industrial Microbiological Culture Collection Management Center (CICC), wherein: Escherichia coli (AS 1.349), Pseudomonas aeruginosa (CGMCC1.860), Enterobacter aerogenes (CGMCC 1.876), Enterobacter cloacae (CGMCC 1.58), Klebsiella pneumoniae subspecies (CGMCC1.716), Acinetobacter baumannii (CICC 22934). The bacterial culture method and MIC determination method are consistent with the method described in Example 2, and the results are shown in Table 2.

[0039] The minimum inhibitory concentration of table 2 antimicrobial peptide derivatives to Gram-negative bacteria

[0040]

[0041] It can be seen from the test results that the antimicrobial peptide derivatives showed a strong selectiv...

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Abstract

The invention relates to an antibacterial peptide derivative from a forest frog and application thereof and belongs to the technical field of biochemical polypeptide medicines. The polypeptide is a small-molecule polypeptide which is designed artificially and synthesized chemically by taking a natural antimicrobial peptide from the skin secretion of a forest frog as a template. The polypeptide comprises 18 amino acid residues and has a sequence as shown in a sequence table. The molecular weight of the polypeptide is 2542.9Da, and the isoelectric point is 12.6. Experiments show that the small-molecule polypeptide has a broad-spectrum effect of killing a variety of bacteria and has no hemolytic activity in an antibacterial and anticancer concentration range, and the bacteria can not become resistant to the small-molecule polypeptide. In addition, Experiments on animals show that the polypeptide has the function of resisting wound infection. The polypeptide has a small molecular weight, can be synthesized artificially and conveniently, is a small-molecule polypeptide with quite high application value and can be used for preparing novel antimicrobial and antiinfectious medicines.

Description

technical field [0001] The invention belongs to the technical field of polypeptide drugs in biochemistry, and specifically relates to an antibacterial peptide derivative with an arbitrary extended structure and the use of the peptide in preparing antibacterial drugs. Background technique [0002] Traditional antibiotics have specific targets when interacting with cells, and then inhibit the growth of bacteria by destroying some specific physiological processes in bacteria. With the large-scale use and abuse of antibiotics, the resistance of bacteria to traditional antibiotics has become a major problem threatening human health. Therefore, it is very urgent to research and develop new antibiotics with novel structures and less resistance. . Antimicrobial peptide derivatives are an important part of the natural defense system of organisms. They are widely found in most animals, plants and microorganisms. Due to their thermal stability and broad-spectrum antibacterial properti...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K7/08A61K38/10A61P31/04A61P17/02
Inventor 尚德静孙玥董维兵
Owner LIAONING NORMAL UNIVERSITY
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