New cationic antimicrobial peptide and application thereof

A cationic and antimicrobial peptide technology, applied in the direction of antibacterial drugs, peptides, antitumor drugs, etc., 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 , significant inhibitory effect, simple and convenient preparation method

Inactive Publication Date: 2015-04-01
LIAONING NORMAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] Of course, antimicrobial peptides from natural sources have immunogenicity due to their large molecular weight, low antibacterial and anticancer activity, cytotoxicity to host cells, or hemolysis, which limits their promotion and application as antibacterial and anticancer drugs. Therefore, while looking for new antimicrobial peptides, scientists have also begun to work on the structural modification or redesign of the original natural antimicrobial peptides, such as replacing some amino acid residues or directly designing the primary structure of the amino acid peptides as needed , in order to obtain antimicrobial peptides with higher activity, more targeted and non-toxic effects on host cells

Method used

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  • New cationic antimicrobial peptide and application thereof
  • New cationic antimicrobial peptide and application thereof
  • New cationic antimicrobial peptide and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Preparation of antimicrobial peptides

[0024] 1. Chemical synthesis method of antimicrobial peptides: 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. Synthetic peptides were purified by reverse HPLC (Vydac218TP1022 column);

[0025] 2. The molecular weight was determined by matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF);

[0026] 3. The purity of the purified antimicrobial peptides is identified by high performance liquid chromatography (HPLC), and the purity used in the test is all greater than 95%. Determine the amino acid sequence structure with an automatic amino acid sequencer;

[0027] The antibacterial peptide of the present invention contains 13 amino acid residues in total, the molecular weight is 1552.13Da, and the isoelectric point is 10.70. The full amino a...

Embodiment 2

[0030] Antibacterial Experiment of Antimicrobial Peptides Against Gram-positive Bacteria

[0031] The pathogenic bacteria in the following examples all come from the General Microbiology Center (CGMCC) of the China Microbiological Culture Collection Management Committee and the China Industrial Microbiological Culture Collection Management Center (CICC), including: Staphylococcus aureus (AS 1.72), grapevine epidermis Coccus (CICC 23664), Bacillus cereus (AS 1.126), Enterococcus faecalis (CGMCC 1.595), Enterococcus faecium (CGMCC 1.2334), Streptococcus mutans (CGMCC 1.2500), Streptococcus salivarius (CGMCC 1.2498), Acidophilus Bacillus (CGMCC 1.3251), Streptococcus lactis (AS 1.1690).

[0032] Minimum inhibitory concentration (minimal inhibitory concentration, MIC): is the lowest sample concentration where no bacterial growth can be detected. The two-fold dilution method was used. The specific method is as follows:

[0033] Bacteria were inoculated on LB solid medium and cul...

Embodiment 3

[0039] Inhibitory Effect of Antimicrobial Peptides on Gram-negative Bacteria

[0040] The pathogenic bacteria in the following examples all come from the General Microbiology Center (CGMCC) of the China Microbiological Culture Collection Management Committee and the China Industrial Microbiology Culture Collection Management Center (CICC), including: Escherichia coli (AS 1.349), Pseudomonas aeruginosa Baumannii (CGMCC 1.860), Acinetobacter baumannii (CICC 22934), Fusobacterium nucleatum (CGMCC 1.2528), Klebsiella pneumoniae subspecies (CGMCC 1.716), Enterobacter cloacae (CGMCC 1.58), Proteus mirabilis ( CICC 22930).

[0041] The bacterial culture method and MIC determination method are consistent with the method described in Example 1.

[0042] The results are shown in Table 2. Antimicrobial peptides showed different degrees of antibacterial effects on Gram-negative bacteria, among which antibacterial effects on Escherichia coli, Pseudomonas aeruginosa, Acinetobacter baumanni...

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Abstract

The invention relates to a new cationic antimicrobial peptide and application thereof, belonging to the technical field of biochemical polypeptide medicines. The new cationic antimicrobial peptide is a small molecule polypeptide which is designed artificially and synthesized chemically by taking a natural antimicrobial peptide in skin secretions of rana temporaria chensinensis david as a template. The small molecule polypeptide comprises 13 amino acid residues, and the C terminal of the small molecule polypeptide is amidated. The small molecule polypeptide has the sequence of IKKILSKIKKLLK-NH2, the molecular weight of 1552.13Da and the isoelectric point of 10.70, and the small molecule polypeptide has 6 positive charges. Tests prove that the small molecule polypeptide has broad-spectrum inhibiting and killing effects on a variety of bacteria and human cancer cells in vitro and has no hemolytic activity and eukaryotic cell toxicity in an antimicrobial and anticancer concentration range, and drug resistance is not easy to produce. In addition, experiments on animals show that the small molecule polypeptide has the effect of resisting wound infection. The small molecule polypeptide has small molecular weight, is synthesized artificially and conveniently, has extremely high application value and can be used for preparing new antibacterial, anti-inflammatory and anti-cancer medicines.

Description

technical field [0001] The invention belongs to the technical field of polypeptide drugs in biochemistry, and specifically relates to a cationic antibacterial peptide with an α-helical structure and the application of the peptide in preparing antibacterial, anti-infection and anticancer drugs. Background technique [0002] With the widespread use and abuse of antibiotics, a large number of drug-resistant bacteria have emerged and spread. In cancer treatment, traditional chemical or natural anticancer drugs also have poor selectivity for normal cells and are prone to drug resistance. Therefore, finding safe and effective new antibacterial and anticancer drugs that are not prone to drug resistance has become a hot spot for scientists all over the world. [0003] Antimicrobial peptides (AMPs) are a class of polypeptides that exist widely in organisms and are used to resist the invasion of foreign microorganisms and some mutant cells in the body. As the first line of defense o...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K7/08A61K38/10A61P31/04A61P17/02A61P35/00
Inventor 尚德静王澈
Owner LIAONING NORMAL UNIVERSITY
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