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Synthetic antibacterial peptide and preparation method and application thereof

A technology of antimicrobial peptides and chemical synthesis, which is applied to the preparation methods of peptides, antibacterial drugs, antifungal agents, etc., can solve the problems of low antibacterial activity and low clinical use value, and achieve strong bactericidal activity and broad-spectrum killing activity Effect

Active Publication Date: 2009-11-04
昆山博青生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The naturally discovered antimicrobial peptide sequence cecropin has relatively low antibacterial activity, and its minimum inhibitory concentrations against Staphylococcus aureus and Pseudomonas aeruginosa are 50 μg / ml and 25 μg / ml, respectively, and its clinical use value is relatively low

Method used

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

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0059] Preparation and separation and purification of embodiment 1 antimicrobial peptide

[0060] In this example, the solid-phase chemical synthesis method was adopted, and the instrument used was the Pioneer polypeptide synthesizer produced by Applied Biosystems, Inc. of the United States. The synthesized peptide was sheared by high-concentration TFA, then purified by reverse column, and the purified peptide was identified by mass spectrometry. The specific test steps are as follows:

[0061] 1. Preparation of antimicrobial peptides (taking the preparation of 0.1 mmol amount as an example)

[0062] All the following reagents for the preparation of antimicrobial peptides were purchased from Applied Biosystems.

[0063] 1.1 Preparation of antimicrobial peptide BQ

[0064] The BQ polypeptide sequence is SEQ ID NO: 2 in the sequence listing:

[0065] BQ: Lys Trp Lys Ser Phe Ile Lys Lys Leu Thr Ser Lys Phe Leu His Ser Ala Lys Lys Lys Phe

[0066] The preparation of the antim...

Embodiment 2

[0082] Example 2 Bactericidal activity detection and in vitro hemolytic activity detection of synthetic antimicrobial peptides

[0083] The strains Pseudomonas aeruginosa CMCC10104 and Staphylococcus aureus CMCC26003 used in the following examples were purchased from China Institute for the Control of Biological Products.

[0084] The bactericidal activity of the synthetic antimicrobial peptides was detected by the 96-well plate method, and the antibacterial peptides cecropin A1 and buforin II synthesized by the solid-phase chemical method were used as controls to evaluate the bactericidal activity of various antimicrobial peptides in the present invention.

[0085] Measure the bactericidal activity of antimicrobial peptides according to the following steps:

[0086] Recover the strains, inoculate the slant and cultivate overnight at 37°C, pick the bacteria in ordinary LB medium, cultivate overnight at 37°C, dilute the bacterial solution to make the bacterial concentration 10 ...

Embodiment 3

[0093] Bactericidal activity detection of embodiment 3 antimicrobial peptide cyclized derivatives

[0094] The modified products after cyclization of polypeptides such as BQ were synthesized by solid phase synthesis. For the synthesis method, see the operating guide for the pioneer peptide synthesizer of Applied Biosystems. After the synthetic product was separated and purified, the minimum inhibitory concentration of the derivative against Pseudomonas aeruginosa was tested, and the bactericidal activity was tested as in Example 2, and the test results are shown in Table 3.

[0095] Table 3 Bactericidal activity detection of antimicrobial peptide cyclized derivatives

[0096]

[0097] The results show that the antibacterial peptide of the present invention still maintains good antibacterial activity after cyclization.

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Abstract

The invention relates to synthetic antibacterial peptide comprising polypeptide selected from SEQ ID NO:1-SEQ IDNO:13 amino acid sequences, and polypeptide of a functional equivalent obtained from the synthetic antibacterial peptide through cyclization, N-terminal and C-terminal modification, translation from L-type amino acid to D-type amino acid, and sequence terminal deletion. The synthetic antibacterial peptide is synthesized by a solid phase chemical method. The synthetic antibacterial peptide has broad-spectrum killing activity to gram negative bacteria, gram positive bacteria and epiphytes, has stronger bactericidal activity than natural antibacterial peptide, has no toxic action on animal and plant cells, and can be used for preparing medicaments for curing infectious diseases.

Description

technical field [0001] The invention relates to a synthetic antibacterial peptide, its preparation method and its application in the preparation of medicines for treating bacterial and fungal infections. The antibacterial peptide has significant bactericidal activity on Gram-positive bacteria, Gram-negative bacteria and fungi. Background technique [0002] Antimicrobial peptides are small molecular polypeptides with biological activity induced by organisms, generally composed of 20-60 amino acids, and the molecular weight is about 2000-7000D. With the rapid development of medical immunology and molecular biology, the study of antimicrobial peptides has increasingly become a hot topic in the field of biotechnology and biomedicine. So far, more than 200 kinds of antimicrobial peptides have been found in many animals (especially insects), plants, microorganisms and humans. These short peptides not only have a broad-spectrum bactericidal effect on bacteria and fungi, but also ha...

Claims

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Application Information

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IPC IPC(8): C07K7/08C07K1/107C07K1/04C12N15/09A61K38/10A61P31/04A61P31/10
Inventor 黄青山
Owner 昆山博青生物科技有限公司
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