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Antimicrobial peptide

A technology of antimicrobial peptides and compositions, applied in the field of antimicrobial peptides, which can solve the problems of toxicity, poor stability, and low antibacterial activity of antimicrobial peptides

Active Publication Date: 2017-03-22
BEIJING PEPMAGIC BIOTECH CORPORATION LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

For example, quite a few natural antimicrobial peptides have low antibacterial activity, poor stability, and high toxicity, such as causing hemolysis of eukaryotic cells; in addition, some natural antimicrobial peptides have poor inhibitory effects on drug-resistant bacteria, which cannot meet the actual application requirements

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Example 1. Synthesis and purification of antimicrobial peptides

[0024] 1. According to the Fmoc solid-phase peptide synthesis method, the antimicrobial peptide BT315 whose X1 is Arg and X2 is Lys is synthesized. The synthesis steps are as follows:

[0025] 1.1 To ensure that the C-terminus of the polypeptide is -CONH 2 Structure, choose Rink Amide resin as polymer solid phase carrier.

[0026] 1.2 Remove the Fmoc protecting group:

[0027] 1.2.1 Put the required polymer solid phase carrier into the synthesis column, add the organic solvent DMF (dimethylformamide) to soak for 10 minutes and then discharge; pass into 20% piperidine / DMF solution to cover the resin surface, pass through N 2 Discharge after reacting for 10 minutes; repeat to feed 20% piperidine / DMF, 2 Discharge after 10 minutes of reaction.

[0028] 1.2.2N 2 Under protection, pass through DMF solvent 5 times to thoroughly wash the above resin.

[0029] 1.3 Condensation reaction:

[0030] Amino Acid...

Embodiment 2

[0052] Embodiment 2. The minimum inhibitory concentration (MIC) and the minimum bactericidal concentration (MBC) of antimicrobial peptide BT315 are measured

[0053] The antimicrobial peptide BT31 prepared in Example 1 was diluted 15 times with sterile culture solution to make 1 mL concentrations of 512, 256, 128, 64, 32, 16, 8, 4, 2, 1, 0.5, 0.25 μg / mL antimicrobial peptide solution. Inoculate the tested strain into 2mL culture medium, culture at 37°C for 8h, and dilute with sterile culture medium to obtain a colony count of approximately 5×10 5 About CFU / mL bacterial liquid. Take 1mL of bacterial liquid and add them to the prepared antimicrobial peptide solutions above. At this time, the concentrations of antimicrobial peptides in each solution are 256, 128, 64, 32, 16, 8, 4, 2, 1, 0.5, 0.25, 0.125 μg / mL. Antibiotics vancomycin and imipenem solutions were prepared in the same way as positive controls; another drug-free group was set as negative controls. Incubate in an o...

Embodiment 3

[0058] The influence of embodiment 3.pH value on the MIC value of antimicrobial peptide BT315

[0059] Adjust the pH of the culture medium containing different sample concentrations to 5.0, 6.0, 7.0, 8.0, 9.0 respectively, and after standing at room temperature for 1 hour, adjust the pH value to 7.0, and then use sterile medium to adjust the volume to be consistent, according to the example The method described in 2 determines the MIC value. The results are shown in Table 2 below.

[0060] The minimum inhibitory concentration (MIC) of table 2 antimicrobial peptide BT315 at different pH

[0061]

[0062] It can be seen from the above table that the antibacterial peptide BT315 has relatively stable antibacterial activity at different pH values, and the activity is the highest at neutral pH. This indicated that the antimicrobial peptide BT315 had better stability.

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Abstract

The invention discloses an antimicrobial peptide, which has the sequence as follow: Tyr-Gly-Arg-Lys-Lys-Arg-Arg-Gln-Arg-Arg-Arg-X1-X2-Arg-Arg-Leu-Trp-Gly-Ala, wherein the X1 is selected from Lys, Arg, Trp, Leu, Phe and Tyr; and the X2 is selected from Trp, Arg, Lys, Phe, Ser and His. The antimicrobial peptide has an obvious inhibiting effect on methicillin-resistant s. aureus, vancomycin-resistant enterococci, carbapenem-resistant pseudomonas aeruginosa and carbapenem-resistant klebsiella pneumoniae, and has lower hemolytic activity, good stability, high antimicrobial activity, and an efficient broad-spectrum bacteriostatic action.

Description

technical field [0001] The invention belongs to the field of biomedicine, and in particular relates to an antibacterial peptide. Background technique [0002] As an important class of clinical drugs, antibiotics have saved countless lives since they were applied. However, in recent years, the abuse of antibiotics and the emergence of drug-resistant bacteria, especially the emergence of multidrug-resistant bacteria (super bacteria) have become one of the major problems facing the world's medical field today, and more and more countries have begun to seek alternatives to antibiotics. Taste. Peptide antibacterial drugs are considered to have broad application prospects in the pharmaceutical industry because of their high antibacterial activity and are not easy to produce drug resistance. They are also the hope for people to solve drug-resistant bacteria and promote human health. [0003] Antimicrobial peptides, also known as antimicrobial peptides or peptide antibiotics, are ...

Claims

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Application Information

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IPC IPC(8): C07K7/08A61K8/64A61Q19/10A61Q5/02A61Q11/00A61K38/10A61P31/04A61P31/10A61P31/12A61P33/00A23L29/00A23K20/147C11D3/48C11D9/50A61F13/00A61F13/15
CPCA61Q5/02A61Q11/00A61Q19/10A61F13/00063A61F13/15203A61K8/64C11D3/48C07K7/08A23V2002/00A61K38/00A61F13/01017A23V2250/55
Inventor 赵树民鲍勇刚石松传周方
Owner BEIJING PEPMAGIC BIOTECH CORPORATION LTD
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