Antibacterial peptide and pharmaceutical composition and application thereof

A technology of antimicrobial peptides and compositions, which is applied in the field of antimicrobial peptides and their pharmaceutical compositions, and can solve the problem that anti-H. etc. to achieve good antibacterial and bactericidal effects, good pH responsiveness, and good antibacterial performance

Active Publication Date: 2021-02-05
SHANGHAI INST OF APPLIED PHYSICS - CHINESE ACAD OF SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The purpose of the present invention is to provide a kind of antibacterial peptide and its pharmaceutical composi

Method used

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  • Antibacterial peptide and pharmaceutical composition and application thereof
  • Antibacterial peptide and pharmaceutical composition and application thereof
  • Antibacterial peptide and pharmaceutical composition and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Example Embodiment

[0032]Example 1: The interaction of four antimicrobial peptides with negatively charged phospholipid membrane

[0033]Experimental Materials:

[0034]The four antimicrobial peptides used in this example were all synthesized by standard solid-phase methods. The purity of the antimicrobial peptide was confirmed by HPLC and ESI-MS. Dioleoylphosphatidylglycerol (1,2-dioleoyl-sn-glycero-3-(phospho-rac-(1-glycerol), DOPG), Dioleoylphosphatidylcholine (1,2-dioleoyl-sn-glycero -3-phosphocholine, DOPC) and cholesterol (cholesterol) were all purchased from Avanti.

[0035]experimental method:

[0036]The molar ratio of DOPC and DOPG is 7:3 to simulate the negatively charged phospholipids of microorganisms. They were mixed and dissolved in chloroform, dried with nitrogen, and the HEPES buffer solution was used to dissolve the phospholipids. The liposomes were prepared by ultrasonic vortexing and incubated on clean mica. , Prepare phospholipid bimolecular membrane, then add 0.1mg / mL antibacterial peptide, ...

Example Embodiment

[0038]Example 2: Minimum inhibitory concentration of four antimicrobial peptides against Helicobacter pylori

[0039]Experimental materials: Helicobacter pylori (Biowell Biotech Co., Ltd.), Brain Heart Extraction Broth Medium (Hangzhou Best Biotech Co., Ltd.), Fetal Bovine Serum (GIBCO).

[0040]experimental method:

[0041]1. Take 9g of brain heart infusion broth medium and dissolve it in 250mL pure water and then sterilize it, add 10% fetal bovine serum after sterilization.

[0042]2. Prepare the concentration of four antimicrobial peptides to 128μg / mL, 64μg / mL, 32μg / mL, 16μg / mL, 8μg / mL, 4μg / mL, 2μg / mL, 1μg / mL, 0.5μg / mL, 0.25μg / mL, 0.125μg / mL.

[0043]3. Take 5mL of the medium in step 1 into a 12mL shaking tube, and add 50μL of pre-dissolved Helicobacter pylori solution, and cover it (do not tighten the tube).

[0044]4. Place the shake tube in an anaerobic bag and seal culture (180 rpm shake overnight). The next day the bacterial solution became mixed. The bacterial solution was diluted 1000 time...

Example Embodiment

[0047]Example 3: The effect of antimicrobial peptides on the growth of E. coli

[0048]Experimental material: LB broth medium (purchased from Qingdao Haibo Biological Technology Co., Ltd.).

[0049]Experimental method: Dilute the E. coli in the mid-log phase to 5*105 / ml, 10μL per well was inoculated into 96-well plates in LB solution containing different concentrations of antimicrobial peptides, 220rpm, at 37℃ in a microplate reader to dynamically monitor the OD600 value (the natural antimicrobial peptide GL22 was used as the positive control ).

[0050]The result isimage 3 As shown, the antimicrobial peptide has a significant inhibitory effect on the growth of E. coli, and is concentration-dependent. The minimum inhibitory concentration (MIC) of GE33 is 4μg / mL, and both GE27 and GD27 are 8μg / mL, which has good bacteriostasis. And bactericidal effect. This shows that the three antibacterial peptides designed in the present invention have good antibacterial properties against Gram-negative ba...

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Abstract

The present invention provides an antibacterial peptide, and a pharmaceutical composition and applications thereof. The antibacterial peptide comprises any one of an antibacterial peptide 1, an antibacterial peptide 2 and an antibacterial peptide 3, the amino acid sequence of the antibacterial peptide 1 is represented by SEQ ID No: 1, the amino acid sequence of the antibacterial peptide 2 is represented by SEQ ID No: 2, and the amino acid sequence of the antibacterial peptide 3 is represented by SEQ ID No: 3. The three antibacterial peptides are formed by modifying natural antibacterial peptides PGLa-AM1 as the basis, and have the characteristics of stronger antibacterial effect and better performance. The inhibitory effect of the three antibacterial peptides on helicobacter pylori is stronger than that of PGLa-AM1, the broad-spectrum antibacterial effect of the antibacterial peptide 3 is the strongest, and the antibacterial peptide 3 has good pH responsiveness, so that the antibacterial peptide 3 is in better contact with helicobacter pylori and inhibits growth of helicobacter pylori. The three antibacterial peptides have advantages of strong antibacterial activity, strong biocompatibility and low cost, and are expected to be widely used.

Description

technical field [0001] The present invention relates to the field of medicine, and more specifically relates to an antimicrobial peptide and its pharmaceutical composition and application. Background technique [0002] Since the human discovery of Helicobacter pylori in 1983, the mystery of Helicobacter pylori was only uncovered after Dr. Marshall in Australia drank a culture solution containing Helicobacter pylori in 1985. Subsequent studies have found that Helicobacter pylori infection can cause 100% chronic gastritis, and there is a 1-2% probability of developing gastric cancer, gastric lymphoma and other terrible diseases. On October 27, 2017, the list of carcinogens released by the International Agency for Research on Cancer of the World Health Organization was initially compiled for reference, and Helicobacter pylori was included in the list of carcinogens. The prevention of Helicobacter pylori infection has attracted widespread attention, and the drug resistance of H...

Claims

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Application Information

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IPC IPC(8): C07K14/46A61K38/17A61P31/04A01N37/46A01P1/00
CPCC07K14/463A61P31/04A01N37/46A61K38/00
Inventor 郭珍张益
Owner SHANGHAI INST OF APPLIED PHYSICS - CHINESE ACAD OF SCI
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