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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  • Summary
  • 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 composition and application, thereby solve the antibacterial effect of anti-H. The question of spectrum antimicrobial activity

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 1

[0032] Example 1: Interaction of four antimicrobial peptides with negatively charged phospholipid membranes

[0033] Experimental Materials:

[0034] The four antimicrobial peptides used in this example were all synthesized by standard solid-phase methods. The purity of antimicrobial peptides 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 purchased from Avanti Company.

[0035] experimental method:

[0036] DOPC and DOPG have a molar ratio of 7:3 to simulate the negatively charged phospholipids of microorganisms, mix them separately, dissolve them in chloroform, dry them with nitrogen, dissolve the phospholipids in HEPES buffer solution, prepare liposomes by ultrasonic vortexing, and incubate on clean mica , to prepare phospholipid bimolecular membranes, and then add 0.1 mg / mL of antimi...

example 2

[0038] Example 2: The minimum inhibitory concentration of four antimicrobial peptides to Helicobacter pylori

[0039] Experimental materials: Helicobacter pylori (Biobowell Biotechnology Co., Ltd.), brain-heart leaching broth medium (Hangzhou Best Biotechnology Co., Ltd.), fetal bovine serum (GIBCO).

[0040] experimental method:

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

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

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

[0044] 4. Place the shaker tube in an anaerobic bag for sealed culture (180-rotation shaker overnight...

example 3

[0047] Example 3: Effect of Antimicrobial Peptides on the Growth of Escherichia coli

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

[0049] Experimental method: Dilute Escherichia coli in mid-log phase to 5*10 5 / ml, 10 μL per hole was inoculated into a 96-well plate in LB solutions containing different concentrations of antimicrobial peptides, at 220 rpm, the value of OD600 was dynamically monitored in a microplate reader at 37°C (wherein the natural antimicrobial peptide GL22 was used as a positive control group ).

[0050] The result is as image 3 As shown, the antimicrobial peptides have obvious inhibitory effect on the growth of Escherichia coli, and it is concentration-dependent. The minimum inhibitory concentration (MIC) of GE33 is 4 μg / mL, and both GE27 and GD27 are 8 μg / mL, which has good antibacterial effect. and bactericidal effect. This shows that the three antimicrobial peptides designed by the present...

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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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