Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Lipopeptide based on melittin and preparation method and application thereof

A technology of melittin and lipopeptide, applied in the preparation method of peptide, botany equipment and method, application, etc., can solve the problems of high hemolysis and low hemolysis, and achieve the effect of reducing solubility

Active Publication Date: 2021-11-23
CHONGQING ACAD OF ANIMAL SCI
View PDF3 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] The present invention intends to provide a melittin-based lipopeptide to overcome the high hemolytic shortcoming of existing melittin modified products, and provide a lipopeptide with broad-spectrum antibacterial activity and low hemolytic property

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Lipopeptide based on melittin and preparation method and application thereof
  • Lipopeptide based on melittin and preparation method and application thereof
  • Lipopeptide based on melittin and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Melittin and melittin-based lipopeptides were artificially synthesized according to the melittin sequence: GIGAVLKVLTTGLPALISWIKRKRQQ-NH2.

[0037] The synthesis steps are as follows:

[0038] (1) Immobilize the carboxyl group of the first amino acid at the carbon end on 2-chloro-trityl chloride resin

[0039]Weigh 2.0g of 2-chloro-trityl chloride resin (the degree of substitution is 0.4mmol·g-1, the content of available chlorine on the resin is 2.0×0.4=0.8mmol), add it to the In the reaction column of amide (DMF), stir for 1 hour to fully swell the resin. DMF was removed, and a DMF solution dissolved in Fmoc-Gln-OH (2×0.8mmol) and diisopropylethylamine (DIEA) (4×0.8mmol) was added to the resin, and the reaction was shaken at room temperature for 2 hours. The reaction solution was removed, and the resin was washed 3 times with DMF, and then a DMF mixed solution containing excess methanol and (4×0.8 mmol) DIEA was added to continue stirring for 1 hour to block the acti...

experiment example 1

[0049] Experimental example 1: Evaluation of antibacterial activity of lipopeptide

[0050] Antibacterial assays were performed using a broth-based microdilution method in accordance with the CLSI Guidelines for Antimicrobial Susceptibility Testing. E.coli ATCC 25922, E.coli ML-35ATCC 43837, K.pneumonia ATCC 700603, P.aeruginosa FADDI-PA070, P.aeruginosa ATCC 27853, S.aureus ATCC 25923, S.aureusATCC 43300, B.subtilis ATCC, 23857 Methicillin-resistant S.aureus 936 was cultured in LB medium, NB medium or MHB medium at 37°C for overnight recovery. The recovered bacterial solution was diluted 40 times in fresh MHB medium and incubated at 37°C for 1.5-2 hours. The working solution of lipopeptide was prepared from the storage solution (vancomycin and colistin sulfate were used as antibiotic control), the concentration gradient was set to 2 times, and the concentration was from 0.015 to 64 μg / mL, in duplicate. The working solution was added to the 96-well plate, and 5 parallels wer...

experiment example 2

[0055] Experimental example 2: Hemolytic evaluation of lipopeptide

[0056] Rabbit red blood cells were separated from rabbit blood, washed with phosphate buffered saline (PBS, pH 7.4), and centrifuged at 4000 rpm for 4 to 5 times. Then, erythrocytes were dispersed in 0.25% (v / v) phosphate buffered saline solution. Prepare stock solutions of melittin and lipopeptide at 512 μM. The lipopeptide stock solution was diluted 2-fold with PBS to obtain working solutions with eight concentration gradients. Subsequently, an equal volume of red blood cell suspension and different concentrations of lipopeptide working solutions were added to the 96-well plate, and incubated at 37°C for 1 hour. After the 96-well plate is centrifuged, carefully transfer the centrifuged supernatant to a new well plate, and set it as a positive control in 490nm Triton X-100 solution (1%, v / v 20μL) to represent 100% cell hemolysis, red blood cells PBS solution (20 μL) was used as a negative control group. ...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention discloses lipopeptide based on melittin and a preparation method and application thereof, and belongs to the field of biological medicine. The lipopeptide is obtained by coupling C10-18 fatty acid at the N end of melittin and performing amidation modification on the C end of melittin. Experiments show that compared with parent melittin, the lipopeptide has a better inhibition effect on gram-positive bacteria and gram-negative bacteria, and has lower hemolytic activity. In an in-vitro stability test, the lipopeptide shows better stability in pepsin, trypsin and serum, is the lipopeptide with great application value, can be used for preparing a novel anti-bacterial infection medicine and the like, and has a very wide application prospect.

Description

technical field [0001] The invention belongs to the field of lipopeptide preparation and relates to the preparation and application of a lipopeptide, in particular to a melittin-based lipopeptide and its preparation method and application. Background technique [0002] Bacterial resistance to traditional antibiotics is a serious threat to human health around the world. Therefore, it is an urgent task in antibacterial research to develop a new class of antibacterial drugs that are not prone to drug resistance. Antimicrobial peptides (AMPs) are naturally occurring polypeptides with broad-spectrum antibacterial functions. Due to their antibacterial mechanism of action different from antibiotics, AMPs are not easy to produce drug resistance and have attracted widespread attention in the industry. [0003] Melittin is a kind of antibacterial peptide, which is the main active ingredient of bee venom and has strong antibacterial activity. The primary structure of melittin is GIGA...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
IPC IPC(8): C07K14/435C07K1/107A61K38/17A61P31/04A01N37/46A01N47/44A01P1/00
CPCC07K14/43572A61P31/04A01N37/46A01N47/44A61K38/00Y02A50/30
Inventor 黄升苏国旗邓余江山陈利李成洪黄金秀付利芝杨飞云
Owner CHONGQING ACAD OF ANIMAL SCI
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com