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Self-assembled dendritic antibacterial peptide Pal3RP, preparation method thereof, self-assembled nano-particles thereof and application of self-assembled dendritic antibacterial peptide Pal3RP and self-assembled nano-particles thereof

A nanoparticle and antimicrobial peptide technology, which is applied in the field of self-assembled dendritic antimicrobial peptide Pal3RP and its preparation, can solve the problems of seldom explored antimicrobial functions, weak peptide resistance to enzymatic hydrolysis, etc., and achieve low hemolytic toxicity and high inhibitory effect , the effect of improving stability

Pending Publication Date: 2022-03-01
NORTHEAST AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] However, the existing self-assembling peptides are mostly used in targeted drugs, gene carriers and immunodiagnostics, and their functions as antibacterial agents are rarely explored
Moreover, self-assembly endows the polypeptide with weak resistance to enzymatic hydrolysis, so the design and preparation of new self-assembled nano-antibacterial polypeptides is imperative

Method used

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  • Self-assembled dendritic antibacterial peptide Pal3RP, preparation method thereof, self-assembled nano-particles thereof and application of self-assembled dendritic antibacterial peptide Pal3RP and self-assembled nano-particles thereof
  • Self-assembled dendritic antibacterial peptide Pal3RP, preparation method thereof, self-assembled nano-particles thereof and application of self-assembled dendritic antibacterial peptide Pal3RP and self-assembled nano-particles thereof
  • Self-assembled dendritic antibacterial peptide Pal3RP, preparation method thereof, self-assembled nano-particles thereof and application of self-assembled dendritic antibacterial peptide Pal3RP and self-assembled nano-particles thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Design of Dendritic Antimicrobial Peptides:

[0022] The amino acid sequence of the dendritic antimicrobial peptide Pal3RP is:

[0023] Pal-GlyGlyGlyLys(ArgProArgProArgPro)Lys(ArgProArgProArgPro)ArgProArgProArgPro

[0024] Using two lysine Lys amino groups and two -NH on the R group 2 As linking sites of the main chain and branch chains of dendrimers respectively, three cationic anti-enzyme polypeptide units (ArgProArgProArgPro) are linked together to design a dendritic antimicrobial peptide with one main chain and two side chains. As shown in the figure, the dendritic structure forms the steric hindrance between the peptide chains, and further uses the flexible amino acid linker GGG to link the dendritic antimicrobial peptide with a n-hexadecanoic acid to provide hydrophobicity and antibacterial properties for it to exert antibacterial activity. The hydrophobic driving force of self-assembly named the dendritic antimicrobial peptide Pal3RP, and its sequence was Pal-G...

Embodiment 2

[0031] Synthesize the above-mentioned dendritic antimicrobial peptides using a peptide synthesizer. The method is a solid-phase chemical synthesis method. The specific steps are:

[0032] 1. The preparation of the polypeptide main chain is carried out one by one from the C-terminal to the N-terminal, and is completed by a polypeptide synthesizer. First, Fmoc-X (X is the first amino acid at the C-terminal of each antimicrobial peptide) is inserted into Wang resin, and then the Fmoc group is removed to obtain X-Wang resin; then Fmoc-Y-Trt-OH (9 -Fmoxy-trimethyl-Y, Y is the second amino acid at the C-terminus of each antimicrobial peptide); according to this procedure, it is synthesized from the C-terminus to the N-terminus until the synthesis is completed, and the side of the Fmoc group is removed chain protection resin;

[0033] 2. Use hydrazine hydrate to remove the Fmoc-Lys(Dde)-OH side chain Dde protecting group, and repeat step 1 to complete the branched-chain amino acid l...

Embodiment 3

[0038] The above-mentioned dendritic antimicrobial peptides are prepared by ultrasonically standing for self-installation of nanoparticles, and the specific steps are:

[0039] 1. Preparation of self-assembled nanoparticles: the dendrimer antimicrobial peptide Pal3RP was dissolved in 10 mM phosphate buffer solution (pH=7.4), with a final concentration of 256 μM. After sonicating for 5 minutes at 28°C, it was placed at 4°C for more than 18 hours to form self-assembled nanoparticles.

[0040] 2. Identification of the self-assembled nanoparticles: the nanoparticles prepared above were observed by a scanning electron microscope (SEM). as attached image 3 and 4 As shown, the dendritic antimicrobial peptide Pal3RP self-assembles into nanoparticles with a diameter of about 20 nm in phosphate buffer.

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Abstract

The invention provides a self-assembled dendritic antibacterial peptide Pal3RP and a preparation method thereof as well as self-assembled nano-particles and application of the self-assembled dendritic antibacterial peptide Pal3RP. The sequence of the gene is Pal-GGGK (RPRPRP) K (RPRPRP) RPRPRP, and Pal is n-hexadecanoic acid. The structural particularity of lysine as branched amino acid is utilized, and meanwhile, a cationic anti-enzymolysis unit and hexadecanoic acid are utilized to form a dendritic structure and steric hindrance between peptide chains, so that the antibacterial peptide has high antibacterial activity, low toxicity and high stability. Further, the dendritic antibacterial peptide is self-assembled into the self-assembled antibacterial peptide nano-particles with the micro-nano size through an ultrasonic standing method. The self-assembled dendritic antibacterial peptide Pal3RP nanoparticles have an obvious inhibition effect on gram negative bacteria and gram positive bacteria, have an efficient inhibition effect on multiple drug-resistant bacteria, and have very low hemolytic toxicity at the same time; the degradation degree of high-concentration protease on the dendritic antibacterial peptide Pal3RP nanoparticles is very weak.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a self-assembled dendritic antibacterial peptide Pal3RP, a preparation method thereof, self-assembled nanoparticles and applications. Background technique [0002] The growing prevalence of drug-resistant bacterial infections, especially Gram-negative drug-resistant infections, is recognized as one of the greatest threats to global public health. In recent years, most small-molecule antibiotics have shown weak therapeutic effects, and infections caused by antibiotic-resistant bacteria require longer treatment courses and higher mortality rates. Therefore, there is an urgent need to develop new antibacterial drugs that can circumvent existing bacterial resistance mechanisms. Antimicrobial peptides (AMPs) have been proposed as an attractive and promising strategy to combat powerful drug-resistant bacterial infections. Unlike the mechanism of action of most antibiotics tha...

Claims

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Application Information

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IPC IPC(8): C07K14/00B82Y5/00B82Y40/00A61K38/16A61K9/14A61P31/04
CPCC07K14/001B82Y5/00B82Y40/00A61K9/14A61P31/04A61K38/00Y02A50/30
Inventor 单安山来振衡陈虹羽
Owner NORTHEAST AGRICULTURAL UNIVERSITY
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