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Micromolecule polypeptide RK12 and application thereof

A technology of small molecule polypeptide, RK12, applied in the field of biomedicine, can solve the problems of complex antibacterial peptide antibacterial mechanism, leakage of bacterial content, permeability change, etc., and achieve good market prospects, small molecular weight, and convenient artificial synthesis.

Active Publication Date: 2019-12-24
佩德生物科技(南通)有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The antibacterial mechanism of antimicrobial peptides is complex, but most of the theories believe that the mechanism involves the interaction between the cationic and hydrophobic properties of antimicrobial peptides and negatively charged microbial cell membranes. After the antimicrobial peptides come into contact with bacterial cell membranes, they cause changes in membrane permeability. , or form a transmembrane hole in the bacterial cell membrane, and finally cause the bacterial content to leak out and die

Method used

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  • Micromolecule polypeptide RK12 and application thereof
  • Micromolecule polypeptide RK12 and application thereof
  • Micromolecule polypeptide RK12 and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0017] The preparation of the small molecule polypeptide RK12 adopts the Fmoc solid-phase synthesis method, and the specific steps are as follows.

[0018] 1. Weigh 0.2g of the resin and place it in a dry and clean reaction tube, add an appropriate amount of N,N-dimethylamide (DMF), activate for 30 minutes, add 1mmol of the first amino acid residue and 150mg of DMAP to the reaction In the tube, react with DMF as a solvent for 3 hours; after the reaction, wash with DMF for 3 to 6 times, add appropriate pyridine and acetic anhydride at a volume ratio of 1:1, and react for 30 minutes; after the reaction, wash with DMF for 3 to 6 times; Then, piperidine was used to elute the protective group Fmoc of the amino acid, twice for 15 min each, followed by four washes with DMF and two washes with methanol.

[0019] 2. Weigh 3mmol of the second amino acid and 3mmol of HBTU into a reaction tube, add 0.5ml of DIEA, react for 40 minutes, wash with DMF for 3 to 6 times, add piperidine solutio...

Embodiment 2

[0023] Antibacterial Experiment of Small Molecule Peptide RK12

[0024] 1. Prepare Acinetobacter baumannii bacteria liquid, incubate at 37°C for 18 hours, and set aside;

[0025] 2. Prepare a small molecule polypeptide RK12 solution with a concentration of 10mg / ml, sterilize and dry a circular qualitative filter with a diameter of 5-7mm, and then immerse it in the above-mentioned small molecule polypeptide RK12 solution with different concentrations;

[0026] 3. Prepare LB medium, sterilize, and reserve;

[0027] 4. Melt LB medium, cool to 50°C, add 1ml of Acinetobacter baumannii bacteria solution, shake evenly, pour into a sterile plate, shake gently to spread the medium evenly on the plate;

[0028] 5. Use sterilized tweezers to neatly and orderly put the filter into the cooled plate, cover with a terracotta cover, mark it, and place it in a constant temperature incubator at 37°C for 24 hours;

[0029] 6. Use a caliper to measure the size of the inhibition zone, and compar...

Embodiment 3

[0035] Therapeutic Effect of Small Molecule Peptide RK12 in Animal Model

[0036] 1) Model construction

[0037] Cultivate Acinetobacter baumannii with LB medium to the logarithmic phase, then centrifuge at 3000rpm for 5min, wash 3 times with sterile normal saline, and finally resuspend with normal saline, and prepare 1×10 7 CFU / ml bacterial suspension, put in a 4°C refrigerator for later use.

[0038] The animal model of systemic infection with Acinetobacter baumannii was selected from male Kunming mice, which were divided into 7 groups, 6 mice in each group, and each group was injected with 1×10 7 The suspension of Acinetobacter baumannii of CFU / ml is grouped as follows: 1. Normal saline control group, RK12 (1,2,4mg / kg) and polymyxin E (0.25,0.5,1mg / kg) treatment group . Before inoculation, the immunosuppressive mouse model should be constructed first, and the construction method is as follows:

[0039] Prepare 20mg / ml cyclophosphamide solution with sterile normal saline...

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Abstract

The invention discloses micromolecule polypeptide RK12. The molecular weight of the micromolecule polypeptide RK12 is 1901.30Da, the isoelectric point of the micromolecule polypeptide RK12 is 12.02, and the amino acid sequence of the micromolecule polypeptide RK12 is as shown in SEQ ID: 1. The invention also discloses an application of the micromolecule polypeptide RK12 to preparation of medicinesfor treating acinetobacter baumannii infection. The polypeptide RK12 disclosed by the invention is small in molecular weight, convenient to artificially synthesize and obvious in bacteriostasis effects on acinetobacter baumannii.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and in particular relates to a small molecular polypeptide RK12 and its application in the preparation of anti-acinetobacter baumannii infection drugs. Background technique [0002] Antimicrobial peptides are small molecule polypeptides with biological activity that are induced in organisms, with a molecular weight of about 1000-7000 and composed of 10-60 amino acid residues. Antimicrobial peptides are high-efficiency, spectral antimicrobial peptide molecules that are rapidly produced by the body after being invaded by microorganisms to participate in the body's immune response. Antimicrobial peptides are widely used as effective defense molecules in the body. Thousands of antimicrobial peptides have been identified from microorganisms, plants, insects, arthropods, amphibians, mammals and even humans. The antibacterial mechanism of antimicrobial peptides is complex, but most of the theories ...

Claims

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Application Information

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IPC IPC(8): C07K7/08A61K38/10A61P31/04
CPCC07K7/08A61P31/04A61K38/00
Inventor 谷徏新
Owner 佩德生物科技(南通)有限公司
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