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Small peptide against clinical multi-drug resistant bacteria

A technology for multidrug-resistant bacteria and drugs, which is applied in the direction of antibacterial drugs, antifungal agents, peptides, etc., can solve the problems of less toxicity research, low antibacterial activity, and unclear mechanism of action, and achieve environmental friendliness. Stable and specific effect

Active Publication Date: 2019-02-26
吴国球
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are few attempts to directly use natural small peptides as antibacterial drugs
The main reasons are: many natural small peptides have low antibacterial activity; the mechanism of action is still unclear; there are few toxicity studies

Method used

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  • Small peptide against clinical multi-drug resistant bacteria
  • Small peptide against clinical multi-drug resistant bacteria
  • Small peptide against clinical multi-drug resistant bacteria

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] Solid Phase Synthesis of Small Peptides Against Clinical Multidrug Resistant Bacteria

[0039] The amino acid sequence of each small peptide is as follows:

[0040] T1.

[0041] Gly-Ser-Lys-Lys-Pro-Val-Pro-Ile-Ile-Tyr-Cys-Gln-Arg-Arg-Thr-Gly-Lys-Cys-Gln-Arg-Met( 12 Asn→ 12 Gln); (GSKKPVPIIYCQRRTGKCQRM) see Seq NO: 1;

[0042] T2.

[0043] Gly-Ser-Lys-Lys-Pro-Val-Pro-Ile-Ile-Tyr-Cys-Asn-Arg-Arg-Ala-Gly-Lys-Cys-Gln-Arg-Met( 15 Thr → 15 Ala); (GSKKPVPIIYCNRRAGKCQRM) see Seq NO: 2;

[0044] T3.

[0045] Gly-Ser-Lys-Lys-Pro-Val-Pro-Ile-Ile-Tyr-Cys-Asn-Arg-Arg-Thr-Ala-Lys-Cys-Gln-Arg-Met( 16 Gly → 16 Ala); (GSKKPVPIIYCNRRTAKCQRM) see Seq NO: 3;

[0046] T4.

[0047] Gly-Ser-Lys-Lys-Pro-Val-Pro-Ile-Ile-Tyr-Cys-Gln-Arg-Arg-Ser-Gly-Lys-Cys-Gln-Arg-Met( 12 Asn→ 12 Gln, 15 Thr → 15 Ser); (GSKKPVPIIYCQRRSGKCQRM) see Seq NO: 4;

[0048] T5.

[0049] Gly-Ser-Lys-Lys-Pro-Val-Pro-Ile-Ile-Tyr-Cys-Gln-Arg-Arg-Thr-Ala-Lys-Cys-Gln-Arg-Met( 12 Asn→ 12 Gln, 16 Gly → 16 A...

Embodiment 2

[0067] Determination of antibacterial activity against clinical drug-resistant bacteria

[0068] 2.1 Source of strain

[0069]All clinical drug-resistant bacteria used in this experiment were isolated from the Bacteria Laboratory of Zhongda Hospital Affiliated to Southeast University. Among them, 4 strains of Escherichia coli, 4 strains of Klebsiella pneumoniae, 4 strains of Enterobacter cloacae, 6 strains of Enterobacter aerogenes, and Klebsiella ornithine 4 strains of Klebsiella ornithinolytica, 3 strains of Pseudomonas aeruginosa, 4 strains of Klebsiella oxytoca, 5 strains of Enterococcus faecium, 3 strains of Candida albicans strain. 所用标准菌株Escherichia coli ATCC 25922,Klebsiella pneumoniae ATCC 700603,Enterobacter cloacae ATCC13047,Enterobacteraerogenes ATCC 49701,Klebsiella ornithinolytica ATCC31898,Klebsiella oxytocaATCC 43086,Enterococcus faecium ATCC 29212,Pseudomonas aeruginosa ATCC 15442,Canidia Albicans ATCC10231由卫生部临检中心提供。

[0070] 2.2 Antibiotic susceptibility te...

Embodiment 3

[0092] Effects of Small Peptide Alone or Combined with Polymyxin on Mice with Pseudomonas Aeruginosa Septicemia

[0093] Establishment of mouse sepsis model: Pseudomonas aeruginosa standard strain (Pseudomonas aeruginosa) ATCC15442 and clinically isolated multidrug-resistant bacteria were cultured in MH medium to the logarithmic growth phase, centrifuged at 1000×g for 15min, discarded the supernatant, and used sterile Resuspended in normal saline to a concentration of 1×10 8 CFU / mL. The mice were anesthetized by intramuscular injection of ketamine 30mg / kg, and intraperitoneal injection of 0.2mL with a concentration of 2.0×10 8 CFU bacteria solution, observed for 72h. The grouping and administration were as follows (10 rats in each group): the control group, normal saline; the first group, 2mg / kg polymyxin; the second group, 2mg / kg master, and the third to fifth groups were respectively 2mg / kg T9, T10, T11; the sixth to eighth groups: 2mg / kg polymyxin and 2mg / kg small peptid...

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Abstract

The invention relates to a small peptide resisting MDR (multiple resistant bacteria). The small peptide is characterized in that the small peptide has the function of resisting clinically isolated MDR. In-vitro activity assay results prove that 11 small peptides have different antibacterial activities for multiple MDR and have the MIC value ranging from 4 mu g / L to 128 mu g / L according to different bacteria. Compared with a mother pattern, the small peptide has enhanced pseudomonas aeruginosa and candida albicans resistant activity. 6 peptides with better antibacterial activity are selected for an in vivo experiment, results prove that the 6 peptides can notably reduce the death rate of a model mouse because of a pseudomonas aeruginosa standard strain and clinical pan-drug-resistant pseudomonas aeruginosa septicemia and have a synergistic effect with polymyxin; no hemolytic reaction is caused when the concentration of 6 small peptides is 10 mg / ml, and the product safety is higher; local stimulation experiment results of rabbit eye conjunctiva show that the small peptide has no acute toxic stimulus reaction.

Description

technical field [0001] The invention belongs to the field of biomedicine, in particular to a group of small peptides against clinical multidrug-resistant bacteria. Background technique [0002] With the use of the first penicillin in the 1940s, the development of antibiotics is of great significance in the history of human medicine, saving countless lives and increasing human life expectancy by more than ten years. But with the widespread use of antibiotics, bacteria have gradually adapted and developed resistance to them. With the passage of time, bacteria that are clinically resistant to multiple antibiotics are becoming more and more common, and clinical multidrug-resistant bacteria (MDR) have emerged. According to data provided by the U.S. Centers for Disease Control (CDC), as early as 1980 to 1992, due to the emergence of drug-resistant bacteria, the average death rate from infectious diseases increased by 58%, making infectious diseases the second largest disease afte...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/00A61K38/16A61P31/04A61P31/10
CPCA61K38/00C07K14/00
Inventor 吴国球
Owner 吴国球
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