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Amyloid hexapeptide and its application in broad-spectrum inhibition of bacterial and fungal biofilms

An amyloid and biofilm technology, which is applied in the direction of antibacterial drugs, antifungal agents, peptides, etc., can solve the problems of long length of peptides, inability to kill, and difficulty in in vitro synthesis, and achieves low cytotoxicity and will not cause bacterial resistance. The effect of medicine

Active Publication Date: 2021-03-05
HOSPITAL OF STOMATOLOGY SUN YAT SEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, most of the peptide inhibitors currently studied, including antibacterial peptides, have a bactericidal effect, and reduce the formation of bacterial biofilms by reducing the number of bacteria. They have low efficiency and cannot kill the "dormant bacteria" inside the biofilm, which may lead to bacterial resistance. drug and other defects
Therefore, current antimicrobial peptides are not ideal antibiofilm agents
[0004] In 2012, Chu et al. reported for the first time in "Science" that the defense peptide 6 (composed of 32 amino acids) secreted by human small intestinal Paneth cells inhibits the formation of biofilm by forming amyloid fibers to wrap bacteria, and defense peptide 6 has no bactericidal effect
However, the length of the above two polypeptides is relatively long, it is difficult to synthesize in vitro, and the cost is high, and the amyloid polypeptide Aβ belongs to the pathogenic protein of Alzheimer's disease, so it is not suitable for clinical application

Method used

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  • Amyloid hexapeptide and its application in broad-spectrum inhibition of bacterial and fungal biofilms
  • Amyloid hexapeptide and its application in broad-spectrum inhibition of bacterial and fungal biofilms
  • Amyloid hexapeptide and its application in broad-spectrum inhibition of bacterial and fungal biofilms

Examples

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Embodiment 1

[0028] This example is five amyloid hexapeptides ASNIVI, TSFVLV, GIDLKI, FAKVDI, and IAVGTF that can be polymerized into amyloid fibers in vitro according to the present invention. Through the prediction of amyloid fibril prediction software, ZipperDB, Tango and Waltz three amyloid fibril prediction software were used to predict the short peptide segments in the C123 sequence that could form amyloid fibrils. The short amyloid peptides to be screened must meet the following requirements: at least two softwares point to each other, and the binding energy corresponding to ZipperDB <-23kcal / mol. And observed by transmission electron microscope in vitro, confirmed that it can be aggregated into amyloid fibrils.

[0029] The specific prediction method is: log in to Pubmed, search for protein, enter streptococcus mutans, Pac, download the amino acid sequence information, and screen out the C123 sequence (1000-1486aa). Log in to ZipperDB (http: / / services.mbi.ucla.edu / zipperdb), Tango...

experiment example 1

[0036] This experimental example is the verification of whether the hexapeptides numbered P1-P13 in Table 1 of the present invention can form amyloid fibrils and the verification of the hexapeptides that can form amyloid fibrils inhibit the formation of biofilms but do not kill bacteria.

[0037]Materials: (1) Storage and use of hexapeptides, hexapeptides are stored in the form of lyophilized powder for a long time, 1 mg / tube in aliquots. Take out 1 mg each time, first dissolve it with 100ul of DMSO, add 900ul of double distilled water, adjust the concentration to 1mg / ml, and store it temporarily in a -20°C refrigerator as a mother solution. When in use, the mother liquor is added to the culture medium and diluted to different concentrations (0.1mg / ml, 0.05mg / ml, 0.025mg / ml and 0.0125mg / ml), and the control group is the medium containing the same concentration of DMSO without adding amyloid hexapeptide , for the cultivation of free bacteria and biofilms. (2) Bacterial culture...

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Abstract

The invention relates to amyloid hexapeptide and its broad-spectrum application of inhibiting bacteria and fungi biofilm, and belongs to the field of biopharmaceuticals. The amyloid hexapeptide provided by the invention can be polymerized into amyloid fibrils in vitro. The present invention also provides the application of the amyloid hexapeptide in the preparation of medicines for inhibiting the formation of biofilms. The amyloid hexapeptide of the present invention can wrap bacteria or fungi by being polymerized into amyloid fibers, and play a broad-spectrum role in inhibiting the formation of bacteria and fungal biofilms, including Gram-positive bacteria, Gram-negative bacteria and fungi, but Amyloid hexapeptide is not bactericidal, has little or no toxicity to cells, and will not cause bacterial drug resistance. It is expected to become a new type of cheap drug with high efficiency against biofilm formation.

Description

technical field [0001] The invention relates to the application of amyloid hexapeptide and its broad-spectrum inhibition of bacterial and fungal biofilms, and belongs to the field of biopharmaceuticals. Background technique [0002] About 60% of human bacterial diseases are caused by biofilms, and bacteria greatly enhance their resistance to the external environment after forming biofilms. At present, the drugs that can be used clinically to control biofilm are all fungicides, including chlorhexidine, antibiotics, etc. Long-term use of fungicides can cause imbalance of flora in the body, make bacteria resistant to drugs, and lead to systemic diseases. And there is no effective anti-biofilm but not bactericidal medicine at present, therefore, it is waiting to find a new anti-biofilm but not bactericidal medicine. [0003] Peptides have the characteristics of simple synthesis, low toxicity, fast drug effect and low immunogenicity, and have received more and more attention in ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/315A61K38/16A61P31/04A61P31/10
CPCA61K38/00A61P31/04A61P31/10C07K14/315Y02A50/30
Inventor 林焕彩陈冬茹
Owner HOSPITAL OF STOMATOLOGY SUN YAT SEN UNIV
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