Human alpha-defensin-5 modifier and applications thereof

A defensin, amino acid technology, applied in animal/human peptides, animal/human proteins, medical preparations containing active ingredients, etc., can solve the problems of reducing antibacterial activity, the impact of antibacterial activity, etc. effect, the effect of improving salt tolerance

Active Publication Date: 2016-11-23
ZHEJIANG LONGTERM MEDICAL TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Although the extraction and artificial synthesis of HD-5 are relatively mature at present, the antibacterial activity of HD-5, which is completely bionic and natural, is easily affected by many environmental factors in the body, such as the presence of physiological concentrations of sodium chloride can greatly reduce its antibacterial activity. Antibacterial activity, which restricts it from becoming a new type of antibacterial drug suitable for industrial development, production and clinical application

Method used

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  • Human alpha-defensin-5 modifier and applications thereof
  • Human alpha-defensin-5 modifier and applications thereof
  • Human alpha-defensin-5 modifier and applications thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] The amino acid sequence of HD-5 is shown in SEQ ID NO.1, specifically:

[0038] Ala 1 -Thr 2 -Cys 3 -Tyr 4 -Cys 5 -Arg 6 -Thr 7 -Gly 8 -Arg 9 -Cys 10 -Ala 11 -Thr 12 -Arg 13 -Glu 14 -Ser 15 -Leu 16 -Ser 17 -Gly 18 -Val 19 -Cys 20 -Glu 21 -Ile 22 -Ser 23 -Gly 24 -Arg 25 -Leu 26 -Tyr 27 -Arg 28 -Leu 29 -Cys 30 -Cys 31 -Arg 32 , where Cys 3 and Cys 31 、Cys 5 and Cys 20 、Cys 10 and Cys 30 Intramolecular disulfide bonds are formed between them.

[0039] The modified HD-5Myr and HD-5Lau of the present invention are obtained by modifying the C-terminus of the HD-5 molecule. The amino acid sequences of HD-5Myr and HD-5Lau are shown in SEQ ID NO.2.

[0040] HD-5Myr consists of 34 amino acid residues with a molecular weight of 3976.8Da.

[0041] The amino acid sequence of HD-5Myr is:

[0042] Ala 1 -Thr 2 -Cys 3 -Tyr 4 -Cys 5 -Arg 6 -Thr 7 -Gly 8 -Arg 9 -Cys 10 -Ala 11 -Thr 12 -Arg 13 -Glu 14 -Ser 15 -Leu 16 -Ser 17 -Gly 1...

Embodiment 2

[0050] The HD-5 and HD-5Myr in the above-mentioned Example 1 were synthesized by "Shanghai Gill Polypeptide Co., Ltd.". The HD-5Lau and MyrHD-5 of the above-mentioned Example 1 were synthesized by "Hangzhou Zhongpei Biochemical Co., Ltd.".

Embodiment 3

[0052] The antibacterial activity of the antimicrobial peptide was measured, and the strength of the antibacterial effect of the HD-5 modified body HD-5Myr, HD-5Lau and MyrHD-5 synthesized in Example 2 was compared with that of the natural HD-5.

[0053] The experiment was carried out according to the method for detecting antibacterial peptide antibacterial activity reported by Ericksen et al. in 2005 (Ericksen B, et al., Antibacterial activity and specificity of the six human{alpha}-defensins. Antimicrob Agents Chemother, 2005).

[0054] Bacteria were inoculated on LB agar plates and cultured at a constant temperature of 37°C until colonies grew. Pick a single colony, inoculate it into 5ml LB medium, and incubate at a constant temperature of 37°C for 4-6 hours with shaking. Wash 2 times with sterile water. Measure the absorbance at 600nm on a multifunctional microplate reader, 1OD 600 =4×10 8 CFU / ml, the test bacteria were diluted to 2×10 with sterile water 6 CFU / ml.

[...

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Abstract

The invention discloses a human alpha-defensin-5 modifier and applications thereof, and belongs to the technical field of biology. The modifier is obtained in the mode that amino acid residues are added at the C tail end of human alpha-defensin-5, and fatty acid is modified and connected to the amino acid residues for modifying; at least one amino acid residue has a free amino group in the added amino acid residues, and the fatty acid with 8-24 carbon atoms is modified and connected on the free amino group. The invention further discloses the applications of the human alpha-defensin-5 modifier to preparing antibacterial medicine and immune adjustment medicine. According to the human alpha-defensin-5 modifier and the applications thereof, the C tail end of the HD-5 extends and is modified, and the antibacterial activity is remarkably enhanced; particularly, Lys extending at the C tail end of the HD-5 is subjected to myristic acid modification, the salt resistance of the modifier can be obviously improved, the limitation of the sensibility of traditional antibacterial peptide salt is broken through, and the human alpha-defensin-5 modifier has important clinical application prospects; macrophage can be remarkably allowed to release inflammatory factors through the modifier, and the human alpha-defensin-5 modifier can be applied to preparing of the immune adjustment medicine.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a modified human α-defensin 5 and its application. Background technique [0002] Antibiotic resistance is a major scientific issue threatening human health. The emergence of broad-spectrum or even ultra-broad-spectrum drug-resistant bacteria such as methicillin-resistant Staphylococcus aureus (MRSA), pan-drug-resistant Acinetobacter baumannii, and carbapenem-resistant Pseudomonas aeruginosa makes the treatment of some infections quite effective. become an important risk factor for death in patients with these infections. At present, the mutation rate of drug-resistant bacteria is faster than the development speed of new antibacterial drugs. Human beings are helpless against some drug-resistant bacterial infections and are facing the severe challenge of the "post-antibiotic era". Therefore, it is imminent to search for new antibacterial drugs, especially new drugs with bactericidal ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/47A61K38/17A61P31/00A61P37/02
CPCC07K14/4723A61K38/00A61P31/00A61P37/02C07K14/435
Inventor 方向明雷如意陈齐兴侯金超程宝莉金悦
Owner ZHEJIANG LONGTERM MEDICAL TECH CO LTD
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