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Ultra-short peptides promoting collagen secretion and application thereof

A collagen-secreting and ultra-short technology, applied in the field of biomedicine, can solve the problems of inability to penetrate the stratum corneum of the skin and act on dermal fibroblasts, the molecular weight of collagen is large, and immune rejection, etc., achieving no hemolytic activity, Low cytotoxicity and the effect of promoting collagen secretion activity

Inactive Publication Date: 2019-01-15
DALIAN UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

This patented technology provides compounds that promote both cellular growth factors like Collagen (which makes up skin) production and antioxidants such as Lutathione peroxydase (LPO), which helps prevent damage from UV light exposure). These compound also enhance other substances involved with enzyme function called Peroxisome Proliferator Activated Receptor-2 α(PPARα)) through their ability to stimulate PGC-1 alpha release during inflammations caused by oxidiative stressors. Overall these technical effectings make this new ingredient useful for various applications related to promotion of body tissue regeneration processes.

Problems solved by technology

This patented technical issue addressed in this patents relating to improving skin quality through prevention from damage due to sunlight exposure during physical exercise. Anti-aged skins lose moisturizing ability overtime causing dryness symptoms like crackling around weatherings. To address these issues, researchers focused on developing new ways to enhance skin's resistance against stress and oxidization without compromising the integrity of the underlying layers of the living environment.

Method used

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  • Ultra-short peptides promoting collagen secretion and application thereof
  • Ultra-short peptides promoting collagen secretion and application thereof
  • Ultra-short peptides promoting collagen secretion and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0017] Chemical synthesis of two ultrashort peptides, Magin-FL and Purin-WH3, which promote collagen secretion:

[0018] (1) Chemical synthesis of two ultra-short peptides, Magin-FL and Purin-WH3, which promote collagen secretion: according to the amino acid sequence of the ultra-short peptide, use an automatic peptide synthesizer (433A, Applied Biosystems) to synthesize its full sequence, and reverse phase by HPLC Column chromatography desalting.

[0019] (2) Molecular weight was determined by matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF).

[0020] (3) The purity of the purified two collagen-promoting ultrashort peptides Magin-FL and Purin-WH3 was identified by high-performance liquid chromatography (HPLC), and the molecular weight was determined by matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF). The isoelectric point was determined by focusing electrophoresis, and the amino acid sequence st...

Embodiment 2

[0024] Hemolytic activity assay:

[0025] The collected healthy human blood was mixed with Alfred's solution for anticoagulation, washed twice with normal saline and resuspended into 10 7 -10 8 cell / ml suspension. The above-mentioned diluted erythrocyte suspension was mixed with the ultrashort peptide sample dissolved in normal saline, incubated at 37°C for 30 minutes, then centrifuged at 1000rpm for 5 minutes, and the supernatant was measured at 540nm for absorbance. Negative control uses physiological saline, positive control uses Triton X-100, and the hemolysis percentage is calculated according to the following formula: hemolysis percentage H%=(A sample-A negative control) / A positive control×100%.

[0026] Table 1. Hemolytic activity of two ultrashort peptides

[0027]

[0028] The results showed that the percentage of hemolysis was 0.41% and 1.62% when the concentration of the two collagen-promoting ultrashort peptides, Magin-FL and Purin-WH3, was 160 μg / ml. It sho...

Embodiment 3

[0030] Cytotoxicity Assay:

[0031] The toxicity of this group of ultrashort peptides to human skin fibroblast HFF-1 was detected by MTT method.

[0032] Human skin fibroblasts HFF-1 were purchased from Shanghai Cell Bank. First culture fibroblasts in DMEM containing 15% fetal bovine serum and double antibodies (penicillin and streptomycin each 100 U / ml), after the cells are congested, digest them with 0.25% trypsin, wash with the above medium Twice, resuspend the cells, and after cell counting, add 100 μl of the cell suspension to a 96-well cell culture plate, so that the number of cells per well reaches 10 5 indivual. Add the sample, and add the same volume of sterilized ultrapure water to the control group, set at 37°C, 5% CO 2 Cultivate in the incubator for 24h. After the culture was over, add 20 μl of 5 mg / ml MTT solution (prepared with cell culture PBS buffer) to each well of the 96-well cell culture plate, continue to cultivate for 5 hours, suck out the liquid in th...

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Abstract

Belonging to the field of biomedical technology, the invention provides ultra-short peptides promoting collagen secretion and application thereof. The two ultra-short peptides promoting collagen secretion Magin-FL and Purin-WH3 have the ability of promoting collagen synthesis, also have the characteristics of extremely strong antioxidant activity, anti-aging activity, no cytotoxicity, no hemolyticactivity, simple preparation method, etc. Therefore, the ultra-short peptides can be used for preparation of drugs treating the repair and regeneration of damaged epidermis, like burns, scalds, skinulcer and the like, replacement of EGF in anti-aging and anti-oxidant cosmetic and skin care product new raw materials for skin repair and stimulation of skin collagen proliferation, and other fields,and the functions of the ultra-short peptides are probably not limited to the above. The ultra-short peptides Magin-FL and Purin-WH3 have small molecular weight and simple structure, and are in low cost by chemical synthesis, and have outstanding advantages compared with EGF costs nearly 2 million dollars per gram.

Description

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Claims

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Application Information

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Owner DALIAN UNIV OF TECH
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