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A class of ultrashort peptides for promoting collagen secretion and its application

A collagen secretion, ultra-short technology, applied in the field of biomedicine, can solve the problems that it cannot penetrate the stratum corneum of the skin and act on the fibroblasts of the dermis, the molecular weight of collagen is large, immune rejection, etc., and achieve no hemolytic activity, Low cytotoxicity, collagen secretion promoting effect

Active Publication Date: 2019-10-11
择天生物科技(大连)有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the collagen-related products on the market all have the main function of directly supplementing collagen. However, the molecular weight of collagen is large, and the application method can hardly make it penetrate the stratum corneum of the skin and act on the fibroblasts of the dermis; The injection method directly supplements collagen, because it will be degraded in the body, multiple injections are required, the process is complicated and the cost is high, and the immune rejection caused by injecting collagen at the same time is a big risk; if it is eaten, collagen will be in the intestinal tract It is decomposed into small molecular polypeptides or single amino acids by digestive juice, so the actual effect of supplementing collagen is limited

Method used

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  • A class of ultrashort peptides for promoting collagen secretion and its application
  • A class of ultrashort peptides for promoting collagen secretion and its application
  • A class of ultrashort peptides for promoting collagen secretion and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] Chemical synthesis of two ultrashort peptides, Magin-FL and Purin-WH3, which promote collagen secretion:

[0019] (μg1 / )ml Chemical synthesis of two ultrashort peptides, Magin-FL and Purin-WH3, which promote collagen secretion: According to the amino acid sequence of the ultrashort peptide, use an automatic peptide synthesizer (433A, Applied Biosystems) to synthesize its full sequence, and use HPLC Desalted by reverse phase column chromatography.

[0020] (2) Molecular weight was determined by matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF).

[0021] (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 seque...

Embodiment 2

[0025] Hemolytic activity assay:

[0026] 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 for absorbance at 540nm. 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%.

[0027] Table 1. Hemolytic activity of two ultrashort peptides

[0028]

[0029] 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

[0031] Cytotoxicity Assay:

[0032] The toxicity of this group of ultrashort peptides to human skin fibroblast HFF-1 was detected by MTT method.

[0033] Human skin fibroblasts HFF-1 were purchased from Shanghai Cell Bank. First culture the 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, and wash Twice, resuspend the cells, and after counting the cells, 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, 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 the wells with ...

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Abstract

The invention provides collagen secretion promotion ultrashort peptides and application thereof, and belongs to the technical field of biomedicine. The two kinds of collagen secretion promotion ultrashort peptides Magin-FL and Purin-WH3 have the capability of promoting the collagen synthesis; in addition, the characteristics of ultra-high oxidization resistance, aging-resistant activity, no cell toxicity, no hemolysis activity, simple preparation method and the like are realized, so that the collagen secretion promotion ultrashort peptides are used for preparing medicine for treating injured skin restoration and regeneration such as medicine for treating burnt injury, scald injury and skin ulceration, and can replace EGF to be used in the fields of anti-aging and anti-oxidization beatification skin care product new raw materials for skin restoration and skin collagen proliferation simulation and the like. In addition, the effects are possibly not limited to the field. The ultrashort peptides Magin-FL and Purin-WH3 have small molecular weight and simple structure; the used chemical synthesis method has very low cost; the ultrashort peptides Magin-FL and Purin-WH3 have the obvious beneficial characteristics through being compared with EFG with the sale price being nearly 2 million U.S dollars per gram.

Description

technical field [0001] The invention relates to a class of collagen secretion-promoting ultrashort peptides and applications thereof, belonging to the technical field of biomedicine. Background technique [0002] Aging is an inevitable law of any biological life process. With the increasingly prominent problem of population aging worldwide, the field of anti-aging has become a global focus. The skin is one of the most obvious organs in the aging process of the body. With age, the skin will undergo degenerative changes, such as loss of elasticity, loss of water content, atrophy, and wrinkles. [0003] Skin aging is a series of complex biological processes in skin tissue, which is the result of the joint action of all endogenous physiological factors and exogenous environmental factors. Aging will not only affect the normal skin tissue structure and physiological functions, but also directly affect the appearance and appearance of the outer layer of the skin. Skin is compose...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K7/06A61K38/08A61K8/64A61Q19/08A61Q19/00A61P17/00
Inventor 于海宁王慧王义鹏
Owner 择天生物科技(大连)有限公司
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