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A Class of Ultrashort Peptides for Promoting Collagen Secretion and Its Application

An ultra-short peptide and glue-promoting technology, applied in the field of biomedicine, can solve the problems of large molecular weight of collagen and limited effect of supplementing collagen, and achieve the effect of low cytotoxicity, no hemolytic activity, and small molecular weight

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 replenishes collagen, so the effect of actual collagen replenishment 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

[0020] Chemical synthesis of anti-aging ultrashort peptide Mapin-WH, Tinin-WH, Orin-G-OT:

[0021] (1) Chemical synthesis method of anti-aging ultrashort peptide Mapin-WH, Tinin-WH, Orin-G-OT: According to the amino acid sequence of the ultrashort peptide, use an automatic peptide synthesizer (433A, Applied Biosystems) to synthesize its full sequence, and pass HPLC reverse phase column chromatography desalting.

[0022] (2) Molecular weight was determined by matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF).

[0023] (3) Three kinds of purified anti-aging ultrashort peptides Mapin-WH, Tinin-WH, Orin-G-OT were identified by high-performance liquid chromatography (HPLC) for their purity, 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 isoelectric focusing electrophoresis, and the amino acid sequence structure was de...

Embodiment 2

[0028] Hemolytic Assay:

[0029] 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%.

[0030] Table 1. Hemolytic activity of three ultrashort peptides

[0031]

[0032] The results showed that the percentage of hemolysis of the three ultrashort peptides Mapin-WH, Tinin-WH and Orin-G-OT was less than 1% when the concentration was 160μg / ml. It shows that Mapin-WH, Ti...

Embodiment 3

[0034] Cytotoxicity Assay:

[0035] The toxicity of this group of ultrashort peptides to human skin fibroblast HFF-1 was detected by MTT method.

[0036] 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 2Cultivate 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 a...

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Abstract

The invention provides a collagen secretion promotion ultrashort peptide and application thereof, and belongs to the technical field of biomedicine. The ultrashort peptide comprises at least one kind of materials from ultrashort peptides Magin-WH, Tinin-WH and Orin-G-OT, has the capability of promoting the collagen synthesis, and has the characteristics of very strong oxidization resistance, anti-aging performance, small molecular weight, simple structure, very low cell toxicity, no hemolytic activity, simple preparation method and the like. The ultrashort peptideis 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. The ultrashort peptide has small molecular weight and simple structure; the used chemical synthesis method has very low cost; the ultrashort peptide has 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 ultrashort peptides for promoting collagen secretion 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 com...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K7/06C07K7/08A61K38/08A61P17/00A61K8/64A61Q19/08A61Q19/00
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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