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Ultrashort peptide for promoting collagen secretion and application of ultrashort peptide

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 obvious clearance rate

Inactive Publication Date: 2019-01-18
DALIAN UNIV OF TECH
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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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  • Ultrashort peptide for promoting collagen secretion and application of ultrashort peptide
  • Ultrashort peptide for promoting collagen secretion and application of ultrashort peptide
  • Ultrashort peptide for promoting collagen secretion and application of ultrashort peptide

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

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

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

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

[0023] (3) The purified three anti-aging ultrashort peptides Mapin-WH, Tinin-WH and Orin-G-OT were identified by high performance liquid chromatography (HPLC), and their molecular weights were determined by matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI- TOF), isoelectric focusing electrophoresis was used to determine the isoelectric point, and an automatic amino acid sequencer was used to det...

Embodiment 2

[0028] Hemolytic assay:

[0029] The collected healthy human blood was mixed with Ashen's solution for anticoagulation, washed twice with normal saline and resuspended to 10 7 -10 8 cell / ml suspension. The above diluted red blood cell suspensions were mixed with ultrashort peptide samples dissolved in physiological saline, incubated at 37° C. for 30 min, and then centrifuged at 1000 rpm for 5 min, and the supernatant was measured for absorbance at 540 nm. Negative control uses normal 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 hemolysis percentages of the three ultrashort peptides Mapin-WH, Tinin-WH and Orin-G-OT were all less than 1% when the concentration was 160μg / ml. It shows that Mapin-WH, Tinin-WH, ...

Embodiment 3

[0034] Cytotoxicity assay:

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

[0036] Human skin fibroblasts HFF-1 were purchased from Shanghai Cell Bank. First, fibroblasts were cultured in DMEM containing 15% fetal bovine serum and double antibody (100U / ml of penicillin and streptomycin), and after the cells were full, they were digested with 0.25% trypsin, and washed with the above medium. Twice, resuspend the cells, add 100 μl of the cell suspension to a 96-well cell culture plate after counting the cells to make 10 cells per well 5 indivual. Add the sample, add the same volume of sterilized ultrapure water to the control group, set it at 37°C, 5% CO 2Cultured in the incubator for 24h. After the culture, 20 μl of 5mg / ml MTT solution (prepared with cell culture PBS buffer) was added to each well of the 96-well cell culture plate, and the culture was continued for 5 hours. time to completely dissolve t...

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Abstract

The invention provides ultrashort peptide for promoting collagen secretion and an application of ultrashort peptide, and belongs to the technical field of biomedicine. The ultrashort peptide containsat least one of ultrashort peptide Mapin-WH, Tinin-WH and Orin-G-OT, has the capability of promoting collagen synthesis, and has the characteristics of extremely high oxidation resistance and anti-aging activity, small molecular weight, simple structure, extremely low cytotoxicity, no hemolytic activity, simple preparation method and the like. The ultrashort peptide is used for preparing medicinesfor repair and regeneration of damaged epidermis such as burns, scalds and skin ulcers and used for replacing EGF in the fields of new raw materials for anti-aging and anti-oxidation beauty skin careproducts for repairing skin and stimulating skin collagen hyperplasia, and functions of the ultrashort peptide is probability not limited to the fields. The ultrashort peptide has small molecular weight and simple structure, is quite low in cost with the adoption of chemical synthesis and has outstanding beneficial characteristics as compared with the EGF costing nearly 2 million US 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 in the world, the field of anti-aging has become a hot spot of global attention. The skin is one of the most obvious organs in the aging process of the body. With the increase of age, the skin will undergo degenerative changes, such as the loss of elasticity, the reduction of water content, atrophy, and the appearance of wrinkles. [0003] Skin aging is a series of complex biological processes in skin tissue, which is the result of the combined action of all endogenous physiological factors and exogenous environmental factors. Aging not only affects normal skin tissue structure and physiological function, but also directly affects the appearanc...

Claims

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Application Information

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IPC IPC(8): C07K7/06C07K7/08A61K38/08A61K38/10A61P17/02A61K8/64A61Q19/08
CPCC07K7/06A61K8/64A61K38/00A61Q19/08C07K7/08
Inventor 于海宁王慧王义鹏
Owner DALIAN UNIV OF TECH
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