Antioxidative small peptides for inhibiting melanin production and preparation method and application thereof

A technology of inhibiting melanin and anti-oxidation, which is applied to the preparation method of peptides, chemical instruments and methods, and cosmetic preparations, etc. It can solve the problems of poor light absorption and unsuitable for daytime use, slow tyrosinase inhibitory effect, and unstable vitamin C. problem, achieve the effect of no hemolytic activity, no cytotoxicity, and simple preparation method

Active Publication Date: 2017-10-20
DALIAN UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, vitamin C is unstable and difficult to be absorbed by the skin, so it is difficult to be widely used; arbutin is a compound containing phenolic groups, although it is safe, it has a slow inhibitory effect on tyrosinase, and its light absorption is not suitable for daytime use; The whitening and freckle removal effect of acid is remarkable, but it has certain toxicity and cannot be used for a long time

Method used

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  • Antioxidative small peptides for inhibiting melanin production and preparation method and application thereof
  • Antioxidative small peptides for inhibiting melanin production and preparation method and application thereof
  • Antioxidative small peptides for inhibiting melanin production and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Chemical synthesis of OGOT, Magin-FL and Ansin-FL:

[0022] (1) Chemical synthesis of OGOT, Magin-FL and Ansin-FL: According to the amino acid sequence of the short peptide, its full sequence was synthesized with an automatic peptide synthesizer (433A, Applied Biosystems), and desalted by HPLC reverse-phase column chromatography.

[0023] (2) Molecular weight was determined by matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF).

[0024] (3) The purity of the purified OGOT, Magin-FL and Ansin-FL was identified by high performance liquid chromatography (HPLC), the molecular weight was determined by matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF), and the isoelectric point was determined by isoelectric focusing electrophoresis , Determining the amino acid sequence structure with an automatic amino acid sequencer.

[0025] OGOT, Magin-FL and Ansin-FL are straight-chain short peptides. Among them...

Embodiment 2

[0027] Antioxidant activity assay of OGOT, Magin-FL and Ansin-FL:

[0028] (1) DPPH radical scavenging activity (DPPH radical scavenging assay)

[0029] Weigh a certain amount of DPPH (2,2-diphenyl-1-picrylhydrazyl hydrate, Sigma, USA), dissolve it in methanol, and make 6×10 -5 The solution of M is prepared and used immediately. Mix 48 μl of DPPH solution with 2 μl of sample (2 mg / ml) (the mass ratio of final sample to DPPH is 3:1), let it stand in the dark for 30 minutes at room temperature, and measure the absorbance at 517 nm. In the blank control group, the sample to be tested was replaced by the sample dissolution medium. The experiment was done in triplicate, and methanol was used when the UV spectrophotometer was zeroed.

[0030] DPPH·clearance rate (%)=(AB-AA) / A B×100 (AB: absorbance value of blank control group; AA: absorbance value of sample group).

[0031] (2)ABTS ·+ Free radical cation scavenging activity

[0032] ABTS (3-ethylbezothiazoline-6-sulfonic acid)...

Embodiment 3

[0041] Tyrosinase half inhibitory concentration IC 50 Determination of value:

[0042] The tyrosinase activity inhibition experiment uses L-DOPA as the reaction substrate, and detects the absorbance at 475nm of the substrate colored substance dopaquinone catalyzed by tyrosinase, which indirectly reflects the inhibitory activity of the sample on tyrosinase.

[0043] Using a 100 μL reaction system, successively add phosphate buffer, samples of different concentrations (10, 25, 50, 100, 200 μM) and mushroom tyrosinase, and then incubate at 37°C for 10min, then add the substrate, and incubate at 37°C After reacting for 10 minutes, measure the OD value at a wavelength of 475nm. Calculate the relative activity inhibition rate of mushroom tyrosinase according to the measured OD value: inhibition rate (%)=(1-OD value of experimental group / OD value of control group)×100%.

[0044] The data were calculated using SPSS software to calculate the half-inhibition concentration IC of each s...

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Abstract

The invention relates to a group of antioxidative small peptides for inhibiting melanin production and a preparation method and application thereof. The small peptides comprise a short peptide OGOT, a short peptide Magin-FL and a short peptide Ansin-FL and belong to the technical field of biomedicine. The three short peptides have strong anti-oxidative activity, can inhibit the activity of tyrosinase, can significantly reduce the production of melanin and have the characteristics of low molecular weight, simple structure, no cytotoxicity, no hemolysis activity, high tissue permeability and simple preparation method. The small peptides can be used to prepare novel skin care raw materials with effects of resisting oxidization and melanin production and fading spots, can be used for preparation of anti-oxidant drugs and food additives and have a good application prospect.

Description

technical field [0001] The invention relates to a group of small peptides for anti-oxidation and inhibition of melanin production, a preparation method and application thereof, and belongs to the technical field of biomedicine and cosmetics. Background technique [0002] Oriental women have always advocated the whitening effect of "skin like snow, condensed like fat", and people have always pursued fair and clean skin. Therefore, the research and development of whitening agents that can inhibit stains and pigmentation caused by sunlight and other reasons have always been the focus of the cosmetics industry. [0003] Skin pigmentation caused by ultraviolet rays mainly occurs in keratinocytes and melanocytes. Ultraviolet radiation triggers DNA damage in keratinocytes, p53 is up-regulated, and finally leads to increased secretion of keratinocyte α-MSH (α-melanocyte stimulating hormone). When α-MSH is combined with After the MC1R (melanocortin 1 receptor) on the melanocyte memb...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K7/08C07K7/06C07K1/20A61K8/64A61Q19/02A61Q19/08A61K38/08A61K38/10A61P39/06A23L33/18
CPCA23L33/18A61K8/64A61K38/00A61K38/08A61K38/10A61K2800/782A61Q19/02A61Q19/08C07K7/06C07K7/08
Inventor 于海宁冯兰王义鹏
Owner DALIAN UNIV OF TECH
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