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Small peptide for inhibiting melanin production and anti-oxidation, preparation method and application thereof

A technology for inhibiting melanin and anti-oxidation, applied in the fields of biomedicine and cosmetics, can solve the problems of good light absorption and unsuitable for daytime use, slow tyrosinase inhibitory effect, unstable vitamin C, etc., and achieves no hemolytic activity and no cytotoxicity. , the preparation method is simple

Active Publication Date: 2020-04-28
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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  • Small peptide for inhibiting melanin production and anti-oxidation, preparation method and application thereof
  • Small peptide for inhibiting melanin production and anti-oxidation, preparation method and application thereof
  • Small peptide for inhibiting melanin production and anti-oxidation, preparation method and application thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] Chemical synthesis of Ansin-2 and Magin-2:

[0020] (1) Chemical synthesis of Ansin-2 and Magin-2: According to the amino acid sequence of the short peptide, the full sequence of the two was synthesized with an automatic peptide synthesizer (433A, Applied Biosystems), and desalted by HPLC reverse-phase column chromatography.

[0021] (2) Molecular weight was determined by matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF).

[0022] (3) Purified Ansin-2 and Magin-2 are identified for their purity by high-performance liquid chromatography (HPLC), the molecular weight is determined by matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF), and the isoelectric point is measured by isoelectric focusing electrophoresis. An automatic amino acid sequencer was used to determine the amino acid sequence structure.

[0023] Ansin-2 and Magin-2 are straight-chain ultrashort peptides. Ansin-2 has a molecular wei...

Embodiment 2

[0025] Ansin-2 and Magin-2 antioxidant activity assay:

[0026] (1) DPPH radical scavenging activity (DPPH radical scavenging assay)

[0027] 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.

[0028] DPPH·clearance rate (%)=(AB-AA) / A B×100 (AB: absorbance value of blank control group; AA: absorbance value of sample group).

[0029] (2)ABTS ·+ Free radical cation scavenging activity

[0030] ABTS (3-ethylbezothiazoline-6-sulfonic acid) (3-ethylbe...

Embodiment 3

[0038] Tyrosinase half inhibitory concentration IC50 Determination of value:

[0039] 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.

[0040] 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%.

[0041] The data were calculated using SPSS software to calculate the half-inhibition concentration IC of each sub...

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Abstract

The invention relates to an anti-oxidization small peptide capable of inhibiting melanogenesis as well as a preparation method and application thereof. The anti-oxidization small peptide comprises a short peptide Ansin-2 and a short peptide Magin-2, and belongs to the technical field of biomedicines. The two short peptides have strong oxidization resistance and can inhibit the activity of tyrosinase and obviously reduce melanogenesis; meanwhile, the small peptide further has the characteristics of small molecular weight, simple structure, no cytotoxicity, no hemolytic activity and high tissue penetration, and the preparation method is simple. The anti-oxidization small peptide can be used for preparing a novel beauty-maintaining and skin-care raw material capable of resisting oxidization, resisting melanogenesis and removing spots, also can be used for preparing an anti-oxidization medicine, a food additive and the like, and has an extremely good application prospect.

Description

technical field [0001] The invention relates to a group of small peptides for inhibiting melanin formation and anti-oxidation, 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 membrane...

Claims

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Application Information

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