CN107641147A

CN107641147A - Antioxidation active peptides and its purposes in beauty treatment fields

Info

Publication number: CN107641147A
Application number: CN201710971828.5A
Authority: CN
Inventors: 张敏; 贾逸童
Original assignee: Yinchuan Li Zhixin Intellectual Property Counseling Service Co Ltd
Current assignee: Yinchuan Li Zhixin Intellectual Property Counseling Service Co Ltd
Priority date: 2017-10-18
Filing date: 2017-10-18
Publication date: 2018-01-30

Abstract

The present invention relates to cosmetic field, and in particular to a kind of antioxidation active peptides and its purposes in beauty treatment fields.The peptide of the present invention can lift skin gloss, remove dark yellow use.Specifically, the invention provides peptide or peptide combination to prepare for lifting skin gloss, removes the purposes in dark yellow composition.The peptide and composition of the present invention can lift the gloss of skin, remove dark yellow use.Specifically, peptide of the invention and composition are suitable to use to remove free radical and protect skin with cosmetic forms.

Description

Antioxidation active peptides and its purposes in beauty treatment fields

Technical field

The present invention relates to cosmetic field, and in particular to a kind of antioxidation active peptides and its application.

Background technology

Relation between free radical and aging is very generally received, its according to be during anaerobic respiration caused by Free radical can cause oxidative damage, and oxidative damage can accumulate and cause the forfeiture of homeostatic mechanism, interference gene expression pattern with And cause cell dysfunction, and then cause aging and death.The generation of oxidation material, the protection of antioxidant and to oxidation damage Exist between the reparation three of wound and connect each other.Many researchs have been carried out to determine whether antioxidant protection can be with the age Increase and decline.These researchs include the analysis to its key component：SOD, CAT, glutathione reductase, glutathione-S- The activity of transferase or expression, and the concentration of the low molecular weight compound with antioxidant properties.For example, Drosophila melanogaster body Life expectancy can be increased by 30% and reduced by damage caused by protein oxidation by interior SOD and CAT overexpression.By epidermis Tissue can produce free radical and other reactive oxygen specieses under the UV lines, cause cellular oxidation stress, this can be obviously promoted Aging.Skin excessively can be caused acute or chronic damage under ultraviolet radiation.Under acute situations, it can produce red Spot or burn, and chronic over-exposure can increase the risk for suffering from cutaneum carcinoma and aging.Furthermore it is known that epidermal cell can pass through increasing Add the expression of various albumen to be reacted to acute or chronic oxidative stress, the albumen for example maintains cell integrity and resistance Enzyme involved by oxidative damage.Oxidation is the biggest threat of skin aging, and diet is unhealthy, and Exposure to Sunlight, pressure, environmental pollution etc. are all Skin free radical can be allowed to spread unchecked, so as to produce the oxidative phenomenas such as complexion dimness, water shortage.These are all that body produces oxidation " crime Stalwart chief culprit ".So no matter from healthy aspects or from skin care aspect, we are required for paying attention in daily life anti-oxidant.

The polyphenoils of human body has what itself was synthesized, also has by food supply.Enzyme and non-enzymatic polyphenoils are being protected Shield is vital due to serving in peroxide injury caused by motion.Supplement polyphenoils is advantageous to Exercise Mechanics reduction The generation of free radical accelerates its removing, with the side effect to Green Tea Extract, thus has to the health of common people and sportsman Benefit, can the generation of delayed motion fatigue and quickening physical efficiency recovery.Older physical exertion person takes antioxidant than year the lighter Effect is more preferable.

Polypeptide is a kind of compound of the molecular structure between amino acid and protein, protein is had certain life Manage function.In the vital movement of the mankind, peptide digesting and assimilating better than free amino acid in vivo, and it is different from amino Delivery system inside acid.Some small peptides are while nutriment necessary to providing growth in humans, development, additionally it is possible to prevent Or treatment disease, function of human body is adjusted, there is the polypeptide of bioactivity to be referred to as biologically active peptide for these.Research finds, a lot The polypeptide of separate sources all has oxidation resistance, such as caseinhydrolysate, soybean protein, bovine serum albumin(BSA), ovalbumin, oil The polypeptide that material Seed Storage Protein, wheat gliadin and zein etc. obtain all has certain inoxidizability.

CN106589068A discloses a kind of madai antioxidation polypeptide and preparation method thereof, and this method is with madai fish-skin or fish scale egg White is raw material, by the enzymolysis to acid protease, isolates and purifies to obtain pure antioxidation polypeptide.The antioxidation polypeptide eliminates day Defect possessed by right antioxidant and worry of the public to artificial synthesized antioxidant is eliminated, food source is based on for exploitation Antioxidation polypeptide and explore its extensive use in food, medicine and established good basis.

CN201210407446 discloses a kind of Antioxidation active synthetic peptide and application thereof, and it is demonstrated experimentally that its synthesized 3 Bar polypeptide has antioxidation activity, and because the anti-oxidation peptide has, molecular weight is small, and activity is high, is easy for workers to synthesize, and cost is low etc. Feature, the large-scale production of the industries such as food, cosmetics, medicine can be widely used in.

But do not allow the anti-oxidation peptide for cosmetic field of consumer satisfaction also on the market at present, especially can Application is reached to cosmetics and produces the peptide of beneficial effect.

The content of the invention

In order to solve the above problems, the present inventor passes through persevering endeavors, have developed with antioxidation activity Pentapeptide combination, and find that it has good application in cosmetics.

Thus, for one of the object of the invention to provide the peptide with antioxidation activity, it has Arg Tyr Pro Leu Asp Glu Lys（PEPTIDE.01）The peptide of sequence.

The present invention also provides the peptide with antioxidation activity and combined, and it includes Arg Tyr Pro Leu Asp Glu Lys （PEPTIDE.01）The peptide and Gln Lys Val Lys Lys Phe Glu Ser of sequence（PEPTIDE.02）The peptide of sequence.

Present invention also offers with skin is promoted, with the dark yellow cosmetic composition of gloss, elimination, it includes having PEPTIDE.01 peptide, or the combination of the peptide comprising PEPTIDE.01 and PEPTIDE.02.

Peptide or the peptide combination of the present invention has antioxidation activity, and then dark yellow with gloss, elimination with promotion skin Effect.Therefore, combined the invention provides PEPTIDE.01 peptide or PEPTIDE.01 and PEPTIDE.02 peptide and preparing use There is the purposes in the dark yellow composition of gloss, elimination in promoting skin.

Peptide and the peptide combination of the present invention has antioxidation activity, and then dark yellow with gloss, elimination with promotion skin Effect.Specifically, peptide of the invention and peptide combination are suitable to use with cosmetic forms.

Therefore, the present invention cosmetic composition is also provided, its include PEPTIDE.01 peptide or PEPTIDE.01 and PEPTIDE.02 peptide combination.

Embodiment

The peptide of the present invention can use the synthetic method of organic chemistry, such as can use solid phase method (Boc methods, Fmoc methods) Or the in general method such as liquid phase method, such as can be automatically synthesized using peptide symthesis device.Reaction condition for peptide symthesis etc. For, can the common technical knowledge based on those skilled in the art arbitrarily set selected synthetic method or appropriate reaction bar Part etc..The purification process of the peptide of chemical synthesis is also well-known to those skilled in the art.The peptide obtained by solid-phase synthesis It can be purified by high pressure liquid chromatography, be freeze-dried and obtain.

In this specification when addressing these peptides, except situation about especially expressing or situation about substantially being excluded on word Outside lower, " peptide " includes their salt., can existing salt in physiological conditions comprising sodium salt, sylvite etc. in this salt.

It is different according to addition form or product form as the present composition, relative to the end article present invention's The content of peptide is the mass % of 0.0001 mass %~1.0, be preferably 0.005 mass %~0.5 mass %, more preferably 0.005 The mass % of quality %~0.010.

Peptide or the peptide combination of the present invention provides in cosmetically acceptable carrier.Carrier can be hydrophobic or close Water.

Appropriate hydrophobic carrier is included for example, waxy nonionic, such as the ester and ether of fatty alcohol and aliphatic acid, carbochain Length is C4 to C22, preferably C8 to C18, most preferably C12 to C18.The example of fatty hydrophobic carrier includes myristic acid isopropyl Ester, isopropyl palmitate, octyl palmitate, wool isopropyl propionate, acetulan, the benzoic ether, pungent of C12-C15 alcohol Sour cetyl, cetin, myristyl myristate, Tetradecyl lactate, acetic acid cetyl, two caprylic/caprics third Diol ester, decyl oleate, acetylated lanolin, enanthic acid stearate, malic acid diisopstearate, octyl hydroxystearate, Octyl hydroxystearate, isopropyl isostearate etc..

Appropriate hydrophilic support solution can be, such as the glycol for the alkoxylate commonly used in glycol and cosmetics, including Ethylene glycol, diethylene glycol, triethylene glycol, propane diols, DPG etc..

The term and statement used herein is used as descriptively rather than restrictive term, and in such a term and statement Use in be not intended to exclude shown in and the feature or part thereof any equivalent, it is appreciated that various modifications are please It is possible for asking in the scope of the present invention of protection.It is therefore understood that although the present invention by preferred embodiment and appoints Feature is selected specifically to disclose, but those skilled in the art can use the modification and transformation of concept disclosed herein, and such repair Decorations and change are considered as in the scope of the present invention such as defined by accessory claim.

To be illustrated more clearly that the present invention, it is described in detail in conjunction with following examples, but these embodiments are only To the exemplary description of the present invention, the limitation to the application should not be construed as.

Embodiment

The following examples are to be used for the explanation present invention in a non limiting manner.

The Arg Tyr Pro Leu Asp Glu Lys of embodiment 1（PEPTIDE.01）The synthesis in solid state of the peptide of sequence

From macromolecule resin, according to amino acid sequence Arg Tyr Pro Leu Asp Glu Lys feature, first by Lys's Carboxyl is connected in the form of covalent bond with resin, and then Lys amino and Glu carboxyl, which shrink, reacts, and after processing, then adds Asp, Asp amino and Leu carboxyl reaction, add amino acid from right to left successively, after having added last Arg amino acid, Resin is cut off again obtains target synthesis heptapeptide.High-efficient liquid phase chromatogram purification obtains product to the end, and purge process uses PhenomenexC 18 （4.6*250mm）Chromatographic column, flow velocity 1.0mL/min.Eluted using two passages, solvent orange 2 A: Water containing 0.1% trifluoroacetic acid；Solvent B:Solution containing 0.09% trifluoroacetic acid（The water of 80% acetonitrile+20%）；Gradient mistake B initial proportions are 39% in journey, and in 20min, B ratio rises to 49%, Detection wavelength 220nm.Synthesis heptapeptide solution is collected, Liquid nitrogen quickly cooling, is then freezed.The product that purity is more than 90% is obtained, and structure is identified through ESI-MS.

The Gln Lys Val Lys Lys Phe Glu Ser of embodiment 2（PEPTIDE.02）The synthesis in solid state of the peptide of sequence

According to scheme same as Example 1, PEPTIDE.02 peptides are synthesized.

The ABTS radicals scavengings effect for the PEPTIDE.01 peptides that embodiment 3 synthesizes

7mmol/L ABTS is prepared with distilled water⁺The potassium persulfate solution of solution and 2.45mmol/L（Must be first in room before Temperature is lower to place 16h）, respectively by ABTS⁺Mixed with potassium persulfate solution by 1: 1 volume ratio, before use with pH7.4,5mmol/L's Mixed liquor is diluted to light absorption value A by phosphate buffer solution₇₃₄For 0.70 ± 0.02.Take 0.5mL various concentrations sample and 0.5mL ABTS free-atom aqueous solutions mixing stands 10min after surveying its light absorption value under 734nm.With deionized water and reduced form paddy The sweet peptide of Guang makees blank control and positive control respectively instead of sample.ABTS radicals scavengings vigor is calculated by following equation：

In formula, A₀：Blank control group light absorption value；A_s：Sample sets light absorption value.

Determined through the present embodiment, PEPTIDE.01 peptides have stronger removing vigor to ABTS free radicals, are 2.5 in concentration Free radical scavenging activity is up to 80%, when concentration increases to 25.0 μ g/mL, the free radical scavenging activity of PEPTIDE.01 peptides during μ g/mL Almost 97%, it is far longer than the positive control GSH of same concentration free radical scavenging activity.

The PEPTIDE.01 peptides and PEPTIDE.01 that embodiment 4 synthesizes combine with PEPTIDE.02 peptide to be surveyed with ORAC methods Determine the antioxidation activity value of anti-oxidation peptide

Carried out based on ORAC reactions in 70mM kaliumphosphate buffers (pH=7.5) environment, fluorescer FL with the buffer into High concentration storing solution, 4 DEG C of preservations after small size packing.Necessary amount is taken out during experiment to be diluted to same 70mM kaliumphosphate buffers Final concentration of 70nM in reaction system.AAPH（Free-radical generating agent）Reaction system end is formulated into 70mM kaliumphosphate buffers Concentration uses for 12mM.Standard polyphenoils Trolox and testing sample also use buffer solution and dilution.AAPH and Trolox Extemporaneous uses.

Specific measure operation is to be separately added into 5 μM in each micropore of 96 orifice plates（Candidate polypeptide solution is the solution of peptide combination When, PEPTIDE.01 and PEPTIDE.01 concentration are 0.25 μM）FL120 μ L are added after the μ L of candidate polypeptide solution 20 37 At DEG C after preset 12min, the μ L of AAPH 60 are added in each hole rapidly with multichannel pipettor and start reaction, and microwell plate is placed in METHOD FOR CONTINUOUS DETERMINATION is carried out with excitation wavelength 485nm, launch wavelength 520nm at 37 DEG C in fluorescence analyser, determined once per 1min Each hole fluorescence intensity, minute 2h.

ORAC experiments need two kinds of controls of setting, i.e., compare (- AAPH) without the FL fluorescence Natural Attenuation of addition free radical (+AAPH) is compareed with the Free Radical in the presence of no antioxidant.It is glimmering under the oxidation resistance and Free Radical of sample Light fall-off curve delays area (NetAUC) directly related.Under fluorescence decline curve under antioxidant action area with The delay portion facet of the difference of area, i.e. fluorescence quenching curve under fluorescence decline curve without Free Radical in the presence of antioxidant Product Net AUC are the protected area of antioxidant.

AUC=1+(f1/f0+f2/f0+f3/f0+……+fn/f0)

Fluorescent value reading when wherein f0 refers to beginning；Fn refers to the fluorescent value reading at n-th minute；Net AUC=AUC antioxidants- AUC is compareed.

Trolox's required for the oxidation resistance that the candidate polypeptide solution that final result is expressed as every micro-molar concentration is worked as Micro-molar concentration.

As a result it is respectively 6.85 ± 0.3 for the ORAC activity values of PEPTIDE.01 antioxidation polypeptides（μmol Trolox/μmol）, Peptide combination for 9.02 ± 0.5（μmol Trolox/μmol）, the positive control VC and GSH that are significantly higher than under equal conditions （ORAC activity values are respectively 0.96 and 1.41）.Illustrate that aforementioned polypeptides and peptide combination are respectively provided with higher antioxidation activity.

Embodiment 5：The preparation of Essence

Composition	Content (weight %)
		The peptide of the present invention（PEPTIDE.01）	0.03
Glutathione	1.5
		1.3 propane diols	3.0
RV	5.0
		Phytosterols oletate	3.0
Fullerene	3.0
		Soybean lecithin	3.0
Carbomer	10.0
		Extractive of perilla	0.1
Flos Rosae Rugosae extract	0.1
		Dendrobium candidum extract	0.1
Purslane extract	0.1
		Hyaluronic acid	0.2
Niacinamide	0.1
		Lauroyl arginine ethyl ester	0.5
Purified water	Residual volume
		Amount to	100

Fullerene, soybean lecithin, carbomer, phytosterols oletate are mixed, stirred after being stirring evenly and then adding into other components Mix and be modulated into Essence.

Embodiment 6：The preparation of Essence

Composition	Content (weight %)
		The peptide combination of the present invention（Mass ratio 1:1 PEPTIDE.01 and PEPTIDE.02）	0.02
Glutathione	2.5
		1.3 propane diols	2.0
RV	6.0
		Phytosterols oletate	4.0
Fullerene	2.0
		Soybean lecithin	2.0
Carbomer	12.0
		Extractive of perilla	0.5
Flos Rosae Rugosae extract	0.2
		Dendrobium candidum extract	0.2
Purslane extract	0.4
		Hyaluronic acid	0.8
Niacinamide	0.5
		Lauroyl arginine ethyl ester	0.1
Purified water	Residual volume
		Amount to	100

Embodiment 7：The preparation of Essence

Composition	Content (weight %)
		The peptide of the present invention（Mass ratio 1:1 PEPTIDE.01 and PEPTIDE.02）	0.04
Glutathione	0.5
		1.3 propane diols	4.0
RV	4.0
		Phytosterols oletate	2.0
Fullerene	4.0
		Soybean lecithin	4.0
Carbomer	8.0
		Extractive of perilla	0.2
Flos Rosae Rugosae extract	0.15
		Dendrobium candidum extract	0.15
Purslane extract	0.2
		Hyaluronic acid	1.0
Niacinamide	0.25
		Lauroyl arginine ethyl ester	0.3
Purified water	Residual volume
		Amount to	100

The activity checking of the Essence of embodiment 8

According to the formula shown in embodiment 5,6 and 7, the composition for external application of Essence is modulated（Skin gloss is lifted, is removed dark Huang is used), and like product purchased in market embodiment as a comparison.For each composition for external application, the female of 10 30-35 year is allowed respectively Property face according to following standards evaluate painting preparation when use feeling.

5 points：Skin surface glossiness is high and glossy effect protrudes；

4 points：Skin surface glossiness is general, and glossy effect is visible；

3 points：There is no visible change；

2 points：Skin surface slightly has rough sensation, without whitening and the effect of showed smooth；

1 point：Skin surface has rough sensation, without whitening and the effect of showed smooth.

Calculate the average mark of above-mentioned evaluation result, the results showed that, the average mark of embodiment 5,6 and 7 more than 4.7 points, Control sample is equally divided into 3.0 points.The effect of embodiment 6 and 7 is most prominent, is equally divided into 4.9.

Although with above embodiments describing the present invention, it should be appreciated that before the spirit without departing substantially from the present invention Put, the present invention further can be modified and changed, and these modification and variation belong to protection scope of the present invention it It is interior.

Sequence table

<110>Yinchuan Li Zhixin intellectual properties Counseling Service Co., Ltd

<120>Antioxidation active peptides and its purposes in beauty treatment fields

<130> HY1701

<160> 2

<170> SIPOSequenceListing 1.0

<210> 1

<211> 7

<212> PRT

<213>Artificial sequence ()

<220>

Claims

1. peptide, it has Arg Tyr Pro Leu Asp Glu Lys（PEPTIDE.01）The amino acid sequence of sequence.

2. peptide according to claim 1, it is made up of PEPTIDE.01 amino acid sequence.

3. peptide combines, it includes Arg Tyr Pro Leu Asp Glu Lys（PEPTIDE.01）The peptide and Gln Lys of sequence Val Lys Lys Phe Glu Ser（PEPTIDE.02）The peptide of sequence.

4. cosmetic composition, it includes the peptide of claim 1 or 2.

5. cosmetic composition, it includes the peptide of claim 3 and combined.

6. the peptide of claim 1 or 2 is being prepared for lifting skin gloss, the use in dark yellow cosmetic composition is removed On the way.

7. the peptide combination of claim 3 is being prepared for lifting skin gloss, remove in dark yellow cosmetic composition Purposes.

8. purposes of the peptide of claim 1 or 2 in anti-oxidation preparation is prepared.

9. the peptide of claim 3 combines the purposes in anti-oxidation preparation is prepared.

10. the purposes of the peptide of claim 1 or 2 or the peptide combination of claim 3 in the preparation for removing free radical is prepared.