CN106265120B - A kind of cytokine-like effect and cosmetic applications of nine Oligopeptide Compositions - Google Patents

A kind of cytokine-like effect and cosmetic applications of nine Oligopeptide Compositions Download PDF

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CN106265120B
CN106265120B CN201610818082.XA CN201610818082A CN106265120B CN 106265120 B CN106265120 B CN 106265120B CN 201610818082 A CN201610818082 A CN 201610818082A CN 106265120 B CN106265120 B CN 106265120B
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skin
oligopeptide compositions
oligopeptide
compositions
tripeptides
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CN106265120A (en
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周守山
何俭
林晓
朱单单
何政红
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Zhuhai Golden Peptide Biotechnology Co Ltd
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Zhuhai Golden Peptide Biotechnology Co Ltd
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00—Cosmetics or similar toiletry preparations
    • A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
    • A61K8/64—Proteins; Peptides; Derivatives or degradation products thereof
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00—Cosmetics or similar toiletry preparations
    • A61K8/02—Cosmetics or similar toiletry preparations characterised by special physical form
    • A61K8/04—Dispersions; Emulsions
    • A61K8/06—Emulsions
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00—Cosmetics or similar toiletry preparations
    • A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
    • A61K8/31—Hydrocarbons
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00—Cosmetics or similar toiletry preparations
    • A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
    • A61K8/33—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing oxygen
    • A61K8/34—Alcohols
    • A61K8/342—Alcohols having more than seven atoms in an unbroken chain
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    • A61K8/345—Alcohols containing more than one hydroxy group
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    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00—Cosmetics or similar toiletry preparations
    • A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
    • A61K8/33—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing oxygen
    • A61K8/37—Esters of carboxylic acids
    • A61K8/375—Esters of carboxylic acids the alcohol moiety containing more than one hydroxy group
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
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    • A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
    • A61K8/40—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing nitrogen
    • A61K8/41—Amines
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00—Cosmetics or similar toiletry preparations
    • A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
    • A61K8/60—Sugars; Derivatives thereof
    • A61K8/602—Glycosides, e.g. rutin
    • A—HUMAN NECESSITIES
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    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
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    • A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
    • A61K8/67—Vitamins
    • A61K8/678—Tocopherol, i.e. vitamin E
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00—Cosmetics or similar toiletry preparations
    • A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/72—Cosmetics or similar toiletry preparations characterised by the composition containing organic macromolecular compounds
    • A61K8/81—Cosmetics or similar toiletry preparations characterised by the composition containing organic macromolecular compounds obtained by reactions involving only carbon-to-carbon unsaturated bonds
    • A61K8/8141—Compositions of homopolymers or copolymers of compounds having one or more unsaturated aliphatic radicals, each having only one carbon-to-carbon double bond, and at least one being terminated by only one carboxyl radical, or of salts, anhydrides, esters, amides, imides or nitriles thereof; Compositions of derivatives of such polymers
    • A61K8/8147—Homopolymers or copolymers of acids; Metal or ammonium salts thereof, e.g. crotonic acid, (meth)acrylic acid; Compositions of derivatives of such polymers
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    • A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
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    • A61K8/84—Cosmetics or similar toiletry preparations characterised by the composition containing organic macromolecular compounds obtained by reactions otherwise than those involving only carbon-carbon unsaturated bonds
    • A61K8/89—Polysiloxanes
    • A61K8/891—Polysiloxanes saturated, e.g. dimethicone, phenyl trimethicone, C24-C28 methicone or stearyl dimethicone
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    • A61K8/92—Oils, fats or waxes; Derivatives thereof, e.g. hydrogenation products thereof
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    • A61Q19/00—Preparations for care of the skin
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    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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    • A61Q19/00—Preparations for care of the skin
    • A61Q19/08—Anti-ageing preparations
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    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
    • A61K2800/40—Chemical, physico-chemical or functional or structural properties of particular ingredients
    • A61K2800/59—Mixtures
    • A61K2800/592—Mixtures of compounds complementing their respective functions
    • A61K2800/5922—At least two compounds being classified in the same subclass of A61K8/18

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Abstract

The present invention relates to cosmetic technical fields; it discloses a kind of for the compound Oligopeptide Compositions of skin repair and its application; according to parts by weight, which includes 1 ~ 30 part of palmityl tripeptides -1,1 ~ 30 part of -1 copper of tripeptides, 1 ~ 50 part of palmityl tripeptides -5,1 ~ 30 part of carnosine, 1 ~ 30 part of acetyl group tetrapeptide -5, -7,0.1 ~ 30 parts of Argirelines of 1 ~ 30 part of palmityl tetrapeptide, 0.1 ~ 30 part of nonapeptide -1,1 ~ 30 part of streptococcus lactis peptide.Oligopeptide Compositions have effects that cytokine-like, and influence the growth of skin flora, are applied in dermatology and beautifying skin, have the function of repairing skin lesion, adjustment metabolism and the growth of antagonism skin condition pathogenic bacteria.

Description

A kind of cytokine-like effect and cosmetic applications of nine Oligopeptide Compositions
Technical field
The present invention relates to cytokine-like effects and beauty of cosmetic technical field, more particularly to a kind of nine Oligopeptide Compositions Hold application.
Background technique
The lesser polypeptide of molecular weight, also referred to as small peptide, oligopeptides, oligopeptide, or be small active peptides, generally by several Amino acid composition.The lesser oligopeptides of molecular weight can directly be absorbed by skin.As medicine, nutrition, Dermatology and molecule are raw The development of object, oligopeptides is by increasingly in-depth study.Scientist's discovery, oligopeptides have various biology to Skin Cell Activity, these biological active agents are just more and more disclosed.
Have cytokine profiles in recent years and be applied to skin injury reparation and cosmetics, He Zheng is red to wait (1999) invention one Kind cosmetic composition (rh-bFGF containing human fibroblastic growth factor, human vascular endothelial growth factor rh-VEGF, people's table Skin cell growth factor rh-EGF, growth factor of human nerve rh-NGF and melanin inhibitor MIF), the composition is in difference Cell metabolism and restoration updating are participated on level and different role site.He Zhenghong etc. (2001) invented a kind of medicine, beauty, And cosmetic composition, with principle blocking, closing or the synthesis for reducing dermal melanin of antisense nucleic acid gene approach, suppression Cutaneous pigmentation processed.Zhou Shoushan etc. (2002) invents the production method and application of a kind of neurotrophic factor (NTF), the factor Wrinkle effect is gone with good skin.
Relative to peptide and proteins such as Porcine HGFs, the small oligopeptides of molecular weight can have higher Cutaneous permeation Property, it is easier to it is absorbed by human skin, has been arrived to a certain degree simultaneously because molecular weight is small, flying for matter just has occurred in bioactivity Jump.Peptide molecular weight is smaller, and " amino acid chain " is shorter, is more easily absorbed and utilized by the body.
To the mechanism of action of oligopeptides active material the study found that each oligopeptides has its specific biological effect. They can not only influence cell Proliferation, differentiation and develop related gene expression with body growth, but also can be to Disease-causing gene Expression generate important adjustment effect.Can gene expression it is all it is horizontal (before transcription, transcription, after transcription, translation and translation Totally 5 levels afterwards) on it is adjusted, it is most few although different oligopeptides respectively have its emphasis or single-minded adjusting horizontal The adjusting of gene expression occurs on transcriptional level for peptide.
Main path has: (1) CAMP or CGMP protein kinase pathway；(2) tyrosine-kinase enzyme system, two above approach master If by phosphorylation and dephosphorylation to some transcription factors and/or confactor, to influence these factors The activity of activated gene transcription；(3) ion channel；(4) the acid mediated approach of inosinyl phosphate inosine；(5) intracellular receptor approach, cell Inner recipient can be the enzyme of catalysis reaction, be also possible to the tonality albumen of gene expression, most of active materials are to gene expression Regulation be to be realized by intracellular receptor approach.
Different action receptors further promotes the synergistic effect generated after growth factor and small molecule victory peptide compounding Skin metabolism, nti-freckle smoothing wrinkle prevent skin aging along with the collective effect of remaining ingredient in cosmetics, repair impaired Skin, improves skin comprehensively, and compounding effect is significantly better than exclusive use.It is reported also very about the research of this respect at present More, but each substance due to using is different, content is inconsistent, and risen effect is also different on the skin for product；So needing Want further research and probe that can provide the Oligopeptide Compositions of effective skin repair comprehensively.
Summary of the invention
The object of the present invention is to provide a kind of compound Oligopeptide Compositions for skin repair.
It is a further object of the present invention to provide the applications of the Oligopeptide Compositions.
In order to achieve the above objectives, the invention adopts the following technical scheme:
A kind of compound Oligopeptide Compositions for skin repair, according to parts by weight, including 1 ~ 30 part of palmityl tripeptides -1, 1 ~ 30 part of -1 copper of tripeptides, 1 ~ 50 part of palmityl tripeptides -5,1 ~ 30 part of carnosine, 1 ~ 30 part of acetyl group tetrapeptide -5,1 ~ 30 part of palmityl Tetrapeptide -7,0.1 ~ 30 part of Argireline, 0.1 ~ 30 part of nonapeptide -1,1 ~ 30 part of streptococcus lactis peptide.
Oligopeptides can be to be modified by chemical method, such as acylated, soda acid.
Further, according to parts by weight, Oligopeptide Compositions include 0.02 part of palmityl tripeptides -1,0.01 part of tripeptides -1 Copper, 0.01 part of palmityl tripeptides -5,0.9 part of carnosine, 0.01 part of acetyl group tetrapeptide -5,0.01 part -7,0.015 parts of palmityl tetrapeptide Argireline, 0.003 part of nonapeptide -1,0.18 part of streptococcus lactis peptide.
Further, the Oligopeptide Compositions further include water, grease, surfactant, moisturizer, antioxidant, anti-corrosion At least one of agent.
Further, the grease be olive oil, rilanit special, caprylic/capric triglyceride, cottonseed oil, soybean oil, Seabuckthorn Oil, jojoba oil, saualane, lanonol, cetostearyl alcohol, dimethyl silicone polymer, polymethylphenylsiloxane, ring-type At least one of polysiloxanes.
Further, the surfactant is polysorbas20, Tween 80, sodium laurate, stearyl alcohol sodium sulphate, polyoxyethylene At least one of aliphatic ester, cetearyl alcohol alcohol ether -2, cetearyl alcohol alcohol ether -21, Phosphoric acid monohexadecyl ester.
Further, the moisturizer be glycerol, Sodium Hyaluronate, glycerine polyacrylate, oligofructose, xylitol, At least one of propylene glycol, butanediol, methylglycine proline, glycine betaine.
Further, the antioxidant is vitamin C, vitamin E, riboflavin, niacin, metallothionein, super oxygen At least one of object mutase and dimethylglycine.
Further, the preservative is methylparaben, ethylparaben, dimethyloldimethylhydantoin, iodine At least one of propilolic alcohol butyl mephenesin Carbamate, DMDM hydantoin, methylisothiazolinone, Phenoxyethanol.
Above-mentioned Oligopeptide Compositions are preparing the application in cosmetics, skin care item or curable product.
Further, the preparation type of the cosmetics, skin care item or curable product is Essence, eye cream, cream, jelly Dry powder, solution, emulsion, creme, gel, facial mask or dressing.
A kind of cream, including 5wt% Oligopeptide Compositions, 61.75wt% water, 5wt% glycerol, 3wt% propylene glycol, 0.15wt% card Wave nurse, 3wt% cetearyl glucoside, 2wt% PEG-100 stearate/stearine, 1wt% cetostearyl alcohol, 5wt% caprylic/capric triglyceride, 2wt% dimethyl silicone polymer, 2wt% Butyrospermum parkii fruit rouge, 0.5wt% tocopherol acetate, 8wt% isohexadecane, 0.5wt% glycerol caprylate, 0.1wt% triethanolamine, 1wt%1,2- pentanediol, wherein Oligopeptide Compositions are such as It is noted earlier.
A kind of emulsion, including 5wt% Oligopeptide Compositions, 69.6wt% water, 3wt% glycerol, 5wt% propylene glycol, 0.1wt% card wave Nurse, 3wt% Glyceryl stearate citrate, 1.5wt% cetostearyl alcohol, 3wt% caprylic/capric triglyceride, the poly- diformazan of 1wt% Radical siloxane, 1wt% Butyrospermum parkii fruit rouge, 0.2wt% tocopherol acetate, 6wt% isohexadecane, 0.5wt% glycerol caprylate, 0.1wt% triethanolamine, 1wt%1,2- pentanediol, wherein Oligopeptide Compositions are as previously described.
The invention has the following advantages:
The physiology of the adjustable Skin Cell of compound Oligopeptide Compositions of the invention, and corresponding Skin Cell can be stimulated to produce Raw endogenic cell factor, such as h-bFGF, h-EGF, h-PDGF, h-aFGF, h-TGF- β, h-NTFs, h-KGF, h-VEGF Deng so as to start skin active repair procedure, repairing various types of skin problems and skin injury, and have to healthy skin There is perfect protective effect.
The Oligopeptide Compositions increase the growth of Skin Cell, enhance skin cell metabolism, as collagen synthesis increases.
The Oligopeptide Compositions have the function of the growth of antagonism skin condition pathogenic bacteria, such as inhibit propionibacterium acnes, grape Coccus, streptococcus growth, and adjust skin Tiny ecosystem.
The Oligopeptide Compositions have the effect of carnitine sample, and mediator transmits between reducing neuromuscular, and has effects that anti-wrinkle.
The Oligopeptide Compositions, which have, reduces inflammatory mediator effect, mitigates inflammatory edema.
The Oligopeptide Compositions improve Lymphatic Circulation and microcirculation, adjust skin condition in normal.
The Oligopeptide Compositions have the function of reducing free radical, glycosylation and lipid oxidation, delay skin aging.
The Oligopeptide Compositions animal skin sensitivity tests, has no stimulation.
The Oligopeptide Compositions are easily obtained and save.
The Oligopeptide Compositions can directly smear face, can also use beauty and the common instrument and method of medical cosmetology, packet It includes the modes such as ultrasonic instrument, ion introducing apparatus, micropin, water laser accunputure, laser, injection and introduces the composition into corresponding skin portion Position.
The Oligopeptide Compositions can be prepared by conventional mixing, dissolution, emulsification, liposomal, freeze drying technology etc..
The mechanism of action of this nine kinds of oligopeptides is:
Palmityl tripeptides -1 are derived from the segment on I-type collagen chain, to skin cell renewal, repair collagen conjunction At having apparent regulation and facilitation, the synthesis of collagen and hyaluronic acid in fibroblast is effectively facilitated, is had Crease-resistant, anti-inflammatory, antiallergic effect.
- 1 copper of tripeptides: copper binding peptide class improves permeability, and original ionic forms is allowed to be difficult to through the inorganic of stratum corneum barrier The deep enough skin of copper ion energy plays effect.Collagen, elastin laminin, hyaluronic acid, glycosaminoglycan and fiber can be improved to connect The synthesis of desmin is used for wound healing and hair growth, and with induced fibroblast secretion bFGF's and TGF-beta1 Effect.It can promote collagen growth, improve the synthesis capability of the skin moisture-keeping factor, slow down the speed of skin lipid peroxidating, improve Skin heart microcirculation.Accelerate the reparation of skin trauma, reduce scar, improves wrinkle, there is powerful reparation function to impaired skin Effect, while compact skin keep skin more aobvious smooth bright；And the synthesis of collagen can be promoted, smooth microgroove and wrinkle.
Palmityl tripeptides -5: the particular sequence having is similar to human body body self mechanism, by activating inactive TGF-β generate collagen, have go wrinkle effect.
Carnosine is the dipeptides that Beta-alanine and histidine form.With strong anti-oxidation, can remove in oxidative stress process The fatty acid excessive oxidation of middle cell membrane and the active oxygen radical and alpha-beta unsaturated aldehyde formed；Significantly inhibit free radical and Lipid oxidation caused by metal ion.
Acetyl group tetrapeptide -5 can improve the microcirculation of lymph and capillary, eliminate oedema, mitigate black eye, improve skin Skin elasticity and smoothness, while there are also the glycosylated effect of collagen is inhibited, collagen aging, aggregation are prevented, to reach To mitigation eye pouch and crease-resistant effect.
Palmityl tetrapeptide -7 is derived from a segment in Immunoglobulin IgG, can significantly reduce inflammatory factor interleukin The level of L-6 has the function of eliminating the potential inflammation of skin, antiallergic, generates to wrinkle is mitigated, restore skin vigor.
Argireline is the N-terminal for imitating SNAP-25 albumen and the oligopeptides that is made of six amino acid, and effect can be with It matches in excellence or beauty with botulinum toxin type A, weakens contraction of muscle, prevent the formation of wrinkle, but avoiding botulinum toxin must inject and make With disadvantage with high costs.
Nonapeptide -1: this is a kind of bionical peptide, it has extraordinary matching with the MC1 receptor on melanocyte, therefore It can be used as the antagonist of melanocyte-stimulating hormone, it is emulative in conjunction with MC1 receptor, prevent tyrosinase from further being swashed It lives and generates melanin.
Streptococcus lactis peptide by Lactococcus lactis or streptococcus lactis secrete it is a kind of be made of 34 amino acid residues it is more Peptide matters, can inhibit most gram-positive bacterias, especially to generating the gram-positive bacteria of gemma such as bacillus subtilis, thermophilic Hot Bacillus stearothermophilus etc. has very strong inhibiting effect.
Specific embodiment
The present invention is described further combined with specific embodiments below:
In following embodiment, palmityl tripeptides -1, -1 copper of tripeptides, acetyl group tetrapeptide -5, palmityl tetrapeptide -7, acetyl group six Peptide -8 is purchased from the pharmacy of Zhuhai federation；Palmityl tripeptides -5, carnosine, nonapeptide -1 are purchased from Shenzhen Wei Qi Biotechnology Co., Ltd； Streptococcus lactis peptide is purchased from Luoyang Qihong Biotechnology Co., Ltd..
Embodiment 1
The preparation of Oligopeptide Compositions
Palmityl tripeptides -1,100ppm solution, 200mL
- 1 copper of tripeptides, 500ppm solution, 20mL
Palmityl tripeptides -5,1000ppm solution, 10mL
Carnosine, powder, 0.9g
Acetyl group tetrapeptide -5,1000ppm solution, 10mL
Palmityl tetrapeptide -7,50ppm solution, 200mL
Argireline, 500ppm solution, 30mL
Nonapeptide -1,100ppm solution, 30mL
Streptococcus lactis peptide, 0.18g.
By above-mentioned oligopeptides according to dissolution properties difference, water is added to be prepared into the solution of 1000mL.
Using above-mentioned solution as mother liquor, it is acceptable to be added to cream, lotion or other cosmetics and skincare products for directly preparation In matrix；10 grams of mannitol can also be added in the solution according to lyophilization, according to 1 ~ 1.5 milliliter/cillin bottle point Freeze-dried powder is made in dress, vacuum freeze drying, gland (sealing).
Embodiment 2
Aging skin is crease-resistant, subtracts wrinkle repair procedure
After conventional cosmetic care program, 70% medicinal alcohol of skin at wrinkle is sterilized, is prepared with embodiment 1 1 addition 1 ~ 3mL physiological saline of freeze-dried powder is dissolved, which is applied to skin, is massaged into and fully absorbs. Beauty and the common instrument and method of medical cosmetology, including ultrasonic instrument, ion introducing apparatus or micropin, water light can also be used The modes such as needle, laser, injection import, and help skin to absorb, to reach optimal skin repair effect.
Embodiment 3
Proliferation of human dermal fibroblasts activity test
Material
The raw material of embodiment 1, be made 0.3 times, 1.0 times, 3.0 times of Oligopeptide Compositions it is spare, human skin fibroblasts (day Gear division research department, this Hokkaido University), α-MEM culture medium (GIBCO), fetal calf serum (GIBCO) mycillin (Invitrogen), Trypan blue solution(Sigma), 0.25% Trypsin-1mM EDTA ˙ 4Na (Invitrogen), IMSO(Sigma), cell number assay kit WST-8(Japan) the full-automatic microplate reader of Infinite500 (Tecan).
Method and result
Human skin fibroblasts are in MEM culture medium (containing 5% fetal calf serum), 5%CO2, 95% humidity, 37 DEG C of CO2Culture It is cultivated in case, changes liquid 1 time/3 days, the cell passage of culture 5 ~ 6 days is sucked out original fluid, washs 2 times through PBS, is added 0.25% At 37 DEG C after 30 seconds, MEM culture medium 10mL is added in Trypsin-1mM EDTA ˙ 4Na solution 0.5mL, and soft pressure-vaccum makes thin Born of the same parents suspend, and move to 50mL centrifuge tube, are centrifuged 2 minutes under 2000rpm, absorb supernatant, appropriate serum-containing media are added, gently Soft pressure-vaccum enables cell even suspension.96 orifice plates are taken, 100 μ L(of cell suspension is added in every hole and contains 1000, cell), it is attached to cell After, the 10 μ L of composition for containing nine kinds of oligopeptides is added by group respectively, and using 10 μ L of equal amount PBS as blank control.Containing cell and 96 orifice plates of tester are cultivated 72 hours in incubator, take out 96 orifice plates, 10 μ L of WST-8 solution are added into each hole, again It is put into incubator to be incubated for 4 hours and develop the color, take out 96 orifice plates, OD value is measured under 450nm, data press mean ± standard deviation table Show, statistical procedures are carried out to data with 11.0 For Windows statistical software of SPSS.Sample room comparison in difference is examined with t. P < 0.05 indicates statistical difference.As a result as follows:
Influence of nine kinds of Oligopeptide Compositions to proliferation of human dermal fibroblasts
Test grouping n OD
Control 4 0.265±0.038
0.3 times of Oligopeptide Compositions 4 0.685±0.057*
1.0 times of Oligopeptide Compositions 4 0.643±0.065*
3.0 times of Oligopeptide Compositions 4 0.626±0.083*
* P < 0.01, the composition test of three groups of oligopeptides are significantly higher than control group, i.e., the composition of oligopeptides, which has, increases into fibre Tie up the effect of cell growth.
Embodiment 4
Collagen test
Material
The Oligopeptide Compositions of embodiment 1, human skin fibroblasts (gear division research department, Hokkaido, Japan university), α-MEM Culture medium (GIBCO), fetal calf serum (GIBCO) mycillin (Invitrogen), Trypan blue solution (Sigma), 0.25% Trypsin-1mM EDTA ˙ 4Na(Invitrogen), IMSO(Sigma), collagen kit (Sircol(tm) Soluble Collagen Assay, Biocolor).
Method and result
96 orifice plates are taken, 1000 cells are added by every hole, change the liquid once within every 3 days, after cell attachment, are added respectively in each group Enter to contain the 10 μ L of composition solution of nine kinds of oligopeptides, and equal amount PBS is added as blank control.After culture 7 days, said by kit Bright book step measures I-type collagen content.All data are indicated with mean ± standard deviation.Using 11.0 For of SPSS Windows statistical software carries out statistical procedures.Sample room comparison in difference is examined with t, and P < 0.05 indicates statistical difference, As a result as follows:
The influence that the composition of nine kinds of oligopeptides secretes human skin fibroblasts I-type collagen
Control group (n=4) Oligopeptide Compositions test group (n=4)
203.56±19.36ng/mL 533.82±32.42ng/mL*
* P < 0.01 two groups of significant difference, the composition group of oligopeptides obviously increase the secretion of I-type collagen.
Embodiment 5
Skin surface bacterial count test
Material
The Oligopeptide Compositions of embodiment 1.Test reference strain: staphylococcus aureus (SA, this laboratory save), hammer Bacterium (SC, this laboratory save), propionibacterium acnes (PA, this laboratory save), escherichia coli (E.coli, this experiment Room saves).(formula: twisting thin fresh beef 500g to Anaerobic Agar culture medium, and triply distilled water 500mL is added, and 1 M NaOH is added 25 mL, stir evenly, and place 4 DEG C overnight, and next day heating hydrolysis 1 hour, multilayer cotton filtering is added into clear filtrate Tryptone 30g, yeast powder 5g, K2HPO45g, 0.025% resazurin 4mL, adds triply distilled water to 1000mL；It boils and cools down Culture medium is placed in anaerobic box (80%N2、10%H2、10%CO2), and add l-cysteine hydrochloric acid 0.5g, protoferriheme solution 1.5%) 10mL, vitamin K1 solution 0.2mL, agar content are set as.Anaerobic culture box (Shanghai).
Method and result
5 people of subject, male 2 (30 years old and 45 years old), women 3 (22 years old, 25 years old and 30 years old).By the subject wing of nose Two sides are denoted as left side blank district and the right side oligopeptides test section of 2 ~ 3cm of diameter.For operator with sterile gloves, use is sterile Exempt from swab and dip sterile saline and Oligopeptide Compositions respectively, uses physiology salt respectively in wing of nose two sides daily with first five day is sampled Water and Oligopeptide Compositions wipe repeatedly (about 10 times), and the cotton swab after the 6th day being wiped again (about 20 times) cuts off shank, put Enter in 5mL physiological saline.Continuous 10 times of dilutions are done to 10 samples of sampling, are diluted to 10--6, 10 are taken respectively-3、10-4、10-5、 10-6Dilution 0.1mL is inoculated on Anaerobic Agar plate respectively with L stick, while with containing golden yellow coccus, streptococcus, acne third The bacterium solution of acidfast bacilli and escherichia coli is positive controls, and being separately added into 10 μ L of Oligopeptide Compositions is positive test group, It is placed in anaerobic culture box.After Anaerobic culturel 3-4 days, observation is as a result, count the clump count of growth.As a result as follows:
Oligopeptide Compositions influence (clump count cfu/mL) to the clump count of skin surface bacterium and Reference Strains
Group/dilution 10-3 10-4 10-5 10-6
Positive group
SA 160 20 3 0
SA+ composition 93** 7 0 0
SC 180 15 2 0
SC+ composition 165 21 2 0
PA 190 25 3 0
PA+ composition 115** 15 0 0
E.coli 205 23 5 0
E.coli+ composition 190 20 4 0
Blank group (n=5) 186.25±28.34 17.00±0.00 - -
Test group (n=5) 103.31±21.36* - - -
* compared with blank group, clump count reduces significant for P < 0.05.SA and PA in * positive control and positive test, Prompt has a degree of clump count to reduce in the presence of composition., clump count is few, and does not do statistical procedures.
Embodiment 6
Cutaneous sensibility test
Material
The Oligopeptide Compositions of embodiment 1.New Zealand White Rabbit (2.36 ± 0.17kg) 8, male and female each 4, Zhongshan University is real Animal center offer is provided.Physiological saline.
Method and result
8 white rabbits are randomly divided into intact skin group and damaged skin group, every group 4, every white rabbit sub-cage rearing, in examination First 24 hours are tested in white rabbit backbone two sides antimere, with operating scissors cropping, with depilatory agent (barium sulfate starch washing powder 3:7:1) Depilation, hair removal section area are each about 50cm in the every side 10%(of body surface area2).In hair removal section row once give tester and repeatedly Tester is given, observes skin wound repair intensity, nine kinds of Oligopeptide Compositions (left side) are done in two sides respectively and physiological saline is (right Side) test.24 hours after skin depilatory, hair-fields skin integrity is removed in inspection, and damaged skin is unfit to do intact skin irritant test, The production of damaged skin is going hair-fields skin (through 75% alcohol disinfecting) to draw " # " word cut using disposable sterilized No. 7 syringe needles, Damage zone area 3cm × 3cm does not hurt corium to stab epidermis, has oozing of blood for degree, and the damaged degree of the left and right sides answers basic one It causes.Left side hair removal section smears the composition of nine kinds of oligopeptides, and physiological saline is smeared on right side, smears position with two layers of gauze covering, uses Nonirritant adhesive plaster fixes gauze, then is fixed with mesh nylon tape.Every white rabbit sub-cage rearing, the complete group of breakage combination are respectively set Single administration and multiple dosing once a day, after giving tested material 24 hours, were removed by upper method successive administration seven days with warm water Tested material is remained, visually observed and records in 1,24,48 and 72 hour and smear that whether there is or not erythema and oedema at position.As a result as follows:
Skin irritation reaction scoring
Erythema: without erythema, score value: 0；Reluctantly as it can be seen that score value: 1；, it is apparent that score value: 2；Moderate is to Severe erythema, and 3 Part；Aubergine erythema simultaneously has eschar to be formed, score value: 4.
Oedema: without oedema, score value: 0；Reluctantly as it can be seen that score value: 1；Slight visible (edge is higher by surrounding skin), score value: 2； Cutaneous protuberance about 1mm is clearly outlined, score value: 3；Cutaneous protuberance about 1mm or more and range expansion, score value: 4.
Erythema/oedema
Grouping 1 hour 24 hours 48 hours 72 hours Scoring
Intact skin group
1(single administration) 0/0,0/0 0/0,0/0 0/0,0/0 0/0,0/0 0
2(single administration) 0/0,0/0 0/0,0/0 0/0,0/0 0/0,0/0 0
3(multiple dosing) 0/0,0/0 0/0,0/0 0/0,0/0 0/0,0/0 0
4(multiple dosing) 0/0,0/0 0/0,0/0 0/0,0/0 0/0,0/0 0
Damaged skin group
1(single administration) 0/0,0/0 1/1,1/1 0/0,0/0 0/0,0/0 0.25
2(single administration) 0/0,0/0 1/1,1/1 0/0,0/0 0/0,0/0 0.25
3(multiple dosing) 0/0,0/0 1/1,1/1 0/0,0/0 0/0,0/0 0.25
4(multiple dosing) 0/0,0/0 1/1,1/1 0/0,0/0 0/0,0/0 0.25
Intact skin and damaged skin are showed no the skin irritatin of Oligopeptide Compositions by standards of grading compared with the control group Property.
Embodiment 7
The preparation (mass percent) of cream:
Water 61.75
Glycerol 5.00
Propylene glycol 3.00
Carbomer 0.15
Cetearyl glucoside 3.00
PEG-100 stearate/stearine 2.00
Cetostearyl alcohol 1.00
Caprylic/capric triglyceride 5.00
Dimethyl silicone polymer 2.00
Butyrospermum parkii fruit rouge 2.00
Tocopherol acetate 0.50
Isohexadecane 8.00
Glycerol caprylate 0.50
Triethanolamine 0.10
The Oligopeptide Compositions 5.00 of embodiment 1
1,2- pentanediol 1.00
Preparation method:
1, Jiang Shui, glycerol, propylene glycol, carbomer stirring are warming up to 85 DEG C, make it completely dissolved, insulated and stirred 30 minutes；
2, by cetearyl glucoside, PEG-100 stearate/stearine, cetostearyl alcohol, the octanoic acid/last of the ten Heavenly stems Acid glycerol three ester, dimethyl silicone polymer, Butyrospermum parkii fruit rouge, tocopherol acetate, isohexadecane, glycerol caprylate stirring rise Temperature makes it completely dissolved to 80 DEG C, and insulated and stirred 20 minutes；
3, the mixture of step 2 is added in the mixture of step 1, is stirred homogeneous 5 minutes, insulated and stirred 10 minutes；
4, stirring is cooled to 45 DEG C, and triethanolamine, 1,2- pentanediol, Oligopeptide Compositions are added and stir 15 minutes, have prepared At.
Embodiment 8
The preparation (mass percent) of emulsion:
Water 69.60
Glycerol 3.00
Propylene glycol 5.00
Carbomer 0.10
Glyceryl stearate citrate 3.00
Cetostearyl alcohol 1.50
Caprylic/capric triglyceride 3.00
Dimethyl silicone polymer 1.00
Butyrospermum parkii fruit rouge 1.00
Tocopherol acetate 0.20
Isohexadecane 6.00
Glycerol caprylate 0.50
Triethanolamine 0.10
1,2- pentanediol 1.00
The Oligopeptide Compositions 5.00 of embodiment 1
Preparation method:
1, Jiang Shui, glycerol, propylene glycol, carbomer stirring are warming up to 85 DEG C, make it completely dissolved, insulated and stirred 30 minutes；
2, by Glyceryl stearate citrate, cetostearyl alcohol, caprylic/capric triglyceride, dimethyl silicone polymer, Butyrospermum parkii fruit rouge, tocopherol acetate, isohexadecane, glycerol caprylate stirring are warming up to 80 DEG C, make it completely dissolved, keep the temperature Stirring 20 minutes；
3, the mixture of step 2 is added in the mixture of step 1, is stirred homogeneous 5 minutes, insulated and stirred 10 minutes；
4, stirring is cooled to 45 DEG C, and triethanolamine, 1,2- pentanediol, Oligopeptide Compositions are added and stir 15 minutes, have prepared At.
Embodiment 9
Cream is prepared according to the formula and method of embodiment 7, according to parts by weight, Oligopeptide Compositions are by 1 part of palmityl three Peptide -1,1 part of -1 copper of tripeptides, 1 part of palmityl tripeptides -5,1 part of carnosine, 1 part of acetyl group tetrapeptide -5,1 part of palmityl tetrapeptide -7,0.1 Part Argireline, 0.1 part of nonapeptide -1,1 part of streptococcus lactis peptide composition.
Embodiment 10
Emulsion is prepared according to the formula and method of embodiment 8, according to parts by weight, Oligopeptide Compositions are by 30 parts of palmityls three Peptide -1,30 parts of -1 copper of tripeptides, 50 parts of palmityl tripeptides -5,30 parts of carnosines, 30 parts of acetyl group tetrapeptides -5,30 parts of palmityl tetrapeptides - 7,30 parts of Argirelines, 30 parts of nonapeptides -1,30 parts of streptococcus lactis peptide compositions.
The above description is merely a specific embodiment, but scope of protection of the present invention is not limited thereto, any Belong to those skilled in the art in the technical scope disclosed by the present invention, any changes or substitutions that can be easily thought of, all answers It is included within the scope of the present invention.Therefore, protection scope of the present invention should be subject to the protection scope in claims.

Claims (6)

1. a kind of compound Oligopeptide Compositions for skin repair, which is characterized in that according to parts by weight, including 0.02 portion of palm - 1,0.01 part of -1 copper of tripeptides of acyl tripeptides, 0.01 part of palmityl tripeptides -5,0.9 part of carnosine, 0.01 part of acetyl group tetrapeptide -5,0.01 - 7,0.015 parts of Argirelines of part palmityl tetrapeptide, 0.003 part of nonapeptide -1,0.18 part of streptococcus lactis peptide.
2. Oligopeptide Compositions according to claim 1, which is characterized in that the Oligopeptide Compositions further include water, grease, table Face activating agent, moisturizer, antioxidant, at least one of the preservatives.
3. described in any item Oligopeptide Compositions are preparing the application in cosmetics or curable product according to claim 1~2.
4. application according to claim 3, which is characterized in that the preparation type of the cosmetics or curable product is essence Magnificent liquid, cream, freeze-dried powder, solution, emulsion, gel, facial mask or dressing.
5. a kind of cream, which is characterized in that including 5wt% Oligopeptide Compositions, 61.75wt% water, 5wt% glycerol, 3wt% the third two Alcohol, 0.15wt% carbomer, 3wt% cetearyl glucoside, 2wt%PEG-100 stearate/stearine, 1wt% cetostearyl alcohol, 5wt% caprylic/capric triglyceride, 2wt% dimethyl silicone polymer, 2wt% Butyrospermum parkii fruit rouge, 0.5wt% tocopherol acetate, 8wt% isohexadecane, 0.5wt% glycerol caprylate, 0.1wt% triethanolamine, 1wt%1,2- Pentanediol, wherein Oligopeptide Compositions are as described in claim 1.
6. a kind of emulsion, which is characterized in that including 5wt% Oligopeptide Compositions, 69.6wt% water, 3wt% glycerol, 5wt% the third two Alcohol, 0.1wt% carbomer, 3wt% Glyceryl stearate citrate, 1.5wt% cetostearyl alcohol, 3wt% caprylic/capric are sweet Oily three esters, 1wt% dimethyl silicone polymer, 1wt% Butyrospermum parkii fruit rouge, 0.2wt% tocopherol acetate, 6wt% different 16 Alkane, 0.5wt% glycerol caprylate, 0.1wt% triethanolamine, 1wt%1,2- pentanediol, wherein Oligopeptide Compositions such as right is wanted It asks described in 1.
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