CN1375276A

CN1375276A - Skin cosmetics containing horny cell growth factor

Info

Publication number: CN1375276A
Application number: CN 01111588
Authority: CN
Inventors: 范开; 张益�; 马素永; 聂李亚
Original assignee: Chongqing Thinking Cosmetics Co Ltd; Fujin Bio-Medicine Co Ltd Chongqing
Current assignee: Chongqing Thinking Cosmetics Co Ltd; Fujin Bio-Medicine Co Ltd Chongqing
Priority date: 2001-03-20
Filing date: 2001-03-20
Publication date: 2002-10-23

Abstract

The present invention belongs to biological ficial cosmetics is a skin cosmetics prepared by KGF, vitamin E, hyaluronic acid etc. additives. This cosmetics can enhance the metabolism of skin horny cell and undertake the functions of crease-resistance and improving one's looks.

Description

The cosmetics for skin that contains keratinocyte growth factor

Skin is a working organ, comprises many tissues, cell type and specialized structure, and existence and health are played many crucial effects.Skin is made up of the two-layer of mutual dependence for existence, the corium of promptly outer field epidermis and internal layer.Wherein epidermal area is that cosmetics for skin is at the zone, in the formation that prevents skin aging, wrinkle and smoothly play an important role aspect tender.

Epidermis is that stratum germinativum and akaryote zone are that horny layer is formed by vigourous cell zone, and stratum germinativum also can be divided into basal layer, spinous layer and granular layer.Every layer of different phase of all representing epidermal keratinocyte differentiation and keratinization develops at last and is positioned at horn cell skin surface, death.The differentiation of epidermis cell relates to fibrin and keratic formation, and the maturation process of epidermis is finished at horny layer, forms the proteic cell of accumulated angle.Stratum basale has the constant germinal cell group of a group, and its offspring all passes through specific atomization.New horn cell approximately needed 14 days to develop into the granular layer cell gradually, then came off the metabolic processes of the epidermal keratinocyte of promptly about 28 days one-period again through 14 days angle of arrival laminar surfaces.Suitably control the breeding of horn cell in its bottom and, can make skin surface gloss and fine and soft to horny layer process of differentiation successively.If all can cause the skin surface out-of-flatness unusually, show as coarse, desquamation and hyperkeratosis because of various factors comes off the change of horn cell division cycle, the defective of cell differentiation or horn cell.Intradermal fibrin such as collagen protein, elasticin and amorphous ground substance are given skin tenacity, elasticity and viscosity.

Along with the variation at age, the various 26S Proteasome Structure and Functions of skin are also changing.The invasion and attack of environment, particularly long-term sun exposure, more much bigger to the damage of skin more than age ageing.

Following variation appears in the structure of skin, physiology, the biochemical increase with the age: (1) 70 years old and comparison in 30 years old, the cell conversion ratio approximately reduces 50% in the epidermis, corium elastic fibers and collagen fiber all reduce, the reason that characteristic that the apparent pattern of the skin of aging that Here it is is thin and Clear ﹠ Transparent and corium blood vessel are easy to rupture.(2) along with the age becomes big, the crosslinked increase of collagen fiber and elastic fibers, but make the corium tropism weaker, thereby lear energy reduces.When skin is pinched time-out, old people's skin slowly restores to the original state.Elastic tissue degeneration causes wrinkle to be deepened.(3) melanocyte enzymatic activity attenuating (per 10 years attenuating 10-20%) causes the irregular pigmentation of skin, and hair becomes canescence.

Therefore, the elasticity of skin depends on the connective tissue of corium and the normal configuration and the function of substrate, the smooth or coarse aquation and the normal SkBF that depends on normal keratinization, horny layer appropriateness of skin.

The hypertrophy of skin surface horn cell, differentiation and fibrinous secretory function are subjected to the adjusting of multiple protein active factors, comprising keratinocyte growth factor (KGF), fibroblast growth factor (FGF), transforming growth factor (TGF),

The somatomedin (PDGF) in epidermal growth factor (EGF) and platelet source.These protein factors are coordinated mutually, safeguard the normal metabolic process of skin jointly.Wherein some protein factor plays a major role in the growth course of skin, and some protein factor plays a major role in the injury repairing of skin.

Humanized's horn cell factor (Keratinocyte growth factor) belongs to FGF family, is made up of 194 aminoacid, and about 24,000 dalton of molecular weight are strand bioactive molecules.Mainly act on the horn cell of skin surface, mucomembranous epithelial cell and fibrocyte are had certain effect.Confirmed that KGF comprises for the function of the horn cell of skin: 1) promote skin keratin hyperplasia; 2) differentiation successively of promotion horn cell; 3) promoting horn cell eccrine fiber albumen such as collagen protein, 4) the shielding angle cell plastid is not subjected to the damage of extraneous factor such as ultra-vioket radiation.

More than these effects for the eubolism of skin, prevent skin aging, keep elasticity and skin smooth and fine and soft very important.

This patent is keratinocyte growth factor (KGF) is applied in the cosmetics for skin, with other biological beauty active matter such as vitamin E and hyaluronic acid etc., corresponding preparation technology by cosmetics, make cream, emulsion or water preparation, reach and prevent skin aging, strengthen elasticity and keep the glossy effect of skin.

The preparation of example one recombinant human horny cell growth factor-2

Human horny cell growth factor-2 (KGF) is made up of 194 aminoacid, and its cDNA template gets with the PCR method amplification.Concrete grammar is; The people KGF cDNA plasmid PUC-hKGF that contains 582bp is as template, synthetic one couple of PCR primers, forward primer: 5 '-CG CAT ATG CAC AAA TGG ATA CTG ACA TGG ATC-3 '-reverse primer 5 '-GC GGA TCC TTA AGT TAT TGC CAT AGG AAG AAA G-3 ', under Tag enzyme and dNTP condition, 94 ℃, 1min, 56 ℃, 1min and 72 ℃, 1min circulation 32 times obtains the PCR fragment of about 600bp.This PCR fragment and P ^ET9mPlasmid reclaims respectively behind Ndc I and Bam HI double digestion.Under the effect of T4 ligase, link transformed into escherichia coli DH32, screening recombiant plasmid called after P ^ET-hkgf(with 0.5mM IPTG abduction delivering, wherein solubility expression reorganization KGF accounts for 30% to this recombinant plasmid transformed BL21 for DE3, Lys) host bacterium.

The engineering bacteria of expressing K GF, through 10 liters of ferment tanks, centrifugal collection thalline.Thalline is crossed Heparin-Sepharose FF and Superdex G75 secondary column chromatography behind pressure homogenate crusher machine, obtain purity at the recombined human KGF more than 90%.The reorganization KGF molecular weight of purification is consistent with design load with N-terminal order-checking confirmation, cultivates to survey through cell in vitro and lives, and its biological activity is 1.0 * 10 ⁶Unit/mg albumen.

Example two contains the preparation of the cosmetics frost of KGF

The KGF that will recombinate is made into every milliliter and contains the 2mg aqueous solution behind determining the protein quantity, make the cosmetics for skin frost with following prescription and method.

Prescription: (quality %)

A, triple pressed stearic acid 4

Octadecanol 4

Glyceryl monostearate 1.5

Paregal O 1

White oil 3

Myristic acid isopropyl ester 3

Squalane 2

DC-200 0.5

Mud is moored golden third fat 0.05

B, distilled water 79

Keratinocyte growth factor 1.0ml (0.002%)

Mud pool tortoise beetle fat 0.18

Vitamin E 0.2

Hyaluronic acid 0.25

C, essence are an amount of

Method for making: A is mixed and heated to 95 ℃ makes oil phase, B is heated to 95 ℃ makes water, A slowly is added among the B,

Even matter 5 minutes constantly stirs adding C when being cooled to 50 ℃, is chilled to the room temperature discharging.

Example three contains the preparation of the cosmetic emulsion of keratinocyte growth factor

Recombinant human horny cell growth factor-2 (KGF) is made every milliliter of aqueous solution that contains 2mg, by following prescription and method preparation.

Prescription (quality %):

A, octadecanol 0.9

Glyceryl monostearate 0.8

Paregal O 1

White oil 3

Myristic acid isopropyl ester 3

Squalane 2

DC-200 0.5

Mud is moored golden third fat 0.05

B, distilled water 85

Keratinocyte growth factor 1.0ml (0.002%)

Vitamin E 0.2

Glycerol 6

QM thickening agent 0.1

Mud pool tortoise beetle fat 0.18

C, triethanolamine 0.15

An amount of method for making of essence: A is heated to 95 ℃ makes oil phase, B part water is added the dissolving of QM thickening agent heated and stirred, temperature to 95 ℃, all the other B are heated to 95 ℃ at large water tank, and the large water tank sucking filtration is advanced reaction pot, and A is filtered into reaction pot, then QM thickening agent water liquid is filtered into reaction pot, start even matter about 5 minutes, and constantly be stirred to 50 ℃ and add C, be chilled to the room temperature discharging.

Claims

1, this patent is characterised in that keratinocyte growth factor (KGF) is added in the substrate of cosmetics for skin, plays wrinkle resistant and U.S. face effect.

2, the additional proportion of keratinocyte growth factor in cosmetics is 0.001-0.01%.

3, Keratiocyte growth mainly refers to recombinant human horny cell growth factor-2 by technique for gene engineering preparation in the clause 2.

4, the cosmetics for skin that contains keratinocyte growth factor in the clause 1 comprises cream, Emulsion and the water preparation of making thus.