WO2002031496A2 - Device for determining skin aging in vitro - Google Patents

Device for determining skin aging in vitro Download PDF

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Publication number
WO2002031496A2
WO2002031496A2 PCT/EP2001/011281 EP0111281W WO0231496A2 WO 2002031496 A2 WO2002031496 A2 WO 2002031496A2 EP 0111281 W EP0111281 W EP 0111281W WO 0231496 A2 WO0231496 A2 WO 0231496A2
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WIPO (PCT)
Prior art keywords
skin
acid
stress
aging
fragments
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PCT/EP2001/011281
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German (de)
French (fr)
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WO2002031496A3 (en
Inventor
Dirk Petersohn
Günter Schmitt
Thomas Förster
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Henkel Kommanditgesellschaft Auf Aktien
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Priority to AU2002213982A priority Critical patent/AU2002213982A1/en
Publication of WO2002031496A2 publication Critical patent/WO2002031496A2/en
Publication of WO2002031496A3 publication Critical patent/WO2002031496A3/en

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    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/5005—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells
    • G01N33/5008—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics
    • G01N33/5082—Supracellular entities, e.g. tissue, organisms
    • G01N33/5088—Supracellular entities, e.g. tissue, organisms of vertebrates
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
    • G01N33/6881—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids from skin

Definitions

  • the present invention relates to methods for determining skin stress and / or skin aging in humans or animals in vitro, test kits for determining skin stress and / or skin aging and the use of spondin 2, cathepsin L, vimentin fragments or actin gamma 1 as a stress and / or aging marker of the skin; furthermore a method for proving the effectiveness of cosmetic or pharmaceutical active substances against skin stress and / or skin aging as well as means for regulating, in particular for maintaining the homeostasis of human or animal skin.
  • the human skin is a very complex organ, which consists of a large number of different cell types.
  • the metabolism of living cells is not static but very dynamic.
  • cells notice changes in their environment (e.g. sunlight) and react to this by changing their RNA and / or protein synthesis performance.
  • Some molecules are increasingly synthesized after a stress stimulus (e.g. sunlight) (e.g. MMP-1), others are produced to a lesser extent (e.g. collagen CH (I)). Furthermore, there will be no significant change in a large number of the synthesis processes (e.g. TIMP-1).
  • a stress stimulus e.g. sunlight
  • MMP-1 e.g. MMP-1
  • others are produced to a lesser extent (e.g. collagen CH (I)).
  • TIMP-1 stress stimulus
  • the macroscopic phenomena of aged skin are based on the one hand on the intrinsic or chronological aging, and on the other hand on the extrinsic aging caused by environmental stress, such as sunlight.
  • the visible times The areas of skin that have aged due to stress are to be understood as the sum of many individual events that accumulate in the skin over a longer period of time.
  • the skin consists of several different cell types (fibroblasts, keratinocytes in different states of differentiation, melanocytes, Langerhans cells, hair follicle cells, sweat gland cells etc.) and the complexity of the protein mixtures produced in the skin is therefore very great.
  • the proteins spondin 2, cathepsin L, actin gamma 1 and fragments of the protein vimentin are secreted into the extracellular space by skin fibroblasts, but not by stressed fibroblasts, but not. Surprisingly, it was also found that fragments of the protein vimentin can help to maintain the homeostasis of the skin.
  • the present invention therefore relates to a method for determining skin stress and / or skin aging in humans or animals in vitro, characterized in that a) a skin sample containing fibroblasts is obtained, b) the fibroblasts are isolated and cultured, and c) the culture supernatant the presence and the amount of the proteins or protein fragments secreted by the fibroblasts Spondin 2, Cathepsin L, Vimentin fragments or Actin Gamma 1 were examined.
  • Another object of the invention is a method for determining skin stress and / or skin aging in humans or animals in vitro, characterized in that a) a mixture of the proteins secreted by the skin cells is obtained by microdialysis and b) the mixture is examined for the presence and the amount of the proteins or protein fragments secreted by the fibroblasts Spondin 2, Cathepsin L, Vimentin fragments or Actin Gamma 1.
  • microdialysis technique is described, for example, in “Microdialysis. A method for measurement of local tissue metabolism ", Nielsen PS, Winge K, Petersen LM; Ugeskr Laeger 1999 Mar 22 161: 12 1735-8; and in” Cutaneous microdialysis for human in vivo dermal absorption studies ", Anderson, C. et al. ; Drugs Pharm. Sci., 1998, 91, 231-244; and also described on the Internet at http://www.microdialysis.se/techniqu.htm, to which reference is hereby made in full.
  • microdialysis When using microdialysis, a probe is typically inserted into the skin and the probe is slowly rinsed with a suitable carrier solution. After the acute reactions have subsided after the puncture, the microdialysis provides proteins which occur in the extracellular space and which can then be isolated and analyzed in vitro, for example by fractionation of the carrier liquid.
  • fibroblasts The cultivation of fibroblasts is known to the person skilled in the art and can be carried out according to the invention in a customary manner, for example as in Langholz, O. et al .; 1997; Experimental Cell Research 235, 22-27; described.
  • the methods according to the invention are preferably carried out in such a way that the examination for the presence and the amount of the proteins or protein fragments secreted by the fibroblasts Spondin 2, Cathepsin L, Vimentin fragments or Actin Gamma 1 by separating the proteins by means of gel electrophoresis and subsequent mass spectrometry, in particular Matrix Assisted Laser Desorption Ionization (MALDI).
  • MALDI Matrix Assisted Laser Desorption Ionization
  • the separation of the proteins is particularly preferably carried out by 2D gel electrophoresis, such as, for example, in LD Adams, Two-dimensional Gel Electrophoresis using the Isodalt System or in LD Adams & SR Gallagher, Two-dimensional Gel Electrophoresis using the O'Farrell System; both in Curent Protocols in Molecular Biology (1997, Eds. FM Ausubel et al.), Unit 10.3.1 - 10.4.13; or in 2-D electrophoresis manual; T. Berkelman, T. Senstedt; Amersham Pharmacia Biotech, 1998 (Order No. 80-6429-60).
  • 2D gel electrophoresis such as, for example, in LD Adams, Two-dimensional Gel Electrophoresis using the Isodalt System or in LD Adams & SR Gallagher, Two-dimensional Gel Electrophoresis using the O'Farrell System; both in Curent Protocols in Molecular Biology (1997, Eds. FM Ausubel et al.), Unit
  • the proteins are obtained from the culture supernatant from dermal fibroblasts preferably by protein precipitation from the culture supernatant, for example with a methanol / chloroform mixture in a ratio of about 4: 1: 4 to the sample and / or by concentration of the proteins, for example by ultrafiltration with a cut off of about 5,000 Da, e.g. using the Vivaspin products from Sartorius.
  • the concentrate or precipitate thus obtained can then be taken up in rehydration buffer in a manner known to those skilled in the art, for example as in “2-D electrophoresis using immobilized pH gradients. Principles and methods ", Berkelman, T. and Stenstedt, T (eds); Amersham Pharmacia Biotech Ine (1998) 80-6429-60;
  • the separation of the proteins preferred according to the invention by 2-dimensional gel electrophoresis can be carried out, for example, by carrying out isoelectric focusing in the first dimension, for example using Immobilin DryStrips with an immobilized pH gradient (IPG) of 4 to 7 from Amersham -Pharmacia or using carrier ampholyte-bearing polyacrylamide gel strips which build up a pH gradient in the electric field (O'Farrell, PH; 1975; J Biol. Chem. 250, 4007-4021) and in the second dimension an SDS-polyacrylamide - Gel electrophoresis, for example with 12.5% polyacrylamide at 30 mA for 3 to 4 h.
  • IPG immobilized pH gradient
  • SDS-polyacrylamide - Gel electrophoresis for example with 12.5% polyacrylamide at 30 mA for 3 to 4 h.
  • Gel staining can be carried out by customary methods, for example by Heukeshoven silver staining or by colloidal Coomassie staining.
  • Spondin 2 is a protein recently identified in normal lung tissue (Manda, R., et al, (1999); Identification of Genes (SPON2 and C20orf2) ...; Genomics 61 (1), 5-14).
  • Spondin 2 a protein with the amino acid sequence (A)
  • amino acid sequence (A) is disclosed in the National Center for Biotechnology Information (NCBI) protein database under number 6172221. This database is accessible on the Internet at the following address: http://www.ncbi.nlm.nih.gov/.
  • pondin 2 also includes those proteins whose amino acid sequence can be obtained from the above-listed sequence (A) by conservative mutations, that is to say by the exchange of structurally or functionally similar amino acids.
  • Cathepsin L was first described as a lysosomal protease. Only later was the identity shown with the "major excretet protein (MEP) of the mouse.
  • Cathepsin L a protein with the amino acid sequence (B)
  • amino acid sequence (B) is in the protein database of the National Center for Biotechnology Information (NCBI) under number 4503155 accessible: http://www.ncbi.nlm.nih.gov/.
  • cathepsin L also includes those proteins whose amino acid sequence can be obtained from the sequence (B) listed above by conservative mutations, that is to say by the exchange of structurally or functionally similar amino acids.
  • Vimentin is an intermediate filament that is involved in the formation of the cytoskeleton in the cell. Surprisingly, it was found that certain vimentin fragments of fibroblasts are secreted after exposure to sunlight. This is a fact that can be used to determine the stress level of human skin. In this way, stress-induced vimentin fragments can be isolated and quantified by microdialysis methods on the test subject and thus serve as a unit of measurement for skin stress.
  • “Vimentin fragments” are protein fragments with the amino acid sequence (C)
  • NCBI National Center for Biotechnology Information
  • vimentin fragment also includes those proteins or protein fragments whose amino acid sequence can be obtained from the sequence (C) listed above by conservative mutations, that is to say by the exchange of structurally or functionally similar amino acids.
  • Actin is a very conserved protein in the organism realm.
  • the actins of mammals are divided into three classes according to their isoelectric point. Actin Alpha is produced by skeletal muscles, Actin Beta by smooth muscles. Actin gamma is synthesized by all other cells, including dermal fibroblasts. Globular actin (G-actin) is arranged in a very dynamic process in rows and thus forms filamentous actin (F-actin). Stressed cells arrange F-actin in complex bundles called "stress fibers". Surprisingly, it was found that actin molecules are secreted by cells after the application of stress.
  • actin Gamma 1 a protein with the amino acid sequence (D)
  • amino acid sequence (D) is disclosed in the National Center for Biotechnology Information (NCBI) protein database under number 4501887. This database is accessible on the Internet at the following address: http://www.ncbi.nlm.nih.gov/.
  • actin gamma 1 also includes those proteins whose amino acid sequence can be obtained from the sequence (D) listed above by conservative mutations, that is to say by the exchange of structurally or functionally similar amino acids.
  • Another object of the invention is a test kit for determining the skin stress and / or skin aging in humans or animals in vitro, which comprises the necessary means for performing the above-mentioned methods according to the invention.
  • Another object of the invention is the use of spondin 2, cathepsin L, vimentin fragments or actin gamma 1 according to the above definitions as a stress and / or aging marker of the skin in humans or animals.
  • the aging markers mentioned are preferably used in one of the methods according to the invention. The aging markers can be used individually or in any combination.
  • a ⁇ further ⁇ Ge "ge ⁇ stand ⁇ de ⁇ _ invention ⁇ is _ a ⁇ Verfahre ⁇ rzum demonstrating the effectiveness of cosmetic or pharmaceutical active agents against skin stress and / or skin aging in vitro, characterized in that a) determines the skin status through one of the methods according to the invention, b) applying an active substance against skin stress and / or skin aging to the skin one or more times, c) determining the skin status again by one of the methods according to the invention and d) determining the effectiveness of the active substance by comparing the results from a) and c).
  • Another object of the invention is a test kit for demonstrating the effectiveness of cosmetic or pharmaceutical active substances against skin stress and / or skin aging in vitro, which comprises the necessary means for carrying out the above-mentioned method according to the invention.
  • Another object of the invention is the use of spondin 2, cathepsin L, vimentin fragments or actin gamma 1 for the detection of the effectiveness of cosmetic or pharmaceutical active ingredients against skin stress and / or skin aging.
  • Another object of the invention is a cosmetic or pharmaceutical agent for regulating, in particular for maintaining the home human or animal skin stasis containing vimentin fragments as defined above.
  • the vimentin fragments can be the same or different.
  • the agents according to the invention are preferably applied topically, in particular in the form of solutions, dispersions (for example emulsions), ointments or creams.
  • Vimentin fragments can also be used in hand washing detergents, hand dishes- ⁇ ülmitte1n ⁇ ⁇ -.äe ⁇ ⁇ Ko ⁇ useful ⁇ to maintain the
  • Homeostasis of human or animal skin can be used.
  • mutually independent objects of the invention are therefore hand washing agents, hand dishwashing agents or personal care products which contain vimentin fragments as defined above.
  • Another object of the invention is the use of vimentin
  • the vimentin fragments are preferably introduced or incorporated as a component in a cosmetic or pharmaceutical preparation or in hand washing detergents, hand dishwashing detergents or personal care products.
  • the pharmaceutical compositions according to the invention can contain at least one further auxiliary or additive, such as, for. B. contain oils, protective colloids, plasticizers, antioxidants and / or emulsifiers.
  • a physiologically compatible oil such as, for example, sesame oil, corn oil, cottonseed oil, soybean oil or peanut oil.
  • ß oil, esters of medium-chain vegetable fatty acids or fish oils such as mackerel, sprat or salmon oil.
  • stabilizers such as a-tocopherol, t-butylated hydroxytoluene, t-butylated hydroxyanisole, ascorbic acid or ethoxyquine.
  • the dosage and duration of use of the vimentin fragments to be used according to the invention or of the agents containing vimentin fragments can be suitably adapted and varied by the person skilled in the art.
  • the hand detergents and hand dishwashing detergents according to the invention and the cosmetic and personal care products such as, for example, hair shampoos, hair lotions, foam baths, shower baths, creams, gels, lotions, alcoholic and aqueous / alcoholic solutions, emulsions, wax / fat masses, stick preparations, powders or ointments can - each by type of formulation - as auxiliary agents and additives, mild surfactants, oil bodies, emulsifiers, superfatting agents, pearlescent waxes, consistency agents, thickeners, polymers, silicone compounds, fats, waxes, stabilizers, biogenic agents, deodorants, antiperspirants, antidandruff agents, film formers, swelling agents, Contain UV protection factors, antioxidants, hydrotropes, preservatives, insect repellents, self-tanners, solubilizers, perfume oils, dyes and the like.
  • auxiliary agents and additives mild surfactants, oil bodies, emulsifiers
  • Suitable mild, ie particularly skin-compatible surfactants are fatty alcohol polyglycol ether sulfates, monoglyceride sulfates, mono- and / or dialkyl sulfosuccinates, fatty acid isethionates, fatty acid sarcosinates, Fettklaretau- ride, fatty acid glutamates, ⁇ -olefinsulfonates, ether carboxylic acids, Alkyloligoglu- coside, fatty acid glucamides, alkylamidobetaines and / or protein fatty acid condensates, the latter preferably based on wheat proteins.
  • Guerbet alcohols based on fatty alcohols having 6 to 18, preferably 8 to 10 carbon atoms, esters of linear C 6 -C 22 fatty acids with linear C 6 -C 22 fatty alcohols, esters of branched C 6 -Ci 3 carboxylic acids are examples of oil bodies with linear C 6 -C 22 fatty alcohols, such as, for example, myristyl myristate, myristyl palmitate, myristyl stearate, myristyl isostearate, myristyl oleate, myristyl behenate, myristyl erucate, cetyl myristate, cetyl palmitate, cetyl stearate, cetyl isyl stearate, cetyl isyl stolate, cetyl isyl styrene "Tatt _ Steäi7lste” afurs ⁇ Stearylisösteärat Steärylöleat, stearyl
  • esters of linear C6-C22 fatty acids with branched alcohols in particular 2-ethylhexanol
  • esters of hydroxycarboxylic acids with linear or branched C 6 -C 22 fatty alcohols in particular dioctyl malates
  • esters of linear and / or branched fatty acids with polyhydric alcohols such as propylene glycol, dimer diol or trimer triol
  • polyhydric alcohols such as propylene glycol, dimer diol or trimer triol
  • Guerbet alcohols triglycerides based on C ⁇ -cio fatty acids
  • esters of C 6 -C 22 - Fatty alcohols and / or Guerbet alcohols with aromatic carboxylic acids especially benzoic acid, esters of C2-Ci 2 -dicarboxylic acids with linear or branched alcohols with 1
  • Suitable emulsifiers are nonionic surfactants from at least one of the following groups:
  • alkyl and / or alkenyl mono- and oligoglycosides with 8 to 22 carbon atoms in the alk (en) yl radical and their ethoxylated analogs;
  • adducts of ethylene oxide and / or of propylene oxide with fatty alcohols, fatty acids, alkylphenols, glycerol mono- and diesters as well as sorbitan mono- and diesters with fatty acids or with castor oil are known, commercially available products.
  • Ci2 / i 8 fatty acid monoesters and diesters of adducts of ethylene oxide with glycerol are known from DE 2024051 PS as refatting agents for cosmetic preparations.
  • Alkyl and / or alkenyl mono- and oligoglycosides their preparation and their use are known from the prior art. They are produced in particular by reacting glucose or oligosaccharides with primary alcohols with 8 to 18 carbon atoms.
  • glycoside residue both monoglycosides in which a cyclic sugar residue is glycosidically bonded to the fatty alcohol and oligomeric glycosides with a degree of oligomerization of up to about 8 are suitable.
  • the degree of oligomerization is a statistical mean value based on a homolog distribution customary for such technical products.
  • polyglycerol esters are polyglyceryl-2 dipolyhydroxystearate (Dehymuls ® PGPH), polyglycerol-3-diisostearate (Lameform ® TGI), polyglyceryl-4 isostearate (Isolan ® Gl 34), polyglyceryl-3 oleate, diisostearoyl polyglyceryl 3 diisostearates (Isolan ® PDI), polyglyceryl-3 methyl glucose distearate (Tego Gare ® 450), polyglyceryl-3 beeswax (Cera Bellina ® ), polyglyceryl-4 caprate (polyglycerol caprate T2010 / 90), polyglyceryl-3 cetyl ether ( Chimexane ® NL), Polyglyceryl-3 Distearate (Cremophor ® GS 32) and Polyglyceryl Polyricinoleate (Admul ® WOL 1403), Polyglyceryl
  • Zwitterionic surfactants can also be used as emulsifiers.
  • Zwitterionic surfactants are surface-active compounds that contain at least one quaternary ammonium group and at least one carboxylate and one sulfonate group in the molecule.
  • Particularly suitable zwitterionic surfactants are the so-called betaines such as the N-alkyl-N, N-dimethylammonium glycinate, for example the coconut alkyldimethylammonium glycinate, N-acylaminopropyl-N, N-dimethylammonium glycinate, for example the coconut acylaminopropyldimethylammonium glycinate, and 2-alkyl-3-carboxylm -hydroxyethylimidazolines each having 8 to 18 carbon atoms in the alkyl or acyl group and the cocoacylaminoethylhydroxyethyl carboxymethylglycinate.
  • betaines such as the N-alkyl-N, N-dimethylammonium glycinate, for example the coconut alkyldimethylammonium glycinate, N-acylaminopropyl-N, N-dimethylammonium glycinate,
  • Suitable emulsifiers are ampholytic surfactants.
  • Ampholytic surfactants are surface-active compounds which, in addition to a Cs / is alkyl or acyl group, contain at least one free amino group and at least one -COOH or -SOsH group in the molecule and are capable of forming internal salts.
  • ampholytic see surfactants are N-alkylglycines, N-alkylpropionic acids, N-alkylaminobutyric acids, N-alkyliminodipropionic acids, N-hydroxyethyl-N-alkylamidopropylglycines, N-alkyltaurines, N-alkylsarcosines, 2-alkylaminopropionic acids and alkylaminoacetic acids each with about 8 up to 18 carbon atoms in the alkyl group.
  • Particularly preferred ampholytic surfactants are N-coconut alkyl aminopropionate, coconut acylaminoethyl aminopropionate and C 1 2 /.
  • quaternary emulsifiers IfTBelraclitT -vöBersoiche of the ester quat type preferably methyl-quaternized difatty acid triethanolamine ester salts, are particularly preferred.
  • Substances such as lanolin and lecithin and, polyethoxylated or aeylated lanolin and lecithin derivatives, polyol fatty acid esters, monoglycerides and fatty acid alkanolamides can be used as superfatting agents, the latter simultaneously serving as foam stabilizers.
  • Pearlescent waxes that can be used are, for example: alkylene glycol esters, especially ethylene glycol distearate; Fatty acid alkanolamides, especially coconut fatty acid diethanolamide; Partial glycerides, especially stearic acid monoglyceride; Esters of polyvalent, optionally hydroxy-substituted carboxylic acids with fatty alcohols having 6 to 22 carbon atoms, especially long-chain esters of tartaric acid; Fatty substances, such as, for example, fatty alcohols, fatty ketones, fatty aldehydes, fatty ethers and fatty carbonates, which have a total of at least 24 carbon atoms, especially lauron and distearyl ether; Fatty acids such as stearic acid, hydroxystearic acid or behenic acid, ring opening products of olefin epoxides with 12 to 22 carbon atoms with fatty alcohols with 12 to 22 carbon atoms and / or polyols with 2 to 15
  • the main consistency agents are fatty alcohols or hydroxy fatty alcohols with 12 to 22 and preferably 16 to 18 carbon atoms and in addition Partial glycerides, fatty acids or hydroxy fatty acids into consideration.
  • Partial glycerides, fatty acids or hydroxy fatty acids into consideration.
  • a combination of these substances with alkyl oligoglucosides and / or fatty acid N-methylglucamides of the same chain length and / or polyglycerol poly-12-hydroxystearates is preferred.
  • Suitable thickeners are, for example, Aerosil types (hydrophilic silicas), polysaccharides, in particular xanthan gum, guar guar, agar agar, alginates and tyloses, carboxymethyl cellulose and hydroxyethyl cellulose, and also higher molecular weight polyethylene glycol mono- and diesters of fatty acids, polyacrylates (eg carbopols) ® from Goodrich or Synthalene ® from Sigma), polyacrylamides, polyvinyl alcohol and polyvinyl pyrrolidone, surfactants such as, for example, ethoxylated fatty acid glycerides, esters of fatty acids with polyols such as pentaerythritol or trimethylol propane, fatty alcohol ethoxylates with restricted homolog distribution or alkyl oligoglucosides and electrolytes such as sodium chloride.
  • Aerosil types hydrophilic silicas
  • polysaccharides in particular
  • Suitable cationic polymers are, for example, cationic cellulose derivatives, such as, for example, a quaternized hydroxyethyl cellulose, which is available under the name Polymer JR 400® from Amerchol, cationic starch, copolymers of diallylammonium salts and acrylamides, quaternized vinylpyrrolidone / vinylimidazole polymers, such as eg Luviquat® (BASF), condensation products of polyglycols and amines, quaternized collagen polypeptides, such as lauryldimonium hydroxypropyl hydrolyzed collagen (Lamequat®L / Grünau), quaternized wheat polypeptides, polyethyleneimine, cationic silicone polymers, _how_z Dimethylaminohydroxypropyldiethylenetriamine (Cartaretine® / Sandoz), copolymers of acrylic acid with dimethyldiallylammonium chloride (Merquat® 550 /
  • Anionic, zwitterionic, amphoteric and nonionic polymers include, for example, vinyl acetate / crotonic acid copolymers, vinyl pyrrolidone / vinyl acrylate copolymers, vinyl acetate / butyl maleate / isobornyl acrylate copolymers, methyl vinyl ether / maleic anhydride copolymers and their polyols and uncrosslinked polyols and their esters , Acrylamidopro- -pyltrimethylammonium chloride / acrylate copolymers, octylacrylamide / methyl methacrylate / tert.butylaminoethyl methacrylate / 2-hydroxyproyl methacrylate copolymers, polyvinyl pyrrolidone, vinyl pyrrolidone ⁇ / inylacetate copolymers, vinylpyrrolidethyl methacrylate and / or dimethylamino methyl
  • Suitable silicone compounds are, for example, dimethylpolysiloxanes, methylphenylpolysiloxanes, cyclic silicones and amino-, fatty acid-, alcohol-, polyether-, epoxy-, fluorine-, glycoside- and / or alkyl-modified silicone compounds which can be both liquid and resinous at room temperature.
  • Simethicones which are mixtures of dimethicones with an average chain length of 200 to 300 dimethylsiloxane units and hydrogenated silicates, are also suitable.
  • a detailed overview of suitable volatile silicones can also be found by Todd et al. in Cosm.Toil. 91, 27 (1976).
  • Typical examples of fats are glycerides
  • waxes include natural waxes such as candelilla wax, carnauba wax, Japanese wax, espresso grass wax, cork wax, guaruma wax, rice-germ oil wax, sugar cane wax.
  • wax ouricury wax, montan wax, beeswax, shellac wax, walrus, lanolin (wool wax), pretzel fat, ceresin, ozokerite (earth wax), petrolatum, paraffin waxes, micro waxes; chemically modified waxes (hard waxes), such as montan ester waxes, Sasol waxes, hydrogenated jojoba waxes and synthetic waxes, such as polyalkylene waxes and polyethylene glycol waxes, are possible.
  • hard waxes such as montan ester waxes, Sasol waxes, hydrogenated jojoba waxes and synthetic waxes, such as polyalkylene waxes and polyethylene glycol waxes
  • Biogenic active substances are, for example, tocopherol, tocopherol acetate, tocopherol paimitat ⁇ ascorbic acid ⁇ deoxyribonucleic acid ⁇ retinol, bisabolol, allantoin, phytantriol, panthenol, AHA acids, amino acids, ceramides, pseudo doceramides, essential oils, plant extracts and vitamin complexes.
  • Cosmetic deodorants counteract, mask or eliminate body odors.
  • Body odors arise from the action of skin bacteria on apocrine sweat, whereby unpleasant smelling breakdown products are formed. Accordingly, deodorants contain active ingredients which act as germ-inhibiting agents, enzyme inhibitors, odor absorbers or odor maskers.
  • all substances which are effective against gram-positive bacteria are suitable as germ-inhibiting agents, which may be added to the cosmetics according to the invention, such as B. 4-hydroxybenzoic acid and its salts and esters, N- (4-chlorophenyl) -N '- (3,4 dichlorophenyl) urea, 2,4,4 ' -Trichlor-2'-hydroxydiphenyl ether (triclosan), 4-chloro -3,5-dimethylphenol, 2,2 'methylene-bis (6-bromo-4-chlorophenol), 3-methyl-4- (1-methylethyl) phenol, 2-benzyl-4-chlorophenol, 3- (4 -Chlorphenoxy) -1, 2-propanediol, 3-iodo-2-propynyl butyl carbamate, Chlorhexidine, 3,4,4'-trichlorocarbanilide (TTC), antibacterial fragrances, Thymol, thyme oil, eugenol
  • Enzyme inhibitors can also be added to the cosmetics according to the invention.
  • Suitable enzyme inhibitors are, for example, esterase inhibitors. These are preferably trialkyl such as trimethyl citrate, tripropyl, triisopropyl, tributyl citrate and especially triethyl citrate (Hydagen® ® CAT, Henkel KGaA, Dusseldorf / FRG). The substances inhibit enzyme activity and thereby reduce odor.
  • esterase inhibitors include sterol sulfates or phosphates, such as, for example, lanosterol, cholesterol, campesteric, stigmasterol and sitosterol sulfate or phosphate, dicarboxylic acids and their esters, such as, for example, glutaric acid, glutaric acid monoethyl ester, glutaric acid diethyl ester, Adipic acid, adipic acid monoethyl ester, adipic acid diethyl ester, malonic acid and malonic acid diethyl ester, hydroxycarboxylic acids and their esters such as citric acid, malic acid, tartaric acid or tartaric acid diethyl ester, and zinc glycinate.
  • dicarboxylic acids and their esters such as, for example, glutaric acid, glutaric acid monoethyl ester, glutaric acid diethyl ester, Adipic acid, adipic acid monoethyl ester,
  • Suitable odor absorbers are substances that absorb odor-forming compounds and can retain them to a large extent. They lower the partial pressure of the individual components and thus also reduce their speed of propagation. It is important that perfumes must remain unaffected. Odor absorbers are not effective against bacteria. They contain, for example, a complex zinc salt of ricinoleic acid or special, largely odorless fragrances, which are known to the person skilled in the art as "fixators", such as, for example, the main component. B. extracts of Labdanum or Styrax or certain abietic acid derivatives. Fragrances or perfume oils act as odor maskers, which in addition to their function as odor maskers Deodorants give their respective fragrance.
  • Perfume oils are, for example, mixtures of natural and synthetic fragrances. Natural fragrances are extracts of flowers, stems and leaves, fruits, fruit peels, roots, woods, herbs and grasses, needles and branches as well as resins and balms. Animal raw materials, such as civet and castoreum, are also suitable. Typical synthetic fragrance compounds are products of the ester, ether, aldehyde, ketone, and alcohol type Type of the esters are, for example
  • the ethers include, for example, benzyl ethyl ether
  • the aldehydes include, for example, the linear alkanals having 8 to 18 carbon atoms, citral, citronellal, citronellyloxyacetaldehyde, cyclamenaldehyde, hydroxycitronellal, lilial and bourgeonal
  • the ketones include, for example, the jonones and methylcedryl ketone
  • the alcohols are anethole, citronellellone Eugenol, isoeugenol, geraniol, linalool, phenylethyl alcohol and terpineol
  • the hydrocarbons mainly include the terpenes and balsams.
  • fragrance oils of lower volatility which are mostly used as aroma components, are also suitable as perfume oils, for example sage oil, chamomile oil, clove oil, lemon balm oil, mint oil, cinnamon leaf oil, linden blossom oil, juniper berry oil, vetiver oil, oliban oil, galbanum oil, labdanum oil and lavandin oil.
  • bergamot oil dihydromyrcenol, lilial, lyral, citronellol, phenylethyl alcohol, ⁇ -hexylcinnamaldehyde, geraniol, benzyl acetone, cyclamen aldehyde, linalool, Boisambrene Forte, Ambroxan, indole, hedione, Sandelice, lemon oil, mandarin oil, orange oil, allyl amyl glycolate, Cyclovertal, lavandin oil, muscatel Sage oil, ß-damascone, geranium oil bourbon, cyclohexyl salicylate, Vertofix Coeur, Iso-E-Super, Fixolide NP, evernyl, iraldein gamma, phenylacetic acid, geranyl acetate, benzyl acetate, rose oxide, romilate, irot
  • auxiliaries such. B. thickeners or complexing agents and / or
  • non-aqueous solvents such as e.g. As ethanol, propylene glycol and / or glycerin.
  • Salts of aluminum, zirconium or zinc are particularly suitable as astringent antiperspirant active ingredients.
  • suitable antiperspirant active ingredients are e.g. Aluminum chloride, aluminum chlorohydrate, aluminum dichlorohydrate, aluminum sesquichlorohydrate and their complex compounds z.
  • B. with amino acids such as glycine.
  • oil-soluble and water-soluble auxiliaries can be present in smaller amounts in antiperspirants.
  • oil soluble aids can e.g. his:
  • water-soluble additives are e.g. Preservatives, water-soluble fragrances, pH adjusters, e.g. Buffer mixtures, water soluble thickeners, e.g. water-soluble natural or synthetic polymers such as e.g. Xanthan gum, hydroxyethyl cellulose, polyvinyl pyrrolidone or high molecular weight polyethylene oxides.
  • Climbazole, octopirox and zinc pyrithione can be used as antidandruff agents.
  • Common film formers are, for example, chitosan, microcrystalline chitosan, quaternized chitosan, polyvinylpyrrolidone, vinylpyrrolidone-vinyl acetate copolymers, polymers of the acrylic acid series, quaternary cellulose derivatives, collagen, hyaluronic acid or its salts and similar compounds.
  • Montmorillonites, clay minerals, pemulene and alkyl-modified carbopol types can serve as swelling agents for aqueous phases. Further suitable polymers or swelling agents can be found in the overview by R. Lochhead in Cosm.Toil. 108, 95 (1993).
  • UV light protection factors are understood to mean, for example, organic substances (light protection filters) which are liquid or crystalline at room temperature and which are able to absorb ultraviolet rays and absorb the energy absorbed in the form of longer-wave radiation, e.g. To give off heat again.
  • UVB filters can be oil-soluble or water-soluble. As oil-soluble substances e.g. to call:
  • 3-benzylidene camphor or 3-benzylidene norcampher and its derivatives for example 3- (4-methylbenzylidene) camphor as described in EP 0693471 B1; • 4-aminobenzoic acid derivatives, preferably 2-ethylhexyl 4-dimethylamino) benzoate, 2-octyl 4- (dimethylamino) benzoate and amyl 4- (dimethylamino) benzoate;
  • esters of cinnamic acid preferably 4-methoxycinnamic acid 2-ethylhexyl ester, 4-methoxycinnamic acid propyl ester, 4-methoxycinnamic acid isoamyl ester 2-cyano-3,3-phenylcinnamic acid 2-ethylhexyl ester (octocrylene);
  • esters of salicylic acid preferably salicylic acid 2-ethylhexyl ester, salicylic acid 4-isopropylbenzyl ester, salicylic acid homomethyl ester;
  • benzophenone preferably 2-hydroxy-4-methoxybenzophenone, 2-hydroxy-4-methoxy-4'-methylbenzophenone, 2,2'-dihydroxy-4-methoxybenzophenone;
  • esters of benzalmalonic acid preferably di-2-ethylhexyl 4-methoxybenzmalonate
  • Triazine derivatives e.g. 2,4,6-trianilino- (p-carbo-2'-ethyl-1'-hexyloxy) -1, 3,5-triazine and octyl triazone, as described in EP 0818450 A1 or di-octyl butamido triazone (Uvasorb ® HEB);
  • Propane-1,3-dione e.g. 1- (4-tert-butylphenyl) -3-4'methoxyphenyl) propane-1,3-dione;
  • Sulfonic acid derivatives of 3-benzylidene camphor such as 4- (2-oxo-3-bomylidene methyl) benzenesulfonic acid and 2-methyl-5- (2-oxo-3-bornylidene) sulfonic acid and its salts.
  • benzoylmethane such as 1- (4'-tert-butylphenyl) -3- (4'-methoxyphenyl) propane-1,3-dione, 4-tert, are particularly suitable as typical UV-A filters.
  • UV-A and UV-B filters can of course also be used in mixtures.
  • insoluble light protection pigments namely finely dispersed metal oxides or salts, are also suitable for this purpose.
  • suitable metal oxides are, in particular, zinc oxide and titanium dioxide and, in addition, oxides of iron, zirconium, silicon, manganese, aluminum and cerium and mixtures thereof.
  • Silicates (talc), barium sulfate or zinc stearate can be used as salts.
  • the oxides and salts are used in the form of the pigments for skin-care and skin-protecting emulsions and decorative cosmetics.
  • the particles should have an average diameter of less than 100 nm, preferably between 5 and 50 nm and in particular between 15 and 30 nm. They can have a spherical shape, but it is also possible to use particles which have an ellipsoidal shape or a shape which differs from the spherical shape in some other way.
  • the pigments can also be surface-treated, ie hydrophilized or hydrophobicized.
  • Typical examples are coated titanium dioxides, such as titanium dioxide T 805 (Degussa) or Eusolex® T2000 (Merck). Silicones, and in particular trialkoxy-octylsilanes or simethicones, are particularly suitable as hydrophobic coating agents. So-called micro- or nanopigments are preferably used in sunscreens. Micronized zinc oxide is preferably used. Further suitable UV light protection filters can be found in the overview by P.Finkel in S ⁇ FW-Journal 122, 543 (1996).
  • secondary light stabilizers of the antioxidant type can also be used, which interrupt the photochemical reaction chain that triggers when UV radiation penetrates the skin.
  • Typical examples are amino acids (e.g. glycine, histidine, tyrosine, tryptophan) and their derivatives, imidazoles (e.g. urocanic acid) and their derivatives, peptides such as D, L-carnosine, D-camosine, L-carnosine and their derivatives (e.g. anserine) , Chlorogenic acid and its derivatives, lipoic acid and its derivatives (e.g.
  • thiols e.g. thioredoxin, glutathione, cysteine, cystine and cystamine and their glycosyl, N-acetyl, methyl, ethyl, Propyl, amyl, butyl and lauryl, palmitoyl, oleyl, ⁇ -linoleyl, cholesteryl and glyceryl esters
  • salts dilauryl thiodipropionate, distearyl thiodipropionate, thiodipropionic acid and their derivatives (esters, ethers, peptides , Lipids, nucleotides, nucleosides and salts) as well as sulfoximine compounds (e.g.
  • citric acid citric acid, lactic acid, malic acid
  • humic acid bile acid, bile extracts, bilirubin, biliverdin, EDTA, EGTA and their derivatives
  • unsaturated fatty acids and their derivatives e.g. ⁇ -linolenic acid, linoleic acid, oleic acid
  • folic acid and its derivatives ubiquinone and ubiquinol and their derivatives
  • vitamin C and derivatives e.g. ascorbyl palmitate, Mg ascorbyl phosphate, ascorbyl acetate
  • tocopherols and derivatives e.g.
  • Vitamin E acetate
  • Vitamin A and derivatives vitamin A palmitate
  • Alkylene glycols such as, for example, ethylene glycol, diethylene glycol, propylene glycol, butylene glycol, hexylene glycol and polyethylene glycols with an average molecular weight of 100 to 1,000 daltons;
  • Methyl compounds such as in particular trimethylolethane, trimethylolpropane, trimethylolbutane, pentaerythritol and dipentaerythritol;
  • Lower alkylglucos.de especially those with 1 to 8 carbons in the alkyl radical, such as methyl and butyl glucoside;
  • Sugar alcohols with 5 to 12 carbon atoms such as sorbitol or mannitol,
  • Aminosugars such as glucamine
  • Dialcohol amines such as diethanolamine or 2-amino-1, 3-propanediol.
  • Suitable preservatives are, for example, phenoxyethanol, formaldehyde solution, parabens, pentanediol or sorbic acid and the other classes of substances listed in Appendix 6, Parts A and B of the Cosmetics Ordinance.
  • insect repellents come N, N-diethyl-m-toluamide, 1, 2-pentanediol or Ethyl butylacetylaminopropionate in question, di-hydroxyacetone is suitable as a self-tanner.
  • Perfume oils include mixtures of natural and synthetic fragrances. Natural fragrances are extracts from flowers (lily, lavender, rose, jasmine, neroli, ylang-ylang), stems and leaves (geranium, Patterson chouli, petitgrain), fruits (anise, coriander, caraway, juniper), fruit schale ⁇ _ (Be ⁇ gramOtt!
  • Synthetic fragrance compounds are products of the ester, ether, aldehyde, ketone, alcohol and hydrocarbon type
  • fragrance compounds of the ester type are, for example, benzyl acetate, phenoxyethyl isobutyrate, p-tert-butyl-cyclohexyl acetate, linalyl acetate, dimethylbenzylcarbinylacetate, phenylethyl acetate at, linalyl benzoate, benzyl formate, ethyl methylphenyl glycinate, allyl cyclohexyl propionate, styrallyl propionate and benzyl salicylate.
  • the ethers include, for example, benzyl ethyl ether
  • the aldehydes include, for example, the linear alkanals with 8 to 18 carbon atoms, citral, citronellal, citronellyloxyacetaldehyde, cyclamaldehyde, hydroxycitronellal, lilial and bourgeonal
  • the ketones include, for example, the jonones, oc-isomethylionone, and methylcedryl ketone the alcohols anethole, citronellol, eugenol, isoeugenol, geraniol, linalool, phenylethyl alcohol and terpineol
  • the hydrocarbons mainly include the terpenes and balsams.
  • fragrance oils of lower volatility which are mostly used as aroma components, are also suitable as perfume oils, for example sage oil, chamomile oil, clove oil, melissa oil, mint oil, cinnamon leaf oil, linden blossom oil, juniper berry oil, vetiver oil, Oliban oil, galbanum oil, labolanum oil and lavandin oil.
  • the dyes which can be used are those substances which are suitable and approved for cosmetic purposes, as compiled, for example, in the publication "Cosmetic Dyes” by the Dye Commission of the German Research Foundation, Verlag Chemie, Weinheim, 1984, pp. 81-106. These dyes are usually used in concentrations of 0.001 to 0.1% by weight, based on the mixture as a whole.
  • the total proportion of auxiliaries and additives can be 1 to 50, preferably 5 to 40,% by weight, based on the composition.
  • the cosmetics and personal care products can be produced by customary cold or hot processes; the phase inversion temperature method is preferably used.

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Abstract

The invention relates to a method for determining skin stress and/or skin aging in humans or animals in vitro. The invention further relates to test kits for determining skin stress and/or skin aging and to the use of spondin 2, cathepsin L, vimentin fragments or actin gamma 1 as a stress and/or aging marker of the skin. The invention also relates to a method for analyzing the effectiveness of cosmetic or pharmaceutical active substances against skin stress and/or skin aging, and to agents for regulating, especially maintaining, homeostasis of human and animal skin.

Description

Verfahren zur in vitro Bestimmung der Hautalterung Method for the in vitro determination of skin aging
Die vorliegende Erfindung betrifft Verfahren zur Bestimmung des Hautstreß und/oder der Hautalterung bei Menschen oder Tieren in vitro, Test-Kits zur Bestimmung des Hautstreß und/oder der Hautalterung sowie die Verwendung von Spondin 2, Cathepsin L, Vimentin-Fragmenten oder Actin Gamma 1 als Streß- und/oder Alterungsmarker der Haut; ferner ein Verfahren zum Nachweis der Wirksamkeit von kosmetischen oder pharmazeutischen Wirkstoffen gegen Hautstreß und/oder Hautalterung sowie Mittel zur Regulierung, insbesondere zur Aufrechterhaltung der Homeostase menschlicher oder tierischer Haut.The present invention relates to methods for determining skin stress and / or skin aging in humans or animals in vitro, test kits for determining skin stress and / or skin aging and the use of spondin 2, cathepsin L, vimentin fragments or actin gamma 1 as a stress and / or aging marker of the skin; furthermore a method for proving the effectiveness of cosmetic or pharmaceutical active substances against skin stress and / or skin aging as well as means for regulating, in particular for maintaining the homeostasis of human or animal skin.
Die menschliche Haut ist ein sehr komplex aufgebautes Organ, welches aus einer Vielzahl verschiedener Zelltypen besteht. Der Metabolismus der lebenden Zellen ist nicht statisch sondern sehr dynamisch. In der Haut bemerken Zellen Veränderungen ihrer Umgebung (z.B. die Einstrahlung von Sonnenlicht) und reagieren darauf mit der Umstellung ihrer RNA- und/oder Proteinsyntheseleistungen.The human skin is a very complex organ, which consists of a large number of different cell types. The metabolism of living cells is not static but very dynamic. In the skin, cells notice changes in their environment (e.g. sunlight) and react to this by changing their RNA and / or protein synthesis performance.
Einige Moleküle werden nach einem Stresstimulus (z.B. Sonnenlicht) vermehrt synthetisiert (z.B. MMP-1), andere wiederum werden in einem geringerem Umfang produziert (z.B. Kollagen CH (I)). Weiterhin wird bei einer Vielzahl der Syntheseprozesse keine signifikante Veränderung erfolgen (z.B. TIMP-1).Some molecules are increasingly synthesized after a stress stimulus (e.g. sunlight) (e.g. MMP-1), others are produced to a lesser extent (e.g. collagen CH (I)). Furthermore, there will be no significant change in a large number of the synthesis processes (e.g. TIMP-1).
Die makroskopischen Phänomene gealterter Haut beruhen zum einen auf der intrinsischen oder chronologischen Alterung, zum anderen auf der extrinsischen Alterung durch Umweltstress, wie z.B. durch Sonnenlicht. Die sichtbaren Zei- chen der durch Stress gealterten Haut sind als Summe vieler Einzelereignisse zu verstehen, die in der Haut über einen längeren Zeitraum akkumulieren.The macroscopic phenomena of aged skin are based on the one hand on the intrinsic or chronological aging, and on the other hand on the extrinsic aging caused by environmental stress, such as sunlight. The visible times The areas of skin that have aged due to stress are to be understood as the sum of many individual events that accumulate in the skin over a longer period of time.
Effektive Anti-Ageprodukte zeigen ihre Wirkung auf ein möglichst breites Spektrum molekularer Einzelereignisse der extrinsischen Hautalterung. Bisher sind jedoch nur wenige solcher Ereignisse in lebenden Hautzellen bekannt, die als Marker gestresster Haut und somit zur Wirksamkeitsüberprüfung von Antiage- Produkten dienen können.Effective anti-aging products show their effect on the broadest possible range of individual molecular events in extrinsic skin aging. So far, however, only a few such events are known in living skin cells, which can serve as a marker for stressed skin and thus for the effectiveness of antiage products.
Die Identifikation neuer stressinduzierter Hautreaktionen ermöglicht es, den komplexen Prozess der Hautalterung und seine kausalen Zusammenhänge zu begreifen.The identification of new stress-induced skin reactions makes it possible to understand the complex process of skin aging and its causal relationships.
Mit diesem Wissen können neue Konzepte für kosmetische oder pharmazeutische Antiage-Produkte entwickelt werden, die ihre Wirkung auf das breite Spektrum der Alterungsprozesse in der Haut ausüben.With this knowledge, new concepts for cosmetic or pharmaceutical anti-aging products can be developed that have an effect on the broad spectrum of aging processes in the skin.
Jeder Zelltyp der Haut synthetisiert ca. 15.000 verschiedene Proteine. Welche Proteine davon für die intrinsische und/oder extrinsische Hautalterung eine Rolle spielen ist bisher weitgehend unklar.Each cell type of the skin synthesizes approximately 15,000 different proteins. So far it is largely unclear which proteins play a role in intrinsic and / or extrinsic skin aging.
Erschwerend kommt hinzu, dass die Haut aus mehreren verschiedenen Zelltypen (Fibroblasten, Keratinozyten in verschiedenen Differenzierungszuständen, Melanozyten, Langerhanszellen Haarfollikelzellen, Schweißdrüsenzellen etc.) besteht und somit die Komplexität in der Haut produzierter Proteingemische sehr groß ist.To make matters worse, the skin consists of several different cell types (fibroblasts, keratinocytes in different states of differentiation, melanocytes, Langerhans cells, hair follicle cells, sweat gland cells etc.) and the complexity of the protein mixtures produced in the skin is therefore very great.
Es ist bisher nicht möglich gewesen, aus dieser immensen Komplexität die Proteine zu identifizieren, die mit der Hautalterung in kausalem Zusammenhang stehen. Ein Verständnis der komplexen Alterungsprozesse in der Haut durch die Identifikation stressregulierter Markerproteine gestattet die gezielte Suche nach Substanzen oder Kombinationen von Substanzen mit einem breiten Antiage- Wirkspektrum.So far it has not been possible to identify the proteins that are causally related to skin aging from this immense complexity. An understanding of the complex aging processes in the skin through the identification of stress-regulated marker proteins enables the targeted search for substances or combinations of substances with a broad spectrum of anti-aging effects.
Produktkonzepte dieser Art konnten jedoch bis zu dem jetzigen Zeitpunkt nicht entwickelt werden, da eine Vielzahl "der Markerproteine für die Hautalterung noch nicht bekannt waren.However, product concepts of this type have not yet been able to be developed because a large number of " marker proteins for skin aging were not yet known.
Überraschenderweise wurde gefunden, daß die Proteine Spondin 2, Cathepsin L, Actin Gamma 1 sowie Fragmente des Proteins Vimentin von Fibroblasten der Haut streßinduzierbar in den extrazellulären Raum sezerniert werden, von nicht gestressten Fibroblasten jedoch nicht. Überraschenderweise wurde außerdem gefunden, daß Fragmente des Proteins Vimentin dazu beitragen können, die Homeostase der Haut aufrecht zu erhalten.Surprisingly, it was found that the proteins spondin 2, cathepsin L, actin gamma 1 and fragments of the protein vimentin are secreted into the extracellular space by skin fibroblasts, but not by stressed fibroblasts, but not. Surprisingly, it was also found that fragments of the protein vimentin can help to maintain the homeostasis of the skin.
Gegenstand der vorliegenden Erfindung ist daher ein Verfahren zur Bestimmung des Hautstreß und/oder der Hautalterung bei Menschen oder Tieren in vitro, dadurch gekennzeichnet, daß man a) eine Fibroblasten enthaltende Hautprobe gewinnt, b) die Fibroblasten isoliert und kultiviert und c) den Kulturüberstand auf das Vorhandensein und die Menge der von den Fibroblasten sezernierten Proteine oder Proteinfragmente Spondin 2, Cathepsin L, Vimentin-Fragmente oder Actin Gamma 1 untersucht.The present invention therefore relates to a method for determining skin stress and / or skin aging in humans or animals in vitro, characterized in that a) a skin sample containing fibroblasts is obtained, b) the fibroblasts are isolated and cultured, and c) the culture supernatant the presence and the amount of the proteins or protein fragments secreted by the fibroblasts Spondin 2, Cathepsin L, Vimentin fragments or Actin Gamma 1 were examined.
Ein weiterer Gegenstand der Erfindung ist ein Verfahren zur Bestimmung des Hautstreß und/oder der Hautalterung bei Menschen oder Tieren in vitro, dadurch gekennzeichnet, daß man a) mittels Mikrodialyse ein Gemisch der von den Hautzellen sezemierten Proteine gewinnt und b) das Gemisch auf das Vorhandensein und die Menge der von den Fibroblasten sezemierten Proteine oder Proteinfragmente Spondin 2, Cathepsin L, Vimentin-Fragmente oder Actin Gamma 1 untersucht.Another object of the invention is a method for determining skin stress and / or skin aging in humans or animals in vitro, characterized in that a) a mixture of the proteins secreted by the skin cells is obtained by microdialysis and b) the mixture is examined for the presence and the amount of the proteins or protein fragments secreted by the fibroblasts Spondin 2, Cathepsin L, Vimentin fragments or Actin Gamma 1.
Die Technik der Mikrodialyse wird beispielsweise in „Microdialysis. A method for measurement of local tissue metabolism", Nielsen PS, Winge K, Petersen LM; Ugeskr Laeger 1999 Mar 22 161 :12 1735-8; sowie in „Cutaneous microdialysis for human in vivo dermal absorption studies", Anderson, C. et al. ; Drugs Pharm. Sei., 1998, 91 , 231-244; und auch im Internet unter http://www.microdialysis.se/techniqu.htm beschrieben, worauf hiermit in vollem Umfang Bezug genommen wird.The microdialysis technique is described, for example, in “Microdialysis. A method for measurement of local tissue metabolism ", Nielsen PS, Winge K, Petersen LM; Ugeskr Laeger 1999 Mar 22 161: 12 1735-8; and in" Cutaneous microdialysis for human in vivo dermal absorption studies ", Anderson, C. et al. ; Drugs Pharm. Sci., 1998, 91, 231-244; and also described on the Internet at http://www.microdialysis.se/techniqu.htm, to which reference is hereby made in full.
Bei der Anwendung der Mikrodialyse führt man typischerweise eine Sonde in die Haut ein und beginnt mit einer geeigneten Trägerlösung die Sonde langsam zu spülen. Nach dem Abklingen der akuten Reaktionen nach dem Einstich liefert die Mikrodialyse Proteine, die im extrazellulären Raum vorkommen und die, beispielsweise durch Fraktionierung der Trägerflüssigkeit, dann in vitro isoliert und analysiert werden können.When using microdialysis, a probe is typically inserted into the skin and the probe is slowly rinsed with a suitable carrier solution. After the acute reactions have subsided after the puncture, the microdialysis provides proteins which occur in the extracellular space and which can then be isolated and analyzed in vitro, for example by fractionation of the carrier liquid.
Die Kultivierung von Fibroblasten ist dem Fachmann bekannt und kann erfindungsgemäß in üblicher Weise durchgeführt werden, beispielsweise wie in Langholz, O. et al.; 1997; Experimental Cell Research 235, 22-27; beschrieben.The cultivation of fibroblasts is known to the person skilled in the art and can be carried out according to the invention in a customary manner, for example as in Langholz, O. et al .; 1997; Experimental Cell Research 235, 22-27; described.
Vorzugsweise werden die erfindungsgemäßen Verfahren so durchgeführt, daß man die Untersuchung auf das Vorhandensein und die Menge der von den Fibroblasten sezemierten Proteine oder Proteinfragmente Spondin 2, Cathepsin L, Vimentin-Fragmente oder Actin Gamma 1 durch Separation der Proteine mittels Gelelektrophorese und nachfolgende Massenspektrometrie, insbesondere Matrix Assistierter Laser Desorptions Ionisation (MALDI) vornimmt. Besonders bevorzugt erfolgt die Separation der Proteine durch 2D- Gelelektrophorese, wie beispielsweise in L.D. Adams, Two-dimensional Gel Electrophoresis using the Isodalt System oder in L.D. Adams & S.R. Gallagher, Two-dimensional Gel Electrophoresis using the O'Farrell System; beide in Cur- rent Protocols in Molecular Biology (1997, Eds. F.M. Ausubel et al.), Unit 10.3.1 - 10.4.13; oder in 2-D Electrophoresis-Manual; T. Berkelman, T. Senstedt; Amersham Pharmacia Biotech, 1998 (Bestell-Nr. 80-6429-60), beschrieben.The methods according to the invention are preferably carried out in such a way that the examination for the presence and the amount of the proteins or protein fragments secreted by the fibroblasts Spondin 2, Cathepsin L, Vimentin fragments or Actin Gamma 1 by separating the proteins by means of gel electrophoresis and subsequent mass spectrometry, in particular Matrix Assisted Laser Desorption Ionization (MALDI). The separation of the proteins is particularly preferably carried out by 2D gel electrophoresis, such as, for example, in LD Adams, Two-dimensional Gel Electrophoresis using the Isodalt System or in LD Adams & SR Gallagher, Two-dimensional Gel Electrophoresis using the O'Farrell System; both in Curent Protocols in Molecular Biology (1997, Eds. FM Ausubel et al.), Unit 10.3.1 - 10.4.13; or in 2-D electrophoresis manual; T. Berkelman, T. Senstedt; Amersham Pharmacia Biotech, 1998 (Order No. 80-6429-60).
Die Gewinnung der Proteine aus dem Kulturüberstand von dermalen Fibroblasten erfolgt vorzugsweise durch Proteinpräzipitation aus dem Kulturüberstand, beispielsweise mit Methanol/Chloroformgemisch im Verhältnis von etwa 4:1 :4 zur Probe und/oder durch Aufkonzentrierung der Proteine, beispielsweise durch Ultrafiltration mit einem Cut off von etwa 5.000 Da, z.B. mittels der Vivaspin-Produkte der Fa. Sartorius.The proteins are obtained from the culture supernatant from dermal fibroblasts preferably by protein precipitation from the culture supernatant, for example with a methanol / chloroform mixture in a ratio of about 4: 1: 4 to the sample and / or by concentration of the proteins, for example by ultrafiltration with a cut off of about 5,000 Da, e.g. using the Vivaspin products from Sartorius.
Das so erhaltene Konzentrat bzw. Präzipitat kann dann in dem Fachmann bekannter Weise in Rehydratisierungspuffer aufgenommen werden, beispielsweise, wie in „2-D Electrophoresis using immobilized pH gradients. Principles and methods", Berkelman, T. and Stenstedt, T (eds); Amersham Pharmacia Biotech Ine (1998) 80-6429-60; beschrieben.The concentrate or precipitate thus obtained can then be taken up in rehydration buffer in a manner known to those skilled in the art, for example as in “2-D electrophoresis using immobilized pH gradients. Principles and methods ", Berkelman, T. and Stenstedt, T (eds); Amersham Pharmacia Biotech Ine (1998) 80-6429-60;
Die erfindungsgemäß bevorzugte Separation der Proteine durch 2- Dimensionale Gelelektrophorese kann beispielsweise so erfolgen, daß man in der ersten Dimension eine isoelektrische Fokussierung vornimmt, beispielsweise unter Verwendung von Immobilin DryStrips mit einem immobilisierten pH- Gradienten (IPG) von 4 bis 7 der Fa. Amersham-Pharmacia oder unter Verwendung von Carrierampholyt tragenden Polyacrylamid-Gelstreifen, die im elektrischen Feld einen pH-Gradienten aufbauen (O'Farrell, P. H.; 1975; J Biol. Chem. 250, 4007-4021) und in der zweiten Dimension eine SDS-Polyacrylamid- Gelelektrophorese, z.B. mit 12.5 %tigem Polyacrylamid bei 30 mA für 3 bis 4 h vornimmt.The separation of the proteins preferred according to the invention by 2-dimensional gel electrophoresis can be carried out, for example, by carrying out isoelectric focusing in the first dimension, for example using Immobilin DryStrips with an immobilized pH gradient (IPG) of 4 to 7 from Amersham -Pharmacia or using carrier ampholyte-bearing polyacrylamide gel strips which build up a pH gradient in the electric field (O'Farrell, PH; 1975; J Biol. Chem. 250, 4007-4021) and in the second dimension an SDS-polyacrylamide - Gel electrophoresis, for example with 12.5% polyacrylamide at 30 mA for 3 to 4 h.
Die Gelfärbung kann nach üblichen Methoden, beispielsweise durch Silberfärbung nach Heukeshoven oder durch Colloidal Coomassieblaufärbung erfolgen.Gel staining can be carried out by customary methods, for example by Heukeshoven silver staining or by colloidal Coomassie staining.
Die massenspektrometrische Charakterisierung der Proteine erfolgt in der -Fachwelt bekannter Weiserbeispielsweise wie in den folgenden Literaturstellen beschrieben:The mass spectrometric characterization of the proteins takes place in the art of well-known Weiser, for example as described in the following literature references:
Methods in Molecular Biology, 1999; Vol 112; 2-D Proteome Analysis Protocols; Editor: A. J. Link; Humana Press; Totowa; New Jersey. Darin insbesondere: Courchesne, P. L. und Patterson, S. D.; S. 487-512.Methods in Molecular Biology, 1999; Vol 112; 2-D Proteome Analysis Protocols; Editor: A. J. Link; Humana Press; Totowa; New Jersey. In particular: Courchesne, P.L. and Patterson, S. D .; Pp. 487-512.
Carr, S. A. und Annan, R. S.; 1997; in: Current Protocols in Molecular Biology; Editor: Ausübet, F. M. et al.; John Wiley and Sons, Inc. 10.2.1-10.21.27.Carr, S.A. and Annan, R. S .; 1997; in: Current Protocols in Molecular Biology; Editor: Ausübet, F. M. et al .; John Wiley and Sons, Inc. 10.2.1-10.21.27.
Es können jedoch erfindungsgemäß auch andere dem Fachmann bekannte Methoden zur Analyse bzw. zum Nachweis der oben genannten Proteine oder Proteinfragmente eingesetzt werden, insbesondere Methoden, die auf der spezifischen Bindung von Antikörpern an Proteine beruhen, beispielsweise Radio- immunoassays und auf Immunofluoreszenz basierende Nachweisverfahren.However, according to the invention, other methods known to the person skilled in the art can also be used for the analysis or detection of the abovementioned proteins or protein fragments, in particular methods which are based on the specific binding of antibodies to proteins, for example radio-immunoassays and detection methods based on immunofluorescence.
Spondin 2 ist ein erst kürzlich im normalen Lungengewebe identifiziertes Protein (Manda, R., et al, (1999); Identification of Genes (SPON2 and C20orf2)...; Genomics 61(1), 5-14). Im Rahmen der vorliegenden Erfindung wird unter „Spondin 2" ein Protein mit der Aminosäurensequenz (A)Spondin 2 is a protein recently identified in normal lung tissue (Manda, R., et al, (1999); Identification of Genes (SPON2 and C20orf2) ...; Genomics 61 (1), 5-14). In the context of the present invention, under "Spondin 2" a protein with the amino acid sequence (A)
N-Term- MENPSPAAAL GKALCALLLA TLGAAGQPLG GESICSARAP AKYSITFTGK WSQTAFPKQY PLFRPPAQWS SLLGAAHSSD YSMWRKNQYV SNGLRDFAER GEAWALMKEI EAAGEALQSV HAVFSAPAVP SGTGQTSAEL EVQRRHSLVS FWRIVPSPD WFVGVDSLDL CDGDRWREQA ALDLYPYDAG TDSGFTFSSP NFATIPQDTV TEITSSSPSH PANSFYYPRL KALPPIARVT LVRLRQSPRA FIPPAPVLPS RDNEIVDSAS VPETPLDCEV SLWSSWGLCG GHCGRLGTKS RTRYVRVQPA NNGSPCPELE EEAECVPDNC V -C-TermN-term- MENPSPAAAL GKALCALLLA TLGAAGQPLG GESICSARAP AKYSITFTGK WSQTAFPKQY PLFRPPAQWS SLLGAAHSSD YSMWRKNQYV SNGLRDFAER GEAWALMKEI EAAGEALQSV HAVFSAPAVP SGTGQTSAEL EVQRRHSLVS FWRIVPSPD WFVGVDSLDL CDGDRWREQA ALDLYPYDAG TDSGFTFSSP NFATIPQDTV TEITSSSPSH PANSFYYPRL KALPPIARVT LVRLRQSPRA FIPPAPVLPS RDNEIVDSAS VPETPLDCEV SLWSSWGLCG GHCGRLGTKS RTRYVRVQPA NNGSPCPELE EEAECVPDNC V -C-term
verstanden, sowie Proteine, die mehr als 40%, insbesondere mehr als 60%, -besonders-bevorzugt~mehrals~~80%~Sequenzhömologie mit dieser Sequenz aufweisen. Die Aminosäuresequenz (A) ist in der Proteindatenbank des National Center for Biotechnology Information (NCBI) unter der Nummer 6172221 offenbart. Diese Datenbank ist im Internet unter folgender Adresse zugänglich: http://www.ncbi.nlm.nih.gov/.having understood, as well as proteins that are more than 40%, in particular more than 60%, -especially-preferably more than ~ 80% ~ ~~ Sequenzhömologie with this sequence. The amino acid sequence (A) is disclosed in the National Center for Biotechnology Information (NCBI) protein database under number 6172221. This database is accessible on the Internet at the following address: http://www.ncbi.nlm.nih.gov/.
Im Rahmen der vorliegenden Erfindung umfaßt „Spondin 2" auch solche Proteine, deren Aminosäuresequenz durch konservative Mutationen, also durch den Austausch strukturell oder funktioneil ähnlicher Aminosäuren, aus der oben aufgeführten Sequenz (A) erhalten werden kann.In the context of the present invention, “spondin 2” also includes those proteins whose amino acid sequence can be obtained from the above-listed sequence (A) by conservative mutations, that is to say by the exchange of structurally or functionally similar amino acids.
Cathepsin L wurde zunächst als eine lysosomale Protease beschrieben. Erst später wurde die Identität mit dem "major excretet protein (MEP) der Maus gezeigt.Cathepsin L was first described as a lysosomal protease. Only later was the identity shown with the "major excretet protein (MEP) of the mouse.
Im Rahmen der vorliegenden Erfindung wird unter „Cathepsin L" ein Protein mit der Aminosäurensequenz (B)In the context of the present invention under “Cathepsin L” a protein with the amino acid sequence (B)
N-Term- MNPTLILAAF CLGIASATLT FDHSLEAQWT KWKAMHNRLY GMNEEGWRRA VWEKNMKMIE LHNQEYREGK HSFTMAMNAF GDMTSEEFRQ VMNGFQNRKP RKGKVFQEPL FYEAPRSVDW REKGYVTPVK NQGQCGSCWA FSATGALEGQ MFRKTGRLIS LSEQNLVDCS GPQGNEGCNG GLMDYAFQYV QDNGGLDSEE SYPYEATEES CKYNPKYSVA NDTGFVDIPK QEKALMKAVA TVGPISVAID AGHESFLFYK EGIYFEPDCS SEDMDHGVLV VGYGFESTES DNNKYWLVKN SWGEEWGMGG YVKMAKDRRN HCGIASAASY PTV -C-TermN-Term MNPTLILAAF CLGIASATLT FDHSLEAQWT KWKAMHNRLY GMNEEGWRRA VWEKNMKMIE LHNQEYREGK HSFTMAMNAF GDMTSEEFRQ VMNGFQNRKP RKGKVFQEPL FYEAPRSVDW REKGYVTPVK NQGQCGSCWA FSATGALEGQ MFRKTGRLIS LSEQNLVDCS GPQGNEGCNG GLMDYAFQYV QDNGGLDSEE SYPYEATEES CKYNPKYSVA NDTGFVDIPK QEKALMKAVA TVGPISVAID AGHESFLFYK EGIYFEPDCS SEDMDHGVLV VGYGFESTES DNNKYWLVKN SWGEEWGMGG YVKMAKDRRN HCGIASAASY PTV -C-Term
verstanden, sowie Proteine, die mehr als 40%, insbesondere mehr als 60%, besonders bevorzugt mehr als 80% Sequenzhomologie mit dieser Sequenz aufweisen. Die Aminosäuresequenz (B) ist in der Proteindatenbank des National Center for Biotechnology Information (NCBI) unter der Nummer 4503155
Figure imgf000009_0001
zugänglich: http://www.ncbi.nlm.nih.gov/.understood, and proteins that have more than 40%, in particular more than 60%, particularly preferably more than 80% sequence homology with this sequence. The amino acid sequence (B) is in the protein database of the National Center for Biotechnology Information (NCBI) under number 4503155
Figure imgf000009_0001
accessible: http://www.ncbi.nlm.nih.gov/.
Im Rahmen der vorliegenden Erfindung umfaßt „Cathepsin L" auch solche Proteine, deren Aminosäuresequenz durch konservative Mutationen, also durch den Austausch strukturell oder funktionell ähnlicher Aminosäuren, aus der oben aufgeführten Sequenz (B) erhalten werden kann.In the context of the present invention, “cathepsin L” also includes those proteins whose amino acid sequence can be obtained from the sequence (B) listed above by conservative mutations, that is to say by the exchange of structurally or functionally similar amino acids.
Vimentin ist ein Intermediärfilament, dass in der Zelle an der Ausbildung des Zytoskeletts beteiligt ist. Überraschenderweise wurde gefunden, daß bestimmter Vimentinfragmente von Fibroblasten nach Stress durch Sonnenlicht sekre- tiert werden. Dies ist eine Tatsache, die man sich für die Bestimmung des Stresszustandes humaner Haut zu Nutze machen kann. So können stressinduzierte Vimentinfragmente durch Mikrodialyseverfahren am Probanden isoliert und quantifiziert werden und so als Maßeinheit für Hautstress dienen.Vimentin is an intermediate filament that is involved in the formation of the cytoskeleton in the cell. Surprisingly, it was found that certain vimentin fragments of fibroblasts are secreted after exposure to sunlight. This is a fact that can be used to determine the stress level of human skin. In this way, stress-induced vimentin fragments can be isolated and quantified by microdialysis methods on the test subject and thus serve as a unit of measurement for skin stress.
Im Rahmen der vorliegenden Erfindung werden unter „Vimentin-Fragmenten" Proteinfragmente mit der Aminosäurensequenz (C)In the context of the present invention, “Vimentin fragments” are protein fragments with the amino acid sequence (C)
N-Term- MSTRSVSSSS YRRMFGGPGT ASRPSSSRSY VTTSTRTYSL GSALRPSTSR SLYASSPGGV YATRSSAVRL RSSVPGVRLL QDSVDFSLAD AINTEFKNTR TNEKVELQEL NDRFANYIDK VRFLEQQNKI LLAELEQLKG QGKSRLGDLY EEEMRELRRQ VDQLTNDKAR VEVERDNLAE DIMRLREKLQ EEMLQREEAE NTLQSFRQDV DNASLARLDL ERKVESLQEE IAFLKKLHEE EIQELQAQIQ EQHVQIDVDV SKPDLTAALR DVRQQYESVA AKNLQEAEEW YKSKFADLSE AANRNNDALR QAKQESTEYR RQVQSLTCEV DALKGTNESL ERQMREMEEN FAVEAANYQD TIGRLQDEIQ NMKEEMARHR EYQDLLNVKM ALDIEIATYR KLLEGEESRI SLPLPNFSSL NLRETNLDSL PLVDTHSKRT LLIKTVETRD GQVINETSQH HDDLE -C-TermN-Term MSTRSVSSSS YRRMFGGPGT ASRPSSSRSY VTTSTRTYSL GSALRPSTSR SLYASSPGGV YATRSSAVRL RSSVPGVRLL QDSVDFSLAD AINTEFKNTR TNEKVELQEL NDRFANYIDK VRFLEQQNKI LLAELEQLKG QGKSRLGDLY EEEMRELRRQ VDQLTNDKAR VEVERDNLAE DIMRLREKLQ EEMLQREEAE NTLQSFRQDV DNASLARLDL ERKVESLQEE IAFLKKLHEE EIQELQAQIQ EQHVQIDVDV SKPDLTAALR DVRQQYESVA AKNLQEAEEW YKSKFADLSE AANRNNDALR QAKQESTEYR RQVQSLTCEV DALKGTNESL ERQMREMEEN FAVEAANYQD TIGRLQDEIQ NMKEEMARHR EYQDLLNVKM ALDIEIATYR KLLEGEESRI SLPLPNFSSL NLRETNLDSL PLVDTHSKRT LLIKTVETRD GQVINETSQH HDDLE -C-term
verstanden, sowie Proteine oder Proteinfragmente, die mehr als 40%, insbe- sσndere"τnehrals 60%7besonders~bevorzugt"mehrals 80% Sequenzhomologie mit dieser Sequenz aufweisen. Die Aminosäuresequenz (C) ist in der Proteindatenbank des National Center for Biotechnology Information (NCBI) unter der Nummer 2119204 offenbart. Diese Datenbank ist im Internet unter folgender Adresse zugänglich: http://www.ncbi.nlm.nih.gov/.understood, as well as proteins or protein fragments which have more than 40%, in particular " τnever than 60% 7 particularly ~ preferably " more than 80% sequence homology with this sequence. The amino acid sequence (C) is disclosed in the National Center for Biotechnology Information (NCBI) protein database under number 2119204. This database is accessible on the Internet at the following address: http://www.ncbi.nlm.nih.gov/.
Im Rahmen der vorliegenden Erfindung umfaßt „Vimentin-Fragment" auch solche Proteine oder Proteinfragmente, deren Aminosäuresequenz durch konservative Mutationen, also durch den Austausch strukturell oder funktionell ähnlicher Aminosäuren, aus der oben aufgeführten Sequenz (C) erhalten werden kann.In the context of the present invention, “vimentin fragment” also includes those proteins or protein fragments whose amino acid sequence can be obtained from the sequence (C) listed above by conservative mutations, that is to say by the exchange of structurally or functionally similar amino acids.
Actin ist ein im Organismenreich sehr konserviertes Protein. Die Actine der Säugetiere werden gemäß ihres isoelektrischen Punkts in drei Klassen eingeteilt. Actin Alpha wird von Skelettmuskulatur, Actin Beta von glatter Muskulatur produziert. Actin Gamma wird von allen übrigen Zellen, also auch von dermalen Fibroblasten synthetisiert. Globuläres Actin (G-Actin) ordnet sich in einem sehr dynamischen Prozess in Reihen an und bildet so filamentöses Actin (F-Actin). Gestresste Zellen ordnen F-Actin in komplexen Bündeln an, die als "Stress Fibers" bezeichnet werden. Überraschenderweise wurde gefunden, daß Actin- moleküle von Zellen nach der Applikation von Stress sekretiert werden. Die Menge der in den extrazellulären Raum der Dermis sekretierten Actinmoleküle kann jedoch ebenso wie die Menge an Vimentinfragmenten für die Beurteilung des Stresszustandes der Haut herangezogen werden. Im Rahmen der vorliegenden Erfindung wird unter „Actin Gamma 1" ein Protein mit der Aminosäurensequenz (D)Actin is a very conserved protein in the organism realm. The actins of mammals are divided into three classes according to their isoelectric point. Actin Alpha is produced by skeletal muscles, Actin Beta by smooth muscles. Actin gamma is synthesized by all other cells, including dermal fibroblasts. Globular actin (G-actin) is arranged in a very dynamic process in rows and thus forms filamentous actin (F-actin). Stressed cells arrange F-actin in complex bundles called "stress fibers". Surprisingly, it was found that actin molecules are secreted by cells after the application of stress. However, the amount of actin molecules secreted into the extracellular space of the dermis, like the amount of vimentin fragments, can be used to assess the state of stress of the skin. In the context of the present invention, under "Actin Gamma 1" a protein with the amino acid sequence (D)
N-Term- MEEEIAALVI DNGSGMCKAG FAGDDAPRAV FPSIVGRPRH QGVMVGMGQK DSYVGDEAQS KRGILTLKYP IEHGIVTNWD DMEKIWHHTF YNELRVAPEE HPVLLTEAPL NPKANREKMT QIMFETFNTP AMYVAIQAVL -SLYÄSGRTTG1VMDSGDGVT-HTVPIYEGYΑ"LPHΑILRLDI-rAGRDLTDYLM KILTERGYSF TTTAEREIVR DIKEKLCYVA LDFEQEMATA ASSSSLEKSY ELPDGQVITI GNERFRCPEA LFQPSFLGME SCGIHETTFN SIMKCDVDIR KDLYANTVLS GGTTMYPGIA DRMQKEITAL APSTMKIKII APPERKYSVW IGGSjLASLS TFQQMWISKQ EYDESGPSIV HRKCF -C-TermN-Term MEEEIAALVI DNGSGMCKAG FAGDDAPRAV FPSIVGRPRH QGVMVGMGQK DSYVGDEAQS KRGILTLKYP IEHGIVTNWD DMEKIWHHTF YNELRVAPEE HPVLLTEAPL NPKANREKMT QIMFETFNTP AMYVAIQAVL -SLYÄSGRTTG1VMDSGDGVT-HTVPIYEGYΑ "LPHΑILRLDI-rAGRDLTDYLM KILTERGYSF TTTAEREIVR DIKEKLCYVA LDFEQEMATA ASSSSLEKSY ELPDGQVITI GNERFRCPEA LFQPSFLGME SCGIHETTFN SIMKCDVDIR KDLYANTVLS GGTTMYPGIA DRMQKEITAL APSTMKIKII APPERKYSVW IGGSjLASLS TFQQMWISKQ EYDESGPSIV HRKCF -C-term
verstanden, sowie Proteine, die mehr als 40%, insbesondere mehr als 60%, besonders bevorzugt mehr als 80% Sequenzhomologie mit dieser Sequenz aufweisen. Die Aminosäuresequenz (D) ist in der Proteindatenbank des National Center for Biotechnology Information (NCBI) unter der Nummer 4501887 offenbart. Diese Datenbank ist im Internet unter folgender Adresse zugänglich: http://www.ncbi.nlm.nih.gov/.understood, and proteins that have more than 40%, in particular more than 60%, particularly preferably more than 80% sequence homology with this sequence. The amino acid sequence (D) is disclosed in the National Center for Biotechnology Information (NCBI) protein database under number 4501887. This database is accessible on the Internet at the following address: http://www.ncbi.nlm.nih.gov/.
Im Rahmen der vorliegenden Erfindung umfaßt „Actin Gamma 1" auch solche Proteine, deren Aminosäuresequenz durch konservative Mutationen, also durch den Austausch strukturell oder funktionell ähnlicher Aminosäuren, aus der oben aufgeführten Sequenz (D) erhalten werden kann.In the context of the present invention, "actin gamma 1" also includes those proteins whose amino acid sequence can be obtained from the sequence (D) listed above by conservative mutations, that is to say by the exchange of structurally or functionally similar amino acids.
Ein weiterer Gegenstand der Erfindung ist ein Test-Kit zur Bestimmung des Hautstreß und/oder der Hautalterung bei Menschen oder Tieren in vitro, daß die erforderlichen Mittel zur Durchführung der obengenannten erfindungsgemäßen Verfahren umfaßt. Ein weiterer Gegenstand der Erfindung ist die Verwendung von Spondin 2, Cathepsin L, Vimentin-Fragmenten oder Actin Gamma 1 gemäß der obengenannten Definitionen als Streß- und/oder Alterungsmarker der Haut bei Menschen oder Tieren. Vorzugsweise werden die genannten Alterungsmarker in einem der erfindungsgemäßen Verfahren verwendet. Die Alterungsmarker können einzeln oder in beliebigen Kombinationen verwendet werden.Another object of the invention is a test kit for determining the skin stress and / or skin aging in humans or animals in vitro, which comprises the necessary means for performing the above-mentioned methods according to the invention. Another object of the invention is the use of spondin 2, cathepsin L, vimentin fragments or actin gamma 1 according to the above definitions as a stress and / or aging marker of the skin in humans or animals. The aging markers mentioned are preferably used in one of the methods according to the invention. The aging markers can be used individually or in any combination.
Ein~weiterer~Ge"geττstand~deτ_Erfindung~ist_ein ~Verfahreτrzum Nachweis der Wirksamkeit von kosmetischen oder pharmazeutischen Wirkstoffen gegen Hautstreß und/oder Hautalterung in vitro, dadurch gekennzeichnet, daß man a) den Hautstatus durch eines der erfindungsgemäßen Verfahren bestimmt, b) einen Wirkstoff gegen Hautstreß und/oder Hautalterung einmal oder mehrmals auf die Haut aufbringt, c) erneut den Hautstatus durch eines der erfindungsgemäßen Verfahren bestimmt und d) die Wirksamkeit des Wirkstoffs durch den Vergleich der Ergebnisse aus a) und c) bestimmt.A ~ further ~ Ge "geττstand ~ deτ _ invention ~ is _ a ~ Verfahreτrzum demonstrating the effectiveness of cosmetic or pharmaceutical active agents against skin stress and / or skin aging in vitro, characterized in that a) determines the skin status through one of the methods according to the invention, b) applying an active substance against skin stress and / or skin aging to the skin one or more times, c) determining the skin status again by one of the methods according to the invention and d) determining the effectiveness of the active substance by comparing the results from a) and c).
Ein weiterer Gegenstand der Erfindung ist ein Test-Kit zum Nachweis der Wirksamkeit von kosmetischen oder pharmazeutischen Wirkstoffen gegen Hautstreß und/oder Hautalterung in vitro, das die erforderlichen Mittel zur Durchführung des obengenannten erfindungsgemäßen Verfahrens umfaßt.Another object of the invention is a test kit for demonstrating the effectiveness of cosmetic or pharmaceutical active substances against skin stress and / or skin aging in vitro, which comprises the necessary means for carrying out the above-mentioned method according to the invention.
Ein weiterer Gegenstand der Erfindung ist die Verwendung von Spondin 2, Cathepsin L, Vimentin-Fragmenten oder Actin Gamma 1 zum Nachweis der Wirksamkeit von kosmetischen oder pharmazeutischen Wirkstoffen gegen Hautstreß und/oder Hautalterung.Another object of the invention is the use of spondin 2, cathepsin L, vimentin fragments or actin gamma 1 for the detection of the effectiveness of cosmetic or pharmaceutical active ingredients against skin stress and / or skin aging.
Ein weiterer Gegenstand der Erfindung ist ein kosmetisches oder pharmazeutisches Mittel zur Regulierung, insbesondere zur Aufrechterhaltung der Homeo- stase menschlicher oder tierischer Haut, das Vimentin-Fragmente gemäß der obengenannten Definition enthält. Die Vimentin-Fragmente können gleich oder voneinander verschieden sein. Die Applikation der erfindungsgemäßen Mittel erfolgt vorzugsweise topisch, insbesondere in Form von Lösungen, Dispersionen (z. B. Emulsionen), Salben oder Cremes,.Another object of the invention is a cosmetic or pharmaceutical agent for regulating, in particular for maintaining the home human or animal skin stasis containing vimentin fragments as defined above. The vimentin fragments can be the same or different. The agents according to the invention are preferably applied topically, in particular in the form of solutions, dispersions (for example emulsions), ointments or creams.
Die Vimentin-Fragmente können auch in Handwaschmitteln, Handgeschirr- ^ ülmitte1n~τ-.äeτ~Kö^^ zweckmäßig~zur Aufrechterhaltung derThe Vimentin fragments can also be used in hand washing detergents, hand dishes- ^ ülmitte1n ~ τ-.äeτ ~ Kö ^^ useful ~ to maintain the
Homeostase menschlicher oder tierischer Haut eingesetzt werden.Homeostasis of human or animal skin can be used.
Weitere, voneinander unabhängige Gegenstände der Erfindung sind daher Handwaschmittel, Handgeschirrspülmittel oder Körperpflegemittel, die Vimentin- Fragmente gemäß der obengenannten Definition enthalten.Further, mutually independent objects of the invention are therefore hand washing agents, hand dishwashing agents or personal care products which contain vimentin fragments as defined above.
Ein weiterer Gegenstand der Erfindung ist die Verwendung von Vimentin-Another object of the invention is the use of vimentin
Fragmenten gemäß der obengenannten Definition in Mitteln zur Regulierung, insbesondere zur Aufrechterhaltung der Homeostase menschlicher oder tierischer Haut sowie zur Herstellung dieser Mittel.Fragments as defined above in agents for regulating, in particular for maintaining the homeostasis of human or animal skin and for producing these agents.
Die Vimentin-Fragmente werden im Sinne der vorliegenden Erfindung vorzugsweise als Komponente in eine kosmetische oder pharmazeutische Zubereitung oder in Handwaschmittel, Handgeschirrspülmittel oder Körperpflegemittel eingebracht bzw. eingearbeitet.For the purposes of the present invention, the vimentin fragments are preferably introduced or incorporated as a component in a cosmetic or pharmaceutical preparation or in hand washing detergents, hand dishwashing detergents or personal care products.
Je nach Art der Formulierung können die erfindungsgemäßen pharmazeutischen Mittel mindestens einen weiteren Hilfs- oder Zusatzstoff, wie z. B. Öle, Schutzkolloide, Weichmacher, Antioxidantien und/oder Emulgatoren enthalten. Im Falle einer Dispersion, insbesondere im Falle einer Suspension oder Emulsion, ist es vorteilhaft, zusätzlich ein physiologisch verträgliches Öl wie beispielsweise Sesamöl, Maiskeimöl, Baumwollsaatöl, Sojabohnenöl oder Erdnu- ßöl, Ester mittelkettiger pflanzlicher Fettsäuren oder Fischöle wie beispielsweise Makrelen-, Sprotten- oder Lachsöl zu verwenden.Depending on the type of formulation, the pharmaceutical compositions according to the invention can contain at least one further auxiliary or additive, such as, for. B. contain oils, protective colloids, plasticizers, antioxidants and / or emulsifiers. In the case of a dispersion, in particular in the case of a suspension or emulsion, it is advantageous to additionally use a physiologically compatible oil such as, for example, sesame oil, corn oil, cottonseed oil, soybean oil or peanut oil. ß oil, esters of medium-chain vegetable fatty acids or fish oils such as mackerel, sprat or salmon oil.
Zur Erhöhung der Stabilität des Wirkstoffes gegen oxidativen Abbau ist es vorteilhaft, Stabilisatoren wie a- Tocopherol, t- Butylhydroxy- toluol, t- Butylhydro- xyanisol, Ascorbinsäure oder Ethoxyquine zuzusetzen.To increase the stability of the active ingredient against oxidative degradation, it is advantageous to add stabilizers such as a-tocopherol, t-butylated hydroxytoluene, t-butylated hydroxyanisole, ascorbic acid or ethoxyquine.
Die Dosierung und Anwendungsdauer der erfindungsgemäß zu verwendenden Vimentin-Fragmente bzw. der Vimentin-Fragmente enthaltenden Mittel kann durch den Fachmann in geeigneter Weise angepaßt und variiert werden.The dosage and duration of use of the vimentin fragments to be used according to the invention or of the agents containing vimentin fragments can be suitably adapted and varied by the person skilled in the art.
Die erfindungsgemäßen Handwaschmittel und Handgeschirrspülmittel sowie die kosmetischen Mittel und Körperpflegemittel wie beispielsweise Haarshampoos, Haarlotionen, Schaumbäder, Duschbäder, Cremes, Gele, Lotionen, alkoholische und wäßrig/alkoholische Lösungen, Emulsionen, Wachs/ Fett-Massen, Stiftpräparate, Puder oder Salben können - je nach Art der Formulierung - als Hilfs- und Zusatzstoffe milde Tenside, Ölkörper, Emulgatoren, Überfettungsmittel, Perlglanzwachse, Konsistenzgeber, Verdickungsmittel, Polymere, Siliconverbindungen, Fette, Wachse, Stabilisatoren, biogene Wirkstoffe, Deodo- rantien, Antitranspirantien, Antischuppenmittel, Filmbildner, Quellmittel, UV- Lichtschutzfaktoren, Antioxidantien, Hydrotrope, Konservierungsmittel, Insek- tenrepellentien, Selbstbräuner, Solubilisatoren, Parfümöle, Farbstoffe und dergleichen enthalten.The hand detergents and hand dishwashing detergents according to the invention and the cosmetic and personal care products such as, for example, hair shampoos, hair lotions, foam baths, shower baths, creams, gels, lotions, alcoholic and aqueous / alcoholic solutions, emulsions, wax / fat masses, stick preparations, powders or ointments can - each by type of formulation - as auxiliary agents and additives, mild surfactants, oil bodies, emulsifiers, superfatting agents, pearlescent waxes, consistency agents, thickeners, polymers, silicone compounds, fats, waxes, stabilizers, biogenic agents, deodorants, antiperspirants, antidandruff agents, film formers, swelling agents, Contain UV protection factors, antioxidants, hydrotropes, preservatives, insect repellents, self-tanners, solubilizers, perfume oils, dyes and the like.
Typische Beispiele für geeignete milde, d.h. besonders hautverträgliche Tenside sind Fettalkoholpolyglycolethersulfate, Monoglyceridsulfate, Mono- und/oder Dialkylsulfosuccinate, Fettsäureisethionate, Fettsäuresarcosinate, Fettsäuretau- ride, Fettsäureglutamate, α-Olefinsulfonate, Ethercarbonsäuren, Alkyloligoglu- coside, Fettsäureglucamide, Alkylamidobetaine und/oder Proteinfettsäurekondensate, letztere vorzugsweise auf Basis von Weizenproteinen. Als Ölkörper kommen beispielsweise Guerbetalkohole auf Basis von Fettalkoholen mit 6 bis 18, vorzugsweise 8 bis 10 Kohlenstoffatomen, Ester von linearen C6-C22-Fettsäuren mit linearen C6-C22-Fettalkoholen, Ester von verzweigten C6-Ci3-Carbonsäuren mit linearen C6-C22-Fettalkoholen, wie z.B. Myristylmy- ristat, Myristylpalmitat, Myristylstearat, Myristylisostearat, Myristyloleat, My- ristylbehenat, Myristylerucat, Cetylmyristat, Cetylpalmitat, Cetylstearat, Cetyli- sostearat, Cetyloleat, Cetylbehenat, Cetylerucat, Stearylmyristat, Stearylpalmi- "tatT_Steäi7lste"afät~Stearylisösteärat Steärylöleat, Stearylbehenat, Stearyleru- cat, Isostearylmyristat, Isostearylpalmitat, Isostearylstearat, Isostearylisostearat, Isostearyloleat, Isostearylbehenat, Isostearyloleat, Oleylmyristat, Oleylpalmitat, Oleylstearat, Oleylisostearat, Oleyloleat, Oleylbehenat, Oleylerucat, Behenylmy- ristat, BeJιenylpalmitat,_Behenylstearat, Behenylisostearat, Behenyloleat, Behe- nylbehenat, Behenylerucat, Erucylmyristat, Erucylpalmitat, Erucylstearat, Eru- cylisostearat, Erucyloleat, Erucylbehenat und Erucylerucat in Betracht.Typical examples of suitable mild, ie particularly skin-compatible surfactants are fatty alcohol polyglycol ether sulfates, monoglyceride sulfates, mono- and / or dialkyl sulfosuccinates, fatty acid isethionates, fatty acid sarcosinates, Fettsäuretau- ride, fatty acid glutamates, α-olefinsulfonates, ether carboxylic acids, Alkyloligoglu- coside, fatty acid glucamides, alkylamidobetaines and / or protein fatty acid condensates, the latter preferably based on wheat proteins. Guerbet alcohols based on fatty alcohols having 6 to 18, preferably 8 to 10 carbon atoms, esters of linear C 6 -C 22 fatty acids with linear C 6 -C 22 fatty alcohols, esters of branched C 6 -Ci 3 carboxylic acids are examples of oil bodies with linear C 6 -C 22 fatty alcohols, such as, for example, myristyl myristate, myristyl palmitate, myristyl stearate, myristyl isostearate, myristyl oleate, myristyl behenate, myristyl erucate, cetyl myristate, cetyl palmitate, cetyl stearate, cetyl isyl stearate, cetyl isyl stolate, cetyl isyl styrene "Tatt _ Steäi7lste" afät ~ Stearylisösteärat Steärylöleat, stearyl, Stearyleru- cat, isostearyl, isostearyl, Isostearylstearat, isostearyl isostearate, Isostearyloleat, isostearyl behenate, Isostearyloleat, oleyl, oleyl palmitate, oleyl stearate, oleyl isostearate, oleate, Oleylbehenat, oleyl, Behenylmy- ristat, BeJιenylpalmitat, _Behenyl stearate, behenyl isostearate, behenyl oleate, behenylbe henate, behenylerucate, erucyl myristate, erucyl palmitate, erucyl stearate, erucyl isostearate, erucyl oleate, erucyl behenate and erucylerucate.
Daneben eignen sich Ester von linearen C6-C22-Fettsäuren mit verzweigten Alkoholen, insbesondere 2-Ethylhexanol, Ester von Hydroxycarbonsäuren mit linearen oder verzweigten C6-C22-Fettalkoholen, insbesondere Dioctyl Malate, Ester von linearen und/oder verzweigten Fettsäuren mit mehrwertigen Alkoholen (wie z.B. Propylenglycol, Dimerdiol oder Trimertriol) und/oder Guerbetalko- holen, Triglyceride auf Basis Cβ-Cio-Fettsäuren, flüssige Mono-/Di- /Triglyceridmischungen auf Basis von C6-Cι8-Fettsäuren, Ester von C6-C22- Fettalkoholen und/oder Guerbetalkoholen mit aromatischen Carbonsäuren, insbesondere Benzoesäure, Ester von C2-Ci2-Dicarbonsäuren mit linearen oder verzweigten Alkoholen mit 1 bis 22 Kohlenstoffatomen oder Polyolen mit 2 bis 10 Kohlenstoffatomen und 2 bis 6 Hydroxylgruppen, pflanzliche Öle, verzweigte primäre Alkohole, substituierte Cyclohexane, lineare und verzweigte C6-C22- Fettalkoholcarbonate, Guerbetcarbonate, Ester der Benzoesäure mit linearen und/oder verzweigten C6-C22-Alkoholen (z.B. Finsolv® TN), lineare oder verzweigte, symmetrische oder unsymmetrische Dialkylether mit 6 bis 22 Kohlen- stoffatomen pro Alkylgruppe, Ringöffnungsprodukte von epoxidierten Fettsäureestern mit Polyolen, Siliconöle und/oder aliphatische bzw. naphthenische Kohlenwasserstoffe, wie z. B. Squalan, Squalen oder Dialkylcyclohexane.In addition, esters of linear C6-C22 fatty acids with branched alcohols, in particular 2-ethylhexanol, esters of hydroxycarboxylic acids with linear or branched C 6 -C 22 fatty alcohols, in particular dioctyl malates, esters of linear and / or branched fatty acids with polyhydric alcohols (such as propylene glycol, dimer diol or trimer triol) and / or Guerbet alcohols, triglycerides based on Cβ-cio fatty acids, liquid mono- / di- / triglyceride mixtures based on C 6 -C 8 fatty acids, esters of C 6 -C 22 - Fatty alcohols and / or Guerbet alcohols with aromatic carboxylic acids, especially benzoic acid, esters of C2-Ci 2 -dicarboxylic acids with linear or branched alcohols with 1 to 22 carbon atoms or polyols with 2 to 10 carbon atoms and 2 to 6 hydroxyl groups, vegetable oils, branched primary Alcohols, substituted cyclohexanes, linear and branched C6-C 22 - fatty alcohol carbonates, Guerbet carbonates, esters of benzoic acid with linear and / or branched C 6 -C 22 alcohols (eg Finsolv ® TN), linear or branched, symmetrical or asymmetrical dialkyl ethers with 6 to 22 carbon atoms Substance atoms per alkyl group, ring opening products of epoxidized fatty acid esters with polyols, silicone oils and / or aliphatic or naphthenic hydrocarbons, such as. B. squalane, squalene or dialkylcyclohexanes.
Als Emulgatoren kommen beispielsweise nichtionogene Tenside aus mindestens einer der folgenden Gruppen in Frage:Examples of suitable emulsifiers are nonionic surfactants from at least one of the following groups:
(1) ÄnTa~geτungsprodukfe~von_2i)Ts^^ oder 0 bis 5 Mol Propylenoxid an lineare Fettalkohole mit 8 bis 22 C-Atomen, an Fettsäuren mit 12 bis 22 C-Atomen, an Alkylphenole mit 8 bis 15 C-Atomen in der Alkylgruppe sowie Alkylamine mit 8 bis 22 Kohlenstoffatomen im Alkylrest;(1) ÄnTa ~ geτungsprodukfe ~ von _ 2i) Ts ^ ^ or 0 to 5 moles of propylene oxide to linear fatty alcohols with 8 to 22 C atoms, to fatty acids with 12 to 22 C atoms, to alkylphenols with 8 to 15 C atoms in the alkyl group and alkylamines having 8 to 22 carbon atoms in the alkyl radical;
(2) Ci2/i8-Fettsäuremon< und -diester von Anlagerungsprodukten von 1 bis 30 Mol Ethylenoxid an Glycerin;(2) Ci 2 / i 8 fatty acid monoesters and diesters of adducts of 1 to 30 moles of ethylene oxide with glycerol;
(3) Glycerinmono- und -diester und Sorbitanmono- und -diester von gesättigten und ungesättigten Fettsäuren mit 6 bis 22 Kohlenstoffatomen und deren Ethylenoxidanlagerungsprodukte;(3) glycerol monoesters and diesters and sorbitan monoesters and diesters of saturated and unsaturated fatty acids having 6 to 22 carbon atoms and their ethylene oxide addition products;
(4) Alkyl- und/oder Alkenylmono- und -oligoglycoside mit 8 bis 22 Kohlenstoffatomen im Alk(en)ylrest und deren ethoxylierte Analoga;(4) alkyl and / or alkenyl mono- and oligoglycosides with 8 to 22 carbon atoms in the alk (en) yl radical and their ethoxylated analogs;
(5) Anlagerungsprodukte von 15 bis 60 Mol Ethylenoxid an Ricinusöl und/oder gehärtetes Ricinusöl;(5) adducts of 15 to 60 moles of ethylene oxide with castor oil and / or hardened castor oil;
(6) Polyol- und insbesondere Polyglycerinester;(6) polyol and especially polyglycerol esters;
(7) Anlagerungsprodukte von 2 bis 15 Mol Ethylenoxid an Ricinusöl und/oder gehärtetes Ricinusöl;(7) adducts of 2 to 15 moles of ethylene oxide with castor oil and / or hardened castor oil;
(8)" Partialester auf Basis linearer, verzweigter, ungesättigter bzw. gesättigter C6/22-Fettsä Liren, " Ricinolsaüre sowie 12-Hydroxystearinsäure und Glycerin, Polyglycerin, Pentaerythrit, Dipentaerythrit, Zuckeralkohole (z.B. Sorbit), Al- kylglucoside (z.B. Methylglucosid, Butylglucosid, Laurylglucosid) sowie Polyglu- coside (z.B. Cellulose);(8) "partial esters based on linear, branched, unsaturated or saturated C6 / 22 Fettsä Liren," Ricinolsaüre and 12-hydroxystearic acid and glycerol, polyglycerol, pentaerythritol, dipentaerythritol, sugar alcohols (eg sorbitol), Al kylglucoside (eg methylglucoside , Butyl glucoside, lauryl glucoside) and polyglucosides (eg cellulose);
(9) Mono-, Di- und Trialkylphosphate sowie Mono-, Di- und/oder Tri-PEG- alkylphosphate und deren Salze; (10) Wollwachsalkohole;(9) mono-, di- and trialkyl phosphates and mono-, di- and / or tri-PEG-alkyl phosphates and their salts; (10) wool wax alcohols;
(11) Polysiloxan-Polyalkyl-Polyether-Copolymere bzw. entsprechende Derivate;(11) polysiloxane-polyalkyl-polyether copolymers or corresponding derivatives;
(12) Mischester aus Pentaerythrit, Fettsäuren, Citronensäure und Fettalkohol gemäß DE 1165574 PS und/oder Mischester von Fettsäuren mit 6 bis 22 Kohlenstoffatomen, Methylglucose und Polyolen, vorzugsweise Glycerin oder Poly- glycerin,(12) mixed esters of pentaerythritol, fatty acids, citric acid and fatty alcohol according to DE 1165574 PS and / or mixed esters of fatty acids with 6 to 22 carbon atoms, methyl glucose and polyols, preferably glycerol or polyglycerol,
(13)~PolyalkylenglycOle~sowie~~ (14) Glycerincarbonat.(13) ~ Polyalkylene glycol ~ and ~~ (14) glycerol carbonate.
Die Anlagerungsprodukte von Ethylenoxid und/oder von Propylenoxid an Fettalkohole, Fettsäuren, Alkylphenole, Glycerinmono- und -diester sowie Sorbi- tanmono- und -diester von Fettsäuren oder an Ricinusöl stellen bekannte, im Handel erhältliche Produkte dar.The adducts of ethylene oxide and / or of propylene oxide with fatty alcohols, fatty acids, alkylphenols, glycerol mono- and diesters as well as sorbitan mono- and diesters with fatty acids or with castor oil are known, commercially available products.
Es handelt sich dabei um Homologengemische, deren mittlerer Alkoxy- lierungsgrad dem Verhältnis der Stoffmengen von Ethylenoxid und/ oder Propylenoxid und Substrat, mit denen die Anlagerungsreaktion durchgeführt wird, entspricht. Ci2/i8-Fettsäuremono- und -diester von Anlagerungsprodukten von Ethylenoxid an Glycerin sind aus DE 2024051 PS als Rückfettungsmittel für kosmetische Zubereitungen bekannt.These are homolog mixtures whose average degree of alkoxylation corresponds to the ratio of the amounts of ethylene oxide and / or propylene oxide and substrate with which the addition reaction is carried out. Ci2 / i 8 fatty acid monoesters and diesters of adducts of ethylene oxide with glycerol are known from DE 2024051 PS as refatting agents for cosmetic preparations.
Alkyl- und/oder Alkenylmono- und -oligoglycoside, ihre Herstellung und ihre Verwendung sind aus dem Stand der Technik bekannt. Ihre Herstellung erfolgt insbesondere durch Umsetzung von Glucose oder Oligosacchariden mit primären Alkoholen mit 8 bis 18 C-Atomen. Bezüglich des Glycosidrestes gilt, daß sowohl Monoglycoside, bei denen ein cyclischer Zuckerrest glycosidisch an den Fettalkohol gebunden ist, als auch oligomere Glycoside mit einem Oligomerisa- tionsgrad bis vorzugsweise etwa 8 geeignet sind. Der Oligomerisierungsgrad ist dabei ein statistischer Mittelwert, dem eine für solche technischen Produkte übliche Homologenverteilung zugrunde liegt.Alkyl and / or alkenyl mono- and oligoglycosides, their preparation and their use are known from the prior art. They are produced in particular by reacting glucose or oligosaccharides with primary alcohols with 8 to 18 carbon atoms. With regard to the glycoside residue, both monoglycosides in which a cyclic sugar residue is glycosidically bonded to the fatty alcohol and oligomeric glycosides with a degree of oligomerization of up to about 8 are suitable. The degree of oligomerization is a statistical mean value based on a homolog distribution customary for such technical products.
Typische Beispiele für geeignete Polyglycerinester sind Polyglyceryl-2 Dipoly- hydroxystearate (Dehymuls® PGPH), Polyglycerin-3-Diisostearate (Lameform® TGI), Polyglyceryl-4 Isostearate (Isolan® Gl 34), Polyglyceryl-3 Oleate, Dii- sostearoyl PolygIyceryl-3 Diisostearate (Isolan® PDI), Polyglyceryl-3 Methylglu- cose Distearate (Tego Gare® 450), Polyglyceryl-3 Beeswax (Cera Bellina®), Polyglyceryl-4 Caprate (Polyglycerol Caprate T2010/90), Polyglyceryl-3 Cetyl Ether (Chimexane® NL), Polyglyceryl-3 Distearate (Cremophor® GS 32) und Polyglyceryl Polyricinoleate (Admul® WOL 1403), Polyglyceryl Dimerate Isostearate sowie deren Gemische.Typical examples of suitable polyglycerol esters are polyglyceryl-2 dipolyhydroxystearate (Dehymuls ® PGPH), polyglycerol-3-diisostearate (Lameform ® TGI), polyglyceryl-4 isostearate (Isolan ® Gl 34), polyglyceryl-3 oleate, diisostearoyl polyglyceryl 3 diisostearates (Isolan ® PDI), polyglyceryl-3 methyl glucose distearate (Tego Gare ® 450), polyglyceryl-3 beeswax (Cera Bellina ® ), polyglyceryl-4 caprate (polyglycerol caprate T2010 / 90), polyglyceryl-3 cetyl ether ( Chimexane ® NL), Polyglyceryl-3 Distearate (Cremophor ® GS 32) and Polyglyceryl Polyricinoleate (Admul ® WOL 1403), Polyglyceryl Dimerate Isostearate and their mixtures.
Weiterhin können als Emulgatoren zwitterionische Tenside verwendet werden. Als zwitterionische Tenside werden solche oberflächenaktiven Verbindungen bezeichnet, die im Molekül mindestens eine quartäre Ammoniumgruppe und mindestens eine Carboxylat- und eine Sulfonatgruppe tragen. Besonders geeignete zwitterionische Tenside sind die sogenannten Betaine wie die N-Alkyl- N,N-dimethylammoniumglycinate, beispielsweise das Kokosalkyldimethylam- moniumglycinat, N-Acylaminopropyl-N,N-dimethylammoniumglycinate, beispielsweise das Kokosacylaminopropyldimethylammoniumglycinat, und 2-Alkyl- 3-carboxylmethyl-3-hydroxyethylimidazoline mit jeweils 8 bis 18 C-Atomen in der Alkyl- oder Acylgruppe sowie das Kokosacylaminoethylhydroxyethyl- carboxymethylglycinat. Besonders bevorzugt ist das unter der CTFA- Bezeichnung Cocamidopropyl Betaine bekannte Fettsäureamid-Derivat. Ebenfalls geeignete Emulgatoren sind ampholytische Tenside. Unter ampholytischen Tensiden werden solche oberflächenaktiven Verbindungen verstanden, die außer einer Cs/is-Alkyl- oder -Acylgruppe im Molekül mindestens eine freie Aminogruppe und mindestens eine -COOH- oder -SOsH-Gruppe enthalten und zur Ausbildung innerer Salze befähigt sind. Beispiele für geeignete ampholyti- sehe Tenside sind N-Alkylglycine, N-Alkylpropionsäuren, N-Alkylaminobutter- säuren, N-Alkyliminodipropionsäuren, N-Hydroxyethyl-N-alkylamidopropylg- lycine, N-Alkyltaurine, N-Alkylsarcosine, 2-Alkylaminopropionsäuren und Alky- laminoessigsäuren mit jeweils etwa 8 bis 18 C-Atomen in der Alkylgruppe. Besonders bevorzugte ampholytische Tenside sind das N-Kokosalkyl- aminopropionat, das Kokosacylaminoethylaminopropionat und das C12/.8- Acylsarcosin. Neben den ampholytischen kommen auch quartäre Emulgatoren IfTBelraclitT -vöBersoiche vom Typ der Esterquats, vorzugsweise methylquater- nierte Difettsäuretriethanolaminester-Salze, besonders bevorzugt sind.Zwitterionic surfactants can also be used as emulsifiers. Zwitterionic surfactants are surface-active compounds that contain at least one quaternary ammonium group and at least one carboxylate and one sulfonate group in the molecule. Particularly suitable zwitterionic surfactants are the so-called betaines such as the N-alkyl-N, N-dimethylammonium glycinate, for example the coconut alkyldimethylammonium glycinate, N-acylaminopropyl-N, N-dimethylammonium glycinate, for example the coconut acylaminopropyldimethylammonium glycinate, and 2-alkyl-3-carboxylm -hydroxyethylimidazolines each having 8 to 18 carbon atoms in the alkyl or acyl group and the cocoacylaminoethylhydroxyethyl carboxymethylglycinate. The fatty acid amide derivative known under the CTFA name Cocamidopropyl Betaine is particularly preferred. Suitable emulsifiers are ampholytic surfactants. Ampholytic surfactants are surface-active compounds which, in addition to a Cs / is alkyl or acyl group, contain at least one free amino group and at least one -COOH or -SOsH group in the molecule and are capable of forming internal salts. Examples of suitable ampholytic see surfactants are N-alkylglycines, N-alkylpropionic acids, N-alkylaminobutyric acids, N-alkyliminodipropionic acids, N-hydroxyethyl-N-alkylamidopropylglycines, N-alkyltaurines, N-alkylsarcosines, 2-alkylaminopropionic acids and alkylaminoacetic acids each with about 8 up to 18 carbon atoms in the alkyl group. Particularly preferred ampholytic surfactants are N-coconut alkyl aminopropionate, coconut acylaminoethyl aminopropionate and C 1 2 /. 8 - acylsarcosine. In addition to the ampholytic, quaternary emulsifiers IfTBelraclitT -vöBersoiche of the ester quat type, preferably methyl-quaternized difatty acid triethanolamine ester salts, are particularly preferred.
Als Überfettungsmittel können Substanzen wie beispielsweise Lanolin und Lecithin sowie, polyethoxylierte oder aeylierte Lanolin- und Lecithinderivate, Polyolfettsäureester, Monoglyceride und Fettsäurealkanolamide verwendet werden, wobei die letzteren gleichzeitig als Schaumstabilisatoren dienen.Substances such as lanolin and lecithin and, polyethoxylated or aeylated lanolin and lecithin derivatives, polyol fatty acid esters, monoglycerides and fatty acid alkanolamides can be used as superfatting agents, the latter simultaneously serving as foam stabilizers.
Als Perlglanzwachse kommen beispielsweise in Frage: Alkylenglycolester, speziell Ethylenglycoldistearat; Fettsäurealkanolamide, speziell Kokosfettsäure- diethanolamid; Partialglyceride, speziell Stearinsäuremonoglycerid; Ester von mehrwertigen, gegebenenfalls hydroxysubstituierte Carbonsäuren mit Fettalkoholen mit 6 bis 22 Kohlenstoffatomen, speziell langkettige Ester der Weinsäure; Fettstoffe, wie beispielsweise Fettalkohole, Fettketone, Fettaldehyde, Fettether und Fettcarbonate, die in Summe mindestens 24 Kohlenstoffatome aufweisen, speziell Lauron und Distearylether; Fettsäuren wie Stearinsäure, Hydrpxystea- rinsäure oder Behensäure, Ringöffnungsprodukte von Olefinepoxiden mit 12 bis 22 Kohlenstoffatomen mit Fettalkoholen mit 12 bis 22 Kohlenstoffatomen und/oder Polyolen mit 2 bis 15 Kohlenstoffatomen und 2 bis 10 Hydroxylgruppen sowie deren Mischungen.Pearlescent waxes that can be used are, for example: alkylene glycol esters, especially ethylene glycol distearate; Fatty acid alkanolamides, especially coconut fatty acid diethanolamide; Partial glycerides, especially stearic acid monoglyceride; Esters of polyvalent, optionally hydroxy-substituted carboxylic acids with fatty alcohols having 6 to 22 carbon atoms, especially long-chain esters of tartaric acid; Fatty substances, such as, for example, fatty alcohols, fatty ketones, fatty aldehydes, fatty ethers and fatty carbonates, which have a total of at least 24 carbon atoms, especially lauron and distearyl ether; Fatty acids such as stearic acid, hydroxystearic acid or behenic acid, ring opening products of olefin epoxides with 12 to 22 carbon atoms with fatty alcohols with 12 to 22 carbon atoms and / or polyols with 2 to 15 carbon atoms and 2 to 10 hydroxyl groups and mixtures thereof.
Als Konsistenzgeber kommen in erster Linie Fettalkohole oder Hydroxyfettalko- hole mit 12 bis 22 und vorzugsweise 16 bis 18 Kohlenstoffatomen und daneben Partialglyceride, Fettsäuren oder Hydroxyfettsäuren in Betracht. Bevorzugt ist eine Kombination dieser Stoffe mit Alkyloligoglucosiden und/oder Fettsäure-N- methylglucamiden gleicher Kettenlänge und/oder Polyglycerinpoly-12- hydroxystearaten.The main consistency agents are fatty alcohols or hydroxy fatty alcohols with 12 to 22 and preferably 16 to 18 carbon atoms and in addition Partial glycerides, fatty acids or hydroxy fatty acids into consideration. A combination of these substances with alkyl oligoglucosides and / or fatty acid N-methylglucamides of the same chain length and / or polyglycerol poly-12-hydroxystearates is preferred.
Geeignete Verdickungsmittel sind beispielsweise Aerosil-Typen (hydrophile Kieselsäuren), Polysaccharide, insbesondere Xanthan-Gum, Guar-Guar, Agar- Agar, Alginate und Tylosen, Carboxymethylcellulose und Hydroxyethylcellulose, ferner höhermolekulare Polyethylenglycolmono- und -diester von Fettsäuren, Polyacrylate, (z.B. Carbopole® von Goodrich oder Synthalene® von Sigma), Polyacrylamide, Polyvinylalkohol und Polyvinylpyrrolidon, Tenside wie bei- spielsweise ethoxylierte Fettsäureglyceride, Ester von Fettsäuren mit Polyolen wie beispielsweise Pentaerythrit oder Trimethylolpropan, Fettalkoholethoxylate mit eingeengter Homologenverteilung oder Alkyloligoglucoside sowie Elektrolyte wie Kochsalz und Ammoniumchlorid.Suitable thickeners are, for example, Aerosil types (hydrophilic silicas), polysaccharides, in particular xanthan gum, guar guar, agar agar, alginates and tyloses, carboxymethyl cellulose and hydroxyethyl cellulose, and also higher molecular weight polyethylene glycol mono- and diesters of fatty acids, polyacrylates (eg carbopols) ® from Goodrich or Synthalene ® from Sigma), polyacrylamides, polyvinyl alcohol and polyvinyl pyrrolidone, surfactants such as, for example, ethoxylated fatty acid glycerides, esters of fatty acids with polyols such as pentaerythritol or trimethylol propane, fatty alcohol ethoxylates with restricted homolog distribution or alkyl oligoglucosides and electrolytes such as sodium chloride.
Geeignete kationische Polymere sind beispielsweise kationische Cellulosederi- vate, wie z.B. eine quaternierte Hydroxyethylcellulose, die unter der Bezeichnung Polymer JR 400® von Amerchol erhältlich ist, kationische Stärke, Copo- lymere von Diallylammoniumsalzen und Acrylamiden, quaternierte Vinylpyrroli- don/Vinylimidazol-Polymere, wie z.B. Luviquat® (BASF), Kondensationsprodukte von Polyglycolen und Aminen, quaternierte Kollagenpolypeptide, wie beispielsweise Lauryldimonium hydroxypropyl hydrolyzed Collagen (Lame- quat®L/Grünau), quaternierte Weizenpolypeptide, Polyethylenimin, kationische Siliconpolymere,_wie_z.B-._AmidomethicDne, _Copolymere_der Adipinsäure und Dimethylaminohydroxypropyldiethylentriamin (Cartaretine®/Sandoz), Copoly- mere der Acrylsäure mit Dimethyldiallylammoniumchlorid (Merquat® 550/Chemviron), Polyaminopolyamide, wie z.B. beschrieben in der FR 2252840 A sowie deren vernetzte wasserlöslichen Polymere, kationische Chitinderivate wie beispielsweise quaterniertes Chitosan, gegebenenfalls mikrokristallin ver- teilt, Kondensationsprodukte aus Dihalogenalkylen, wie z.B. Dibrombutan mit Bisdialkylaminen, wie z.B. Bis-Dimethylamino-1 ,3-propan, kationischer Guar- Gum, wie z.B. Jaguar® CBS, Jaguar® C-17, Jaguar® C-16 der Firma Cela- nese, quaternierte Ammoniumsalz-Polymere, wie z.B. Mirapol® A-15, Mirapol® AD-1 , Mirapol® AZ-1 der Firma Miranol.Suitable cationic polymers are, for example, cationic cellulose derivatives, such as, for example, a quaternized hydroxyethyl cellulose, which is available under the name Polymer JR 400® from Amerchol, cationic starch, copolymers of diallylammonium salts and acrylamides, quaternized vinylpyrrolidone / vinylimidazole polymers, such as eg Luviquat® (BASF), condensation products of polyglycols and amines, quaternized collagen polypeptides, such as lauryldimonium hydroxypropyl hydrolyzed collagen (Lamequat®L / Grünau), quaternized wheat polypeptides, polyethyleneimine, cationic silicone polymers, _how_z Dimethylaminohydroxypropyldiethylenetriamine (Cartaretine® / Sandoz), copolymers of acrylic acid with dimethyldiallylammonium chloride (Merquat® 550 / Chemviron), polyaminopolyamides, as described for example in FR 2252840 A and their crosslinked water-soluble polymers, cationic chitin derivatives such as quaternier tes chitosan, optionally microcrystalline divides, condensation products from dihaloalkylene, such as dibromobutane with bisdialkylamines, such as bis-dimethylamino-1, 3-propane, cationic guar gum, such as Jaguar® CBS, Jaguar® C-17, Jaguar® C-16 from Cela- nese, quaternized ammonium salt polymers, such as Mirapol® A-15, Mirapol® AD-1, Mirapol® AZ-1 from Miranol.
Als anionische, zwitterionische, amphotere und nichtionische Polymere kommen beispielsweise Vinylacetat/Crotonsäure-Copolymere, Vinylpyrroli- don/Vinylacrylat-Copolymere, Vinylacetat/Butylmaleat/ Isobornylacrylat-Co- polymere, Methylvinylether/Maleinsäureanhydrid-Copolymere und deren Ester, unvemetzte und mit Polyolen vernetzte Polyacrylsäuren, Acrylamidopro- -pyltrimethylammoniumchlorid/Acrylat-Copolymere, Octylacrylamid/Methylmeth- acrylat/tert.Butylaminoethylmethacrylat/2-Hydroxyproyl-methacrylat-Copolyme- re, Polyvinylpyrrolidon, VinylpyrrolidonΛ/inylacetat-Copolymere, Vinylpyrrolidon/ Dimethylaminoethylmethacrylat/Vinylcaprolactam-Terpolymere sowie gegebenenfalls derivatisierte Celluloseether und Silicone in Frage.Anionic, zwitterionic, amphoteric and nonionic polymers include, for example, vinyl acetate / crotonic acid copolymers, vinyl pyrrolidone / vinyl acrylate copolymers, vinyl acetate / butyl maleate / isobornyl acrylate copolymers, methyl vinyl ether / maleic anhydride copolymers and their polyols and uncrosslinked polyols and their esters , Acrylamidopro- -pyltrimethylammonium chloride / acrylate copolymers, octylacrylamide / methyl methacrylate / tert.butylaminoethyl methacrylate / 2-hydroxyproyl methacrylate copolymers, polyvinyl pyrrolidone, vinyl pyrrolidoneΛ / inylacetate copolymers, vinylpyrrolidethyl methacrylate and / or dimethylamino methyl acrylate / derivates Silicones in question.
Geeignete Siliconverbindungen sind beispielsweise Dimethylpolysiloxane, Me- thylphenylpolysiloxane, cyclische Silicone sowie amino-, fettsäure-, alkohol-, polyether-, epoxy-, fluor-, glykosid- und/oder alkylmodifizierte Siliconverbindungen, die bei Raumtemperatur sowohl flüssig als auch harzförmig vorliegen können. Weiterhin geeignet sind Simethicone, bei denen es sich um Mischungen aus Dimethiconen mit einer durchschnittlichen Kettenlänge von 200 bis 300 Dimethylsiloxan-Einheiten und hydrierten Silicaten handelt. Eine detaillierte Übersicht über geeignete flüchtige Silicone findet sich zudem von Todd et al. in Cosm.Toil. 91, 27 (1976).Suitable silicone compounds are, for example, dimethylpolysiloxanes, methylphenylpolysiloxanes, cyclic silicones and amino-, fatty acid-, alcohol-, polyether-, epoxy-, fluorine-, glycoside- and / or alkyl-modified silicone compounds which can be both liquid and resinous at room temperature. Simethicones, which are mixtures of dimethicones with an average chain length of 200 to 300 dimethylsiloxane units and hydrogenated silicates, are also suitable. A detailed overview of suitable volatile silicones can also be found by Todd et al. in Cosm.Toil. 91, 27 (1976).
Typische Beispiele für Fette sind Glyceride, als Wachse kommen u.a. natürliche Wachse, wie z.B. Candelillawachs, Carnaubawachs, Japanwachs, Espar- tograswachs, Korkwachs, Guarumawachs, Reis-keimölwachs, Zuckerrohr- wachs, Ouricurywachs, Montanwachs, Bienenwachs, Schellackwachs, Walrat, Lanolin (Wollwachs), Bürzelfett, Ceresin, Ozokerit (Erdwachs), Petrolatum, Paraffinwachse, Mikrowachse; chemisch modifizierte Wachse (Hartwachse), wie z.B. Montanesterwachse, Sasolwachse, hydrierte Jojobawachse sowie synthetische Wachse, wie z.B. Polyalkylenwachse und Polyethylenglycolwach- se in Frage.Typical examples of fats are glycerides, waxes include natural waxes such as candelilla wax, carnauba wax, Japanese wax, espresso grass wax, cork wax, guaruma wax, rice-germ oil wax, sugar cane wax. wax, ouricury wax, montan wax, beeswax, shellac wax, walrus, lanolin (wool wax), pretzel fat, ceresin, ozokerite (earth wax), petrolatum, paraffin waxes, micro waxes; chemically modified waxes (hard waxes), such as montan ester waxes, Sasol waxes, hydrogenated jojoba waxes and synthetic waxes, such as polyalkylene waxes and polyethylene glycol waxes, are possible.
Ais-Stabilisatoren -können~Metalls"alze~voτrFettsäurenτ~wie z.B. Magnesium-, Aluminium- und/oder Zinkstearat bzw. -ricinoleat eingesetzt werden.Ais be stabilizers ~ -can metal "Alze voτrFettsäurenτ ~ ~ such as magnesium, aluminum and / or zinc stearate or ricinoleate used.
Unter biogenen Wirkstoffen sind beispielsweise Tocopherol, Tocopherolacetat, Tocopherolpaimitat^Ascorbinsäure^ Desoxyribonucleinsäure^Retinol, Bisabolol, Allantoin, Phytantriol, Panthenol, AHA-Säuren, Aminosäuren, Ceramide, Pseu- doceramide, essentielle Öle, Pflanzenextrakte und Vitaminkomplexe zu verstehen.Biogenic active substances are, for example, tocopherol, tocopherol acetate, tocopherol paimitat ^ ascorbic acid ^ deoxyribonucleic acid ^ retinol, bisabolol, allantoin, phytantriol, panthenol, AHA acids, amino acids, ceramides, pseudo doceramides, essential oils, plant extracts and vitamin complexes.
Kosmetische Deodorantien (Desodorantien) wirken Körpergerüchen entgegen, überdecken oder beseitigen sie. Körpergerüche entstehen durch die Einwirkung von Hautbakterien auf apokrinen Schweiß, wobei unangenehm riechende Abbauprodukte gebildet werden. Dementsprechend enthalten Deodorantien Wirkstoffe, die als keimhemmende Mittel, Enzyminhibitoren, Geruchsabsorber oder Geruchsüberdecker fungieren.Cosmetic deodorants counteract, mask or eliminate body odors. Body odors arise from the action of skin bacteria on apocrine sweat, whereby unpleasant smelling breakdown products are formed. Accordingly, deodorants contain active ingredients which act as germ-inhibiting agents, enzyme inhibitors, odor absorbers or odor maskers.
Als keimhemmende Mittel, die gegebenenfalls den erfindungsgemäßen Kosme- tika zugesetzt werden, sind grundsätzlich alle gegen grampositive Bakterien wirksamen Stoffe geeignet, wie z. B. 4-Hydroxybenzoesäure und ihre Salze und Ester, N-(4-Chlorphenyl)-N'-(3,4 dichlorphenyl)harnstoff, 2,4,4'-Trichlor-2'- hydroxydiphenylether (Triclosan), 4-Chlor-3,5-dimethylphenol, 2,2'-Methylen- bis(6-brom-4-chlorphenol), 3-Methyl-4-(1-methylethyl)phenol, 2-Benzyl-4- chlorphenol, 3-(4-Chlorphenoxy)-1 ,2-propandiol, 3-lod-2-propinylbutylcarbamat, Chlorhexidin, 3,4,4'-Trichlorcarbanilid (TTC), antibakterielle Riechstoffe, Thy- mol, Thymianöl, Eugenol, Nelkenöl, Menthol, Minzöl, Farnesol, Phenoxyetha- nol, Glycerinmonolaurat (GML), Diglycerinmonocaprinat (DMC), Salicylsäure-N- alkylamide wie z. B. Salicylsäure-n-octylamid oder Salicylsäure-n-decylamid.In principle, all substances which are effective against gram-positive bacteria are suitable as germ-inhibiting agents, which may be added to the cosmetics according to the invention, such as B. 4-hydroxybenzoic acid and its salts and esters, N- (4-chlorophenyl) -N '- (3,4 dichlorophenyl) urea, 2,4,4 ' -Trichlor-2'-hydroxydiphenyl ether (triclosan), 4-chloro -3,5-dimethylphenol, 2,2 'methylene-bis (6-bromo-4-chlorophenol), 3-methyl-4- (1-methylethyl) phenol, 2-benzyl-4-chlorophenol, 3- (4 -Chlorphenoxy) -1, 2-propanediol, 3-iodo-2-propynyl butyl carbamate, Chlorhexidine, 3,4,4'-trichlorocarbanilide (TTC), antibacterial fragrances, Thymol, thyme oil, eugenol, clove oil, menthol, mint oil, farnesol, phenoxyethanol, glycerol monolaurate (GML), diglycerol monocaprinate (DMC), salicylic acid-N - alkylamides such as B. salicylic acid-n-octylamide or salicylic acid-n-decylamide.
Auch Enzyminhibitoren können den erfindungsgemäßen Kosmetika zugesetzt werden. Geeignete Enzyminhibitoren sind beispielsweise Esteraseinhibitoren. Hierbei handelt es sich vorzugsweise um Trialkylcitrate wie Trimethylcitrat, Tripropylcitrat, Triisopropylcitrat, Tributylcitrat und insbesondere Triethylcitrat (Hydagen® CAT, Henkel KGaA, Düsseldorf/FRG). Die Stoffe inhibieren die Enzymaktivität und reduzieren dadurch die Geruchsbildung. Weitere Stoffe, die als Esteraseinhibitoren in Betracht kommen, sind Sterolsulfate oder -phospha- te, wie beispielsweise Lanosterin-, Cholesterin-, Campesterin-, Stigmasterin- und Sitosterinsulfat bzw -phosphat, Dicarbonsäuren und deren Ester, wie beispielsweise Glutarsäure, Glutarsäuremonoethylester, Glutarsäurediethylester, Adipinsäure, Adipinsäuremonoethylester, Adipinsäurediethylester, Malonsäure und Malonsäurediethylester, Hydroxycarbnonsäuren und deren Ester wie beispielsweise Citronensäure, Äpfelsäure, Weinsäure oder Weinsäurediethylester, sowie Zinkglycinat.Enzyme inhibitors can also be added to the cosmetics according to the invention. Suitable enzyme inhibitors are, for example, esterase inhibitors. These are preferably trialkyl such as trimethyl citrate, tripropyl, triisopropyl, tributyl citrate and especially triethyl citrate (Hydagen® ® CAT, Henkel KGaA, Dusseldorf / FRG). The substances inhibit enzyme activity and thereby reduce odor. Other substances which can be considered as esterase inhibitors are sterol sulfates or phosphates, such as, for example, lanosterol, cholesterol, campesteric, stigmasterol and sitosterol sulfate or phosphate, dicarboxylic acids and their esters, such as, for example, glutaric acid, glutaric acid monoethyl ester, glutaric acid diethyl ester, Adipic acid, adipic acid monoethyl ester, adipic acid diethyl ester, malonic acid and malonic acid diethyl ester, hydroxycarboxylic acids and their esters such as citric acid, malic acid, tartaric acid or tartaric acid diethyl ester, and zinc glycinate.
Als Geruchsabsorber eignen sich Stoffe, die geruchsbildende Verbindungen aufnehmen und weitgehend festhalten können. Sie senken den Partialdruck der einzelnen Komponenten und verringern so auch ihre Ausbreitungsgeschwindigkeit. Wichtig ist, daß dabei Parfüms unbeeinträchtigt bleiben müssen. Geruchsabsorber haben keine Wirksamkeit gegen Bakterien. Sie enthalten beispielsweise als Hauptbestandteil ein komplexes Zinksalz der Ricinolsäure oder spezielle, weitgehend geruchsneutrale Duftstoffe, die dem Fachmann als "Fixateure" bekannt sind, wie z. B. Extrakte von Labdanum bzw. Styrax oder bestimmte Abietinsäurederivate. Als Geruchsüberdecker fungieren Riechstoffe oder Parfümöle, die zusätzlich zu ihrer Funktion als Geruchsüberdecker den Deodorantien ihre jeweilige Duftnote verleihen. Als Parfümöle seien beispielsweise genannt Gemische aus natürlichen und synthetischen Riechstoffen. Natürliche Riechstoffe sind Extrakte von Blüten, Stengeln und Blättern, Früchten, Fruchtschalen, Wurzeln, Hölzern, Kräutern und Gräsern, Nadeln und Zweigen sowie Harzen und Balsamen. Weiterhin kommen tierische Rohstoffe in Frage, wie beispielsweise Zibet und Castoreum. Typische synthetische Riechstoffverbindungen sind Produkte vom Typ der Ester, Ether, Aldehyde, Ketone, Alkohole
Figure imgf000024_0001
Typ der Ester sind z.B.Suitable odor absorbers are substances that absorb odor-forming compounds and can retain them to a large extent. They lower the partial pressure of the individual components and thus also reduce their speed of propagation. It is important that perfumes must remain unaffected. Odor absorbers are not effective against bacteria. They contain, for example, a complex zinc salt of ricinoleic acid or special, largely odorless fragrances, which are known to the person skilled in the art as "fixators", such as, for example, the main component. B. extracts of Labdanum or Styrax or certain abietic acid derivatives. Fragrances or perfume oils act as odor maskers, which in addition to their function as odor maskers Deodorants give their respective fragrance. Perfume oils are, for example, mixtures of natural and synthetic fragrances. Natural fragrances are extracts of flowers, stems and leaves, fruits, fruit peels, roots, woods, herbs and grasses, needles and branches as well as resins and balms. Animal raw materials, such as civet and castoreum, are also suitable. Typical synthetic fragrance compounds are products of the ester, ether, aldehyde, ketone, and alcohol type
Figure imgf000024_0001
Type of the esters are, for example
Benzylacetat, p-tert.-Butylcyclohexylacetat, Linalylacetat, Phenylethylacetat, Linalylbenzoat, Benzylformiat, Allylcyclohexylpropionat, Styrallylpropionat und Benzylsalicylat. Zu den Ethern zählen beispielsweise Benzylethylether, zu den Aldehyden z.B. die linearen Alkanale mit 8 bis 18 Kohlenstoffatomen, Citral, Citronellal, Citronellyloxyacetaldehyd, Cyclamenaldehyd, Hydroxycitronellal, Lilial und Bourgeonal, zu den Ketonen z.B. die Jonone und Methylcedrylketon, zu den Alkoholen Anethol, Citronellol, Eugenol, Isoeugenol, Geraniol, Linalool, Phenylethylalkohol und Terpineol, zu den Kohlenwasserstoffen gehören hauptsächlich die Terpene und Balsame. Bevorzugt werden jedoch Mischungen verschiedener Riechstoffe verwendet, die gemeinsam eine ansprechende Duftnote erzeugen. Auch ätherische Öle geringerer Flüchtigkeit, die meist als Aromakomponenten verwendet werden, eignen sich als Parfümöle, z.B. Salbeiöl, Kamillenöl, Nelkenöl, Melissenöl, Minzenöl, Zimtblätteröl, Lindenblütenöl, Wa- cholderbeerenöl, Vetiveröl, Olibanöl, Galbanumöl, Labdanumöl und Lavandinöl. Vorzugsweise werden Bergamotteöl, Dihydromyrcenol, Lilial, Lyral, Citronellol, Phenylethylalkohol, α-Hexylzimtaldehyd, Geraniol, Benzylaceton, Cyclamenaldehyd, Linalool, Boisambrene Forte, Ambroxan, Indol, Hedione, Sandelice, Citronenöl, Mandarinenöl, Orangenöl, Allylamylglycolat, Cyclovertal, Lavandinöl, Muskateller Salbeiöl, ß-Damascone, Geraniumöl Bourbon, Cyclohexylsali- cylat, Vertofix Coeur, Iso-E-Super, Fixolide NP, Evernyl, Iraldein gamma, Phe- nylessigsäure, Geranylacetat, Benzylacetat, Rosenoxid, Romilat, Irotyl und Floramat allein oder in Mischungen, eingesetzt. Antitranspirantien (Antiperspirantien) reduzieren durch Beeinflussung der Aktivität der ekkrinen Schweißdrüsen die Schweißbildung, und wirken somit Achselnässe und Körpergeruch entgegen. Wässrige oder wasserfreie Formulierungen von Antitranspirantien enthalten typischerweise folgende Inhaltsstoffe:Benzyl acetate, p-tert-butylcyclohexyl acetate, linalyl acetate, phenylethyl acetate, linalyl benzoate, benzyl formate, allyl cyclohexyl propionate, styrallyl propionate and benzyl salicylate. The ethers include, for example, benzyl ethyl ether, the aldehydes include, for example, the linear alkanals having 8 to 18 carbon atoms, citral, citronellal, citronellyloxyacetaldehyde, cyclamenaldehyde, hydroxycitronellal, lilial and bourgeonal, the ketones include, for example, the jonones and methylcedryl ketone, and the alcohols are anethole, citronellellone Eugenol, isoeugenol, geraniol, linalool, phenylethyl alcohol and terpineol, the hydrocarbons mainly include the terpenes and balsams. However, preference is given to using mixtures of different fragrances which together produce an appealing fragrance. Essential oils of lower volatility, which are mostly used as aroma components, are also suitable as perfume oils, for example sage oil, chamomile oil, clove oil, lemon balm oil, mint oil, cinnamon leaf oil, linden blossom oil, juniper berry oil, vetiver oil, oliban oil, galbanum oil, labdanum oil and lavandin oil. Preferably, bergamot oil, dihydromyrcenol, lilial, lyral, citronellol, phenylethyl alcohol, α-hexylcinnamaldehyde, geraniol, benzyl acetone, cyclamen aldehyde, linalool, Boisambrene Forte, Ambroxan, indole, hedione, Sandelice, lemon oil, mandarin oil, orange oil, allyl amyl glycolate, Cyclovertal, lavandin oil, muscatel Sage oil, ß-damascone, geranium oil bourbon, cyclohexyl salicylate, Vertofix Coeur, Iso-E-Super, Fixolide NP, evernyl, iraldein gamma, phenylacetic acid, geranyl acetate, benzyl acetate, rose oxide, romilate, irotyl and floramate alone or in mixtures, used. Antiperspirants (antiperspirants) reduce sweat formation by influencing the activity of the eccrine sweat glands and thus counteract armpit wetness and body odor. Aqueous or anhydrous formulations of antiperspirants typically contain the following ingredients:
(a) adstringierende Wirkstoffe,(a) astringent active ingredients,
(b) ÖlkömpOπerrteπ(b) ÖlkömpOπerrteπ
(c) nichtionische Emulgatoren,(c) nonionic emulsifiers,
(d) Coemulgatoren,(d) co-emulsifiers,
(e) Konsistenzgeber,(e) consistency generator,
(f) Hilfsstoffe wie z. B. Verdicker oder Komplexierungsmittel und/oder(f) auxiliaries such. B. thickeners or complexing agents and / or
(g) nichtwässrige Lösungsmittel wie z. B. Ethanol, Propylenglykol und/oder Glycerin.(g) non-aqueous solvents such as e.g. As ethanol, propylene glycol and / or glycerin.
Als adstringierende Antitranspirant-Wirkstoffe eignen sich vor allem Salze des Aluminiums, Zirkoniums oder des Zinks. Solche geeigneten antihydrotisch wirksamen Wirkstoffe sind z.B. Aluminiumchlorid, Aluminiumchlorhydrat, Alumini- umdichlorhydrat, Aluminiumsesquichlorhydrat und deren Komplexverbindungen z. B. mit Propylenglycol-1 ,2. Aluminiumhydroxyallantoinat, Aluminiumchlorid- tartrat, Aluminium-Zirkonium-Trichlorohydrat, Aluminium-Zirkonium-tetrachloro- hydrat, Aluminium-Zirkonium-pentachlorohydrat und deren Komplexverbindungen z. B. mit Aminosäuren wie Glycin.Salts of aluminum, zirconium or zinc are particularly suitable as astringent antiperspirant active ingredients. Such suitable antiperspirant active ingredients are e.g. Aluminum chloride, aluminum chlorohydrate, aluminum dichlorohydrate, aluminum sesquichlorohydrate and their complex compounds z. B. with propylene glycol-1, 2nd Aluminum hydroxyallantoinate, aluminum chloride tartrate, aluminum zirconium trichlorohydrate, aluminum zirconium tetrachlorohydrate, aluminum zirconium pentachlorohydrate and their complex compounds z. B. with amino acids such as glycine.
Daneben können in Antitranspirantien übliche öllösliche und wasserlösliche Hilfsmittel in geringeren Mengen enthalten sein. Solche öllöslichen Hilfsmittel können z.B. sein:In addition, customary oil-soluble and water-soluble auxiliaries can be present in smaller amounts in antiperspirants. Such oil soluble aids can e.g. his:
• entzündungshemmende, hautschützende oder wohlriechende ätherische Öle,• anti-inflammatory, skin-protecting or fragrant essential oils,
• synthetische hautschützende Wirkstoffe und/oder • öllösliche Parfümöle.• synthetic skin-protecting agents and / or • oil-soluble perfume oils.
Übliche wasserlösliche Zusätze sind z.B. Konservierungsmittel, wasserlösliche Duftstoffe, pH-Wert-Stellmittel, z.B. Puffergemische, wasserlösliche Verdickungsmittel, z.B. wasserlösliche natürliche oder synthetische Polymere wie z.B. Xanthan-Gum, Hydroxyethylcellulose, Polyvinylpyrrolidon oder hochmolekulare Polyethylenoxide.Common water-soluble additives are e.g. Preservatives, water-soluble fragrances, pH adjusters, e.g. Buffer mixtures, water soluble thickeners, e.g. water-soluble natural or synthetic polymers such as e.g. Xanthan gum, hydroxyethyl cellulose, polyvinyl pyrrolidone or high molecular weight polyethylene oxides.
Als Antischuppenmittel können Climbazol, Octopirox und Zinkpyrithion eingesetzt werden.Climbazole, octopirox and zinc pyrithione can be used as antidandruff agents.
Gebräuchliche Filmbildner sind beispielsweise Chitosan, mikrokristallines Chi- tosan, quaterniertes Chitosan, Polyvinylpyrrolidon, Vinylpyrrolidon-Vinylacetat- Copolymerisate, Polymere der Acrylsäurereihe, quatemäre Cellulose-Derivate, Kollagen, Hyaluronsäure bzw. deren Salze und ähnliche Verbindungen.Common film formers are, for example, chitosan, microcrystalline chitosan, quaternized chitosan, polyvinylpyrrolidone, vinylpyrrolidone-vinyl acetate copolymers, polymers of the acrylic acid series, quaternary cellulose derivatives, collagen, hyaluronic acid or its salts and similar compounds.
Als Quellmittel für wäßrige Phasen können Montmorillonite, Clay Mineralstoffe, Pemulen sowie alkylmodifizierte Carbopoltypen (Goodrich) dienen. Weitere geeignete Polymere bzw. Quellmittel können der Übersicht von R.Lochhead in Cosm.Toil. 108. 95 (1993) entnommen werden.Montmorillonites, clay minerals, pemulene and alkyl-modified carbopol types (Goodrich) can serve as swelling agents for aqueous phases. Further suitable polymers or swelling agents can be found in the overview by R. Lochhead in Cosm.Toil. 108, 95 (1993).
Unter UV-Lichtschutzfaktoren sind beispielsweise bei Raumtemperatur flüssig oder kristallin vorliegende organische Substanzen (Lichtschutzfilter) zu verstehen, die in der Lage sind, ultraviolette Strahlen zu absorbieren und die aufgenommene Energie in Form längerwelliger Strahlung, z.B. Wärme wieder abzugeben. UVB-Filter können öllöslich oder wasserlöslich sein. Als öllösliche Substanzen sind z.B. zu nennen:UV light protection factors are understood to mean, for example, organic substances (light protection filters) which are liquid or crystalline at room temperature and which are able to absorb ultraviolet rays and absorb the energy absorbed in the form of longer-wave radiation, e.g. To give off heat again. UVB filters can be oil-soluble or water-soluble. As oil-soluble substances e.g. to call:
• 3-Benzylidencampher bzw. 3-Benzylidennorcampher und dessen Derivate, z.B. 3-(4-Methylbenzyliden)campher wie in der EP 0693471 B1 beschrieben; • 4-Aminobenzoesäurederivate, vorzugsweise 4-Dimethylamino)benzoesäure- 2-ethylhexylester, 4-(Dimethylamino)benzoesäure-2-octylester und 4-(Di- methylamino)benzoesäureamylester;3-benzylidene camphor or 3-benzylidene norcampher and its derivatives, for example 3- (4-methylbenzylidene) camphor as described in EP 0693471 B1; • 4-aminobenzoic acid derivatives, preferably 2-ethylhexyl 4-dimethylamino) benzoate, 2-octyl 4- (dimethylamino) benzoate and amyl 4- (dimethylamino) benzoate;
• Ester der Zimtsäure, vorzugsweise 4-Methoxyzimtsäure-2-ethylhexylester, 4-Methoxyzimtsäurepropylester, 4-Methoxyzimtsäureisoamylester 2-Cyano- 3,3-phenylzimtsäure-2-ethylhexylester (Octocrylene);• esters of cinnamic acid, preferably 4-methoxycinnamic acid 2-ethylhexyl ester, 4-methoxycinnamic acid propyl ester, 4-methoxycinnamic acid isoamyl ester 2-cyano-3,3-phenylcinnamic acid 2-ethylhexyl ester (octocrylene);
• Ester der Salicylsäure, vorzugsweise Salicylsäure-2-ethylhexylester, Salicyl- säure-4-isopropylbenzylester, Salicylsäurehomomenthylester;• esters of salicylic acid, preferably salicylic acid 2-ethylhexyl ester, salicylic acid 4-isopropylbenzyl ester, salicylic acid homomethyl ester;
• Derivate des Benzophenons, vorzugsweise 2-Hydroxy-4-methoxybenzo- phenon, 2-Hydroxy-4-methoxy-4'-methylbenzophenon, 2,2'-Dihydroxy-4- methoxybenzophenon;• Derivatives of benzophenone, preferably 2-hydroxy-4-methoxybenzophenone, 2-hydroxy-4-methoxy-4'-methylbenzophenone, 2,2'-dihydroxy-4-methoxybenzophenone;
• Ester der Benzalmalonsäure, vorzugsweise 4-Methoxybenzmalonsäuredi-2- ethylhexylester;• Esters of benzalmalonic acid, preferably di-2-ethylhexyl 4-methoxybenzmalonate;
• Triazinderivate, wie z.B. 2,4,6-Trianilino-(p-carbo-2'-ethyl-1'-hexyloxy)-1 ,3,5- triazin und Octyl Triazon, wie in der EP 0818450 A1 beschrieben oder Di- octyl Butamido Triazone (Uvasorb® HEB);Triazine derivatives, e.g. 2,4,6-trianilino- (p-carbo-2'-ethyl-1'-hexyloxy) -1, 3,5-triazine and octyl triazone, as described in EP 0818450 A1 or di-octyl butamido triazone (Uvasorb ® HEB);
• Propan-1 ,3-dione, wie z.B. 1-(4-tert.Butylphenyl)-3-4'methoxyphenyl)propan- 1,3-dion;Propane-1,3-dione, e.g. 1- (4-tert-butylphenyl) -3-4'methoxyphenyl) propane-1,3-dione;
• Ketotricyclo(5.2.1.0)decan-Derivate, wie in der EP 0694521 B1 beschrieben.• Ketotricyclo (5.2.1.0) decane derivatives, as described in EP 0694521 B1.
Als wasserlösliche Substanzen kommen in Frage:Possible water-soluble substances are:
• 2-Phenylbenzimidazol-5-sulfonsäure und deren Alkali-, Erdalkali-, Ammonium-, Alkylammonium-, Alkanolammonium- und Glucammoniumsalze;• 2-phenylbenzimidazole-5-sulfonic acid and its alkali, alkaline earth, ammonium, alkylammonium, alkanolammonium and glucammonium salts;
• Sulfonsäurederivate von Benzophenonen, vorzugsweise 2-Hydroxy-4- methoxybenzophenon-5-sulfonsäure und ihre Salze;• Sulfonic acid derivatives of benzophenones, preferably 2-hydroxy-4-methoxybenzophenone-5-sulfonic acid and its salts;
• Sulfonsäurederivate des 3-Benzylidencamphers, wie z.B. 4-(2-Oxo-3- bomylidenmethyl)benzolsulfonsäure und 2-Methyl-5-(2-oxo-3-bornyliden)- sulfonsäure und deren Salze. Als typische UV-A-Filter kommen insbesondere Derivate des Benzoylmethans in Frage, wie beispielsweise 1-(4'-tert.Butylphenyl)-3-(4'-methoxyphenyl)pro- pan-1 ,3-dion, 4-tert.-Butyl-4'-methoxydibenzoylmethan (Parsol 1789), 1-Phenyl- 3-(4'-isopropylphenyl)-propan-1 ,3-dion sowie Enaminverbindungen, wie beschrieben in der DE 19712033 A1 (BASF). Die UV-A und UV-B-Filter können selbstverständlich auch in Mischungen eingesetzt werden. Neben den genannten löslichen Stoffen kommen für diesen Zweck auch unlösliche Lichtschutzpigmente, nämlich feindisperse Metalloxide bzw. Salze in Frage. Beispiele für geeignete Metalloxide sind insbesondere Zinkoxid und Titandioxid und daneben Oxide des Eisens, Zirkoniums, Siliciums, Mangans, Aluminiums und Cers sowie deren Gemische. Als Salze können Silicate (Talk), Bariumsulfat oder Zinkstea- rat eingesetzt werden. Die Oxide und Salze werden in Form der Pigmente für hautpflegende und hautschützende Emulsionen und dekorative Kosmetik verwendet. Die Partikel sollten dabei einen mittleren Durchmesser von weniger als 100 nm, vorzugsweise zwischen 5 und 50 nm und insbesondere zwischen 15 und 30 nm aufweisen. Sie können eine sphärische Form aufweisen, es können jedoch auch solche Partikel zum Einsatz kommen, die eine ellipsoide oder in sonstiger Weise von der sphärischen Gestalt abweichende Form besitzen. Die Pigmente können auch oberflächenbehandelt, d.h. hydrophilisiert oder hydrophobiert vorliegen. Typische Beispiele sind gecoatete Titandioxide, wie z.B. Titandioxid T 805 (Degussa) oder Eusolex® T2000 (Merck). Als hydrophobe Coatingmittel kommen dabei vor allem Silicone und dabei speziell Trialkoxy- octylsilane oder Simethicone in Frage. In Sonnenschutzmitteln werden bevorzugt sogenannte Mikro- oder Nanopigmente eingesetzt. Vorzugsweise wird mikronisiertes Zinkoxid verwendet. Weitere geeignete UV-Lichtschutzfilter sind der Übersicht von P.Finkel in SÖFW-Journal 122, 543 (1996) zu entnehmen.• Sulfonic acid derivatives of 3-benzylidene camphor, such as 4- (2-oxo-3-bomylidene methyl) benzenesulfonic acid and 2-methyl-5- (2-oxo-3-bornylidene) sulfonic acid and its salts. Derivatives of benzoylmethane, such as 1- (4'-tert-butylphenyl) -3- (4'-methoxyphenyl) propane-1,3-dione, 4-tert, are particularly suitable as typical UV-A filters. -Butyl-4'-methoxydibenzoylmethane (Parsol 1789), 1-phenyl-3- (4'-isopropylphenyl) -propane-1, 3-dione and enamine compounds as described in DE 19712033 A1 (BASF). The UV-A and UV-B filters can of course also be used in mixtures. In addition to the soluble substances mentioned, insoluble light protection pigments, namely finely dispersed metal oxides or salts, are also suitable for this purpose. Examples of suitable metal oxides are, in particular, zinc oxide and titanium dioxide and, in addition, oxides of iron, zirconium, silicon, manganese, aluminum and cerium and mixtures thereof. Silicates (talc), barium sulfate or zinc stearate can be used as salts. The oxides and salts are used in the form of the pigments for skin-care and skin-protecting emulsions and decorative cosmetics. The particles should have an average diameter of less than 100 nm, preferably between 5 and 50 nm and in particular between 15 and 30 nm. They can have a spherical shape, but it is also possible to use particles which have an ellipsoidal shape or a shape which differs from the spherical shape in some other way. The pigments can also be surface-treated, ie hydrophilized or hydrophobicized. Typical examples are coated titanium dioxides, such as titanium dioxide T 805 (Degussa) or Eusolex® T2000 (Merck). Silicones, and in particular trialkoxy-octylsilanes or simethicones, are particularly suitable as hydrophobic coating agents. So-called micro- or nanopigments are preferably used in sunscreens. Micronized zinc oxide is preferably used. Further suitable UV light protection filters can be found in the overview by P.Finkel in SÖFW-Journal 122, 543 (1996).
Neben den beiden vorgenannten Gruppen primärer Lichtschutzstoffe können auch sekundäre Lichtschutzmittel vom Typ der Antioxidantien eingesetzt werden, die die photochemische Reaktionskette unterbrechen, welche ausgelöst wird, wenn UV-Strahlung in die Haut eindringt. Typische Beispiele hierfür sind Aminosäuren (z.B. Glycin, Histidin, Tyrosin, Tryptophan) und deren Derivate, Imidazole (z.B. Urocaninsäure) und deren Derivate, Peptide wie D,L-Carnosin, D-Camosin, L-Carnosin und deren Derivate (z.B. Anserin), Chlorogensäure und deren Derivate, Liponsäure und deren Derivate (z.B. Dihydroliponsäure), Au- rothioglucose, Propylthiouracil und andere Thiole (z.B. Thioredoxin, Glutathion, Cystein, Cystin, Cystamin und deren Glycosyl-, N-Acetyl-, Methyl-, Ethyl-, Pro- pyl-, Amyl-, Butyl- und Lauryl-, Palmitoyl-, Oleyl-, γ-Linoleyl-, Cholesteryl- und Glycerylester) sowie deren Salze, Dilaurylthiodipropionat, Distearylthiodi- propionat, Thiodipropionsäure und deren Derivate (Ester, Ether, Peptide, Lipide, Nukleotide, Nukleoside und Salze) sowie Sulfoximinverbindungen (z.B. Buthio- ninsulfoximine, Homocysteinsulfoximin^ ButioninsulfoneL Penta^ Hexa-, Hep- tathioninsulfoximin) in sehr geringen verträglichen Dosierungen (z.B. pmol bis μmol/kg), ferner (Metall)-Chelatoren (z.B. α-Hydroxyfettsäuren, Palmitinsäure, Phytinsäure, Lactoferrin), α-Hydroxysäuren (z.B. Citronensäure, Milchsäure, Äpfelsäure), Huminsäure, Gallensäure, Gallenextrakte, Bilirubin, Biliverdin, EDTA, EGTA und deren Derivate, ungesättigte Fettsäuren und deren Derivate (z.B. γ-Linolensäure, Linolsäure, Ölsäure), Folsäure und deren Derivate, Ubi- chinon und Ubichinol und deren Derivate, Vitamin C und Derivate (z.B. Ascor- bylpalmitat, Mg-Ascorbylphosphat, Ascorbylacetat), Tocopherole und Derivate (z.B. Vitamin-E-acetat), Vitamin A und Derivate (Vitamin-A-palmitat) sowie Ko- niferylbenzoat des Benzoeharzes, Rutinsäure und deren Derivate, α- Glycosylrutin, Ferulasäure, Furfurylidenglucitol, Camosin, Butylhydroxytoluol, Butylhydroxyanisol, Nordihydroguajakharzsäure, Nordihydroguajaretsäure, Trihydroxybutyrophenon, Harnsäure und deren Derivate, Mannose und deren Derivate, Superoxid-Dismutase, Zink und dessen Derivate (z.B. ZnO, ZnSO4) Selen und dessen Derivate (z.B. Selen-Methionin), Stilbene und deren Derivate (z.B. Stilbenoxid, trans-Stilbenoxid) und die erfindungsgemäß geeigneten Derivate (Salze, Ester, Ether, Zucker, Nukleotide, Nukleoside, Peptide und Lipide) dieser genannten Wirkstoffe. Zur Verbesserung des Fließverhaltens können ferner Hydrotrope, wie beispielsweise Ethanol, Isopropylalkohol, oder Polyole eingesetzt werden. Polyole, die hier in Betracht kommen, besitzen vorzugsweise 2 bis 15 Kohlenstoffatome und mindestens zwei Hydroxylgruppen. Die Polyole können noch weitere funktioneile Gruppen, insbesondere Aminogruppen, enthalten bzw. mit Stickstoff modifiziert sein. Typische Beispiele sindIn addition to the two aforementioned groups of primary light stabilizers, secondary light stabilizers of the antioxidant type can also be used, which interrupt the photochemical reaction chain that triggers when UV radiation penetrates the skin. Typical examples are amino acids (e.g. glycine, histidine, tyrosine, tryptophan) and their derivatives, imidazoles (e.g. urocanic acid) and their derivatives, peptides such as D, L-carnosine, D-camosine, L-carnosine and their derivatives (e.g. anserine) , Chlorogenic acid and its derivatives, lipoic acid and its derivatives (e.g. dihydrolipoic acid), aurothioglucose, propylthiouracil and other thiols (e.g. thioredoxin, glutathione, cysteine, cystine and cystamine and their glycosyl, N-acetyl, methyl, ethyl, Propyl, amyl, butyl and lauryl, palmitoyl, oleyl, γ-linoleyl, cholesteryl and glyceryl esters) and their salts, dilauryl thiodipropionate, distearyl thiodipropionate, thiodipropionic acid and their derivatives (esters, ethers, peptides , Lipids, nucleotides, nucleosides and salts) as well as sulfoximine compounds (e.g. buthioninesulfoximine, homocysteine sulfoximine ^ Butioninsulfone L Penta ^ hexa-, heptathionine sulfoximine) in very low tolerable dosages (e.g. pmol to μmol / kg), also (metal) chelato ren (e.g. α-hydroxy fatty acids, palmitic acid, phytic acid, lactoferrin), α-hydroxy acids (e.g. citric acid, lactic acid, malic acid), humic acid, bile acid, bile extracts, bilirubin, biliverdin, EDTA, EGTA and their derivatives, unsaturated fatty acids and their derivatives (e.g. γ-linolenic acid, linoleic acid, oleic acid), folic acid and its derivatives, ubiquinone and ubiquinol and their derivatives, vitamin C and derivatives (e.g. ascorbyl palmitate, Mg ascorbyl phosphate, ascorbyl acetate), tocopherols and derivatives (e.g. vitamin E acetate) ), Vitamin A and derivatives (vitamin A palmitate) as well as coniferyl benzoate of benzoin, rutinic acid and its derivatives, α-glycosyl rutin, ferulic acid, furfurylidene glucitol, camosin, butyl hydroxytoluene, butylated hydroxyanisole, nordihydroguajakh resin acid, nordihydrophihydroxybenzate, trihydro acid, Mannose and its derivatives, superoxide dismutase, zinc and its derivatives (e.g. ZnO, ZnSO 4 ) selenium and its derivatives (e.g. Selen-M ethionine), stilbenes and their derivatives (for example stilbene oxide, trans-stilbene oxide) and the derivatives suitable according to the invention (salts, esters, ethers, sugars, nucleotides, nucleosides, peptides and lipids) of these active substances. Hydrotropes such as ethanol, isopropyl alcohol or polyols can also be used to improve the flow behavior. Polyols that come into consideration here preferably have 2 to 15 carbon atoms and at least two hydroxyl groups. The polyols can also contain further functional groups, in particular amino groups, or be modified with nitrogen. Typical examples are
• Glycerin;• glycerin;
• Alkylenglycole, wie beispielsweise Ethylenglycol, Diethylenglycol, Propy- lenglycol, Butylenglycol, Hexylenglycol sowie Polyethylenglycole mit einem durchschnittlichen Molekulargewicht von 100 bis 1.000 Dalton;Alkylene glycols, such as, for example, ethylene glycol, diethylene glycol, propylene glycol, butylene glycol, hexylene glycol and polyethylene glycols with an average molecular weight of 100 to 1,000 daltons;
• technische Oligoglyceringemische mit einem Eigenkondensationsgrad von 1 ,5 bis 10 wie etwa technische Diglyceringemische mit einem Diglyceringe- halt von 40 bis 50 Gew.-%;Technical oligoglycerol mixtures with a degree of self-condensation of 1.5 to 10, such as technical diglycerol mixtures with a diglycerol content of 40 to 50% by weight;
• Methyolverbindungen, wie insbesondere Trimethylolethan, Trimethylolpropan, Trimethylolbutan, Pentaerythrit und Dipentaerythrit;• Methyl compounds, such as in particular trimethylolethane, trimethylolpropane, trimethylolbutane, pentaerythritol and dipentaerythritol;
• Niedrigalkylglucos.de, insbesondere solche mit 1 bis 8 Kohlenstoffen im Alkylrest, wie beispielsweise Methyl- und Butylglucosid;• Lower alkylglucos.de, especially those with 1 to 8 carbons in the alkyl radical, such as methyl and butyl glucoside;
• Zuckeralkohole mit 5 bis 12 Kohlenstoffatomen, wie beispielsweise Sorbit oder Mannit,Sugar alcohols with 5 to 12 carbon atoms, such as sorbitol or mannitol,
• Zucker mit 5 bis 12 Kohlenstoffatomen, wie beispielsweise Glucose oder Saccharose;• Sugar with 5 to 12 carbon atoms, such as glucose or sucrose;
• Aminozucker, wie beispielsweise Glucamin;Aminosugars, such as glucamine;
• Dialkoholamine, wie Diethanolamin oder 2-Amino-1 ,3-propandiol.• Dialcohol amines, such as diethanolamine or 2-amino-1, 3-propanediol.
Als Konservierungsmittel eignen sich beispielsweise Phenoxyethanol, Formal- dehydlösung, Parabene, Pentandiol oder Sorbinsäure sowie die in Anlage 6, Teil A und B der Kosmetikverordnung aufgeführten weiteren Stoffklassen. Als Insekten-Repellentien kommen N,N-Diethyl-m-toluamid, 1 ,2-Pentandiol oder Ethyl Butylacetylaminopropionate in Frage, als Selbstbräuner eignet sich Di- hydroxyaceton.Suitable preservatives are, for example, phenoxyethanol, formaldehyde solution, parabens, pentanediol or sorbic acid and the other classes of substances listed in Appendix 6, Parts A and B of the Cosmetics Ordinance. As insect repellents come N, N-diethyl-m-toluamide, 1, 2-pentanediol or Ethyl butylacetylaminopropionate in question, di-hydroxyacetone is suitable as a self-tanner.
Als Parfümöle seien genannt Gemische aus natürlichen und synthetischen Riechstoffen. Natürliche Riechstoffe sind Extrakte von Blüten (Lilie, Lavendel, Rosen, Jasmin, Neroli, Ylang-Ylang), Stengeln und Blättern (Geranium, Pat- chouli, Petitgrain), Früchten (Anis, Koriander, Kümmel, Wacholder), Frucht- schaleττ_(BeτgramOtt ! Zitrσne "Ora"ngen Wατzeln~(Macis, Angelica; Sellerie, Kardamon, Costus, Iris, Calmus), Hölzern (Pinien-, Sandel-, Guajak-, Zedern-, Rosenholz), Kräutern und Gräsern (Estragon, Lemongras, Salbei, Thymian), Nadeln und Zweigen (Fichte, Tanne, Kiefer, Latschen), Harzen und Balsamen (Galbanum, Elemi, Benzoe, Myrrhe, Olibanum, Opoponax). Weiterhin kommen tierische Rohstoffe in Frage, wie beispielsweise Zibet und Castoreum. Typische synthetische Riechstoffverbindungen sind Produkte vom Typ der Ester, Ether, Aldehyde, Ketone, Alkohole und Kohlenwasserstoffe. Riechstoffverbindungen vom Typ der Ester sind z.B. Benzylacetat, Phenoxyethylisobutyrat, p-tert.-Bu- tyleyclohexylacetat, Linalylacetat, Dimethylbenzylcarbinylacetat, Phenylethyla- cetat, Linalylbenzoat, Benzylformiat, Ethylmethylphenylglycinat, Allylcyclohe- xylpropionat, Styrallylpropionat und Benzylsalicylat. Zu den Ethern zählen beispielsweise Benzylethylether, zu den Aldehyden z.B. die linearen Alkanale mit 8 bis 18 Kohlenstoffatomen, Citral, Citronellal, Citronellyloxyacetaldehyd, Cycla- menaldehyd, Hydroxycitronellal, Lilial und Bourgeonal, zu den Ketonen z.B. die Jonone, oc-lsomethylionon und Methylcedrylketon, zu den Alkoholen Anethol, Citronellol, Eugenol, Isoeugenol, Geraniol, Linalool, Phenylethylalkohol und Terpineol, zu den Kohlenwasserstoffen gehören hauptsächlich die Terpene und Balsame. Bevorzugt werden jedoch Mischungen verschiedener Riechstoffe verwendet, die gemeinsam eine ansprechende Duftnote erzeugen. Auch ätherische Öle geringerer Flüchtigkeit, die meist als Aromakomponenten verwendet werden, eignen sich als Parfümöle, z.B. Salbeiöl, Kamillenöl, Nelkenöl, Melis- senöl, Minzenöl, Zimtblätteröl, Lindenblütenöl, Wacholderbeerenöl, Vetiveröl, Olibanöl, Galbanumöl, Labolanumöl und Lavandinöl. Vorzugsweise werden Bergamotteöl, Dihydromyrcenol, Lilial, Lyral, Citronellol, Phenylethylalkohol, α- Hexylzimtaldehyd, Geraniol, Benzylaceton, Cyclamenaldehyd, Linalool, Boi- sambrene Forte, Ambroxan, Indol, Hedione, Sandelice, Citronenöl, Mandari- nenöl, Orangenöl, Allylamylglycolat, Cyclovertal, Lavandinöl, Muskateller Salbeiöl, ß-Damascone, Geraniumöl Bourbon, Cyclohexylsalicylat, Vertofix Coeur, Iso-E-Super, Fixolide NP, Evernyl, Iraldein gamma, Phenylessigsäure, Gerany- lacetat, Benzylacetat, Rosenoxid, Romilllat, Irotyl und Floramat allein oder in Mischungen, eingesetzt.Perfume oils include mixtures of natural and synthetic fragrances. Natural fragrances are extracts from flowers (lily, lavender, rose, jasmine, neroli, ylang-ylang), stems and leaves (geranium, Patterson chouli, petitgrain), fruits (anise, coriander, caraway, juniper), fruit schaleττ _ (BeτgramOtt! Citrσne " Ora " ningen Wατzeln ~ (Macis, Angelica; Celery, Cardamom, Costus, Iris, Calmus), woods (pine, sandal, guaiac, cedar, rosewood), herbs and grasses (tarragon, lemongrass , Sage, thyme), needles and twigs (spruce, fir, pine, mountain pine), resins and balsams (galbanum, elemi, benzoin, myrrh, olibanum, opoponax). Animal raw materials, such as civet and castoreum, are also suitable Synthetic fragrance compounds are products of the ester, ether, aldehyde, ketone, alcohol and hydrocarbon type, and fragrance compounds of the ester type are, for example, benzyl acetate, phenoxyethyl isobutyrate, p-tert-butyl-cyclohexyl acetate, linalyl acetate, dimethylbenzylcarbinylacetate, phenylethyl acetate at, linalyl benzoate, benzyl formate, ethyl methylphenyl glycinate, allyl cyclohexyl propionate, styrallyl propionate and benzyl salicylate. The ethers include, for example, benzyl ethyl ether, the aldehydes include, for example, the linear alkanals with 8 to 18 carbon atoms, citral, citronellal, citronellyloxyacetaldehyde, cyclamaldehyde, hydroxycitronellal, lilial and bourgeonal, the ketones include, for example, the jonones, oc-isomethylionone, and methylcedryl ketone the alcohols anethole, citronellol, eugenol, isoeugenol, geraniol, linalool, phenylethyl alcohol and terpineol, the hydrocarbons mainly include the terpenes and balsams. However, preference is given to using mixtures of different fragrances which together produce an appealing fragrance. Essential oils of lower volatility, which are mostly used as aroma components, are also suitable as perfume oils, for example sage oil, chamomile oil, clove oil, melissa oil, mint oil, cinnamon leaf oil, linden blossom oil, juniper berry oil, vetiver oil, Oliban oil, galbanum oil, labolanum oil and lavandin oil. Bergamot oil, dihydromyrcenol, lilial, lyral, citronellol, phenylethyl alcohol, α-hexylcinnamaldehyde, geraniol, benzylacetone, cyclamenaldehyde, linalool, boi sambrene forte, ambroxan, indole, hedione, sandelice, lemon oil, mandarinamine, allyl glycol, orange oil, orange oil, are preferred , Lavandin oil, muscatel sage oil, ß-damascone, geranium oil bourbon, cyclohexyl salicylate, Vertofix Coeur, Iso-E-Super, Fixolide NP, evernyl, iraldein gamma, phenylacetic acid, geranyl acetate, benzyl acetate, rose oxide, romilllate, irotyl and floramate alone Mixtures used.
Als Farbstoffe können die für kosmetische Zwecke geeigneten und zugelassenen Substanzen verwendet werden, wie sie beispielsweise in der Publikation "Kosmetische Färbemittel" der Farbstoffkommission der Deutschen Forschungsgemeinschaft, Verlag Chemie, Weinheim, 1984, S.81-106 zusammengestellt sind. Diese Farbstoffe werden üblicherweise in Konzentrationen von 0,001 bis 0,1 Gew.-%, bezogen auf die gesamte Mischung, eingesetzt.The dyes which can be used are those substances which are suitable and approved for cosmetic purposes, as compiled, for example, in the publication "Cosmetic Dyes" by the Dye Commission of the German Research Foundation, Verlag Chemie, Weinheim, 1984, pp. 81-106. These dyes are usually used in concentrations of 0.001 to 0.1% by weight, based on the mixture as a whole.
Der Gesamtanteil der Hilfs- und Zusatzstoffe kann 1 bis 50, vorzugsweise 5 bis 40 Gew.-% - bezogen auf die Mittel - betragen. Die Herstellung der Kosmetika und Körperpflegemittel kann durch übliche Kalt - oder Heißprozesse erfolgen; vorzugsweise arbeitet man nach der Phaseninversionstemperatur-Methode. The total proportion of auxiliaries and additives can be 1 to 50, preferably 5 to 40,% by weight, based on the composition. The cosmetics and personal care products can be produced by customary cold or hot processes; the phase inversion temperature method is preferably used.

Claims

Patentansprüche: claims:
1. Verfahren zur Bestimmung des Hautstreß und/oder der Hautalterung bei Menschen oder Tieren in vitro, dadurch gekennzeichnet, daß man a) eine Fibroblasten enthaltende Hautprobe gewinnt, b) die Fibroblasten isoliert und kultiviert und c) den Kulturüberstand auf das Vorhandensein und die Menge der von deπ~Fibroblasten sezernierten Proteine oder Proteinfragmente Spondin 2, Cathepsin L, Vimentin-Fragmente oder Actin Gamma 1 untersucht.1. A method for determining skin stress and / or skin aging in humans or animals in vitro, characterized in that a) a skin sample containing fibroblasts is obtained, b) the fibroblasts are isolated and cultured, and c) the culture supernatant for the presence and the amount secreted by fibroblasts deπ ~ proteins or protein fragments spondin 2, cathepsin L, vimentin fragments or actin gamma examined. 1
2. Verfahren zur Bestimmung des Hautstreß und/oder der Hautalterung bei Menschen oder Tieren in vitro, dadurch gekennzeichnet, daß man a) mittels Mikrodialyse ein Gemisch der von den Hautzellen sezemierten Proteine gewinnt und b) das Gemisch auf das Vorhandensein und die Menge der von den Fibroblasten sezemierten Proteine oder Proteinfragmente Spondin 2, Cathepsin L, Vimentin-Fragmente oder Actin Gamma 1 untersucht.2. A method for determining skin stress and / or skin aging in humans or animals in vitro, characterized in that a) a mixture of the proteins secreted by the skin cells is obtained by microdialysis and b) the mixture on the presence and the amount of the fibroblasts secreted proteins or protein fragments Spondin 2, Cathepsin L, Vimentin fragments or Actin Gamma 1 were examined.
3. Verfahren nach Anspruch 1 oder 2, dadurch gekennzeichnet, daß man die Untersuchung in Schritt c) gemäß Anspruch 1 bzw. in Schritt b) gemäß Anspruch 2 durch Separation der Proteine mittels Gelelektrophorese und nachfolgende Massenspektrometrie, insbesondere Matrix Assistierter Laser Desorpti- ons Ionisation (MALDI), durchführt.3. The method according to claim 1 or 2, characterized in that the investigation in step c) according to claim 1 or in step b) according to claim 2 by separation of the proteins by means of gel electrophoresis and subsequent mass spectrometry, in particular matrix-assisted laser desorption ionization (MALDI).
4. Verfahren nach einem der Ansprüche 1 bis 3, dadurch gekennzeichnet, daß man in Schritt c) gemäß Anspruch 1 bzw. in Schritt b) gemäß Anspruch 2 auf das Vorhandensein und die Menge von Spondin 2 oder einem Fragment davon untersucht, das durch folgende Aminosäuresequenz (A) gekennzeichnet ist: N-Term- MENPSPAAAL GKALCALLLA TLGAAGQPLG GESICSARAP AKYSITFTGK WSQTAFPKQY PLFRPPAQWS SLLGAAHSSD YSMWRKNQYV SNGLRDFAER GEAWALMKEI EAAGEALQSV HAVFSAPAVP SGTGQTSAEL EVQRRHSLVS FWRIVPSPD WFVGVDSLDL CDGDRWREQA ALDLYPYDAG TDSGFTFSSP NFATIPQDTV TEITSSSPSH PANSFYYPRL KALPPIARVT LVRLRQSPRA FIPPAPVLPS RDNEIVDSAS VPETPLDCEV SLWSSWGLCG GHCGRLGTKS RTRYVRVQPA NNGSPCPELE EEAECVPDNC V -C-Term4. The method according to any one of claims 1 to 3, characterized in that one examines in step c) according to claim 1 or in step b) according to claim 2 for the presence and the amount of spondin 2 or a fragment thereof, by the following Amino acid sequence (A) is labeled: N-Term MENPSPAAAL GKALCALLLA TLGAAGQPLG GESICSARAP AKYSITFTGK WSQTAFPKQY PLFRPPAQWS SLLGAAHSSD YSMWRKNQYV SNGLRDFAER GEAWALMKEI EAAGEALQSV HAVFSAPAVP SGTGQTSAEL EVQRRHSLVS FWRIVPSPD WFVGVDSLDL CDGDRWREQA ALDLYPYDAG TDSGFTFSSP NFATIPQDTV TEITSSSPSH PANSFYYPRL KALPPIARVT LVRLRQSPRA FIPPAPVLPS RDNEIVDSAS VPETPLDCEV SLWSSWGLCG GHCGRLGTKS RTRYVRVQPA NNGSPCPELE EEAECVPDNC V -C-term
5. Verfahren nach einem der Ansprüche 1 bis 3, dadurch gekennzeichnet, daß man in Schritt c) gemäß Anspruch 1 bzw. in Schritt b) gemäß Anspruch 2 auf das Vorhandensein und die Menge eines Vimentin-Fragmentes untersucht, aas durch folgende Aminosäuresequenz (C) gekennzeichnet ist:5. The method according to any one of claims 1 to 3, characterized in that one examines in step c) according to claim 1 or in step b) according to claim 2 for the presence and the amount of a vimentin fragment, aas by the following amino acid sequence (C ) is marked:
N-Term- MSTRSVSSSS YRRMFGGPGT ASRPSSSRSY VTTSTRTYSL GSALRPSTSR SLYASSPGGV YATRSSAVRL RSSVPGVRLL QDSVDFSLAD AINTEFKNTR TNEKVELQEL NDRFANYIDK VRFLEQQNKI LLAELEQLKG QGKSRLGDLY EEEMRELRRQ VDQLTNDKAR VEVERDNLAE DIMRLREKLQ EEMLQREEAE NTLQSFRQDV DNASLARLDL ERKVESLQEE IAFLKKLHEE EIQELQAQIQ EQHVQIDVDV SKPDLTAALR DVRQQYESVA AKNLQEAEEW YKSKFADLSE AANRNNDALR QAKQESTEYR RQVQSLTCEV DALKGTNESL ERQMREMEEN FAVEAANYQD TIGRLQDEIQ NMKEEMARHR EYQDLLNVKM ALDIEIATYR KLLEGEESRI SLPLPNFSSL NLRETNLDSL PLVDTHSKRT LLIKTVETRD GQVINETSQH HDDLE -C-TermN-Term MSTRSVSSSS YRRMFGGPGT ASRPSSSRSY VTTSTRTYSL GSALRPSTSR SLYASSPGGV YATRSSAVRL RSSVPGVRLL QDSVDFSLAD AINTEFKNTR TNEKVELQEL NDRFANYIDK VRFLEQQNKI LLAELEQLKG QGKSRLGDLY EEEMRELRRQ VDQLTNDKAR VEVERDNLAE DIMRLREKLQ EEMLQREEAE NTLQSFRQDV DNASLARLDL ERKVESLQEE IAFLKKLHEE EIQELQAQIQ EQHVQIDVDV SKPDLTAALR DVRQQYESVA AKNLQEAEEW YKSKFADLSE AANRNNDALR QAKQESTEYR RQVQSLTCEV DALKGTNESL ERQMREMEEN FAVEAANYQD TIGRLQDEIQ NMKEEMARHR EYQDLLNVKM ALDIEIATYR KLLEGEESRI SLPLPNFSSL NLRETNLDSL PLVDTHSKRT LLIKTVETRD GQVINETSQH HDDLE -C term
6. Verfahren nach einem der Ansprüche 1 bis 3, dadurch gekennzeichnet, daß man in Schritt c) gemäß Anspruch 1 bzw. in Schritt b) gemäß Anspruch 2 auf das Vorhandensein und die Menge von Actin gamma 1 oder einem Fragment davon untersucht, das durch folgende Aminosäuresequenz (D) gekennzeichnet ist: N-Term- MEEEIAALVI DNGSGMCKAG FAGDDAPRAV FPSIVGRPRH QGVMVGMGQK DSYVGDEAQS KRGILTLKYP IEHGIVTNWD DMEKIWHHTF YNELRVAPEE HPVLLTEAPL NPKANREKMT QIMFETFNTP AMYVAIQAVL SLYASGRTTG IVMDSGDGVT HTVPIYEGYA LPHAILRLDL AGRDLTDYLM KILTERGYSF TTTAEREIVR DIKEKLCYVA LDFEQEMATA ASSSSLEKSY ELPDGQVITI GNERFRCPEA LFQPSFLGME SCGIHETTFN SIMKCDVDIR KDLYANTVLS GGTTMYPGIA DRMQKEITAL APSTMKIKII APPERKYSVW IGGSILASrS FαQMWISKQ-EYDESGPSIVΗRKCF^C-Term6. The method according to any one of claims 1 to 3, characterized in that one examines in step c) according to claim 1 or in step b) according to claim 2 for the presence and the amount of actin gamma 1 or a fragment thereof, which by the following amino acid sequence (D) is labeled: N-Term MEEEIAALVI DNGSGMCKAG FAGDDAPRAV FPSIVGRPRH QGVMVGMGQK DSYVGDEAQS KRGILTLKYP IEHGIVTNWD DMEKIWHHTF YNELRVAPEE HPVLLTEAPL NPKANREKMT QIMFETFNTP AMYVAIQAVL SLYASGRTTG IVMDSGDGVT HTVPIYEGYA LPHAILRLDL AGRDLTDYLM KILTERGYSF TTTAEREIVR DIKEKLCYVA LDFEQEMATA ASSSSLEKSY ELPDGQVITI GNERFRCPEA LFQPSFLGME SCGIHETTFN SIMKCDVDIR KDLYANTVLS GGTTMYPGIA DRMQKEITAL APSTMKIKII APPERKYSVW IGGSILASrS FαQMWISKQ-EYDESGPSIVΗRKCF ^ C-Term
7. Verfahren nach einem der Ansprüche 1 bis 3, dadurch gekennzeichnet, daß man in Schritt c) gemäß Anspruch 1 bzw. in Schritt b) gemäß Anspruch 2 auf das Vorhandensein und die Menge, von Cathepsin L oder einem Fragment davon untersucht, das durch folgende Aminosäuresequenz (B) gekennzeichnet ist:7. The method according to any one of claims 1 to 3, characterized in that in step c) according to claim 1 or in step b) according to claim 2 for the presence and the amount of cathepsin L or a fragment thereof examined by the following amino acid sequence (B) is labeled:
N-Term- MNPTLILAAF CLGIASATLT FDHSLEAQWT KWKAMHNRLY GMNEEGWRRA VWEKNMKMIE LHNQEYREGK HSFTMAMNAF GDMTSEEFRQ VMNGFQNRKP RKGKVFQEPL FYEAPRSVDW REKGYVTPVK NQGQCGSCWA FSATGALEGQ MFRKTGRLIS LSEQNLVDCS GPQGNEGCNG GLMDYAFQYV QDNGGLDSEE SYPYEATEES CKYNPKYSVA NDTGFVDIPK QEKALMKAVA TVGPISVAID AGHESFLFYK EGIYFEPDCS SEDMDHGVLV VGYGFESTES DNNKYWLVKN SWGEEWGMGG YVKMAKDRRN HCGIASAASY PTV -C-TermN-Term MNPTLILAAF CLGIASATLT FDHSLEAQWT KWKAMHNRLY GMNEEGWRRA VWEKNMKMIE LHNQEYREGK HSFTMAMNAF GDMTSEEFRQ VMNGFQNRKP RKGKVFQEPL FYEAPRSVDW REKGYVTPVK NQGQCGSCWA FSATGALEGQ MFRKTGRLIS LSEQNLVDCS GPQGNEGCNG GLMDYAFQYV QDNGGLDSEE SYPYEATEES CKYNPKYSVA NDTGFVDIPK QEKALMKAVA TVGPISVAID AGHESFLFYK EGIYFEPDCS SEDMDHGVLV VGYGFESTES DNNKYWLVKN SWGEEWGMGG YVKMAKDRRN HCGIASAASY PTV -C-term
8. Test-Kit zur Bestimmung des Hautstreß und/oder der Hautalterung bei Menschen oder Tieren in vitro, umfassend Mittel zur Durchführung der Verfahren nach einem der Ansprüche 1 bis 7. 8. Test kit for determining skin stress and / or skin aging in humans or animals in vitro, comprising means for carrying out the method according to one of claims 1 to 7.
9. Verwendung von Spondin 2, Cathepsin L, Vimentin-Fragmenten oder Actin Gamma 1 als Streß- und/oder Alterungsmarker der Haut bei Menschen oder Tieren.9. Use of spondin 2, cathepsin L, vimentin fragments or actin gamma 1 as a stress and / or aging marker of the skin in humans or animals.
10. Verfahren zum Nachweis der Wirksamkeit von kosmetischen oder pharmazeutischen Wirkstoffen gegen Hautstreß und/oder Hautalterung in vitro, dadurch gekennzeichnet, daß man a) den Hautstatus durch ein Verfahren nach einem der Ansprüche 1 bis 7 bestimmt, b) einen Wirkstoff gegen Hautstreß und/oder Hautalterung einmal oder mehrmals auf die Haut aufbringt, c) erneut den Hautstatus durch ein Verfahren nach einem der Ansprüche 1 bis 7 bestimmt und d) die Wirksamkeit des Wirkstoffs durch den Vergleich der Ergebnisse aus a) und c) bestimmt.10. A method for proving the effectiveness of cosmetic or pharmaceutical active substances against skin stress and / or skin aging in vitro, characterized in that a) the skin status is determined by a method according to one of claims 1 to 7, b) an active substance against skin stress and / or applies skin aging to the skin one or more times, c) the skin status is determined again by a method according to one of claims 1 to 7 and d) the effectiveness of the active ingredient is determined by comparing the results from a) and c).
11. Test-Kit zum Nachweis der Wirksamkeit von kosmetischen oder pharmazeutischen Wirkstoffen gegen Hautstreß und/oder Hautalterung in vitro, umfassend Mittel zur Durchführung des Verfahrens nach Anspruch 10.11. Test kit for demonstrating the effectiveness of cosmetic or pharmaceutical active substances against skin stress and / or skin aging in vitro, comprising means for carrying out the method according to claim 10.
12. Verwendung von Spondin 2, Cathepsin L, Vimentin-Fragmenten oder Actin Gamma 1 zum Nachweis der Wirksamkeit von kosmetischen oder pharmazeutischen Wirkstoffen gegen Hautstreß und/oder Hautalterung.12. Use of Spondin 2, Cathepsin L, Vimentin fragments or Actin Gamma 1 to demonstrate the effectiveness of cosmetic or pharmaceutical active ingredients against skin stress and / or skin aging.
13. Kosmetisches oder pharmazeutisches Mittel zur Regulierung, insbesondere zur Aufrechterhaltung der Homeostase menschlicher oder tierischer Haut, enthaltend Vimentin-Fragmente.13. Cosmetic or pharmaceutical agent for regulation, in particular for maintaining the homeostasis of human or animal skin, containing vimentin fragments.
14. Handwaschmittel, Handgeschirrspülmittel oder Körperpflegemittel, enthaltend Vimentin-Fragmente. 14. Hand wash, hand dishwashing detergent or personal care product containing vimentin fragments.
15. Verwendung von Vimentin-Fragmenten in Mitteln zur Regulierung, insbesondere zur Aufrechterhaltung der Homeostase menschlicher oder tierischer Haut.15. Use of vimentin fragments in agents for regulating, in particular for maintaining the homeostasis of human or animal skin.
16. Verwendung von Vimentin-Fragmenten zur Herstellung von Mitteln zur Regulierung, insbesondere zur Aufrechterhaltung der Homeostase menschlicher odef ierischer Haut". 16. Use of vimentin fragments for the production of regulating agents, in particular for maintaining the homeostasis of human or animal skin " .
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WO2002053774A2 (en) * 2001-01-03 2002-07-11 Henkel Kommanditgesellschaft Auf Aktien Method for determining homeostasis of the skin
WO2002053774A3 (en) * 2001-01-03 2003-07-17 Henkel Kgaa Method for determining homeostasis of the skin
WO2004005542A2 (en) * 2002-07-03 2004-01-15 Henkel Kommanditgesellschaft Auf Aktien Method for identifying infection-specific regulated genes of the skin
WO2004005542A3 (en) * 2002-07-03 2004-05-13 Henkel Kgaa Method for identifying infection-specific regulated genes of the skin
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FR2923610A1 (en) * 2007-11-14 2009-05-15 Galderma Res & Dev NON-INVASIVE METHOD OF COLLECTING BIOLOGICAL DATA FOR THE ESTABLISHMENT OF DIAGNOSIS OF SKIN PATHOLOGY.
WO2009068825A1 (en) * 2007-11-14 2009-06-04 Galderma Research & Development Non-invasive method for collecting biological data for establishing a diagnosis of a skin pathology
JP2015517995A (en) * 2012-04-02 2015-06-25 モデルナ セラピューティクス インコーポレイテッドＭｏｄｅｒｎａ Ｔｈｅｒａｐｅｕｔｉｃｓ，Ｉｎｃ． Modified polynucleotides for the production of cosmetic proteins and peptides
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US9878056B2 (en) 2012-04-02 2018-01-30 Modernatx, Inc. Modified polynucleotides for the production of cosmetic proteins and peptides

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