DE102004025805A1 - Cosmetic compound for incorporation in pharmaceutical products for treatment of e.g. human skin, hair and nails - Google Patents

Abstract

A cosmetic compound for the treatment of materials incorporating keratin incorporates a polypeptide sequence (i) in a cosmetic-compatible medium. The polypeptide sequence (i) has a binding affinity to human hair, nails and skin keratin. The polypeptide sequences are: SEQ ID NO: (a) 1 Positions 2193-2482; 2606-2871; a polypeptide sequence in which up to 60% of the amino-acids are changed with respect to (a); a polypeptide sequence in which up to 50% of the amino-acids are changed with respect to (b). An independent claim is also included for synthesis of the polypeptide sequence.

Description

vertebrate contain filaments of which a group composed of keratins is. These keratins, which also occur in hair, skin and nails, bind specific proteins such as desmoplakin by means of a special Sequenzmotifs, a sg. Keratin-binding domain (Fontao L, Favre B, Riou S, Geerts D, Jaunin F, Saurat JH, Green KJ, Sonnenberg A, Borradori L., Interaction of the bullous pemphigoid antigen 1 (BP230) and desmoplakin with intermediate filaments is mediated by distinct sequences in their COOH terminus., Mol Biol Cell. 2003 May; 14 (5): 1978-92. Epub 2003 Jan 26; Hopkinson SB, Jones JC., The N terminus of the transmembrane protein BP180 interacts with the N-terminal domain of BP230, which is mediating keratin cytoskeleton anchorage to the cell surface at the site of the hemidesmosome, Mol Biol Cell. 2000 Jan; 11 (1): 277-86).

task

task The object of the present invention was to provide novel keratin-binding effector molecules which a binding peptide with high affinity have keratin or keratin-containing substances such as skin or hair, and to which certain effector molecules are bound. Such keratin-binding effector allow a high local concentration of effector molecules on the keratin, i.e. a so-called "targeting or a long duration of action on the keratin.

description the invention

• (i) mindestens einer Polypeptidsequenz, die eine Bindungsaffinität zu einem Keratin besitzt,
• (ii) einem Effektormolekül, das natürlicherweise nicht mit der Polypeptidsequenz (i) verknüpft ist.

The invention relates to keratin-binding effector molecules consisting of

• (i) at least one polypeptide sequence having a binding affinity to a keratin,
• (ii) an effector molecule that is not naturally linked to the polypeptide sequence (i).

Polypeptide sequences (i)

The Polypeptide sequence (i) has a binding affinity for one Keratin. The binding of the polypeptide sequence (i) to a keratin can be tested under the conditions described in Examples 1 and 2 become.

Especially well-suited keratin-binding polypeptides are such sequences, that are contained in the human desmoplakin or by alteration human desmoplakin polypeptide sequences such as amino acid insertions, Substitutions or deletions derived therefrom.

The Polypeptide sequence of the human desmoplakin is shown in SEQ ID NO: 1. A suitable keratin-binding domain (domain B) is the polypeptide sequence SEQ ID NO: 1 position 2269 to 2508 and their functional equivalents. Another keratin-binding domain (domain C) is the polypeptide sequence SEQ ID NO: 1 position 2606 to 2871 and their functional equivalents.

The keratin-binding domains are in 1 shown.

Preferred polypeptide sequences (i) comprise an amino acid sequence according to SEQ ID NO: 1, in particular the positions:
Domain B: polypeptide sequence SEQ ID NO: 1 position 2315 to 2450
Domain C: polypeptide sequence SEQ ID NO: 1 position 2645 to 2661
Domain C: polypeptide sequence SEQ ID NO: 1 position 2774 to 2788
Domain C: Polypeptide Sequence SEQ ID NO: 1 Position 2844 to 2854

Includes according to the invention are also "functional equivalents" of the specifically disclosed Polypeptide sequences (i) and the use of these in the inventive method.

"Functional equivalents" or analogues of the specifically disclosed polypeptides are, in the context of the present invention, different polypeptides, which furthermore have the desired biological activity, such as keratin binding, for example, by "functional equivalents" polypep tide sequences which show at least 10% binding according to one of the binding tests described in Example 1 or 2, preferably at least 50%, particularly preferably 75%, very particularly preferably 90%, of the binding comprising a polypeptide with domain B or domain C of SEQ ID NO: 1 in the binding test according to Example 1 or 2.

"Functional equivalents" are understood according to the invention in particular also muteins, which in at least one sequence position of the above mentioned amino acid sequences but have a different than the specified amino acid but still possess one of the above biological activities. "Functional equivalents" thus include the by one or more amino acid additions, Substitutions, deletions and / or inversions available Mutants, said changes in any sequence position may occur, as long as they become a mutant with the property profile according to the invention to lead. Functional equivalence is especially given when the patterns of reactivity between mutein and unchanged Polypeptide match qualitatively, i.e. for example binding to an organism-specific keratin with comparable specificity, but with different affinity.

Examples of suitable amino acid substitutions are shown in the following table: Original rest Examples of substitution Ala Ser bad Lys Asn Gln; His Asp Glu Cys Ser Gln Asn Glu Asp Gly Per His Asn; Gln Ile Leu; Val Leu Ile; Val Lys Arg; Gln; Glu mead Leu; Ile Phe Met; Leu; Tyr Ser Thr, Gly, Ala Thr Ser Trp Tyr Tyr Trp; Phe Val Ile; Leu

Known is that in SEQ ID No: 1 naturally present at position 2849 Serine e.g. can be exchanged for glycine to phosphorylation to get around this position (Fontao L, Favre B, Riou S, Geerts D, Jaunin F, Saurat JH, Green KJ, Sonnenberg A, Borradori L., Interaction of the bullous pemphigoid antigen 1 (BP230) and desmoplakin with intermediate filaments is mediated by distinct sequences within their COOH terminus., Mol Biol Cell. 2003 May; 14 (5): 1978-92. Epub 2003 Jan 26).

"Functional equivalents" are understood according to the invention in particular also muteins, which in at least one sequence position of the above mentioned amino acid sequences but have a different than the specified amino acid but still possess one of the above biological activities. "Functional equivalents" thus include the by one or more amino acid additions, Substitutions, deletions and / or inversions available Muteins, with the ge called changes can occur in any sequence position as long as they become one Mutein with the property profile according to the invention to lead.

"Functional equivalents" in the above sense are also "precursors" of the described Polypeptides and "functional Derivatives "and" salts "of the polypeptides.

"Precursors" are natural or synthetic precursors of the polypeptides with or without the desired biological activity.

Salts are understood as meaning both salts of carboxyl groups and acid addition salts of amino groups of the protein molecules of the invention in known manner and include inorganic salts such as sodium, calcium, ammonium, iron and zinc salts, as well as salts with organic bases such as amines such as triethanolamine, arginine, lysine, piperidine and the like. Acid addition salts, such as, for example, salts with mineral acids, such as hydrochloric acid or sulfuric acid, and salts with organic acids, such as acetic acid and oxalic acid, are likewise provided by the invention.

"Functional Derivatives of polypeptides according to the invention can on functional amino acid side groups or at their N- or C-terminal end by known techniques also be prepared. Such derivatives include, for example aliphatic esters of carboxylic acid groups, Amides of carboxylic acid groups, available by reaction with ammonia or with a primary or secondary amine; N-acyl derivatives free amino groups, prepared by reaction with acyl groups; or O-acyl derivatives of free hydroxy groups prepared by reaction with acyl groups.

Of course, "functional equivalents" also include Polypeptides which are accessible from other organisms, as well as naturally occurring Variants. For example, areas can be identified by sequence comparison homologous sequence regions and based on the concrete Specifications of the invention equivalent Determine enzymes.

"Functional equivalents" also include Fragments, preferably single domains or sequence motifs, the polypeptides according to the invention, which e.g. the desired have biological function.

"Functional equivalents" are also fusion proteins, which is one of the above-mentioned polypeptide sequences or derived therefrom functional equivalents and at least one other, functionally distinct, heterologous Sequence in functional N- or C-terminal linkage (i.e. without mutual essential functional impairment the fusion protein parts). Nonlimiting examples for such heterologous sequences are e.g. Signal peptides or enzymes.

According to the invention, "functional equivalents" encompassed are homologs to the specifically disclosed proteins. These have at least 60 %, preferably at least 75%, in particular at least 85%, such as e.g. 90%, 95% or 99%, homology to one of the specifically disclosed Amino acid sequences, calculated according to the algorithm of Pearson and Lipman, Proc. Natl. Acad, Sci. (USA) 85 (8), 1988, 2444-2448. A percentage homology a homologous invention Polypeptide means in particular percentage identity of the amino acid residues based on the total length one of the amino acid sequences specifically described herein.

in the Trap of a possible Protein glycosylation includes "functional equivalents" of the invention above designated type in deglycosylated or glycosylated form and by changing the Glycosylation pattern available modified forms.

homologous the polypeptides of the invention (i) can produced by mutagenesis, e.g. by point mutation or shortening of the Protein.

homologous of the polypeptides of the invention can by screening combinatorial libraries of mutants, e.g. Truncation mutants, be identified. For example, a variegated bank of Protein variants by combinatorial mutagenesis at the nucleic acid level be generated, such. by enzymatic ligation of a mixture synthetic oligonucleotides. There are a variety of procedures to produce banks of potential homologs from a degenerate oligonucleotide sequence can be used. The Chemical synthesis of a degenerate gene sequence can be performed in a DNA synthesizer carried out and the synthetic gene can then be transformed into a suitable expression vector be ligated. The use of a degenerate set of sentences allows the Provision of all Sequences in a mixture containing the desired set of potential Encode protein sequences. Process for the synthesis of degenerate Oligonucleotides are known in the art (e.g., Narang, S.A. (1983) Tetrahedron 39: 3; Itakura et al. (1984) Annu. Rev. Biochem. 53: 323; Itakura et al., (1984) Science 198: 1056; Ike et al. (1983) Nucleic Acids Res. 11: 477).

Several techniques for screening gene products of combinatorial libraries made by point mutations or truncation and for screening cDNA libraries for gene products having a selected property are known in the art. These techniques can be adapted to the rapid screening of gene libraries prepared by combinatorial mutagenesis according to the invention Homologer have been generated. The most commonly used techniques for screening large libraries that are subject to high throughput analysis include cloning the library into replicable expression vectors, transforming the appropriate cells with the resulting vector library, and expressing the combinatorial genes under conditions that demonstrate the desired activity isolation of the vector encoding the gene whose product was detected is facilitated. Recursive ensemble mutagenesis (REM), a technique that increases the frequency of functional mutants in the banks, can be used in combination with the screening assays to identify homologs (Arkin and Yourvan (1992) PNAS 89: 7811-7815; Delgrave et al., (1993) Protein Engineering 6 (3): 327-331).

• i. die Polypeptidsequenz (Domäne B)
• ii. die Polypeptidsequenz (Domäne C)
• iii. eine Polypeptidsequenz, die gegenüber (a) in bis zu 70% der Aminosäuren verändert ist,
• iv. eine Polypeptidsequenz, die gegenüber (b) in bis zu 70% der Aminosäuren verändert ist,

mit der Massgabe, dass die Keratin-Bindung der Polypeptidsequenz (c) oder (d) mindestens 10 % des Wertes beträgt, den die Polypeptidsequenz (a) oder (b) aufweist, gemessen in dem Test gemäß Beispiel 1 oder 2. Mit Domäne B bzw. C sind hier die oben beschriebenen Keratinbindedomänen des humanen Desmoplakins (SEQ ID NO:1) gemeint. Mit Veränderung von Aminosäuren sind hiermit Aminosäuresubstitutionen, -insertionen und -deletionen oder beliebige Kombinationen aus diesen drei Möglichkeiten gemeint.A particularly advantageous embodiment are keratin-binding effector molecules wherein the polypeptide sequence (i) comprises at least one of the following polypeptide sequences,

• i. the polypeptide sequence (domain B)
• ii. the polypeptide sequence (domain C)
• iii. a polypeptide sequence that is altered from (a) in up to 70% of the amino acids,
• iv. a polypeptide sequence that is altered from (b) in up to 70% of the amino acids,

with the proviso that the keratin binding of the polypeptide sequence (c) or (d) is at least 10% of the value of the polypeptide sequence (a) or (b) measured in the assay of Example 1 or 2. With domain B or C is meant herein the above-described keratin binding domains of human desmoplakin (SEQ ID NO: 1). By altering amino acids is meant amino acid substitutions, insertions and deletions or any combination of these three possibilities.

Prefers polypeptide sequences (i) are used which are highly specific for the desired organism affinity have. For Accordingly, applications in dermal cosmetics become polypeptide sequences (i) is preferably used which is a particular to the human skin keratin high affinity to have. For applications in hair cosmetics such polypeptide sequences are preferred, which have a particularly high affinity for human hair keratin.

For applications in the pet field, such polypeptide sequences will accordingly become (i) preferred to the corresponding keratin, for example Dog keratin or cat keratin have a particularly high affinity.

It can but also more than one polypeptide sequence (i) used in the effector molecule according to the invention For example, a sequence (i) that has a high binding affinity to human Skin keratin, in conjunction with a sequence (i); the one high affinity to human hair keratin. There may also be multiple copies of the same polypeptide sequence (i) are connected in series, for example, a higher To achieve binding.

suitable Keratin-binding polypeptide sequences (i) are known. For example contain desmoplakins and plectins keratin-binding domains. (Fontao L, Favre B, Riou S, Geerts D, Jaunin F, Saurat JH, Green KJ, Sonnenberg A, Borradori L., Interaction of the bullous pemphigoid antigen 1 (BP230) and desmoplakin with intermediate filaments is mediated by distinct sequences in their COOH terminus., Mol Biol Cell. 2003 May; 14 (5): 1978-92. Epub 2003 Jan 26; Hopkinson SB, Jones JC., The N terminus of the transmembrane protein BP180 interacts with the N-terminal domain of BP230, which mediating keratin cytoskeleton anchorage to the cell surface at the site of the hemidesmosome, Mol Biol Cell. 2000 Jan; 11 (1): 277-86).

By Alignments of such known protein sequences, for example with a computer program like Vector NTI 8 (version of September 25th 2002) of the company InforMax Inc. such areas are mapped and identified.

Further suitable polypeptide sequences (i) with good binding to human Keratin are sequence regions that in a high homology alignment or sequence identity and interpreted as consensus sequences of the keratinid domains can be.

Particularly preferred among these sequence regions are the following:
Domain B: polypeptide sequence SEQ ID NO: 1 position 2315 to 2450
Domain C: polypeptide sequence SEQ ID NO: 1 position 2645 to 2661
Domain C: polypeptide sequence SEQ ID NO: 1 position 2774 to 2788
Domain C: Polypeptide Sequence SEQ ID NO: 1 Position 2844 to 2854

If it wanted is that the polypeptide sequences (i) to a keratin from a non-human organism have a particularly good binding, are preferred as suitable sequence motifs from the keratin-binding Protein, e.g. Desmoplakin or plectin, the corresponding organism selected.

2 shows an alignment of keratin-binding molecules.

Effector molecules (ii)

When effector (ii) below are molecules understood, which have a certain predictable effect. This can either proteinaceous molecules such as enzymes or non-proteinogenic molecules such as dyes, Light stabilizers, vitamins and fatty acids, or metal ion-containing compounds.

Under the proteinaceous effector molecules are enzymes and antibodies, prefers. Among the enzymes are the following as effector molecules (ii) preferred oxidases, peroxidases, proteases, tyrosinases, metal-binding Enzymes, lactoperoxidase, lysozyme, amyloglycosidase, glucose oxidase, Superoxide dismutase, photolyase, calalase.

Well suitable as proteinaceous effector molecules (ii) are also hydrolysates of proteins from plant and animal sources, for example Hydrolysates of proteins of marine origin or silk hydrolyzates.

Under the non-proteinaceous effector molecules (ii) are, for example, dyes semi-permanent dyes or oxidation dyes are preferred. at The reactive dyes is preferably a component as effector molecule (ii) with the keratin-binding polypeptide sequence (i) linked and then at the site of action, i.e. after attachment to the hair, oxidative with the second dye component coupled.

When Dyes are all common Hair Dyes for the inventive molecules suitable. Suitable dyes are those skilled in the handbooks of For example, Schrader, Basics and Formulations of cosmetics Cosmetics, Hüthig Verlag, Heidelberg, 1989, ISBN 3-7785-1491-1 known.

Further suitable effector molecules (ii) are carotenoids. Carotenoids according to the invention are the following To understand compounds: β-carotene, Lycopene, lutein, astaxanthin, zeaxanthin, cryptoxanthin, citranaxanthin, Canthaxanthin, bixin, β-apo-4-carotenal, β-apo-8-carotenal, β-apo-8-carotenoic acid ester, individually or as a mixture. Preferred carotenoids are β-carotene, Lycopene, lutein, astaxanthin, zeaxanthin, citranaxanthin and canthaxanthin.

Under Retinoids are in the context of the present invention vitamin A alcohol (Retinol) and its derivatives such as vitamin A aldehyde (retinal), vitamin A acid (Retinoic acid) and Vitamin A esters (e.g., retinyl acetate, retinyl propionate and retinyl palmitate) meant. The term retinoic acid includes both all-trans retinoic acid and 13-cis retinoic acid. The Terms retinol and retinal preferably include the all-trans compounds. When Preferred retinoid is used for the suspensions according to the invention all-trans retinol, hereinafter referred to as retinol.

Further preferred effector molecules (ii) are vitamins, especially vitamin A and their esters.

Vitamins, provitamins and vitamin precursors from groups A, C, E and F, in particular 3,4-didehydroretinol, β-carotene (provitamin of vitamin A), ascorbic acid (vitamin C), and the palmitic acid esters, glucosides or phosphates of ascorbic acid, tocopherols , in particular α-tocopherol and its esters, for example the acetate, the nicotinate, the phosphate and the succinate; vitamin F, which is understood as meaning essential fatty acids, especially linoleic acid, linolenic acid and arachidonic acid;
Vitamin A and its derivatives and provitamins advantageously show a particular skin-smoothing effect.

The vitamins, provitamins or vitamin precursors of the vitamin B group or derivatives thereof which are preferably to be used according to the invention and the derivatives of 2-furanone include, inter alia:
Vitamin B ₁ , trivial name thiamine, chemical name 3 - [(4'-amino-2'-methyl-5'-pyrimidinyl) methyl] -5- (2-hydroxyethyl) -4-methylthiazolium chloride.

Vitamin B ₂ , trivial name riboflavin, chemical name 7,8-dimethyl-10- (1-D-ribityl) benzo [g] pteridine-2,4 (3H, 10H) -dione. In free form, riboflavin occurs, for example, in whey, other riboflavin derivatives can be isolated from bacteria and yeasts. A stereoisomer of riboflavin which is likewise suitable according to the invention is the lyxoflavin which can be isolated from fishmeal or liver and carries a D-arabityl residue instead of the D-ribityl.

Vitamin B ₃ . Under this name, the compounds nicotinic acid and nicotinamide (niacinamide) are often performed. According to the invention, the nicotinic acid amide is preferred.

Vitamin B ₅ (pantothenic acid and panthenol). Panthenol is preferably used. Derivatives of panthenol which can be used according to the invention are, in particular, the esters and ethers of panthenol and also cationically derivatized panthenols. In a further preferred embodiment of the invention, it is also possible to use derivatives of 2-furanone in addition to pantothenic acid or panthenol. Particularly preferred derivatives are the commercially available substances dihydro-3-hydroxy-4,4-dimethyl-2 (3H) -furanone with the trivial name pantolactone (Merck), 4 hydroxymethyl-γ-butyrolactone (Merck), 3,3-dimethyl 2-hydroxy-γ-butyrolactone (Aldrich) and 2,5-dihydro-5-methoxy-2-furanone (Merck), expressly including all stereoisomers.

advantageously, these compounds impart moisturizing to the keratin-binding effector molecules according to the invention as well as skin-soothing properties.

Vitamin B ₆ , which is understood hereunder no uniform substance, but the known under the common names pyridoxine, pyridoxamine and pyridoxal derivatives of 5 hydroxymethyl-2-methylpyridin-3-ols.

Vitamin B ₇ (biotin), also known as vitamin H or "skin vitamin". Biotin is (3aS, 4S, 6aR) -2-oxohexahydrothienol [3,4-d] imidazole-4-valeric acid.

panthenol, Pantolactone, nicotinic acid amide and biotin are very particular to the invention prefers.

According to the invention suitable Derivatives (salts, esters, sugars, nucleotides, nucleosides, peptides and Lipids). As lipophilic, oil-soluble antioxidants from this group are tocopherol and its derivatives, gallic acid esters, Flavonoids and carotenoids as well as butylhydroxytoluene / anisole are preferred. As water-soluble antioxidants are amino acids, e.g. Tyrosine and cysteine and their derivatives as well as tannins, especially those vegetable origin preferred.

triterpenes in particular triterpenic acids like ursolic acid, Rosemary acid, betulinic acid, boswellic and bryonic acid.

Further preferred effector molecules (ii) are UV photoprotective filters. These include organic substances to understand that are able to absorb ultraviolet rays and the absorbed energy in the form of longer wavelength radiation, e.g. Warmth, to give up again. The organic substances may be oil-soluble or water-soluble.

As oil-soluble UV-B filters, for example, the following substances can be used:
3-benzylidene camphor and its derivatives, eg 3- (4-methylbenzylidene) camphor;
4-aminobenzoic acid derivatives, preferably 2-ethylhexyl 4- (dimethylamino) benzoate, 2-octyl 4- (dimethylamino) benzoate and 4- (dimethylamino) benzoic acid ester;
Esters of cinnamic acid, preferably 4-methoxycinnamic acid 2-ethylhexyl ester, 4-propyl methoxycinnamate, isoamyl 4-methoxycinnamate, 4-isopentyl methoxycinnamate, 2-cyano-3-phenylcinnamic acid 2-ethylhexyl ester (octocrylene);
Esters of salicylic acid, preferably 2-ethylhexyl salicylate, 4-isopropylbenzyl salicylate, homomenthyl salicylate;
Derivatives of 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, such as 2,4,6-trianilino- (p-carbo-2'-ethyl-1'-hexyloxy) -1,3,5-triazine (Octyltriazone) and Dioctyl Butamido Triazone (Uvasorb HEB ^®):
Propane-1,3-diones such as 1- (4-tert-butylphenyl) -3- (4'-methoxyphenyl) propane-1,3-dione.

Suitable water-soluble substances are:
2-phenylbenzimidazole-5-sulfonic acid and its alkali, alkaline earth, ammonium, alkylammonium, alkanolammonium and glucammonium salts;
Sulfonic acid derivatives of benzophenones, preferably 2-hydroxy-4-methoxybenzophenone-5-sulfonic acid and its salts;
Sulfonic acid derivatives of 3-Benzylidencamphers, such as 4- (2-oxo-3-bomylidenemethyl) benzenesulfonic acid and 2-methyl-5- (2-oxo-3-bomylidene) sulfonic acid and salts thereof.

Especially preferred is the use of esters of cinnamic acid, preferably 4-methoxycinnamic acid 2-ethylhexyl ester, 4-methoxycinnamate, 2-cyano-3-phenyl-cinnamic acid 2-ethylhexyl ester (Octocrylene).

Of Another is the use of derivatives of benzophenone, in particular 2-hydroxy-4-methoxybenzophenone, 2-hydroxy-4-methoxy-4'-methylbenzophenone, 2,2'-dihydroxy-4-methoxybenzophenone and the use of propane-1,3-diones, e.g. 1- (4-tert-butylphenyl) -3- (4-methoxy-phenyl) -propane-1,3-dione prefers.

Typical UV-A filters are:
Derivatives of benzoylmethane, such as 1- (4'-tert-butylphenyl) -3- (4'-methoxyphenyl) propane-1,3-dione, 4-tert-butyl-4'-methoxydibenzoylmethane or 1-phenyl-3 - (4'-isopropylphenyl) -propane-1,3-dione;
Amino-hydroxy-substituted derivatives of benzophenones such as N, N-diethylaminohydroxybenzoyl-n-hexylbenzoate.

The UV-A and UV-B filters can Of course also be used in mixtures.

When more sunscreen filters can but also other insoluble ones Pigments, e.g. finely dispersed metal oxides or salts such as Titanium dioxide, iron oxide, aluminum oxide, cerium oxide, zirconium oxide, silicates (talc), Barium sulfate and zinc stearate are used. The particles should while a mean diameter of less than 100 nm, preferably between 5 and 50 nm and in particular between 15 and 30 nm.

Next The two aforementioned groups of primary photoprotective agents can also secondary Sunscreens of the antioxidant type are used, which interrupt the photochemical reaction chain that is triggered when UV radiation penetrates the skin. Typical examples are superoxide dismutase, Catalase, tocopherols (vitamin E) and ascorbic acid (vitamin C).

A Another group are anti-irritants that are anti-inflammatory Effect on damaged by UV light Skin possessed. Such substances are, for example, bisabolol, phytol and phytantriol.

Linkage of the effector molecules (ii) with the keratin-binding polypeptide sequence (i)

The effector (ii) are linked to a polypeptide sequence (i) that has a binding affinity to a Keratin possesses, connected. The connection between (i) and (ii) can be both covalent and on ionic or van-der Waals interactions based.

Prefers is a covalent link. This can be over, for example the side chains of the polypeptide sequence (i) take place, in particular via amino functions or carboxyl functions or thiol functions. Preferred is a Association over the Amino functions of one or more lysine residues, one or more multiple thiol group of cysteine residues or via the N-terminal or C-terminal Function of the polypeptide (i). The Linkage of the Effector Molecules (ii) with the polypeptide sequence (i) may be either directly, i. as covalent shortcut two chemical functions already present in (i) and (ii) take place, for example, an amino function of (i) is with a Carboxylate function of (ii) linked to the acid amide. The shortcut but can also over a so-called linker, i. an at least bifunctional molecule, which binds with a function with (i) and with a other function associated with (ii) becomes.

If the effector molecule (ii) also consists of a polypeptide sequence, the linkage of (i) and (ii) by a so-called fusion protein, i. a consistent Polypeptide sequence consisting of the two partial sequences (i) and (ii) consists.

It is also possible to incorporate so-called spacer elements between (i) and (ii), for example polypeptide sequences which have a potential cleavage site for a protease, lipase, esterase, phosphatase, Have hydrolase or polypeptide sequences that allow easy purification of the fusion protein, for example, so-called His-tags, ie oligohistidine residues.

The shortcut in the case of a non-proteinaceous effector molecule with the Polypeptide sequence (i) is preferably carried out by functionalizable Residues (side groups) on the polypeptide (i) that have a chemical function of the effector molecule make a covalent bond.

Prefers Here is the bond over one Amino, thiol or hydroxy function of the polypeptide (i), for example with a carboxyl function of the effector molecule (ii), optionally after activation, can undergo a corresponding amide, thioester or ester bond.

A further preferred linkage the polypeptide sequence (i) with an effector molecule (ii) the use of a customized linker. Such a linker has two or more so-called anchor groups with which he has the polypeptide sequence (i) and one or more effector molecules (ii) can link. For example can be an anchor group for (i) be a thiol function by means of which the linker with a Cysteine residue of the polypeptide (i) can enter into a disulfide bond. An anchor group for (ii) may be, for example, a carboxyl function by means of which the linker with a hydroxyl function of the effector molecule (ii) can undergo an ester bond.

The Use of such custom linker allows the exact Adaptation of the link the wished Effector molecule. It also makes it possible several effector molecules with a polypeptide sequence (i) defined to link.

• – Sulfhydryl-reaktiven Gruppen z.B. Maleimide, Pydridyldisulfide, α-Haloacetyle, Vinylsulfone) Amin-reaktive Gruppen (z.B. Succinimidylester, Carbodiimde, Hydroxymethyl-Phosphin, Imidoester, PFP-Ester etc.)
• – Zucker bzw. oxidierte Zucker-reaktive Gruppen (z.B. Hydrazide etc.)
• – Carboxy-reaktive Gruppen (z.B. Carbodiimide etc.)
• – Hydroxyl-reaktive Gruppen (z.B. Isocyanate etc.)
• – Thymin-reaktive Gruppen (z.B. Psoralen etc.)
• – unselektive Gruppen (z.B. Arylazide etc.)
• – photoaktivierbare Gruppen (z.B. Perfluorphenylazid etc.)
• – Metallkomplexierenden Gruppen (z.B. EDTA, Hexahis, Ferritin)
• – Antikörper und -fagmente (z.B. single-chain antibodies, F(ab)-Fragmente von Antikörpern, katalytische Antikörper).

The linker used depends on the functionality to be coupled. Examples of suitable molecules are those which couple to polypeptides (i) by means of sulfhydryl-reactive groups, for example maleimides, pyridyl disulfides, α-haloacetyls, vinyl sulfone and to effector molecules (ii)

• Sulfhydryl-reactive groups, for example maleimides, pydridyl disulfides, α-haloacetyls, vinyl sulfones) amine-reactive groups (for example succinimidyl esters, carbodiimde, hydroxymethyl-phosphine, imidoesters, PFP esters, etc.)
• - sugars or oxidized sugar-reactive groups (eg hydrazides etc.)
• Carboxy-reactive groups (eg carbodiimides etc.)
• - Hydroxyl-reactive groups (eg isocyanates etc.)
• - thymine-reactive groups (eg psoralen etc.)
• - unselective groups (eg aryl azides etc.)
• - photoactivatable groups (eg perfluorophenyl azide etc.)
• Metal-complexing groups (eg EDTA, hexahis, ferritin)
• - Antibodies and -fagments (eg single-chain antibodies, F (ab) fragments of antibodies, catalytic antibodies).

alternative For example, a direct coupling between active / effect substance and the keratin binding domain can be achieved e.g. by means of Carbodiimides, glutaric dialdehyde or others known to those skilled in the art Crosslinkers are performed.

Of the Linker can be stable, thermo gapable, photocleavable or enzymatic be cleavable (especially by lipases, esterases, proteases, phosphatases, Hydrolases etc.). Appropriate chemical structures are those skilled in the art and are integrated between the moieties (i) and (ii).

Examples for enzymatic cleavable linkers which can be used in the novel molecules are For example, in WO 98/01406 called on the entire contents hereby express reference is taken.

The invention Keratin-binding effector molecules can also - if desired - again easy be separated from the keratin. For this purpose, for example, a rinse with Keratin, whereby the keratin-binding effector molecules from their existing binding to the keratin are displaced and with the keratin be saturated from the rinsing solution. This is a reversible adhesion of a variety of effector molecules to keratin possible.

The invention Keratin-binding effector molecules have a wide field of application in human cosmetics, in particular Skin and hair care, animal care, leather care and leather processing.

Prefers become the inventive Keratin-binding effector molecules for the Skin, nail and hair cosmetics applied. They allow a high Concentration and long duration of action of skin, nail and hair care or skin, nail, and hair protection Effector substances.

suitable Auxiliaries and additives for the production of hair cosmetic or skin cosmetic preparations are familiar to the expert and can from manuals cosmetics, such as Schrader, bases and recipes the cosmetics, Hüthig Verlag, Heidelberg, 1989, ISBN 3-7785-1491-1 be removed.

To a further embodiment serves this hair cosmetic or skin cosmetic preparation of Care or protection of the skin or hair and is in the form of a Emulsion, a dispersion, a suspension, an aqueous Surfactant preparation, a milk, a lotion, a cream, a Balsams, an ointment, a gel, a granule, a powder, a pencil preparation, such as. a lipstick, a foam, an aerosol or of a spray. Such formulations are well suited for topical Preparations. As emulsions come oil-in-water emulsions and Water-in-oil emulsions or microemulsions in question.

in the As a rule, the hair cosmetic or skin cosmetic preparation used for application on the skin (topically) or hair. Under Topical preparations are to be understood as meaning those preparations which are suitable, the active ingredients in fine distribution and preferred Apply to the skin in a resorbable form through the skin. Therefor are suitable e.g. aqueous and watery-alcoholic Solutions, sprays, foams, Foam aerosols, ointments, watery Gels, O / W or W / O type emulsions, microemulsions or cosmetic Stick preparations.

To a preferred embodiment of the cosmetic according to the invention Contains means the agent is a carrier. Preferred as a carrier is water, a gas, a water-based fluid, an oil, a gel, a Emulsion or microemulsion, a dispersion or a mixture from that. The named carriers show a good skin compatibility. Especially advantageous for topical preparations are aqueous Gels, emulsions or microemulsions.

When Emulsifiers can nonionic surfactants, zwitterionic surfactants, ampholytic surfactants or anionic emulsifiers. The emulsifiers can be used in the composition of the invention in amounts of 0.1 to 10, preferably 1 to 5 wt .-%, based on the composition should be included.

As a nonionic surfactant, for example, a surfactant of at least one of the following groups may be used:
Addition products of 2 to 30 moles of ethylene oxide and / or 0 to 5 moles of propylene oxide to linear fatty alcohols having 8 to 22 carbon atoms, to fatty acids having 12 to 22 carbon atoms and alkylphenols having 8 to 15 carbon atoms in the alkyl group;
C _12/18 fatty acid _mono- and diesters of addition products of 1 to 30 moles of ethylene oxide with glycerol;
Glycerol mono- and diesters and sorbitan mono- and diesters of saturated and unsaturated fatty acids having 6 to 22 carbon atoms and their ethylene oxide addition products;
Alkyl mono- and oligoglycosides having 8 to 22 carbon atoms in the alkyl radical and their ethoxylated analogs;
Addition products of 15 to 60 moles of ethylene oxide with castor oil and / or hydrogenated castor oil;
Polyol and in particular polyglycerol esters, such as Polyglycerinpolyricinoleat, Polygl ycerinpoly-12-hydroxystearate or Polyglycerindimerat. Also suitable are mixtures of compounds of several of these classes of substances;
Addition products of 2 to 15 moles of ethylene oxide with castor oil and / or hydrogenated castor oil;
Partial esters based on linear, branched, unsaturated or saturated C _6/22 fatty acids, ricinoleic acid and 12-hydroxystearic acid and glycerol, polyglycerol, pentaerythritol, dipentaerythritol, sugar alcohols (eg sorbitol), alkyl glucosides (eg methyl glucoside, butyl glucoside, lauryl glucoside ) as well as polyglucosides (eg cellulose);
Mono-, di- and trialkyl phosphates and mono-, di- and / or tri-PEG-alkyl phosphates and their salts;
Lanolin alcohol;
Polysiloxane-polyalkyl-polyether copolymers or corresponding derivatives;
Mixed esters of pentaerythritol, fatty acids, citric acid and fatty alcohol according to DE PS 1165574 and / or mixed esters of fatty acids having 6 to 22 carbon atoms, methyl glucose and polyols, preferably glycerol or polyglycerol and
polyalkylene glycols;
Betaine.

Farther can as emulsifiers zwitterionic surfactants are used. As zwitterionic Surfactants become such surface active Compounds referred to in the molecule at least one quaternary ammonium group and at least one carboxylate or sulfonate group. Particularly suitable zwitterionic surfactants are the so-called Betaines such as the N-alkyl-N, N-dimethylammonium glycinates, for example cocoalkyldimethylammonium glycinate, N-acylamino-propyl-N, N dimethylammonium glycinates, for example cocoacylaminopropyldimethylammonium glycinate, and 2-alkyl-3-carboxylmethyl-3-hydroxyethylimidazolines each 8 to 18 carbon atoms in the alkyl or acyl group and the Kokosacylaminoethylhydroxyethyl-carboxymethylglycinat. This is particularly preferred under the CTFA name Cocamidopropyl Betaine known fatty acid amide derivative.

Also suitable emulsifiers are ampholytic surfactants. Ampholytic surfactants are understood as meaning those surface-active compounds which, apart from a C _8,18 -alkyl or -acyl group in the molecule, contain at least one free amino group and at least one -COOH or -SO ₃ H group and which are capable of forming internal salts. Examples of suitable ampholytic surfactants are N-alkylglycines, N-alkylpropionic acids, N-alkylamino-butanoic acids, N-alkyliminodipropionic acids, N-hydroxyethyl-N-alkylamido-propylglycines, N-alkyltaurines, N-alkylsarcosines, 2-alkylaminopropionic acids and alkylaminoacetic acids each having about 8 to 18 C atoms in the alkyl group.

Particularly preferred ampholytic surfactants are N-cocoalkylaminopropionate, cocoacylaminoethylaminopropionate and C _12/18 acylsarcosine. In addition to the ampholytic, quaternary emulsifiers are also suitable, those of the esterquat type, preferably methyl-quaternized difatty acid triethanolamine ester salts, being particularly preferred. In addition, alkyl ether sulfates, monoglyceride sulfates, fatty acid sulfates, sulfosuccinates and / or ether carboxylic acids can be used as anionic emulsifiers.

Guerbet alcohols based on fatty alcohols containing 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 -C ₁₃ carboxylic acids are included as oil bodies linear C ₆ -C ₂₂ -fatty alcohols, esters of linear C ₆ -C ₂₂ -fatty acids with branched alcohols, especially 2-ethylhexanol, esters of linear and / or branched fatty acids with polyhydric alcohols (such as propylene glycol, dimerdiol or trimer triol) and / or Guerbet alcohols, triglycerides based on C ₆ -C ₁₀ fatty acids, liquid mono- / di-, triglyceride mixtures based on C ₆ -C ₁₈ fatty acids, esters of C ₆ -C ₂₂ fatty alcohols and / or Guerbet alcohols with aromatic carboxylic acids, in particular benzoic acid, esters of C ₂ -C ₁₂ dicarboxylic acids with linear or branched alcohols having 1 to 22 carbon atoms or polyols having 2 to 10 carbon atoms and 2 to 6 hydroxyl groups, vegetable oils, branched te primary alcohols, substituted cyclohexanes, linear C ₆ -C ₂₂ fatty alcohol, Guerbetcarbonate, esters of benzoic acid with linear and / or branched C ₆ -C ₂₂ alcohols (eg Finsolv ^® TN), dialkyl ethers, ring-opening products of epoxidized ring-opening products of epoxidized fatty acid esters with polyols, silicone oils and / or aliphatic or naphthenic hydrocarbons into consideration. As oil body can also be used also silicone compounds, for example dimethylpolysiloxanes, Methylphenylpolysiloxane, cyclic silicones and amino, fatty acid, alcohol, polyether, epoxy, fluorine, alkyl and / or glycoside-modified silicone compounds which are both liquid at room temperature may be resinous. The oil bodies may be present in the compositions according to the invention in amounts of from 1 to 90, preferably from 5 to 80, and in particular from 10 to 50,% by weight, based on the composition.

By Coupling of corresponding compounds to a keratin-binding Polypeptide (i) can be significantly extended the duration of action on the skin. The coupling takes place as described above, formulation and application is carried out by methods known to those skilled in the art. Especially for deodorants are suitable effector molecules (ii): perfume oils, Cyclodextrins, ion exchangers, zinc ricinoleate, antimicrobial / bacteriostatic Compounds (e.g., DCMX, Irgasan DP 300, TCC).

For antiperspirants suitable are: tannins, and zinc / aluminum salts.

• – virale Erkrankungen (z.B. Herpes, Coxsackie, Varicella zoster, Cytomegalovirus etc)
• – bakterielle Erkrankungen (z.B. TB, Syphilis etc.)
• – pilzliche Erkrankungen (z.B. Candida, Cryptococcus, Histoplasmosis, Aspergillus, Mucormycosis etc.)
• – Tumorerkrankungen (z.B. Melanome, Adenome etc.)
• – Autoimmunerkrankungen (z.B. PEMPHIGUS VULGARIS, BULLOUS PEMPHIGOID, SYSTEMIC LUPUS ERYTHEMATOSIS etc.)
• – Sonnenbrand
• – parasitärer Befall (z.B. Zecken, Milben, Flöhe etc.)
• – Insektenkontakt (z.B. blutsaugende Insekten wie Anopheles etc.)

Another field of application for the novel substances is the therapeutic or prophylactic use in certain diseases of the skin and mucous membranes. Especially in the oral, pharyngeal and nasal areas, it is advantageous to strengthen active ingredients for therapy / prophylaxis via a keratin binding domain and to bind them for a longer time. Areas of application for this are in particular:

• - viral diseases (eg herpes, coxsackie, varicella zoster, cytomegalovirus etc)
• - bacterial diseases (eg TB, syphilis etc.)
• - fungal diseases (eg Candida, Cryptococcus, Histoplasmosis, Aspergillus, Mucormycosis etc.)
• - Tumor diseases (eg melanomas, adenomas etc.)
• Autoimmune diseases (eg PEMPHIGUS VULGARIS, BULLOUS PEMPHIGOID, SYSTEMIC LUPUS ERYTHEMATOSIS, etc.)
• - Sunburn
• - parasitic infestation (eg ticks, mites, fleas etc.)
• Insect contact (eg blood-sucking insects such as Anopheles etc.)

The suitable for therapy or prophylaxis (e.g., corticoids, immunosuppressant compounds, antibiotics, antifungals, anti-viral Compounds, insect repellent, etc.) can be described above Linker (depending on the functionality to be linked, a linker to be optimized) to the keratin-binding polypeptides (i).

Experimental part

example 1

bound to hair

• – Zugabe von 1 ml Ethanol zur Entfettung
• – Zentrifugation, Entfernung von Ethanol und Trocknung der Haare
• – Zugabe von 1 ml 1 % BSA in PBS und Inkubation während 1 h/RT, leichte Schwenkbewegungen
• – Zentrifugation, Entfernung des Überstands
• – Inkubation mit der zu testenden Keratinbindedomäne (als Standardreferenz „Domäne B" bzw. „Domäne C", an die N-terminal His₆ tag angekoppelt ist) in PBS/0,05 % Tween 20 während 4 h/RT
• – Zentrifugation, Entfernung des Überstands
• – 3× Waschen mit PBS/0,05% Tween 20
• – Inkubation mit monoklonalen Maus anti-His6-Antikörper (Sigma H-1029; 1:1000 in PBS/0,05 % Tween 20) während 2 h/RT
• – 3× Waschen mit PBS/0,05 % Tween 20
• – Inkubation mit Peroxidase gekoppeltem anti-Maus Antikörper (Sigma A-9917; 1:5000 in PBS/0,05 % Tween 20) während 2 h/RT, leichte Schwenkbewegungen
• – 3× Waschen mit PBS/0,05 % Tween 20
• – Zugabe von Peroxidasesubstrat (900 μl/Eppendorfgefäß): 0,1 ml TMB-Lösung (42 mM TMB in DMSO) und 10 ml Substratpuffer (0,1 M Natriumacetat pH 4,9) miteinander mischen und 14,7 μl H₂O₂ 3 %ig zusetzten
• – Mit 100 μl 2 M H₂SO₄ die Reaktion abstoppen
• – Die Absorption wird bei 450 nm gemessen

10 mg head hair (freshly washed, from Caucasians) are cut into 5 mm long pieces and transferred to Eppendorf tubes to perform the binding detection:

• - Add 1 ml of ethanol for degreasing
• - Centrifugation, removal of ethanol and drying of the hair
• Add 1 ml of 1% BSA in PBS and incubate for 1 h / RT, swirl lightly
• - Centrifugation, removal of the supernatant
• Incubation with the keratin binding domain to be tested (as standard reference "Domain B" or "Domain C" to which N-terminal His ₆ tag is coupled) in PBS / 0.05% Tween 20 for 4 h / RT
• - Centrifugation, removal of the supernatant
• - Wash 3 times with PBS / 0.05% Tween 20
• Incubation with monoclonal mouse anti-His6 antibody (Sigma H-1029, 1: 1000 in PBS / 0.05% Tween 20) for 2 h / RT
• - Wash 3 times with PBS / 0.05% Tween 20
• Incubation with peroxidase-coupled anti-mouse antibody (Sigma A-9917; 1: 5000 in PBS / 0.05% Tween 20) for 2 h / RT, slight panning movements
• - Wash 3 times with PBS / 0.05% Tween 20
• Addition of peroxidase substrate (900 μl / Eppendorf tube): Mix together 0.1 ml TMB solution (42 mM TMB in DMSO) and 10 ml substrate buffer (0.1 M sodium acetate pH 4.9) and 14.7 μl H ₂ O ₂ 3% ig zusetzten
• - Stop the reaction with 100 μl 2 MH ₂ SO ₄
• The absorption is measured at 450 nm

Example 2

Binding to skin

• – zur Inkubation mit den verschiedenen Reagentien, Transfer in ein Falcongefäß
• – Zugabe von Ethanol zur Entfettung, Entfernung von Ethanol und Trocknung
• – Inkubation mit 1 % BSA in TBS und Inkubation während 1 h/RT
• – Abblocken der unspezifischen Bindung mittels 1 % BSA in TBS-Puffer (1 h/RT)
• – Entfernung des Überstands
• – Inkubation mit der zu testenden Keratinbindedomäne (als Standardreferenz „Domäne B" bzw. „Domäne C", an die N-terminal His₆ tag angekoppelt ist) in TBS/0,05 % Tween 20 während 4 h/RT
• – Entfernung des Überstands
• – 3× Waschen mit TBS/0,05 % Tween 20
• – Inkubation mit monoklonalen Maus anti-His6-Antikörper (Sigma H-1029; 1:1000 in TBS/0,05 % Tween 20) während 2 h/RT
• – 3× Waschen mit TBS/0,05 % Tween 20
• – Inkubation mit Peroxidase gekoppeltem anti-Maus Antikörper (Sigma A-9917; 1:5000 in TBS/0,05 % Tween 20) während 2 h/RT
• – 3× Waschen mit TBS/0,05 % Tween 20
• – Zugabe von Peroxidasesubstrat (4,5 ml/Falcongefäß): 0,1 ml TMB-Lösung (42 mM TMB in DMSO) und 10 ml Substratpuffer (0,1 M Natriumacetat pH 4,9) miteinander mischen und 14,7 μl H₂O₂ 3 %ig zusetzten
• – Mit 0,5 ml 2 M H₂SO₄ die Reaktion abstoppen
• – Die Absorption wird bei 450 nm gemessen

The first step is the transfer of the outer keratin layer from the skin to a stable carrier. For this purpose, a Tesa ^™ adhesive tape is applied firmly to depilated human skin (freshly washed inside of the forearms of Caucasians). The tesa strip is fixed to a glass slide and binding detected as follows:

• - for incubation with the different reagents, transfer to a Falcon vessel
• - Addition of ethanol for degreasing, removal of ethanol and drying
• Incubation with 1% BSA in TBS and incubation for 1 h / RT
• Blocking of non-specific binding by means of 1% BSA in TBS buffer (1 h / RT)
• - Removal of the supernatant
• Incubation with the keratin binding domain to be tested (as standard reference "Domain B" or "Domain C" to which N-terminal His ₆ tag is coupled) in TBS / 0.05% Tween 20 for 4 h / RT
• - Removal of the supernatant
• 3x washing with TBS / 0.05% Tween 20
• Incubation with mouse monoclonal anti-His6 antibody (Sigma H-1029; 1: 1000 in TBS / 0.05% Tween 20) during 2 h / RT
• 3x washing with TBS / 0.05% Tween 20
• Incubation with peroxidase-coupled anti-mouse antibody (Sigma A-9917; 1: 5000 in TBS / 0.05% Tween 20) for 2 h / RT
• 3x washing with TBS / 0.05% Tween 20
• Addition of peroxidase substrate (4.5 ml / Falcon vessel): Mix together 0.1 ml TMB solution (42 mM TMB in DMSO) and 10 ml substrate buffer (0.1 M sodium acetate pH 4.9) and add 14.7 μl H ₂ O ₂ 3% ig zusetzten
• - Stop the reaction with 0.5 ml of 2 MH ₂ SO ₄
• The absorption is measured at 450 nm

Example 3

bound to synthetic skin

Human epidermis reconstituted in vitro on different carrier materials can be obtained from different manufacturers (eg SkinEthic ^™ , EpiDerm ^™ , cultivation according to the manufacturer's instructions). The evaluation of the binding activity of keratin binding domains is carried out analogously to Example 2: the synthetic skin preparations are incubated with the test keratin binding domains (tag-bound), anti-tag antibodies and streptavidin-alkaline phosphatase. After staining (eg with NBT-BCIP) the specimens are examined either directly or better after making thin sections in cross-section by means of a microscope.

It follows a sequence listing according to WIPO St. 25. This can from the official publication platform downloaded from the DPMA.

Claims (9)

Keratin-binding effector molecules consisting out (i) at least one polypeptide sequence having a binding affinity to a Has keratin, (ii) an effector molecule not naturally associated with the Polypeptide sequence (i) linked is. Keratin-binding effector molecules according to claim 1, wherein the Polypeptide sequence (i) binding affinity to human hair, nail or skin keratin. Keratin-binding effector molecules according to claim 1, wherein the Polypeptide sequence (i) at least one of the following polypeptide sequences includes, (a) the polypeptide sequence SEQ ID NO: 1 position 2269 until 2508 (b) the polypeptide sequence SEQ ID NO: 1 position 2606 until 2871 (c) a polypeptide sequence which is resistant to (a) in up to 70% of the amino acids changed is (d) a polypeptide sequence which is resistant to (b) in up to 70% of the amino acids changed is with the proviso in that the keratin binding of the polypeptide sequence (c) or (d) is at least 10% of the value, having the polypeptide sequence (a) or (b) measured in the Test according to example 1 or 2. Keratin-binding effector molecules according to claim 1, wherein the effector (ii) comprises a polypeptide sequence. Keratin-binding effector molecules according to claim 1, wherein the effector (ii) an enzymatic activity has. Keratin-binding effector molecules according to claim 1, wherein the effector (ii) is a dye or a dye component. Keratin-binding effector molecules according to claim 1, wherein the effector (ii) is a fungicide, insecticide or biocide. Keratin-binding effector molecules according to claim 1, wherein the effector (ii) is a vitamin or provitamin. Use of keratin-binding effector molecules after Claim 1 for the preparation of compositions for the skin and hair cosmetics.