AU9153998A - Enzymatic foam compositions for dyeing keratinous fibres - Google Patents

Description

WO 99/15137 PCT/DK98/00406 TITLE: ENZYMATIC FOAM COMPOSITIONS FOR DYEING KERATINOUS FIBRES FIELD OF THE INVENTION The present invention relates to enzymatic foam compositions for bleaching or 5 dyeing keratinous fibres such as hair, fur, hide and wool, a method for dyeing such fibres, and the use of a composition containing an oxidation enzyme and a foaming agent for dyeing hair and other keratinous fibres. BACKGROUND OF THE INVENTION 10 Hair is often dyed to cover grey hair that appears with age. Further, during the last few decades, hair dyeing has become more and more popular in the western world for purely cosmetic purposes, i.e. the hair is dyed a different colour than its natural colour in order to obtain a desired "look". In general, hair dyeing compositions on the market today can be divided into three main 15 groups: - temporary hair dyes, - semi-permanent hair dyes, and - permanent oxidative hair dyes. The temporary hair dyes are only intended to change the natural hair colour for a short 20 period of time and usually function by depositing dyes on the surface of the hair. Such hair dyes are easy to remove with normal shampooing. When using semi-permanent hair dyes the dyed colour can survive for five or more shampooings. This is achieved by using dyes having a high affinity for hair keratin and which are able to penetrate into the interior of the hair shaft. 25 Permanent hair dyes are very resistant to sunlight, shampooing and other hair treatments and need only be refreshed about once a month as new hair grows out. With these dyeing systems, the dyes are created directly in and on the hair. Small aromatic dye precursors (e.g. p-phenylene-diamine and o-aminophenol) penetrate deep into the hair, where they are oxidised by an oxidising agent to coloured polymeric compounds. These 30 coloured compounds are larger than the dye precursors and cannot be washed out of the hair. By including compounds referred to as modifiers (or couplers) in the hair dyeing composition, a number of hair colour tints can be obtained. Catechol and Resorcinol are examples of such modifiers.

WO 99/15137 PCT/DK98/00406 2 Traditionally,

H

2 0 2 (hydrogen peroxide) has been used as the oxidising agent (colour builder), but also as a bleaching agent. Dyeing compositions comprising H 2 0 2 are often referred to as "lightening dyes" due to the bleaching effect of H 2 0 2 . The use of H 2 0 2 in dye compositions has some disadvantages, however, as H 2 0 2 5 damages the hair. Further, oxidative dyeing often demands high pH (normally around pH 9-10), which also inflicts damage on the hair and irritation to the scalp. Consequently, it is not recommendable to dye the hair too often with dye compositions comprising H 2 0 2 . To overcome the disadvantages of using H 2 0 2 it has been suggested to use oxidation enzymes to replace H 2 0 2 . 10 US patent No. 3,251,742 (Revlon) describes a method for dyeing human hair by dye formation in situ. An oxidative enzyme is used for colour formation reactions at a substantially neutral pH (pH 7-8.5). Laccases, tyrosinases, polyphenolases and catacolases are mentioned as suitable oxidation enzymes. EP patent No. 0504005 (Perma S.A.) concerns compositions for dyeing hair which do 15 not require the presence of H 2 0 2 . The compositions comprise an enzyme capable of catalyzing the formation of polymeric dyes and also dye precursors, such as bases and couplers, in a buffer solution wherein the pH of the composition is between 6.5 and 8, the enzyme having an optimal activity in this pH range. Rhizoctonia praticola laccase and Rhus vernicifera laccase have a pH-optimum between 6.5 and 8 and can be used to form the 20 polymeric dyes according to this patent. The problem arises, however, when using such oxidation enzymes to dye keratinous fibrous such as human hair, that oxygen is the limiting factor in the enzymatic reaction, since the reaction proceeds relatively quickly, especially for laccases. This is especially the case with compositions having a relatively high viscosity, such as gels and 25 mousses. As a result, the reaction proceeds extensively at the parts of the hair lying at the hair/air interface, since the laccase at the interface has access to an adequate supply of oxygen from the air. In contrast, the major portion of the hair lying below this inter face, i.e. between the air and the scalp, does not have access to adequate oxygen for the enzymatic oxidation to proceed after exhaustion of oxygen present in the reaction 30 media, whereby the reaction proceeds slower (or comes to a complete stop) and this part of the hair is dyed relatively little compared to the surface hair fibres. Thus, when using oxidative enzymes for dyeing of hair, it would be desirable to obtain a more uniform and faster dyeing effect throughout the hair than that which can be obtained with the presently available hair dyeing compositions containing oxidative 35 enzymes.

WO 99/15137 PCT/DK98/00406 3 SUMMARY OF THE INVENTION The object of the present invention is to provide improved enzyme-based compositions for bleaching or dyeing of hair and other keratinous fibres, in particular 5 compositions which provide the advantages made possible by use of oxidative enzymes, e.g. reduced damage to hair, but which at the same time result in an improved uniformity of the hair dyeing effect. It has now surprisingly been found that hair dyeing compositions comprising an oxidation enzyme and a foaming agent result in improved uniformity of the dyeing effect, 10 in particular an improved dyeing of the parts of the hair removed from the air interface. Such foam compositions of the invention serve to provide oxygen for the enzymatic oxidation process to parts of the hair that would otherwise rapidly suffer from an insufficient supply of oxygen, thereby ensuring that oxidation can proceed more uniformly throughout the hair. As a result, a more uniform hair dyeing is obtained, with 15 reduced variation in hair colour intensity between different areas of the hair. In a first aspect, the invention thus relates to an enzymatic foam composition adapted for dyeing of keratinous fibres, comprising 1) at least one oxidation enzyme, 2) at least one foaming agent, 3) at least one dye precursor and optionally 4) at least one modifier. 20 Another aspect of the invention relates to a method for dyeing keratinous fibres, comprising contacting the fibres with the enzymatic foam composition for a period of time and under conditions sufficient to permit oxidation of the dye precursor to a coloured compound A further aspect of the invention relates to the use of the enzymatic foam 25 composition for oxidative dyeing of keratinous fibres, in particular hair, fur, hide or wool. BRIEF DESCRIPTION OF THE DRAWING Figure 1 shows the effect on colour uniformity by dyeing with a foam formulation of the invention (See Example 1) The first and second columns shows the Coefficient of 30 Variance (CV) (determined as AL* and AE*) for a dyeing composition comprising foam SDS-Betaine-pPD-Laccase; the third and fourth columns show the CV of a dyeing composition comprising foam SDS-pPDP-Laccase; the fifth and sixth columns (references) show the CV of a dyeing composition comprising pPD-laccase-water. Figure 2 shows the effect on colour uniformity by dyeing with a foam formulation 35 of the invention (see Example 1). The first column shows the Coefficient of Variance WO 99/15137 PCT/DK98/00406 4 (CV) (determined as AL* and AE*) for a dyeing composition comprising foam SDS Betaine-oAP-Laccase; the second column shows the CV of a dyeing composition comprises foam SDS-OAP-Laccase; the third column (reference) shows the CV of a dyeing composition comprising oAP-laccase-water. 5 Figure 3 shows the effect of hair dyeing using tyrosinase as the oxidation enzyme. DETAILED DESCRIPTION OF THE INVENTION The composition of the invention comprising 1) at least one oxidation enzyme and 2) at least one foaming agent is in particular a composition adapted for dyeing keratinous 10 fibres, e.g. hair, fur, hide or wool. For the dyeing of keratinous fibres, the composition of the invention further comprises 3) at least one dye precursor and optionally 4) at least one modifier. A preferred use of the composition is as a permanent dye for the dyeing of human hair. The oxidation enzyme is an oxidoreductase, i.e. an enzyme classified under the 15 Enzyme Classification number E.C. 1 (Oxidoreductases) in accordance with the Recommendations (1992) of the International Union of Biochemistry and Molecular Biology (IUBMB)) which catalyses oxidoreduction reactions. Within the group of oxidoreductases enzymes are preferred which catalyse the oxidation of a substrate (an electron or hydrogen donor) by acting on oxygen (02) 20 and/or a peroxide as the acceptor. Such enzymes include enzymes classified within the enzyme classes comprising oxidases, including E.C. 1.1.3. E.C. 1.2.3, E.C. 1.3.3, E.C. 1.4.3, E.C. 1.5.3, E.C. 1.7.3, E.C. 1.8.3 and E.C. 1.9.3, laccases and related enzymes in E.C. 1.10.3, and peroxidases in E.C. 1.11. According to the invention three types of oxidoreductases are specifically 25 contemplated: a) Laccases or related enzymes, which act on molecular oxygen and yield water (H 2 0) without any need for peroxide (e.g. H 2 0 2 ), b) Oxidases, which act on molecular oxygen (02) and yield peroxide (H 2 0 2 ), and c) Peroxidases, which act on peroxide (e.g. H 2 0 2 ) and yield water (H 2 0). 30 Also, enzyme systems which comprise a combination of more than one enzyme from a single group or from different groups among the three types of enzymes are contemplated. In the present specification, although reference will often be made to a single enzyme for the sake of simplicity, it is to be understood that the description is generally applicable to such combinations of more than one enzyme. Further, although 35 the invention is generally described in terms of the preferred aspect relating to the WO 99/15137 PCT/DK98/00406 5 dyeing of hair, it is to be understood that the description is generally applicable to foam compositions according to the invention adapted for dyeing of other types of keratinous fibres. Particularly preferred enzymes are laccases and related enzymes, the term 5 "laccases and related enzymes" including enzymes comprised by the enzyme classi fication E.C. 1.10.3.2 (laccases) and catechol oxidase enzymes comprised by E.C. 1.10.3.1, bilirubin oxidase enzymes comprised by the enzyme classification E.C. 1.3.3.5 and mono-phenol mono-oxygenase enzymes comprised by the enzyme classification E.C. 1.14.99.1. Laccases are multi-copper containing enzymes that 10 catalyze the oxidation of phenols and aromatic amines. Laccase-mediated oxidations result in the production of aryloxy-radical intermediates from suitable phenolic substrates; the ultimate coupling of the intermediates so produced provides a combination of dimeric, oligomeric, and polymeric reaction products. Certain reaction products can be used to form dyes suitable for dyeing hair. 15 The laccase may be derived from a microorganism, e.g. a fungus or a bacteria, or a plant. Preferably, the laccase employed is derived from a fungus. More preferably, it is derived from a strain of Polyporus sp., in particular a strain of P. pinsitus or P. versicolor, a strain of Myceliophthora sp., e.g. M. thermophila, a strain of Rhizoctonia sp., in particular a strain of Rh. praticola or Rh. solani, a strain of Pyricularia sp, in 20 particular P. oryzae, or a strain of Scytalidium, such as S. thermophilium. The laccase may also be from a plant such as Rhus sp., e.g. Rhus vernicifera, In specific embodiments of the invention the oxidoreductase is a laccase such as a Polyporus sp. laccase, especially the Polyporus pinisitus laccase (also called Trametes villosa laccase) described in WO 96/00290 (from Novo Nordisk Biotec Inc.) or a 25 Myceliophthora sp. laccase, especially the Myceliophthora thermophila laccase described in WO 95/33836 (from Novo Nordisk Biotech Inc.). Further, the laccase may be a Scytalidium sp. laccase such as the S. thermophilium laccase described in WO 95/33837 and WO 97/19998 (from Novo Nordisk Biotech Inc.), the contents of which is incorporated herein by reference, or a Pyricularia sp. 30 laccase, such as the Pyricularia oryzae laccase which can be purchased from SIGMA under the trade name SIGMA No. L5510, or a Coprinus sp. laccase, such as a C. cinereus laccase, especially a C. cinereus IFO 30116 laccase, or a Rhizoctonia sp. laccase, such as a Rh. solani laccase, especially the neutral Rh. solani laccase described in WO 95/07988 (from Novo Nordisk A/S) having a pH optimum in the range 35 of from 6.0 to 8.5.

WO 99/15137 PCT/DK98/00406 6 The laccase may also be derived from a fungus such as Collybia, Fomes, Lentinus, Pleurotus, Aspergillus, Neurospora, Podospora, Phlebia, e.g. P. radiata (WO 92/01046), Coriolus sp., e.g. C. hirsitus (JP 2-238885), or Botrytis. Bilirubin oxidase may preferably be derived from a strain of Myrothecium sp., such 5 as M. verrucaria. Oxidases yielding peroxide (H 2 0 2 ) are typically used in combination with a peroxidase to remove or at least reduce the peroxide produced. Suitable oxidases include glucose oxidase (E.C. 1.1.3.4), hexose oxidase (E.C. 1.1.3.5), L-amino-acid oxidase (E.C. 1.4.3.2), xylitol oxidase, galactose oxidase (E.C. 10 1.1.3.9), pyranose oxidase (E.C. 1.1.3.10) and alcohol oxidase (E.C. 1.1.3.13). If an L-amino acid oxidase is used it may be derived from a Trichoderma sp. such as Trichoderma harzianum, such as the L-amino acid oxidase described in WO 94/25574 (from Novo Nordisk A/S), or Trichoderma viride. A suitable glucose oxidase may originate from Aspergillus sp., such as a strain of 15 Aspergillus niger, or from a strain of Cladosporium sp. in particular Cladosporium oxysporum. Hexose oxidases from the red sea-weed Chondrus crispus (commonly known as Irish moss)(Sullivan and Ikawa, (1973), Biochim. Biophys. Acts, 309, p. 11-22; Ikawa, (1982), Meth. in Enzymol. 89, Carbohydrate Metabolism Part D, 145-149) oxidise a 20 broad spectrum of carbohydrates, such as D-glucose, D-galactose, maltose, cellobiose, lactose, D-glucose 6-phosphate, D-mannose, 2-deoxy-D-glucose, 2-deoxy-D-galactose, D-fructose, D-glucuronic acid, and D-xylose. The foaming agent may be present in the form of a single agent or a combination of two or more agents. The foaming agent is typically selected from soaps and ani 25 onic, cationic, non-ionic, amphoteric, sugar surfactants and/or zwitterionic surfactants and mixtures thereof. The foaming agent(s) may be present at levels of from 0.1% to 15%, preferably from 0.2 to 13%, more preferably from 0.25 to 10%, e.g. from 0.5 to 8% by weight of the final composition. Examples of anionic surfactants suitable for use as the foaming agent are soaps, e.g. in the form of alkali 30 or ethanol amine, isopropanol 2-methyl-2-amino-1,3-propandiol salts of fatty acids such as laurate, myristate, palmitate, stearate, isostearate, behenate, oleate, linoleate, etc.; fatty alcohol ether sulfates such as sodium lauryl ether sulfate; fatty alcohol sulfates such as sodium lauryl sulfate (SLS and SDS); sulfo succinates, e.g. dioctyl sodium sulfo succinate; a,-olefin sulfonates; alkyl amide ether sulfates; fatty 35 acid condensation products; alkyl ether phosphates and monoglyceride sulfates.

WO 99/15137 PCT/DK98/00406 7 Examples of non-ionic surfactants suitable for use as the foaming agent are especially the nonionic fatty acids and fatty amines that often are used as foam stabilisers, thickeners and boosters, e.g. fatty acid alkanol amides and dialkanol amides and fatty acid alkanol amide polyglycol ethers and fatty amine oxides. 5 Examples of amphoteric surfactants suitable for use in combination with anionic surfactants as the foaming agent are alkyl betaines, alkyl imidazolinium betaines, alkyl sulfo betaines, amidoalkyl betaines, N-alkyl-13-amino propionates, etc. Examples of foaming agents in the form of sugar surfactants include (a) alkyl and/or alkenyloligoglycosides and/or (b) fatty acid-N-alkylpolyhydroxyalkylamides. 10 The alkyl- and/or alkenyloligoglycoside (a) may have the formula (I): R'-O-[G]p (1) in which R' = a C 4

-

2 2 alkyl and/or alkenyl group, G = a sugar residue with 5 or 6 carbon atoms, and p = 1-10. The fatty acid-N-alkylpolyhydroxyalkylamide (b) may have the formula: 15

R

2

CO-N(R

3 )-[Z] (11) in which R 2 CO = a C- 6

-

22 aliphatic acyl residue, R 3 = H, alkyl or hydroxyalkyl with 1-4 carbon atoms, and [Z] = a linear or branched polyhydroxyalkyl residue with 3-12 carbon atoms and 3-10 OH groups. It also includes a) alkyl and alkenyl oligoglycosides of the formula R 1 -O-[G]p and 20 b) alkali and/or alkali metal salts of C- 12

-

2 2 secondary 2,3-alkyl sulphates, where R1 = C- 4

.

22 alkyl and/or alkenyl; G = a sugar residue with 5 or 6 carbon atoms, and p = 1-10. The weight ratio (1):(Il) is preferably 1:99-99:1. Also included are fatty acid-N-alkyl polyhydroxyalkyl amides; and sugar surfactants of: saccharose esters, sorbitan esters and/or polysorbates. 25 A sugar surfactant may also comprise 10-40% by weight of alkyl and/or alkenyl oligoglucoside of the formula R'-O-[G]p, 10-40% by weight of an alkyl- and/or alkenyl-oligoglucoside of the formula R 2 -O-(G)p and 80-20% by weight of an alkyl ether sulphate of the formula R-(OCH 2

CH

2 ), O-SO 3 M, in which R' = C.

1 1 alkyl or alkenyl; (G) = a glucose group.; p = 1-10, preferably 1-3; R 2 = C- 12

-

22 alkyl or 30 alkenyl; R 3 = C-6- 22 alkyl or alkenyl; M = an alkali or alkaline earth metal, or an ammonium or alkanolammonium ion, preferably Na or Mg; and n = 1-20, preferably 2-7. Preferably, R 2 and R 3 are independently a C- 1 2 -1 4 alkyl. Another example of a sugar surfactant is a polyglycerin fatty acid ester polyoxyalkylene ether R R 1

R

2

R

3 N +-CH(Y)-CH 2

-O-CH

2

-C(CH

3

)

2

-C(OH)(H)-C(=O)-NH-

WO 99/15137 PCT/DK98/00406 8

CH

2

-CH

2 -OH X- (I) where R, R 1 , R 2 = C- 1

-

24 alkyl or C- 8

.

24 alkenyl; R 3 = C- 1

.

18 alkylene; X = a monovalent organic or inorganic anion; and Y = OH or H. A sugar surfactant may also include (A) 1-5 wt.% of a fatty alcohol polyglycol ether, 1-5% of a Guerbet alcohol, and 1-5% of a polyol partial ester, (B) 1-5% of a 5 anionic polymer, (C) 15-30% of a fatty alcohol polyglycol ether sulphate, (D) 15-30% of an alkyloligoglycoside; and a sulphated product of fatty acid-N alkylpolyhydroxyalkyl amides of the formula R' CO- N(R 2 )-Z, where R 1 CO = C-6-22 aliphatic acyl; R 2 = H, C- 1

-

4 alkyl or C 1

.

4 hydroxyalkyl; Z = C-_'-1 2 polyhydroxyalkyl containing 3-10 hydroxy groups. 10 Sugar surfactants can also be selected from alkyl oligoglycosides of formula (I) and carboxylic acid N-polyhydroxyalkylamides of formula (II): R'-O(G) (I)

R

2

CO-NR

3 -Z (II) where R' = optionally hydroxylated

C-

1

.

8 alkyl; G = a sugar residue with 5 or 6 15 carbon atoms; p = 1-10; R 2 CO = C- 1

.

8 aliphatic acyl; R 3 = H, C- 1

.

8 alkyl or C- 1

.

8 hydroxyalkyl; Z = C- 3 -1 2 polyhydroxyalkyl containing 3-10 OH groups. Examples of preferred foaming agents are SDS (sodium dodecyl sulfate), sodium dodecyl ether sulfate and soaps. It may also be desired to add other additives that function as stabilisers, boosters 20 and thickeners, for example one or more compounds selected from fatty acid alkanol amides, dialkanol amides or fatty alkanol amides, polyglycol ethers such as ethoxylated lauric acid monoethanol amide, or fatty amine oxides such as alkyl dimethyl amine oxide. Compositions of the invention may furthermore include, as a stabiliser, a water soluble polymer capable of stabilising the foam, e.g. one or more of a cellulose derivative 25 such as cationized cellulose, hydroxyethyl cellulose, hydroxymethyl cellulose, hydroxypropylmethyl cellulose, an amphoteric resin, vinylethylether maleate polymer, polyvinyl pyrrolidone and acrylic resin alkanol amine. To obtain dyeing of the keratinous fibres such as hair, the dyeing composition of the invention also comprises at least one dye precursor which is converted into a 30 coloured compound (i.e. a dye) by the oxidation enzyme. Without being limited thereto, the dye precursor may be an aromatic compound belonging to one of three major che mical families: the aromatic diamines, aminophenols (or aminonaphthols) and the phenols. Examples of isatin derivative dye precursors can be found in DE 4,314,317 Al. Further, a number of indole or indoline derivative dye precursors are disclosed in WO 99/15137 PCT/DK98/00406 9 WO 94/00100. The dye precursors mentioned in these documents are hereby incorpo rated herein by reference. Examples of suitable dye precursors include compounds from the group comprising p-phenylene-diamine (pPD), p-toluylene-diamine, chloro-p-phenylenediamine, p 5 aminophenol, o-aminophenol, 3,4-diaminotoluene, 2,5-diaminotoluene, 2-methyl-1,4 diaminobenzene, 4-methyl-o-phenylenediamine, 2-methoxy-p-phenylenediamine, 2 chloro-1,4-diamino-benzene, 4-amino diphenylamine, 1-amino-4-p-methoxyethylamino benzene, 1 -amino-4-bis-(P-hydroxyethyl) -a m in o b e n z e n e , 1-3-diamino-benzene, 2 methyl-1,3-diamino-benzene, 2,4-diaminotoluene, 2,6-diaminopyridine, 1-hydroxy-2 10 amino-benzene, 1-hydroxy-3-amino-benzene, 1-methyl-2-hydroxy-4-amino-benzene, 1 methyl-2-hydroxy-4-p-hydroxyethylamino-benzene, 1-hydroxy-4-amino-benzene, 1 hydroxy-4-methylamino-benzene, 1-methoxy-2,4-diamino-benzene, 1-ethoxy-2,3 diamino-benzene, 1-p-hydroxyethyloxy-2,4-diamino-benzene, 3,4-diamino benzhydrazide, 3,5-diamino-benzhydrazide, 3-hydroxytyramine and catechin; phen 15 azines, such as 4,7-phenazinedicarboxylic acid, 2,7-phenazinedicarboxylic acid, 2 phenazinecarboxylic acid, 2,7-diaminophenazine, 2,8-diaminophenazine, 2,7-diamino 3,8-dimethoxyphenazine, 2,7-diamino-3-methoxyphenazine, 2,7-diamino 3 methoxyphenazine, 3-dimethyl 2,8-phenazinediamine, 2,2'-[(8-amino-7-methyl-2 phenazinyl)iminolbis-ethanol, 2,2'-[(8-amino-7-methoxy-2-phenazinyl)imino]bis-ethanol, 20 2,2'-[(8-amino-7-chloro-2-phenazinyl)imino]lbis-ethanol, 2-[(8-amino-7-methyl-2-phena zinyl)amino]-ethanol, 2,2'-[(8-amino-2-phenazinyl)imino]bis-ethanol, 3-amino-7 (dimethylamino)-2,8-dimethyl-5-phenyl-chloride, 9-(diethylamino)- benzol[alphenazine 1,5-diol, N-[8-(diethylamino)-2-phenazinyll-methanesulfonamide, N-(8-methoxy-2 phenazinyl)-methanesulfonamide, N,N,N',N'-tetramethyl-2,7-phenazinediamine, 3,7-di 25 methyl-2-phenazinamine; p-amino benzoic acids, such as p-amino benzoic acid ethyl, p amino benzoic acid glyceride, p-amino benzoic acid isobutyl, p-dimethylamino benzoic acid amil, p-dimethylamino benzoic acid octyl, p-diethoxyamino benzoic acid amil, p dipropoxyamino benzoic acid ethyl; acetylsalicylic acid, and isatin derivatives, such as 2,3-diamino benzoic acid. 30 Examples of other dye precursors include amino benzoic acid compounds of the general formula 1 WO 99/15137 PCT/DK98/00406 10 COR Z X

NH

2 Y wherein R is an amino, a mono - or disubstituted amino or OR', where R' is H, alkyl, 5 alkenyl, alkynyl, halogenalkyl, nitro, benzyl, phenyl or substituted phenyl, X, Y and Z may each independently be hydrogen, alkyl, alkenyl, alkynyl, halogenalkyl, nitro, benzyl, phenyl, substituted phenyl, amino, hydroxy or mercapto with the proviso that at least one of the groups X, Y and Z is an amino group or a salt thereof. Compounds where R' is a methyl, ethyl or 10 isopropyl ester are preferred. In one embodiment, the oxidoreductase is used with the dye precursor directly to oxidise it into a coloured compound. The dye precursor may be used alone or in combination with other dye precursors. However, it is believed that when using a diamine or an aminophenol as the dye 15 precursor, at least one of the intermediates in the copolymerisation must be an ortho or para-diamine or aminophenol. Examples of such are described in US patent No. 3,251,742 (Revlon), the contents of which are incorporated herein by reference. Optionally, the dyeing composition of the invention (especially hair dyeing compositions) also comprises a modifier (coupler) by means of which a number of 20 colour tints can be obtained. Modifiers are normally used in hair dyeing compo sitions, as the colours resulting from hair dyeing compositions without a modifier are usually unacceptable for most people. Modifiers are typically m-aromatic diamines, m-aminophenols, polyphenols, amino naphthalines or naphthols. The modifier (coupler) reacts with the dye precur 25 sor in the presence of the oxidative enzyme, converting it into a coloured compound. Examples of modifiers (couplers) include m-phenylene-diamine, 2,4-diaminoanisole, 1 hydroxynaphthalene(a-naphthol), 1,4-dihydroxybenzene(hydroquinone), 1,5 dihydroxynapthalene, 1,2-dihydroxybenzene(pyrocatechol), 1,3-dihydroxybenzene (re sorcinol), 1,3-dihydroxy-2-methylbenzene, 1,3-dihydroxy-4-chlorobenzene(4 30 chlororesorcinol), 1,2,3-trihydroxybenzene, 1,2,4-trihydroxybenzene, 1,2,4-trihydroxy 5-methylbenzene, and 1,2,4-trihydroxytoluene.

WO 99/15137 PCT/DK98/00406 11 Compositions according to the invention may in addition contain additives and excipients commonly used in hair-dyeing compositions and other cosmetic products, e.g. humectants, oils, thickening agents, antioxidants, buffers, preservatives, fragrances, colours, etc. 5 In a second aspect, the invention relates to a method for dyeing keratinous fibres, in particular hair, fur, hide and wool, using a composition as described above. The dyeing method can be conducted with one or more dye precursors and optionally in combination with one or more modifiers. The amount of dye precursor(s) and other ingredients used in the composition of the invention for this purpose are in accordance 10 with usual commercial amounts. Hair dyeing is typically carried out at or near room temperature, preferably around the optimum temperature of the enzyme being used, and at a pH in the range of from 3.0 to 9.0, preferably 4.0 to 8.5, especially 6.0 to 8.0. Suitable dye precursors and optional modifiers are described above. Although it is contemplated that the method of the invention typically will be 15 performed using a single pre-mixed composition applied to the hair or other fibres as a foam comprising the oxidation enzyme, the foaming agent, the dye precursor and the optional modifier in order to obtain the most uniform result, it is also contemplated that the method may alternatively be performed by mixing two or more separate compositions and/or ingredients in order to result in the desired foam composition 20 containing all of the necessary ingredients. The invention is further illustrated in the following non-limiting examples. MATERIALS AND METHODS Except where otherwise stated, the materials and methods used were the following. 25 Materials Hair: 6" De Meo Virgin Natural White Hair (De Meo Brothers Inc., USA) 30 Enzyme: Laccase from Myceliophtora termophilia described in WO 95/33837 (available from Novo Nordisk Biotech, Inc.) Dosed as 0.1 mg enzyme per ml of reaction liquid. Dye precursors: 35 The dye precursors used were selected from the following: WO 99/15137 PCT/DK98/00406 12 p-phenylene-diamine (pPD) o-aminophenol (oAP) Buffer: 5 0.1 M borate buffer, pH 7.0 with 60 ppm Ca ++ Analytical Equipment: Minolta CR200 Chroma Meter Daylight bulb: 1000 LUX (D65) 10 Methods Preparation of foam compositions Foams are made using foam dispensers or a blender, the foam compositions being gently blended by stirring during production to avoid foam formation at that stage. The 15 following formulations were used: "Minimal type SDS 0.4% Dye precursor 0.5% 20 Buffer up to 100% The enzyme is added to the formulation as 0.1 mg enzyme protein per ml dye solution. "Simple type A" 25 SDS 2.0% Betaine phosphate 2.0% Dye precursor 0.5% Buffer up to 100% The enzyme is added to the formulation as 0.1 mg enzyme protein per ml dye 30 solution Reference: "still water" Because foam can only be applied in a maximum amount of about 2.5 g per g of hair, the precursor concentration in the reference is reduced by 50%: WO 99/15137 PCT/DK98/00406 13 Dye precursor 0.25% Buffer up to 100% The enzyme is added to the formulation as 0.1 mg enzyme protein per ml dye solution. 5 A total of 5 ml of reference dye solution is added to the hair (Still water). Assessment of hair colour The quantitative colour of the hair wisps is determined on a Minolta CR200 Chroma Meter using the parameters L* ("0"= black and "100"= white), a* ("-"= green and 10 "+" =red) and b* ("-" blue and " + " yellow). AL*, Aa* and Ab* are the delta values of L*, a* and b* respectively compared to L*, a* and b* of untreated hair (e.g. AL* = L*%ie - L*,aten air), AE* is calculated as AE* = sq.root(AL* 2 + Aa* 2 + Ab* 2 ) and is an expression for the total quantitative colour change. 15 EXAMPLES Example 1 Hair dyeing composition comprising laccase formulated as a foam 20 1 gram hair wisps were dyed with foam formulations (prepared as described above) while hanging on a sloped metal plate. The foamed dye solutions were massaged into the hair wisps, whereas the reference solution was poured onto the wisps in a 150 ml glass beaker. 1-2 g of dye formulation was administered on each hair wisp. The dyeing was performed for 30 minutes at a temperature of 30'C, two wisps being used for each 25 formulation. After administration of the dye formulation, the wisps were hung on the mount and placed in a heating cabinet during the dyeing period. The reference was left untouched. After dyeing, the wisps were rinsed and washed once and then dried. The results of dyeing with two different dye precursors using the two foam formulations mentioned above are shown in the Figures 1 and 2. 30 Figure 1 shows the effect on colour uniformity by dyeing with a foam formulation containing the precursor pPD. The two above-mentioned foam formulations are each tested on two hair wisps, the dye uniformity being expressed as the "CV" (coefficient of variance) of AE* and AL* after measuring each hair wisp 12 times from tip to root.

WO 99/15137 PCT/DK98/00406 14 Figure 1 clearly shows that the colour uniformity is better for the hair wisps dyed with the foam formulations. Their CV is significantly lower than for the hair wisps dyed in the "still water" reference solution. The same effect is seen in Figure 2, where the dye precursor is oAP. In this case, 5 only one hair wisp is shown for each formulation, and again the foam improves colour uniformity. Example 2 Hair dyeing composition comprising tyrosinase formulated as a foam 10 Materials Hair: De Meo Virgin Natural White Hair, 1 gram hair wisps. Enzyme: Tyrosinase from Sigma. Conc: 0.9 mg e.p/ml formulation (e.p=active enzyme protein) 15 Dye precursor: p-phenylene-diamine (PPD) or p-toluene-diamine (PTD) Conc: 0.1% formulation Buffer: 0.1M Borate buffer, pH 7.0 with 60 ppm Ca" + Foam formulation: 0.02 g MiniRiskTM shampoo per ml formulation MiniRisk T M contains 15% SDS 20 Methods 1) The precursor is dissolved in 0.1M borate buffer for 15 min. under stirring. 2) MiniRisk

TM

, 0.1M borate buffer and enzyme are mixed gently. 3) The presursor is then added to the formulation and mixed gently. 25 4) The formulation foam is made using a foam dispenser. 5) The foam is applied in a maximum amount of about 2.5 g per g of hair by massaging it into the hair wisps. 6) The wisps are hung on a metal plate for 30 minutes at a temperature of 30 0 C for dyeing. 30 7) After 30 minutes the hair is rinsed for 3 minutes, washed with MiniRisk T M shampoo for 15 sec., then rinsed again and air-dried overnight. 8) After drying the hair color is measured using a Minolta Chroma Meter, an untreated hair wisp being used as a reference.

WO 99/15137 PCT/DK98/00406 15 The results are shown in Figure 3, in which it may be seen that hair dyeing is obtained with tyrosinase when combined with either PPD or PTD as a dye precursor. Although the AE value for PPD is somewhat higher than for PTD, it is still at an acceptable level, in particular in light of the fact that tyrosinase is generally not 5 regarded as an enzyme that is suitable for hair colouring purposes. This example thus shows that when used according to the present invention, acceptable hair dyeing may be obtained with tyrosinase.

Claims (14)

1. A composition adapted for dyeing of keratinous fibres, comprising 1) at least one oxidation enzyme, 2) at least one foaming agent, 3) at least one dye precursor and 5 optionally 4) at least one modifier.

2. The composition of claim 1, wherein the oxidation enzyme is an oxidoreductase selected from laccases and related enzymes, oxidases and peroxidases.

3. The composition of claim 2, wherein the oxidation enzyme is a laccase.

4. The composition of claim 3, wherein the laccase is derived from a fungus. 10

5. The composition of claim 4, wherein the laccase is derived from a strain of Polyporus sp., in particular a strain of P. pinsitus or P. versicolor, a strain of Myceliophthora sp., in particular M. thermophila, a strain of Rhizoctonia sp., in particu lar Rh. praticola or Rh. solani, or a strain of Scytalidium, in particular S. thermophilium, a strain of Pyricularia sp., in particular P. oryzae; or from a strain of Collybia, Fomes, 15 Lentinus, Pleurotus, Aspergillus, Neurospora, Podospora, Phlebia, e.g. P. radiata, Coriolus sp., e.g. C. hirsitus., Botrytis , or Coprinus sp.

6. The composition of any of claims 1-5 wherein the foaming agent is selected from soaps and anionic, non-ionic, amphoteric and zwitterionic surfactants.

7. The composition of claim 6 comprising at least one foaming agent selected from 20 sodium dodecyl sulfate and sodium dodecyl ether sulfate.

8. The composition of any of the preceding claims, comprising at least one dye precursor selected from aromatic diamines, aminophenols, phenols, aminonaphthalines and naphthols.

9. The composition of any of the preceding claims, comprising at least one modifier 25 selected from m-diamines, m-aminophenols and polyphenols.

10. A method for dyeing keratinous fibres, comprising contacting the fibres with a foam composition comprising 1) at least one oxidation enzyme, 2) at least one foaming agent, 3) at least one dye precursor and optionally 4) at least one modifier for a period of time and under conditions sufficient to permit oxidation of the dye precursor to a 30 coloured compound.

11. The method of claim 10, wherein the dyeing is carried out at a pH in the range from 3.0 to 9.0, preferably 4.0 to 8.5, especially 6.0 to 8.0.

12. The method of claim 10 or 11, wherein the foam composition is as defined in any of claims 2-9. WO 99/15137 PCT/DK98/00406 17

13. Use of a foam composition comprising 1) at least one oxidation enzyme, 2) at least one foaming agent, 3) at least one dye precursor and optionally 4) at least one modifier for oxidative dyeing of keratinous fibres, in particular hair, fur, hide or wool.

14. Use according to claim 13, wherein the foam composition is as defined in 5 any of claims 2-9.