WO2015104713A1

WO2015104713A1 - Coloring compositions and methods

Info

Publication number: WO2015104713A1
Application number: PCT/IL2015/050039
Authority: WO
Inventors: Amihay Freeman; Noa Hadar
Original assignee: Ramot at Tel- Aviv University Ltd.
Priority date: 2014-01-11
Filing date: 2015-01-11
Publication date: 2015-07-16

Abstract

Methods of coloring a surface such as hair, utilizing a dye composition comprising a macromolecule having at least one dye molecule associated therewith, are disclosed herein. The method may further comprise applying a priming composition comprising a cationic polymer to the surface, such that the dye composition selectively colors the cationic polymer applied on the surface. Further disclosed are compositions- of-matter comprising a macromolecule and at least one oxidative dye associated therewith, as well as processes for preparing a composition-of-matter which comprises a macromolecule and at least one dye molecule associated therewith. Kits comprising a priming composition and at least one dye composition, as well as applicators configured for applying a priming composition and/or at least one dye composition, are further disclosed.

Description

COLORING COMPOSITIONS AND METHODS

FIELD AND BACKGROUND OF THE INVENTION

The present invention, in some embodiments thereof, relates to coloring compositions and methods and more particularly, but not exclusively, to cosmetic hair coloring compositions and methods utilizing same.

Hair color products represent a major and rapidly growing category of salon products. In 2011, sales of hair color products in the U.S. alone were estimated at $730 million dollars for salon hair color products, with annual growth of 3.5 %, and $1.9 billion for home hair color products, with annual growth of 3.1 %. Sales of home hair color products are expected to reach $2.2 billion in 2016.

On average a woman dyes her hair at a mean frequency of 6-8 times per year, spends 2-3 hours and more at a salon to dye her hair, at a cost of over $100 per visit, and spends $18, 000- $24, 000 during her lifetime on hair dying. Most hair dye users are 30- 60 years old.

Most hair dyes can be classified in one of three categories, which differ notably in their permanence and chemistry [Ghosh & Sinha, Analytical Lett 2008, 41:2291-2321; Corbett, Soc Dyers Colourists August 1976, 285-303].

Temporary hair dyes are adsorbed to the surface of hair follicles, and can therefore be easily washed out with shampoo. A wide variety of dyes can be used as temporary hair dyes, including azo-compounds, diazo-compounds, xanthanes, triphenyl methanes, nitro compounds, anthraquinones and aminoanthraquinines. Temporary hair dyes are relatively non-toxic. However, there is a limited market for hair dyes with low fastness properties.

Semi-permanent hair dyes are relatively small molecules, which can penetrate the hair shaft to some degree, thereby providing a moderate degree of fastness. Examples include nitrophenylenediamines, nitroaminophenols and aminoanthraquinones. Small amounts of alkaline agent and/or oxidizing agent are sometimes used. Semi-permanent dyes generally last for 4-7 washes with shampoo. In the absence of bleaching, the hair cannot be dyed to a lighter color with such dyes.

Permanent hair dyes are the most commonly used hair dyes, representing 80 % of the market share for hair color products, and are commonly referred to as oxidative dyes due to the mechanism of their color formation. Dye precursors are mixed with an oxidizing agent such as hydrogen peroxide immediately before being applied to the hair. An alkaline agent, typically ammonia, is used to swell the hair, so as to facilitate entry of the reagents into the hair shaft. Oxidation of the dye precursors results in coupling of the precursors to form dyes. Because the dye precursors are relatively small molecules (such as phenylenediamines, aminophenols and resorcinol) they penetrate the hair shaft, whereas the dye formed by oxidation in the hair comprise larger molecules and cannot readily escape, especially after removal of the alkaline agent. The oxidizing agent also bleaches the natural color of the hair, such that the oxidative dye is the only coloring agent remaining in the hair. This is essential when the hair is being dyed to a lighter color.

Both the alkaline agent and oxidizing agent in oxidative dye compositions cause damage to hair [Nagai et al., Electrophoresis 1991, 12:451-453]. In particular, oxidizing agents cause morphological degradation of the hair surface, and a dramatic decrease in hydrophobicity of the hair surface, apparently due to oxidation of cysteine residues and removal of the 18-methyleicosanoic acid lipid layer on the hair surface [Baghdadli & Luengo, Phys 2008, Conference Series 100:052034; Jeong et al., Dermatol 2010, 37:882-887; Habe et al., Surf Interface Anal 2011, 43:410-412].

In addition to damaging hair, the reactive agents in oxidative hair dye compositions are also associated with irritation, unpleasant smells and allergic reactions. In addition, the necessity of performing oxidative chemistry on the head is inconvenient and introduces uncertainty as to what the final hair color will be. However, it has been very difficult to develop a commercially successful permanent hair dye composition which does not utilize alkaline agents and oxidative chemical reactions.

Lee et al. [Int J Nanomedicine 2011, 6:2879-2888] describe incorporation of phenylenediamine in nanoparticles by ion complex formation with polyglutamic acid, and suggest the use of such nanoparticles as a means for delivering phenylenediamine with reduced adverse effects. Delivery of phenylenediamine in nanoparticles presumably does not alter the mechanism by which phenylenediamine is used as a dye precursor for an oxidative dye.

Colored silica nanoparticles can diffuse into bleached hair fibers and dye the hair, but do not diffuse into non-bleached hair [Sampaio et al., Color Technol 2010, 127:55-61]. Hence, this technique cannot be used to dye hair without using oxidizing agents to bleach the hair.

U.S. Patent No. 7,968,084 describes dye molecules covalently bonded to an amine-containing polymer or oligomer such as polyethyleneimine, polyallylamine or polylysine, which are applied to the hair. To set or cure the polymer, a polymer with negatively charged groups is added, resulting in a complex with low solubility. Alternatively, the dye molecules may be bonded to the polymer with negatively charged groups, which can be cured by addition of an amine-containing polymer.

The use of enzymes to covalently cross-link agents such as coloring agents to proteins in hair has been proposed.

U.S. Patent No. 5,490,980 describes a use of transglutaminase to cross-link amine groups in an active agent to glutamine in skin, hair or nails.

International Patent Application Publication No. WO 01/07009 describes a use of lysine oxidase to attach agents to body tissue, including hair, by converting amine groups to aldehyde groups, which then cross-link with other amine groups.

Several strategies for dying hair have been developed based on the concept of conjugating a dye to a molecule which binds hair.

U.S. Patent Nos. 7,220,405 and 7,285.264 describe peptides identified as binding to hair, skin and nails with high affinity, and hair colorants consisting of a coloring agent coupled to one or more hair-binding peptides.

U.S. Patent Application Publication Nos. 2009/0156485 and 2009/0098074 describe conjugation of an effector molecule, such as a dye, to a keratin-binding polypeptide.

U.S. Patent No. 5,597,386 describes a hair dye consisting of an anti-hair antibody immobilized on a bulky coloring material.

Cationic polymers are well known to have affinity to hair, due to the net negative charge of hair, and have been used in conditioners, shampoo, hair mousse, hair spray and hair dye compositions to bind to hair and/or to neutralize negative charges on the hair.

U.S. Patent No. 7,731,761 describes polymeric hair dyes comprising cationic dye moieties, for providing affinity to negatively charged hair. U.S. Patent No. 4,182,612 describes water soluble polymer dyes comprising at least one primary, secondary or tertiary amino group per repeating unit, such as polyethyleneimine or polydiallylamine with attached chromophores. The polymer dyes are claimed therein to be resistant to shampooing, and to produce minimal skin staining after being flushed from the skin with water.

U.S. Patent No. 4,228,259 describes water soluble cationic polymer dyes comprising secondary, tertiary or quaternary ammonium groups in the polymer backbone.

U.S. Patent No. 5,827,330 describes melanin-like polymers formed by polymerization of dihydroxyindole derivatives comprising quaternary ammonium groups.

Cationic cellulose derivatives have been reported to enhance the fastness of semi-permanent dyes. This effect was interpreted as being associated more with the hydrophobic nature of the polymers than with their ionic charge [Ballarin et al., Int J Cosmetic Sci 2010, 1-7].

U.S. Patent No. 8,192,505 describes positively charged lipid vesicles comprising alkyl trimonium salts as carriers for direct dyes, for enhancing the permanence of direct hair dyes. The dye can be in the lipid membrane of the vesicle or in the vesicle interior. U.S. Patent No. 8,152,860 describes positively charged vesicles made from hydrophobized polysaccharides as carriers for direct dyes.

U.S. Patent No. 3,489,686 describes that cationic polyethyleneimine and alkoxylated polyethyleneimine enhance the deposition and retention of sparingly soluble particulate substances, such as antimicrobial particles in shampoo, on washed surfaces. It is suggested therein that the polymer attaches to the particles and imparts a net positive charge which provides affinity of the particle to a negatively charged surface.

Additional background art includes Chandrashekara & Ranganathaiah [Colloids and Surfaces B: Biointerfaces 2009, 69: 129-134]; Da Chow [Textile Res J 1971, 41:444- 450]; Hannan et al. [Textile Res J 1978, 48:57-58]; Lewis [Int J Cosmetic Sci 1996, 18: 123-135]; Lewis [Rev Prog Coloration 1998, 28: 12-17]; Reuveny et al. [Biotechnology and Bioengineering 1983, 25:2969-2980]; Reuveny et al. [Biotechnology and Bioengineering 1983, 25:469-480]; Rouviere et al. [Biotechnol J 2012, 7: 1-10]; Tang et al. [Dyes and Pigments 2006, 68:69-73]; Wang et al. ["Application of Polymeric Dyes in Textile Fields", The Proceedings of the 3rd International Conference on Functional Molecules]; International Patent Application Publication No. WO 93/05445; International Patent Application Publication No. WO 01/45652; U.S. Patent No. 3,489,686; U.S. Patent No. 3,917,817; U.S. Patent No. 4,013,787; U.S. Patent No. 4,314,808; U.S. Patent No. 4,946,472; U.S. Patent No. 5,733,343; U.S. Patent No. 7,306,632; U.S. Patent No. 6,264,933; U.S. Patent No. 6,277,588; U.S. Patent No. 6,506,374; U.S. Patent No. 7,300,473; U.S. Patent No. 7,364,594; U.S. Patent No. 7,381,423; U.S. Patent No. 7,833,519; U.S. Patent No. 7,896,932; U.S. Patent Application Publication No. 2002/0122780; U.S. Patent Application Publication No. 2003/0229947; U.S Patent Application Publication No. 2006/0140889; U.S. Patent Application Publication No. 2009/0099075; U.S. Patent Application Publication No. 2009/0211593; U.S. Patent Application Publication No. 2010/0015076; U.S. Patent Application Publication No. 2010/0132133. SUMMARY OF THE INVENTION

In order to overcome the drawbacks of oxidative dyes, the present inventors have devised and successfully prepared and practiced hair coloring methodology which exhibits a high degree of permanence, yet does not involve either exposure of the user to harsh, unpleasant and allergenic chemical reagents, or performance of time-consuming, messy and unpredictable chemical reactions in the hair. This methodology is based on priming hair by applying thereon a layer of a cationic polymer, and then applying a pre- prepared composition-of-matter comprising macromolecules associated with dyes onto the primed hair. The dye of the composition-of-matter, is a "ready-to-use" dye, such as, for example, an oxidative dye pre-prepared "in factory" from reaction between its precursors in the presence of an oxidizing agent. The composition-of-matter can then selectively color the cationic polymer.

According to an aspect of some embodiments of the invention, there is provided a composition-of-matter comprising a macromolecule and at least one oxidative dye associated with the macromolecule, the oxidative dye being a reaction product of at least one oxidative dye precursor and an oxidizing agent.

According to an aspect of some embodiments of the invention, there is provided a process for preparing a composition-of-matter which comprises a macromolecule and at least one dye molecule being in association with the macromolecule, the process comprising contacting the macromolecule with a reaction mixture that comprises at least one oxidative dye precursor of the oxidative dye and an oxidizing agent.

According to an aspect of some embodiments of the invention, there is provided a kit for coloring hair, the kit comprising:

a priming composition comprising a cationic polymer; and

at least one dye composition comprising a macromolecule having at least one dye molecule associated therewith,

the cationic polymer and the dye composition being packaged individually within the kit and being such that upon application of the priming composition on a hair of a subject, the dye composition selectively colors the cationic polymer.

According to an aspect of some embodiments of the invention, there is provided an applicator for coloring hair, the applicator being configured for contacting a portion of the hair with at least one dye composition.

According to an aspect of some embodiments of the invention, there is provided an applicator for priming hair for coloring, the applicator being configured for applying a composition comprising a cationic polymer onto a portion of the hair.

According to an aspect of some embodiments of the invention, there is provided a method of coloring hair, the method comprising contacting the hair with a dye composition comprising a composition-of-matter described herein, thereby coloring the hair.

According to an aspect of some embodiments of the invention, there is provided a method of coloring a surface, the method comprising:

applying a priming composition comprising a cationic polymer onto at least a portion of the surface, to thereby obtain a cationic polymer coating a surface; and

subsequent to the applying, contacting the surface with a dye composition comprising a macromolecule having at least one dye molecule associated therewith, thereby coloring the surface,

the cationic polymer and the dye composition being such that upon application of the cationic polymer on the surface, the dye composition selectively colors the cationic polymer. According to an aspect of some embodiments of the invention, there is provided a method of coloring hair, the method comprising:

applying a priming composition comprising a cationic polymer onto the hair, to thereby obtain a cationic polymer coating hair strands; and

subsequent to the applying, contacting the hair with a dye composition comprising a macromolecule having at least one dye molecule associated therewith, thereby coloring the hair,

the cationic polymer and the dye composition being such that upon application of the cationic polymer on a hair of a subject, the dye composition selectively colors the cationic polymer.

According to some embodiments of the invention the oxidative dye is a reaction product of the at least one oxidative dye precursor and the oxidizing agent in an alkaline aqueous solution.

According to some embodiments of the invention, the reaction mixture further comprises an alkaline aqueous solution.

According to some embodiments of the invention, the process further comprises isolating the composition-of-matter by filtration.

According to some embodiments of the invention, the composition-of-matter is prepared according to a process described herein.

According to some embodiments of the invention, the composition-of-matter is water-soluble.

According to some embodiments of the invention, the macromolecule associated with the at least one dye molecule is water-soluble.

According to some embodiments of the invention, the dye composition is water- soluble.

According to some embodiments of the invention, the macromolecule has a molecular weight of at least 10 kDa.

According to some embodiments of the invention, the macromolecule comprises a polypeptide.

According to some embodiments of the invention, the macromolecule has a neutral or negative net charge in aqueous solution. According to some embodiments of the invention, the macromolecule associated with the at least one dye molecule has a neutral or negative net charge in aqueous solution.

According to some embodiments of the invention, the at least one oxidative dye precursor is selected from the group consisting of 1-naphthol, l-acetoxy-2- methylnaphthalene, 2,7-naphthalenediol, 1,5-naphthalenediol, 2-aminopyridine, 2- amino-3-hydroxypyridine, 2,4,5, 6-tetraaminopyrimidine, 2,6-diaminopyridine, 2,6- dimethoxy-3,5-pyridinediamine, 2,5,6-triamino-4-pyrimidinol, l-hydroxyethyl-4,5- diaminopyrazole, hydroquinone, pyrocatechol, 4-amino-2-hydroxytoluene, 4- aminophenol, p-methylaminophenol, 3-aminophenol, 3-amino-2,4-dichlorophenol, 5- amino-6-chloro-o-cresol, 2-methyl-5-hydroxyethylaminophenol, 2-aminophenol, 2- amino-5-ethylphenol, 6-amino-m-cresol, 6-amino-2,4-dichloro-m-cresol, o- phenylenediamine, m-phenylenediamine, p-phenylenediamine, hydroxyethyl-p- phenylenediamine, N,N-bis(2-hydroxyethyl)-p-phenylenediamine, N-phenyl-p- phenylenediamine, 2-methoxy-p-phenylenediamine, 2-chloro-p-phenylenediamine, 2,6- diaminotoluene, 2,6-dihydroxyethylaminotoluene, 2,5-diaminotoluene, 2,5-diamino- hydroxyethylbenzene, 2,4-diaminoanisole, 2,4-diaminophenoxyethanol, 2-amino-4- hydroxyethylaminoanisole, benzodioxoles, 1,2,4-trihydroxybenzene, resorcinol, 4- chlororesorcinol, 2-methylresorcinol, 1 ,3-bis(2,4-diaminophenoxy)propane, hydroxyethyl-3,4-methylenedioxyaniline, and phenyl-methyl-pyrazolone.

According to some embodiments of the invention, the at least one oxidative dye precursor is selected from the group consisting of 4-aminophenol, 3-aminophenol, p- phenylenediamine and resorcinol.

According to some embodiments of the invention, the dye composition is washable from hair upon which the priming composition is not applied, within a time period of less than 5 minutes under a water shower stream of 20 liters per minutes and a water temperature of 37 °C, wherein the hair has a length of 5 cm.

According to some embodiments of the invention, the dye composition comprises a solution of the macromolecule associated with the at least one dye molecule.

According to some embodiments of the invention, the dye composition comprises a composition-of-matter described herein. According to some embodiments of the invention, the cationic polymer is colorless.

According to some embodiments of the invention, the cationic polymer comprises primary alkylamine groups.

According to some embodiments of the invention, the cationic polymer is a cross -linked polymer.

According to some embodiments of the invention, the cationic polymer is porous, and a porosity of the cationic polymer is characterized by an exclusion limit in a range of from 50 to 200 kDa.

According to some embodiments of the invention, the cationic polymer comprises a polyacrylamide derivative.

According to some embodiments of the invention, the cationic polymer is in a form of beads.

According to some embodiments of the invention, the kit further comprises a conditioner selected to be capable of adhering to the cationic polymer.

According to some embodiments of the invention, the kit comprises a plurality of dye compositions, each dye composition being individually packaged within the kit and comprising a macromolecule associated with at least one dye molecule, and each dye composition of the plurality of dye compositions having a different shade.

According to some embodiments of the invention, the kit further comprises instructions for applying one of the dye compositions to one portion of the hair, and another of the dye compositions to another portion of the hair, so as to obtain hair colored with a plurality of shades.

According to some embodiments of the invention, the kit further comprises instructions for applying the priming composition to hair prior to applying the dye composition to hair.

According to some embodiments of the invention, the kit further comprises instructions for applying the priming composition to a portion of the hair at which coloring is desired.

According to some embodiments of the invention, the kit further comprises an applicator configured for applying at least one of the at least one dye composition to hair. According to some embodiments of the invention, the kit further comprises an applicator configured for applying the priming composition to hair.

According to some embodiments of the invention, the applicator is in communication with a plurality of reservoirs for comprising different dye compositions, the applicator being configured for contacting the abovementioned portion of the hair with any of the dye compositions.

According to some embodiments of the invention, the applicator is further configured for applying a composition comprising a cationic polymer onto a portion of the hair.

According to some embodiments of the invention, the method further comprises contacting the coated hair with a conditioner selected to be capable of adhering to the cationic polymer.

According to some embodiments of the invention, applying a cationic polymer is effected by an applicator configured for applying to hair a composition comprising the cationic polymer.

According to some embodiments of the invention, the method comprises contacting a portion of the hair with a first dye composition comprising a macromolecule having at least one dye molecule associated therewith, and contacting another portion of the hair with a second dye composition comprising a macromolecule having at least one dye molecule associated therewith, the first dye composition and the second dye composition having different shades, thereby coloring the hair with a plurality of shades.

According to some embodiments of the invention, the contacting is effected by an applicator configured for applying to hair at least one dye composition comprising a macromolecule having at least one dye molecule associated therewith.

According to some embodiments of the invention, the dye composition comprises a composition-of-matter described herein.

Unless otherwise defined, all technical and/or scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which the invention pertains. Although methods and materials similar or equivalent to those described herein can be used in the practice or testing of embodiments of the invention, exemplary methods and/or materials are described below. In case of conflict, the patent specification, including definitions, will control. In addition, the materials, methods, and examples are illustrative only and are not intended to be necessarily limiting.

BRIEF DESCRIPTION OF THE DRAWINGS

Some embodiments of the invention are herein described, by way of example only, with reference to the accompanying images. With specific reference now to the images in detail, it is stressed that the particulars shown are by way of example and for purposes of illustrative discussion of embodiments of the invention. In this regard, the description taken with the images makes apparent to those skilled in the art how embodiments of the invention may be practiced.

In the images:

FIG. 1 is a scheme depicting the result of external hair dying according to some embodiments of the invention, by coloring a surface ("printing") of the hair fiber;

FIG. 2 is a scheme depicting the coloring of a hair fiber according to some embodiments of the invention, by addition of sponge-like coating absorbing material to a surface of a hair fiber, followed by addition of purified, dyed macromolecules;

FIG. 3 is a scheme depicting products prepared "in-factory" in accordance with some embodiments of the invention;

FIG. 4 is a scheme depicting a dyed macromolecule carrier prepared by admixing macromolecules with oxidative dye precursors/couplers, followed by purification of a dye-macromolecule conjugate by ultrafiltration; sponge-like coating absorbing material in a form of particles (spherical or flat) of less than 1 μιη in size; and a hair fiber coated by sponge-like coating particles dyed by the dye-macromolecule conjugate, in accordance with some embodiments of the invention;

FIG. 5 presents images of 8 compositions comprising a bovine serum albumin macromolecule reacted with oxidative dye precursors for 0, 10 and 25 minutes, according to some embodiments of the invention;

FIG. 6 is an images of 8 compositions comprising dyed bovine serum albumin purified by ultrafiltration, prepared according to some embodiments of the invention (1 - dark brown; 2 - light brown; 3 - medium dark brown; 4 - dark yellow; 5 - red-brown; 6 - yellow; 7 - dark red; 8 - light reddish brown); FIGs. 7 A and 7B are microscopic images of hair shafts coated with colorless microbeads comprising polyacrylamide derivatized with n-butylamine side chains, according to some embodiments of the invention;

FIGs. 8 A and 8B are images of previously untreated white hair treated with the medium dark brown (FIG. 8A) and light reddish brown (FIG. 8B) dyed bovine serum albumin shown in FIG. 6, with (right) and without (left) prior treatment of the hair with the microbeads shown in FIGs. 7 A and 7B;

FIGs. 9 A and 9B are images of partially bleached hair treated with the medium dark brown (FIG. 9A) and light reddish brown (FIG. 9B) dyed bovine serum albumin shown in FIG. 6, with (right) and without (left) prior treatment of the hair with the microbeads shown in FIGs. 7 A and 7B; and

FIG. 10 is a microscopic image of hair shafts coated with derivatized polyacrylamide microbeads, following treatment with the medium dark brown dyed bovine serum albumin shown in FIG. 6. DESCRIPTION OF SPECIFIC EMBODIMENTS OF THE INVENTION

The present invention, in some embodiments thereof, relates to coloring compositions and more particularly, but not exclusively, to cosmetic hair coloring compositions.

Before explaining at least one embodiment of the invention in detail, it is to be understood that the invention is not necessarily limited in its application to the details set forth in the following description or exemplified by the Examples. The invention is capable of other embodiments or of being practiced or carried out in various ways.

In order to overcome the drawbacks of current methodologies for coloring hair, the present inventors sought hair dyes which exhibit a high degree of permanence, yet do not involve either exposure of the user to harsh, unpleasant and allergenic chemical reagents, or performance of time-consuming, messy and unpredictable chemical reactions in the hair. In addition, the present inventors envisioned that a permanent dye adhered to the surface of the hair can overcome the necessity of bleaching in order to obscure the prior hair color, e.g. natural hair color, so as to obtain shades that are lighter than, or otherwise considerably different than, the prior hair color. The present inventors have envisioned that a convenient dye composition is such that carries an ex-vivo prepared dye, and have designed to this effect, and successfully practiced, a dye composition in which a dye is associated with a macromolecule, and which can be pre-prepared (e.g., "in factory"), so as to avoid performance of chemical reactions on the hair. In order to assure adhesion of such a dye composition to the hair, the present inventors have utilized a priming composition which is based on cationic polymers, which are selected so as to allow absorption of the dye composition, in such a way that the dye composition selectively colors the polymer.

While reducing the present invention to practice, the present inventors have demonstrated that a hair color exhibiting excellent fastness and ease of application, as well as the ability to obtain shades that are lighter than the prior hair color, can be obtained by priming hair by applying a layer of a cationic polymer, and then applying a dye composition comprising macromolecules associated with dyes onto the primed hair. The dye composition can then selectively color the cationic polymer.

The methodology designed by the present inventors is especially advantageous in that essentially any dye can be used while being associated with the macromolecule in a controlled, e.g. industrial production ("in factory") prior to application of the hair color, which allows for a wide variety of potential shades with highly predictable results, and eliminates the need for harsh chemical reagents and messy reactions performed in or on the hair of a user. In addition, the macromolecule does not need to exhibit any general adhesive properties, except for adherence to the priming polymer, which greatly reduces the risk of inadvertent staining of skin, clothes and the like as a result of the coloring procedure.

Referring now to the drawings, Figures 1 and 2 present an overview of hair dying in accordance with some embodiments of the invention, wherein the hair dye colors the surface of the hair fibers, which may be considered analogous to printing on the hair fiber surface. Figures 2-4 depict the use, in accordance with some embodiments of the invention, of a coating material, e.g. sponge-like particles, on the surface of the hair fibers for facilitating coloring of the hair fiber surface by dyes. The coating material and dyes may be products pre-fabricated in a factory. Figure 4 further depicts the preparation and use, in accordance with some embodiments of the invention, of a purified dye-macromolecule conjugate as a dye for use in combination with sponge-like hair-coating particles.

Figures 5 and 6 show the coloring of bovine serum albumin by an oxidative dye, to obtain various shades. Figures 7A and 7B shows how hair is coated by microbeads composed of a colorless cationic polyacrylamide derivative. Figures 8A-10 show how the colored macromolecule colors hair by coloring the 3 -dimensional cationic polymer layer attached to the surface of the hair. Figure 9B further shows hair colored so as to obtain a lighter shade.

These results demonstrate the efficacy of embodiments of the invention in coloring hair. It is expected that many surfaces other than hair can be colored by embodiments of the invention.

The dye composition:

As discussed hereinabove, the present inventors have designed and successfully practiced a dye composition which is based on a macromolecule having a dye molecule associated therewith.

Thus, according to the present embodiments, there is provided a dye composition which comprises a macromolecule having a dye molecule associated therewith.

According to an aspect of some embodiments of the invention, there is provided a composition-of-matter comprising a macromolecule and at least one dye molecule associated with the macromolecule, as described herein. In some embodiments, the composition-of-matter consists of a macromolecule and at least one dye molecule associated with the macromolecule, as described herein.

For the purpose of brevity, a composition-of-matter comprising a macromolecule and at least one dye molecule associated with the macromolecule is also referred to herein simply as the "composition-of-matter".

As defined herein, the composition-of-matter may comprise a single species of a macromolecule with at least one dye molecule associated therewith, or the composition of matter may comprise a mixture of different macro molecules with associated dye molecules. The mixture may comprise different species of macromolecules and/or different dye molecules associated with the macromolecules.

According to the present embodiments, a dye composition comprises a composition-of-matter as described herein. Herein, the term "dye composition" encompasses a composition-of-matter described herein per se, as well as compositions comprising the composition-of-matter and one or more additional ingredients, such as a carrier as described herein.

In some embodiments, the dye composition is consisted of a composition-of- matter as described herein. In some embodiments, a dye composition comprises a composition-of-matter as described herein and a carrier, as described herein.

I. The macromolecule:

Herein, the term "macromolecule" refers to a compound consisting of molecules having a molecular weight of at least 1 kDa, as well as to the individual molecules of the compound, depending on the context. Thus, references to a macromolecule (in the singular) are to be understood as relating to a compound consisting of a plurality of like macromolecules .

The macromolecule of a composition-of-matter may be associated with the dye molecule(s) via chemical and/or physical interactions. When associated via chemical interactions, the association may be effected, for example, by one or more covalent bonds and/or by one or more non-covalent interactions. Examples of non-covalent interactions include hydrogen bonds, electrostatic interactions, Van der Waals interactions and hydrophobic interactions. When associated via physical interactions, the association may be effected, for example, via absorption, entrapment, and the like.

In some embodiments, a non-covalent or physical association of a dye molecule with the macromolecule is characterized by a dissociation constant of less than 10^"5 M. In some embodiments, the dissociation constant is less than 10^"6 M. In some embodiments, the dissociation constant is less than 10^" M. In some embodiments, the dissociation constant is less than 10 -^"8 M. In some embodiments, the dissociation constant is less than 10^"9 M. In some embodiments, the dissociation constant is less than 10^"10 M.

In some embodiments, a macromolecule is associated with a dye molecule via a hydrophobic interaction, for example, a water-insoluble dye molecule adheres to a hydrophobic region, e.g. a hydrophobic core, of a macromolecule.

In some embodiments, a dye molecule is covalently bound to the macromolecule. In some embodiments, a dye molecule is bound, e.g. covalently, to an amine group in a macromolecule e.g. a lysine residue and/or N-terminus of a polypeptide.

In some embodiments of any of the aspects described herein, the macromolecule has a molecular weight of at least 2 kDa. In some embodiments, the macromolecule has a molecular weight of at least 4 kDa. In some embodiments, the macromolecule has a molecular weight of at least 6 kDa. In some embodiments, the macromolecule has a molecular weight of at least 8 kDa. In some embodiments, the macromolecule has a molecular weight of at least 10 kDa. In some embodiments, the macromolecule has a molecular weight of at least 20 kDa. In some embodiments, the macromolecule has a molecular weight of at least 30 kDa. In some embodiments, the macromolecule has a molecular weight of at least 40 kDa. In some embodiments, the macromolecule has a molecular weight of at least 50 kDa. Any integer between 1-50 kDa is also contemplated in these embodiments.

In some embodiments, the macromolecule has a molecular weight of 200 kDa or less. In some embodiments, the macromolecule has a molecular weight of 150 kDa or less. In some embodiments, the macromolecule has a molecular weight of 100 kDa or less. In some embodiments, the macromolecule has a molecular weight of 75 kDa or less. Any integer between 50-200 kDa is also contemplated in these embodiments.

In some embodiments, the macromolecule has a molecular weight in a range of from 4 to 200 kDa, including any integer therebetween. In some embodiments, the macromolecule has a molecular weight in a range of from 10 to 200 kDa. In some embodiments, the macromolecule has a molecular weight in a range of from 20 to 150 kDa. In some embodiments, the macromolecule has a molecular weight in a range of from 30 to 100 kDa. In some embodiments, the macromolecule has a molecular weight in a range of from 50 to 75 kDa. In exemplary embodiments, the macromolecule has a molecular weight of about 66 kDa. Any integer between the indicated values is also contemplated.

In some embodiments, the composition-of-matter has a molecular weight which is only slightly greater than that of the macromolecule per se (when not associated with the dye molecule(s)). In some embodiments, the molecular weight of the composition- of-matter is less than 20 % greater than that of the macromolecule. In some embodiments, the molecular weight of the composition-of-matter is less than 10 % greater than that of the macromolecule.

Herein, a molecular weight of a composition-of-matter refers to the molecular weight of the macromolecule with the associated dye molecules, regardless of whether the dye molecules are covalently bound to the macromolecule.

In some embodiments, the composition-of-matter has a molecular weight of at least 2 kDa. In some embodiments, the composition-of-matter has a molecular weight of at least 4 kDa. In some embodiments, the composition-of-matter has a molecular weight of at least 6 kDa. In some embodiments, the composition-of-matter has a molecular weight of at least 8 kDa. In some embodiments, the composition-of-matter has a molecular weight of at least 10 kDa. In some embodiments, the composition-of-matter has a molecular weight of at least 20 kDa. In some embodiments, the composition-of- matter has a molecular weight of at least 30 kDa. In some embodiments, the composition-of-matter has a molecular weight of at least 40 kDa. In some embodiments, the composition-of-matter has a molecular weight of at least 50 kDa. Any integer between 1-50 kDa is also contemplated in these embodiments.

In some embodiments, the composition-of-matter has a molecular weight of 200 kDa or less. In some embodiments, the composition-of-matter has a molecular weight of 150 kDa or less. In some embodiments, the composition-of-matter has a molecular weight of 100 kDa or less. In some embodiments, the composition-of-matter has a molecular weight of 80 kDa or less. Any integer between 50-200 kDa is also contemplated in these embodiments.

In some embodiments, the composition-of-matter has a molecular weight in a range of from 4 to 200 kDa. In some embodiments, the composition-of-matter has a molecular weight in a range of from 10 to 200 kDa. In some embodiments, the composition-of-matter has a molecular weight in a range of from 20 to 150 kDa. In some embodiments, the composition-of-matter has a molecular weight in a range of from 30 to 100 kDa. In some embodiments, the composition-of-matter has a molecular weight in a range of from 50 to 80 kDa. Any integer between the indicated values is also contemplated.

As exemplified herein, compositions-of-matter described herein can be readily isolated from smaller molecules such as free dye molecules and/or dye precursor molecules and/or breakdown products of the macromolecule, due to the considerable size of the macromolecule in the composition-of-matter, using techniques such as filtration, e.g. ultrafiltration.

In some embodiments, the macromolecule, i.e., the macromolecule per se, without the associated dye molecule(s), is water-soluble.

In some embodiments, the composition-of-matter is water-soluble. As dye molecules are often water-insoluble (a property which enhances the permanence of the dye), in some embodiments, a water-solubility of the macromolecule provides water- solubility to the composition-of-matter.

Without being bound by any particular theory, it is believed that a water-soluble composition-of-matter for coloring hair is advantageous in that it can be readily spread throughout hair without resorting to use of carriers such as alcohol, oils, silicones, and the like, which may be irritating, allergenic and/or damaging to the hair, and/or require removal from the hair, e.g., by shampooing, to remove oils or other hydrophobic carriers. It is further believed that a water-soluble composition-of-matter is easily removed, by washing with water, from areas which are not intended to be colored, e.g. skin, clothes and/or furniture, that is, they do not easily stain surfaces.

In some embodiments, the composition-of-matter is provided or utilized in a form of a solution, in which the macromolecule having the dye associated therewith is soluble. In some embodiments, the solution is an aqueous solution. In some of these embodiments, a dye composition comprises a composition-of-matter as described herein and an aqueous solution as carrier, as described herein. In some of these embodiments, a composition-of-matter as described herein is provided in a form of a solution.

In some embodiments, the composition-of-matter has a neutral or negative net charge in aqueous solution.

In some embodiments, the macromolecule per se has a neutral or negative net charge in aqueous solution.

Herein, the phrase "net charge" refers to the net electric charge, e.g. total positive charges minus total negative charges, of a molecule or molecules, e.g. a macromolecule and one or more dye molecules, not including the charge of any ions associated with the molecule(s) by non-covalent bonds. The net charge can be determined as being positive, neutral or negative, by various techniques, for example, by observing a direction of movement of a molecule, if any, in an electric field.

The aqueous solution may have any pH in a range of from 4-10. Thus, a molecule having a negative charge in aqueous solution at a pH of 10 and a positive charge in aqueous solution at a pH of 4 is considered as having a negative charge in aqueous solution.

In some embodiments, the neutral or negative net charge is in an aqueous solution having a pH in a range of from 5-9. In some embodiments, the neutral or negative net charge is in an aqueous solution having a pH in a range of from 6-8. In some embodiments, the neutral or negative net charge is in an aqueous solution having a pH of 7.

Generally, the macromolecule is selected such that when it is associated with the dye, a net charge of the composition-of-matter is neutral or negative, as described herein (such that it can interact with the cationic polymer when utilized, as described hereinafter), and the composition-of-matter exhibits at least some hydrophobicity (e.g., some hydrophobic or partially hydrophobic portions).

In some embodiments, the macromolecule comprises at least one amine group. In some embodiments, the macromolecule comprises a plurality of amine groups.

In some embodiments, the macromolecule comprises a biomolecule, either naturally-occurring biomolecule or synthetic biomolecule. Naturally-occurring biomolecules can be isolated from a natural source, or synthetically prepared. Commercially available biomolecules are also contemplated.

In some embodiments, the macromolecule comprises a polypeptide. In some embodiments, the macromolecule comprises a water-soluble polypeptide.

In some embodiments, the polypeptide is a naturally occurring protein, or a homolog thereof (e.g., exhibiting at least 80 %, at least 85 %, at least 90 %, or at least 95 % homology, including any value from 80 to 99.99 %).

In some embodiments, the polypeptide has a specific tertiary structure (e.g. a "folded" polypeptide). Significantly, naturally occurring proteins typically have a specific tertiary structure. In some embodiments, the polypeptide is a globular protein. In some embodiments, the polypeptide is a water-soluble globular protein. In some embodiments, the polypeptide is an albumin protein. Serum albumin, e.g. bovine serum albumin, is an exemplary albumin.

In some embodiments, the polypeptide is a monomeric polypeptide, that is, the macromolecule contains one polypeptide chain.

In some embodiments, the macromolecule contains a plurality of polypeptide chains. In some embodiments, the polypeptide chains are associated in a form of a specific quaternary structure. The polypeptide chains in the macromolecule may be the same or different. Thus, the polypeptide may be a dimeric polypeptide, a trimeric polypeptide, a tetrameric polypeptide, and so forth.

The term "polypeptide" as used herein encompasses native peptide macromolecules (either degradation products, synthetically synthesized peptides or recombinant peptides) and peptidomimetic macromolecules (typically, synthetically synthesized peptides), as well as peptoid and semipeptoid macromolecules which are peptide analogs, which may have, for example, modifications rendering the polypeptides more stable. Such modifications include, but are not limited to N-terminus modification, C-terminus modification, peptide bond modification, backbone modifications, and residue modification. Methods for preparing peptidomimetic compounds are well known in the art and are specified, for example, in Quantitative Drug Design, C.A. Ramsden Gd., Chapter 17.2, F. Choplin Pergamon Press (1992), which is incorporated by reference as if fully set forth herein. Further details in this respect are provided herein below.

Peptide bonds (-CO-NH-) within the polypeptide may be substituted, for example, by N-methylated amide bonds (-N(CH3)-CO-), ester bonds (-C(=0)-0-), ketomethylene bonds (-CO-CH₂-), sulfinylmethylene bonds (-S(=0)-CH₂-), a-aza bonds (-NH-N(R)-CO-), wherein R is any alkyl (e.g. methyl), amine bonds (-CH₂-NH-), sulfide bonds (-CH₂-S-), ethylene bonds (-CH₂-CH₂-), hydroxyethylene bonds (- CH(OH)-CH₂-), thioamide bonds (-CS-NH-), olefinic double bonds (-CH=CH-), fluorinated olefinic double bonds (-CF=CH-), retro-amide bonds (-NH-CO-), peptide derivatives (-N(R)-CH₂-CO-), wherein R is the "normal" side chain, naturally present on the carbon atom. These modifications can occur at any of the bonds along the polypeptide chain and even at several (2-3) bonds at the same time.

Natural aromatic amino acids, Trp, Tyr and Phe, may be substituted by non- natural aromatic amino acids such as l,2,3,4-tetrahydroisoquinoline-3-carboxylic acid (Tic), naphthylalanine, ring-methylated derivatives of Phe, halogenated derivatives of Phe or O-methyl-Tyr.

The polypeptides of some embodiments of the invention may also include one or more modified amino acids or one or more non-amino acid monomers e.g. fatty acids, complex carbohydrates etc.

The term "amino acid" is understood to include the 20 naturally occurring amino acids; those amino acids often modified post-translationally in vivo, including, for example, hydroxyproline, phosphoserine and phospho threonine; and other unusual amino acids including, but not limited to, 2-aminoadipic acid, hydroxylysine, isodesmosine, nor-valine, nor-leucine and ornithine. Furthermore, the term "amino acid" includes both D- and L- amino acids.

Tables 1 and 2 below list naturally occurring amino acids (Table 1) and non- conventional or modified amino acids e.g. synthetic (Table 2) which can be used with some embodiments of the invention.

Table 1

Amino Acid Three-Letter Abbreviation One-letter Symbol

Alanine Ala A

Arginine Arg R

Asparagine Asn N

Aspartic acid Asp D

Cysteine Cys C

Glutamine Gin Q

Glutamic Acid Glu E

Glycine Gly G

Histidine His H

Isoleucine He I

Leucine Leu L Lysine Lys K

Methionine Met M

Phenylalanine Phe F

Proline Pro P

Serine Ser S

Threonine Thr T

Tryptophan Trp w

Tyrosine Tyr Y

Valine Val V

Any amino acid as above Xaa X

Table 2

Non-conventional amino Code Non-conventional amino Code acid acid

ornithine Orn hydroxyproline Hyp a-aminobutyric acid Abu aminonorbornyl- Norb carboxylate

D-alanine Dala aminocyclopropane- Cpro carboxylate

D-arginine Darg N-(3- Narg guanidinopropyl)glycine

D-asparagine Dasn N-(carbamylmethyl)glycine Nasn

D-aspartic acid Dasp N- (c arboxymethyl)glycine Nasp

D-cysteine Dcys N-(thiomethyl)glycine Ncys

D-glutamine Dgln N-(2-carbamylethyl)glycine Ngln

D-glutamic acid Dglu N-(2-carboxyethyl)glycine Nglu

D-histidine Dhis N-(imidazolylethyl)glycine Nhis

D-isoleucine Dile N-( 1 -methylpropyl)glycine Nile

D-leucine Dleu N-(2-methylpropyl)glycine Nleu

D-lysine Dlys N-(4-aminobutyl)glycine Nlys

D-methionine Dmet N-(2-methylthioethyl)glycine Nmet D-ornithine Dorn N-(3-aminopropyl)glycine Norn

D-phenylalanine Dphe N-benzylglycine Nphe

D-proline Dpro N-(hydroxymethyl)glycine Nser

D- serine Dser N-( 1 -hydroxyethyl)glycine Nthr

D-threonine Dthr N-(3-indolylethyl) glycine Nhtrp

D-tryptophan Dtrp N-( ?-hydroxyphenyl)glycine Ntyr

D-tyrosine Dtyr N-( 1 -methylethyl)glycine Nval

D-valine Dval N-methylglycine Nmgly

D-N-methylalanine Dnmala L-N-methylalanine Nmala

D-N-methylarginine Dnmarg L-N-methylarginine Nmarg

D-N-methylasparagine Dnmasn L-N-methylasparagine Nmasn

D-N-methylasparatate Dnmasp L-N-methylaspartic acid Nmasp

D-N-methylcysteine Dnmcys L-N-methylcysteine Nmcys

D-N-methylglutamine Dnmgln L-N-methylglutamine Nmgln

D-N-methylglutamate Dnmglu L-N-methylglutamic acid Nmglu

D-N-methylhistidine Dnmhis L-N-methylhistidine Nmhis

D-N-methylisoleucine Dnmile L-N-methylisolleucine Nmile

D-N-methylleucine Dnmleu L-N-methylleucine Nmleu

D-N-methyllysine Dnmlys L-N-methyllysine Nmlys

D-N-methylmethionine Dnmmet L-N-methylmethionine Nmmet

D-N-methylornithine Dnmorn L-N-methylornithine Nmorn

D-N-methylphenylalanine Dnmphe L-N-methylphenylalanine Nmphe

D-N-methylproline Dnmpro L-N-methylproline Nmpro

D-N-methylserine Dnmser L-N-methylserine Nmser

D-N-methylthreonine Dnmthr L-N-methylthreonine Nmthr

D-N-methyltryptophan Dnmtrp L-N-methyltryptophan Nmtrp

D-N-methyltyrosine Dnmtyr L-N-methyltyrosine Nmtyr

D-N-methylvaline Dnmval L-N-methylvaline Nmval

L-norleucine Nle L-N-methylnorleucine Nmnle

L-norv aline Nva L-N-methylnorv aline Nmnva

L-ethylglycine Etg L-N-methyl-ethylglycine Nmetg L-t-butylglycine Tbug L-N-methyl-t-butylglycine Nmtbug

L-homophenylalanine Hphe L-N-methyl- Nmhphe homophenylalanine

a-naphthylalanine Anap N-methyl-a-naphthylalanine Nmanap penicillamine Pen N-methylpenicillamine Nmpen γ-aminobutyric acid Gabu N-methyl-y-aminobutyrate Nmgabu cyclohexylalanine Chexa N-methyl-cyclohexylalanine Nmchexa cyclopentylalanine Cpen N-methyl-cyclopentylalanine Nmcpen a-amino-a-methylbutyrate Aabu N-methyl-a-amino-a- Nmaabu methylbutyrate

a-aminoisobutyric acid Aib N-methyl-a- Nmaib aminoisobutyrate

D-a-methylarginine Dmarg L- a-methylarginine Marg

D-a-methylasparagine Dmasn L-a-methylasparagine Masn

D-a-methylaspartate Dmasp L-a-methylaspartate Masp

D-a-methylcysteine Dmcys L-a-methylcysteine Mcys

D-a-methylglutamine Dmgln L- a-methylglutamine Mgln

D-a-methyl glutamic acid Dmglu L- a-methylglutamate Mglu

D-a-methylhistidine Dmhis L-a-methylhistidine Mhis

D-a-methylisoleucine Dmile L-a-methylisoleucine Mile

D-a-methylleucine Dmleu L- a-methylleucine Mleu

D-a-methyllysine Dmlys L-a-methyllysine Mlys

D-a-methylmethionine Dmmet L- a-methylmethionine Mmet

D-a-methylornithine Dmorn L- a-methylornithine Morn

D-a-methylphenylalanine Dmphe L- a-methylphenylalanine Mphe

D-a-methylproline Dmpro L- a-methylproline Mpro

D-a-methylserine Dmser L-a-methylserine Mser

D-a-methylthreonine Dmthr L- a-methylthreonine Mthr

D-a-methyltryptophan Dmtrp L- a-methyltryptophan Mtrp

D-a-methyltyrosine Dmtyr L- a-methyltyro sine Mtyr D-a-methylvaline Dmval L-a-methylvaline Mval

N-cyclobutylglycine Ncbut L-a-methylnorv aline Mnva

N-cycloheptylglycine Nchep L- a-methylethylglycine Metg

N-cyclohexylglycine Nchex L- a-methyl- i-butylglycine Mtbug

N-cyclodecylglycine Ncdec L-a-methyl- Mhphe

homophenylalanine

N-cyclododecylglycine Ncdod a-methyl-a-naphthylalanine Manap

N-cyclooctylglycine Ncoct a-methylpenicillamine Mpen

N-cyclopropylglycine Ncpro a-methyl-y-aminobutyrate Mgabu

N-cycloundecylglycine Ncund a-methyl-cyclohexylalanine Mchexa

N- (2- aminoethyl)glycine Naeg a-methyl-cyclopentylalanine Mcpen

N-(2,2- Nbhm N-(N-(2,2-diphenylethyl) Nnbhm diphenylethyl)glycine carbamylmethyl-glycine

N-(3,3- Nbhe N-(N-(3,3-diphenylpropyl) Nnbhe diphenylpropyl)glycine carbamylmethyl-glycine

1 -carboxy- 1 -(2,2-diphenyl Nmbc 1,2,3,4- Tic

ethylamino)cyclopropane tetrahydroisoquinoline-3- carboxylic acid

phospho serine pSer pho spho threonine pThr phospho tyro sine pTyr O-methyl-tyrosine

2-aminoadipic acid hydroxylysine

The polypeptides of some embodiments of the invention are preferably utilized in a linear form, although it will be appreciated that in cases where cyclization does not severely interfere with polypeptide characteristics, cyclic forms of the polypeptide can also be utilized.

In some embodiments wherein a polypeptide is water-soluble, as described herein, the polypeptide preferably includes one or more non-natural or natural polar amino acids, including but not limited to serine and threonine which are capable of increasing polypeptide solubility due to their hydroxyl-containing side chain. The polypeptides of some embodiments of the invention may be synthesized by any techniques that are known to those skilled in the art of peptide synthesis. For solid phase peptide synthesis, a summary of the many techniques may be found in J. M. Stewart and J. D. Young, Solid Phase Peptide Synthesis, W. H. Freeman Co. (San Francisco), 1963 and J. Meienhofer, Hormonal Proteins and Peptides, vol. 2, p. 46, Academic Press (New York), 1973. For classical solution synthesis see G. Schroder and K. Lupke, The Peptides, vol. 1, Academic Press (New York), 1965.

In general, these methods comprise the sequential addition of one or more amino acids or suitably protected amino acids to a growing polypeptide chain. Normally, either the amino or carboxyl group of the first amino acid is protected by a suitable protecting group. The protected or derivatized amino acid can then either be attached to an inert solid support or utilized in solution by adding the next amino acid in the sequence having the complimentary (amino or carboxyl) group suitably protected, under conditions suitable for forming the amide linkage. The protecting group is then removed from this newly added amino acid residue and the next amino acid (suitably protected) is then added, and so forth. After all the desired amino acids have been linked in the proper sequence, any remaining protecting groups (and any solid support) are removed sequentially or concurrently, to afford the final polypeptide compound. By simple modification of this general procedure, it is possible to add more than one amino acid at a time to a growing chain, for example, by coupling (under conditions which do not racemize chiral centers) a protected tripeptide with a properly protected dipeptide to form, after de-protection, a pentapeptide and so forth. Further description of polypeptide synthesis is disclosed in U.S. Pat. No. 6,472,505.

A preferred method of preparing a polypeptide some embodiments of the invention involves solid phase polypeptide synthesis.

Large scale polypeptide synthesis is described by Andersson Biopolymers 2000; 55(3):227-50.

In some embodiments, the macromolecule is selected so as to chemically resemble the chemical structure of human hair.

An exemplary such macromolecule is BSA (bovine serum albumin).

In some embodiments, the macromolecule comprises a synthetic polymer. In some embodiments, the synthetic polymer is a co-polymer. The properties of such a polymer or co-polymer may be determined by appropriate selection of monomers and/or proportions of different monomers. In some embodiments, a co-polymer is selected so as to comprise a variety of functional groups, similar to the variety of functional groups in polypeptides.

In some embodiments, a synthetic polymer or co-polymer is selected or designed so as to exhibit a neutral or negative net charges when associated with the dye molecule and/or to exhibit hydrophobicity when associated with the dye molecule (e.g., at some portions thereof).

Exemplary synthetic macromolecules are co-polymers formed by copolymerization of two or more types of monomers selected from the following: one or more types of monomers generally form negatively-charged polymers, one or types of monomers generally form positively-charged polymers and one or more types of monomers form hydrophobic polymers. Any combination of the above types of monomers is contemplated for providing a synthetic polymer.

In some embodiments, the synthetic polymer is formed by polymerization of one or more monomers comprising allylamine, (meth)acrylic acid, (meth)acrylamide and/or derivatives thereof.

Herein, the term "(meth) acrylic acid" refers to acrylic acid and/or methacrylic acid.

Similarly, the term "(meth)acrylamide" refers to acrylamide and/or methacrylamide .

Examples of monomers which may be used to provide a polymer with negatively charged functional groups include (meth)acrylic acid.

Examples of monomers which may be used to provide a polymer with positively charged functional groups include allylamine and N-aminoalkyl-(meth)acrylamide. Examples of N-aminoalkyl-acrylamide residues are described herein.

Examples of monomers which may be used to provide a polymer with non- charged and relatively hydrophobic residues include (meth) acrylamide and hydrophobic (meth)acrylic acid esters such as alkyl (meth)acrylic acid esters.

Examples of monomers which may be used to provide a polymer with non- charged and relatively hydrophilic residues include (meth)acrylic acid and/or (meth)acrylamide derivatized with a hydroxy-containing group, such as hydroxyethyl (meth)acrylate.

The net charge of the polymer may be determined according to the ratio of monomers with positively charged functional groups to monomers with positively charged functional groups. For example, a ratio of 1: 1 may provide a polymer with neutral net charge, and a polymer with negative net charge may be obtained by including more monomers with negatively charged functional groups than with positively charged functional groups.

The hydrophobicity/hydrophilicity of the polymer may be determined according to the amount of charged functional groups, the net charge, and/or the hydrophobicity/hydrophilicity of the monomers.

All other factors being equal, the less charged functional groups are present, the more hydrophobic the polymer is likely to be.

All other factors being equal, the lower the net charge (neutral net charge being the lowest net charge possible), the more hydrophobic the polymer is likely to be.

All other factors being equal, methacrylic acid, methacrylamide and derivatives thereof will likely provide more hydrophobicity than the equivalent acrylic acid, acrylamide and derivatives thereof.

All other factors being equal, the larger an alkyl group in a monomer, e.g., an alkyl group in (meth) acrylic acid esters, the more hydrophobic the polymer is likely to be.

In some embodiments, a polymer with hydrophobic regions and charged functional groups is provided by co-polymerizing monomers which provide a polymer with relatively hydrophobic residues, such as described herein, and monomers which provide charged functional groups, such as described herein. In some embodiments, the monomers which provide charged functional groups are included in similar amounts, e.g. in a ratio from 1: 1.25 to 1.25: 1, such that the net charge of the polymer is neutral or low, e.g. weakly negative. In some embodiments, monomers which provide non- charged but relatively hydrophilic residues may be further included, for example, to enhance a water- solubility of a polymer while preserving hydrophobic regions in the polymer. II. The Dye

Herein, the phrase "dye molecule" refers to any molecule which absorbs visible light so as to produce a color detectable by the human eye. It is to be appreciated that in embodiments wherein a "dye molecule" is covalently bound to the macromolecule as described herein, the phrase "dye molecule" refers to a moiety of the macromolecule- dye conjugate. A "dye" in the context of the present embodiments therefore does not encompass oxidative dye precursors or any other dye precursor which do not produce a detectable color per se, but only when undergoing a chemical reaction.

In some embodiments, the dye molecule has a molecular weight of less than 1000 Da. In some embodiments, the dye molecule has a molecular weight of less than 750 Da. In some embodiments, the dye molecule has a molecular weight of less than 500 Da.

In some embodiments, the at least one dye molecules comprises more than one species of dye molecule i.e., a plurality of dye compounds. In such embodiments, different dye molecules may be associated with a single macromolecule and/or with separate macromolecules e.g. separate molecules of the same species of macromolecule. The combination of a plurality of dye molecules allows for a wider variety of shades to be obtained, and is particularly useful for obtaining shades that are especially desirable for hair coloring, such as "natural" hair color shades.

The dye molecule(s) may comprise any dye molecule used in hair coloring e.g. temporary dyes, semi-permanent dyes, as well as any dye molecule used in other coloring methodologies e.g. textile dying, food coloring, painting, printing.

Without being bound by any particular theory, it is believed that the properties of the composition-of-matter described herein will depend primarily on the properties of the large macromolecule, rather than that of the smaller dye molecule, and that consequently, dye molecules with essentially any chemical properties can be used in embodiments of the invention, with satisfactory results. It is further believed that dye molecules generally considered as unsuitable for applying onto a subject, e.g. due to toxicity, can be used when associated with a macromolecule in accordance with embodiments of the invention.

Examples of dye molecules which may be used in embodiments of the invention include, without limitation, azo compounds such as Acid Red 33, Food Red 1, Basic Brown 16, Basic Brown 17, Acid Orange 7; diazo compounds such as Direct Black 51, Acid Orange 24, D&C Brown No. 1; xanthanes such as D&C Red No. 22; triphenylmethanes such as FDC Blue No. 1; nitro compounds such as nitroaminophenols (e.g. 2-amino-4-nitrophenol, 2-amino-5-nitrophenol, 4- hydroxypropylamino-3-nitrophenol, 3-nitro-p-hydroxyethylaminophenol), nitrophenylenediamines (e.g. 4-nitro-o-phenylenediamine, 4-nitro-m-phenylenediamine, 2-nitro-p-phenylenediamine, N-(2-hydroxyethyl)-2-nitro-p-phenylenediamine), D&C Yellow No. 7; anthraquinones and aminoanthraquinines such as D&C Green No. 5, 1,2,4,5-tetraaminoanthraquinone, 1,4-diaminoanthraquinone; naphthoquinones such as 2-hydroxy-l,4-naphthoquinone (lawsone, Natural Orange 6) and 5 -hydroxy- 1,4- naphthoquinone juglone, Natural Brown 7); HC orange 2, HC orange 3, HC yellow 4, HC yellow 9, HC red 1, HC red 3, HC red 11, HC red 13, HC red 14, HC violet 2, HC blue 22, Disperse violet 1, Disperse violet 4, Disperse blue 3, Disperse red 11 and Disperse red 15. 2-Hydroxy-l,4-naphthoquinone is an exemplary dye molecule.

In some embodiments, a dye molecule associated with the macromolecule is a molecule of an oxidative dye.

Herein, the phrase "oxidative dye" refers to dye compounds formed by oxidation of precursor compounds (also referred to herein as "oxidative dye precursors") - for example, by reaction of at least one oxidative dye precursor and an oxidizing agent - which results in formation of covalent bonds linking two or more precursor compound molecules.

In some embodiments, the oxidative dye is formed by oxidizing at least one precursor in an alkaline aqueous solution.

The formation of oxidative dyes from precursors is well known in the art of hair coloring, and suitable precursors and the types of dye molecules which are formed therefrom have been studied to a considerable extent and are well understood by those of skill in the art. As the oxidation of precursors typically results in a mixture of dye compounds, it is common practice to characterize oxidative dyes by the precursors from which they are formed, rather than by a molecular structure of the dye itself.

Herein, the term "precursor" encompasses compounds referred to the art as

"primary intermediates" as well as compounds referred to in the art as "couplers" or "secondary intermediates". Examples of oxidative dye precursors include, without limitation, 1-naphthol, 1- acetoxy-2-methylnaphthalene, 2,7-naphthalenediol, 1,5-naphthalenediol, 2- aminopyridine, 2-amino-3-hydroxypyridine, 2,4,5,6-tetraaminopyrimidine, 2,6- diaminopyridine, 2,6-dimethoxy-3,5-pyridinediamine, 2,5,6-triamino-4-pyrimidinol, 1- hydroxyethyl-4,5-diaminopyrazole, hydroquinone, pyrocatechol, 4-aminophenol, p- methylaminophenol, 3-aminophenol, 3-amino-2,4-dichlorophenol, 4-amino-2- hydroxytoluene (5-amino-o-cresol), 5-amino-6-chloro-o-cresol, 2-methyl-5- hydroxyethylaminophenol, 2-aminophenol, 2-amino-5-ethylphenol, 6-amino-m-cresol, 6-amino-2,4-dichloro-m-cresol, o-phenylenediamine, m-phenylenediamine, p- phenylenediamine, hydroxyethyl-p-phenylenediamine, N,N-bis(2-hydroxyethyl)-p- phenylenediamine, N-phenyl-p-phenylenediamine, 2-methoxy-p-phenylenediamine, 2- chloro-p-phenylenediamine, 2,6-diaminotoluene, 2,6-dihydroxyethylaminotoluene, 2,5- diaminotoluene, 2,5-diamino-hydroxyethylbenzene, 2,4-diaminoanisole, 2,4- diaminophenoxyethanol, 2-amino-4-hydroxyethylaminoanisole, benzodioxoles, 1,2,4- trihydroxybenzene, resorcinol, 4-chlororesorcinol, 2-methylresorcinol, l,3-bis(2,4- diaminophenoxy)propane, hydroxyethyl-3,4-methylenedioxyaniline, and phenyl- methyl-pyrazolone. Dye precursors used in exemplary embodiments include 4- aminophenol, 3-aminophenol, p-phenylenediamine and resorcinol.

In some embodiments, the oxidative dye is formed from at least one primary intermediate.

In some embodiments, a primary intermediate comprises an aromatic (or heteroaromatic) ring substituted by an amine group at a para or ortho position with respect to a hydroxy group or another amine group. In some embodiments, the amine group is at a para position with respect to the hydroxy group or other amine group.

Examples of such primary intermediates include, without limitation, o- phenylenediamine, p-phenylenediamine, 4-aminophenol, 2-aminophenol, 2,5- diaminopyrimidine and substituted derivatives thereof, i.e., any of the aforementioned compounds with one or more substituents added, such as hydroxyethyl-p- phenylenediamine, N,N-bis(2-hydroxyethyl)-p-phenylenediamine, 2-methoxy-p- phenylenediamine, N-phenyl-p-phenylenediamine, 2-chloro-p-phenylenediamine, 2,5- diaminotoluene, 2,5-diamino-hydroxyethylbenzene, p-methylaminophenol, 2-amino-5- hydroxytoluene, 2-amino-5-ethylphenol, 6-amino-m-cresol, 6-amino-2,4-dichloro-m- cresol, 2,4,5, 6-tetraaminopyrimidine and 2,5,6-triamino-4-pyrimidinol.

Additional examples of primary intermediates include, without limitation, 4,5- diaminopyrazole derivatives such as l-hydroxyethyl-4,5-diaminopyrazole.

In some embodiments, the oxidative dye is formed from at least one primary intermediate and at least one coupler.

The coupler is typically an aromatic or heteroaromatic compound having 1 or 2 aromatic or heteroaromatic rings. Many couplers will be known to the skilled person.

In some embodiments, a coupler comprises an aromatic (or heteroaromatic) ring substituted by at least two amine and/or hydroxy groups at positions which are at meta positions with respect to one another.

Examples of such couplers include, without limitation, m-phenylenediamine, 3- aminophenol, m-diaminopyridines (e.g. 2,6-diaminopyridine, 3,5-diaminopyridine), resorcinol, and substituted derivatives thereof (i.e., any of the aforementioned compounds with one or more substituents added), such as 4-amino-2-hydroxytoluene (5-amino-o-cresol), 5-amino-6-chloro-o-cresol, 2-methyl-5 hydroxyethylaminophenol, 3-amino-2,4-dichlorophenol, hydroxyethyl-3,4-methylenedioxyaniline, 2,6- diaminotoluene, 2,6-dihydroxyethylaminotoluene, 2,4-diaminoanisole, 2,4- diaminophenoxyethanol, 2-amino-4-hydroxyethylaminoanisole, 1 ,3-bis(2,4- diaminophenoxy)propane, 2,6-dimethoxy-3,5-pyridinediamine, 4-chlororesorcinol and 2-methylresorcinol.

Additional examples of couplers include, without limitation, naphthols (e.g. 1- naphthol) and substituted derivatives thereof, such as l-acetoxy-2-methylnaphthalene, 2,7-naphthalenediol and 1,5-naphthalenediol; benzodioxoles (e.g. 1,3-benzodioxole); as well as phenyl methyl pyrazolone, 2-aminopyridine, 2-amino-3-hydroxypyridine, 1,2,4- trihydroxybenzene, hydroquinone and pyrocatechol.

Oxidative dye precursors and combination thereof used in exemplary embodiments include:

4-aminophenol (4AMP), p-phenylenediamine (p-PND) as primary intermediates, and 3-aminophenol (3AMP) as coupler (which may be reacted in a molar ratio of about 1: 1: 1); 4 AMP and p-PND as primary intermediates, and resorcinol as coupler (which may be reacted in a molar ratio of about 1: 1: 1);

4AMP as primary intermediate, and 3AMP as coupler (which may be reacted in a molar ratio of about 1: 1);

4AMP as primary intermediate, and resorcinol as coupler (which may be reacted in a molar ratio of about 1: 1);

p-PND as primary intermediate, and 3AMP as coupler (which may be reacted in a molar ratio of about 1: 1);

p-PND as primary intermediate, and resorcinol as coupler (which may be reacted in a molar ratio of about 1: 1); and

p-PND without any additional precursor.

Typically, an oxidative dye is formed by oxidation of a primary intermediate to a quinonediimine or quinoneimine form, which than initiates electrophilic aromatic substitution of another precursor molecule (primary intermediate or coupler), followed by further oxidation to form a dye molecule which is dimeric i.e., formed from two linked precursor molecules. Further electrophilic aromatic substitution may occur, resulting in a larger, e.g. trimeric or tetrameric, dye molecule.

An example of covalent linking of two precursor molecules to form a dimeric dye molecule is depicted in Scheme I.

Scheme I

[0]=oxidation

An example of covalent linking of three precursor molecules to form a trimeric dye molecule is depicted in Scheme II. Scheme II

X=0 or NH; [0]=oxidation

In view of the above, many other structures of oxidative dye molecules will be apparent, based on different precursor molecules, e.g. as described herein and/or in the art, and/or additional reactions e.g. reactions which form tetrameric dye molecules, pentameric dye molecules, and so forth.

Without being bound by any particular theory, it is believed that the use of oxidative dyes is advantageous because of the especially wide variety of shades which may be obtained with various combinations of oxidative dye precursors, as well as the fact that such combinations have been thoroughly studied and skilled practitioners will therefore know how to obtain a desired shade by selecting a suitable combination of precursors. It is further believed that separation of a composition-of-matter comprising an oxidative dye molecule from free dye molecules and precursor molecules substantially eliminates the risk of adverse effects associated with oxidative dyes and their precursors.

According to an aspect of some embodiments of the present invention, there is provided a composition-of-matter as described herein, wherein the dye molecule is a reaction product of an oxidative dye precursor as described herein (including a combination of a primary intermediate and a coupler) and an oxidizing agent, as described herein.

In some embodiments, the composition-of-matter exhibits little or no adhesion to hair (except for hair primed by a cationic polymer as described herein).

In some embodiments, the composition-of-matter is readily removable from untreated, e.g. un-primed, hair by washing with water.

In some embodiments, the composition-of-matter is washable from hair (which has not been primed as described herein), within a time period of less than 5 minutes under a water shower stream of 20 liters per minutes and a water temperature of 37 °C, wherein the hair has a length of 5 cm.

In some embodiments, the composition-of-matter is washable within a time period of less than 4 minutes, less than 3 minutes, less than 2 minutes, less than 1 minutes and even less than 30 seconds or less than 20 seconds, under the above- indicated conditions.

It is to be understood that while the composition-of-matter described herein may require longer time periods for being washed off hair at higher or shorter lengths, the washability is determined by the above-indicated conditions.

It is further to be understood that the phrase "water shower stream" describes stream of water coming out from all types of available shower heads, including fixed, positionable and handheld shower heads, shower heads equipped with a selector for multiple spray patterns, as long as the indicated conditions of water stream and temperature are maintained for determining the time period required for washing the formulation from the hair.

It is further to be understood that by "washable" it is meant that at least 90 % of a composition that has been applied to the hair has been washed off the hair.

In some embodiments, at least 99 % of the composition-of-matter is washed off during a time period described herein.

III. Process of preparation:

As exemplified herein, a composition-of-matter such as described herein can be prepared in a relatively simple manner by reacting a macromolecule with a dye molecule or oxidative dye precursor(s).

Hence, according to another aspect of embodiments of the invention, there is provided a process for preparing a composition-of-matter which comprises a macromolecule and at least one dye molecule being in association with the macromolecule. The process comprises contacting a macromolecule such as described herein with a reaction mixture that comprises at least one dye molecule or oxidative dye precursor such as described herein.

It is to be understood that any composition-of-matter prepared according to the process described herein is within the scope of embodiments of the invention. In some embodiments, the reaction mixture comprises at least one oxidative dye precursor (of an oxidative dye molecule to be formed by the process) and an oxidizing agent. As exemplified herein, such a reaction mixture may be used to produce a composition-of-matter that comprises a macromolecule associated with an oxidative dye molecule, as described herein.

In some embodiments, a concentration of oxidative dye precursors in the reaction mixture is in a range of from 1 mM to 100 mM. In some embodiments, the concentration is in a range of from 3 mM to 30 mM. In exemplary embodiments, the concentration is about 10 mM.

Hydrogen peroxide is an exemplary oxidizing agent. In some embodiments, a concentration of hydrogen peroxide in the reaction mixture is in a range of from 0.05 % to 5 %. In some embodiments, the concentration is in a range of from 0.1 % to 2.5 %. In some embodiments, the concentration is in a range of from 0.2 % to 1.5 %. In some embodiments, the concentration is in a range of from 0.3 % to 1 %. In exemplary embodiments, the concentration is about 0.5 %. Other oxidizing agents that are suitable for use for preparing oxidative dyes can also be used in the context of the present embodiments.

In some embodiments, the reaction mixture comprises a dye molecule e.g. a dye molecule as described herein.

In some embodiments, a concentration of oxidative dye precursors in the reaction mixture is in a range of from 0.5 mM to 50 mM. In some embodiments, the concentration is in a range of from 1.5 mM to 15 mM.

In some embodiments, the dye molecule is a naphthoquinone dye such as 2- hydroxy-l,4-naphthoquinone (lawsone, Natural Orange 6) and 5 -hydroxy- 1,4- naphthoquinone juglone, Natural Brown 7). In exemplary embodiments, a concentration of 2-hydroxy-l,4-naphthoquinone in the reaction mixture is about 5 mM.

In some embodiments, a concentration of macromolecule in the reaction mixture is in a range of from 1 mg/ml mM to 100 mg/ml. In some embodiments, the concentration is in a range of from 3 mg/ml to 30 mg/ml. In exemplary embodiments, the concentration is about 10 mg/ml. In some embodiments, the reaction mixture further comprises an alkaline aqueous solution. In some embodiments, the alkaline solution is a carbonate solution (e.g. a K₂CO₃ and/or Na₂C0₃ solution). In some embodiments, a concentration of carbonate in the solution is in a range of from 0.2 M to 2 M. In some embodiments, the concentration is in a range of from 0.3 M to 1.5 M. In some embodiments, the concentration is in a range of from 0.4 M to 1 M. In exemplary embodiments, the concentration is about 0.6 M. Other alkaline solutions and/or alkalizing agents within a solution, that are suitable for use for preparing oxidative dyes can also be used in the context of the present embodiments.

In some embodiments, a pH of the reaction mixture is in a range of from 8 to 12. In some embodiments, a pH of the reaction mixture is in a range of from 8.5 to 11. In some embodiments, a pH of the reaction mixture is in a range of from 9 to 10. In exemplary embodiments, the pH of the reaction mixture is about 9.5.

In some embodiments, the reaction is terminated after a time period in a range of from about 5 minutes to about 60 minutes. In some embodiments, the reaction is terminated after a time period in a range of from about 10 minutes to about 40 minutes. In exemplary embodiments, the reaction is terminated after about 30 minutes.

In exemplary embodiments, bovine serum albumin (at a concentration of about 10 mg/ml) is reacted with about 10 mM of oxidative dye precursors or about 5 mM of 2-hydroxy-l,4-naphthoquinone. The oxidative dye precursors may comprise:

4-aminophenol (4AMP), p-phenylenediamine (p-PND) as primary intermediates, and 3-aminophenol (3AMP) as coupler, reacted in a molar ratio of about 1: 1: 1;

4 AMP and p-PND as primary intermediates, and resorcinol as coupler, reacted in a molar ratio of about 1: 1: 1;

4AMP as primary intermediate, and 3AMP as coupler, reacted in a molar ratio of about 1: 1;

4AMP as primary intermediate, and resorcinol as coupler, reacted in a molar ratio of about 1: 1;

p-PND as primary intermediate, and 3 AMP as coupler, reacted in a molar ratio of about 1: 1;

p-PND as primary intermediate, and resorcinol as coupler, reacted in a molar ratio of about 1 : 1 ; or p-PND without any additional precursor.

Exemplary reaction conditions comprise aqueous K₂CO₃ (pH 9.5) with H₂O₂ at a concentration of 0.5 %, the reaction being terminated after about 30 minutes.

In some embodiments, the process further comprises isolating the composition- of-matter. In some embodiments, isolation of the composition-of-matter is used to terminate the reaction described herein. In some embodiments, the isolating is by filtration e.g. by ultrafiltration. The skilled person will be capable of selecting a suitable molecular weight cut-off for filtration, based on the molecular weight of the macromolecule and the molecular weight of free dye molecules and/or dye precursors and/or breakdown products of the macromolecule.

Methods of coloring:

According to another aspect of embodiments of the invention, there is provided a method of coloring a surface, the method comprising contacting the surface with a dye composition that comprises a macromolecule and a dye molecule in association with the macromolecule, as described herein.

In some embodiments, the dye composition comprises any of the compositions- of-matter as described herein. In some embodiments, the composition-of-matter comprises an oxidative dye molecule (in association with the macromolecule) which is a reaction product of an oxidative dye precursor and an oxidizing agent, as described herein.

In exemplary embodiments, the dye composition comprises a composition-of- matter which comprises BSA as a macromolecule, being is association with any of the exemplary oxidative dyes exemplified herein.

The surface can be a bodily surface or any other inanimate surface.

Since, as described herein, a composition-of-matter or a dye composition comprising same as described can be water soluble and washable, suitable surfaces include those that are not subjected to washing.

In any of the embodiments described herein, when a surface to be colored is likely to be subjected to washing or to be contacted with water, the method further comprises, applying a priming composition, which comprises a cationic polymer such as described herein, onto a least a portion of the surface, to thereby obtain a cationic polymer coating a surface. The cationic polymer and the dye composition are such that upon application of the priming composition, e.g. the cationic polymer, on a surface, the dye selectively colors the cationic polymer. Thus, the surface becomes colored by coloring the cationic polymer applied thereon. In some embodiments, the cationic polymer is applied prior to contacting the surface with the dye composition, that is, contacting the surface with the dye composition is effected subsequent to applying the priming composition.

Herein, the phrase "selectively colors" refers to a substrate, e.g. a surface having the cationic polymer described herein applied thereon, being colored to a greater degree than another substrate e.g. a surface without the cationic polymer. By a "greater degree" it is meant that a greater concentration of a coloring agent, e.g. a dye described herein, is present after application of the dye composition, prior to and/or subsequent to washing of the substrates with water.

In some embodiments, "selectively colors" refers to non-washable coloring of the substrate. That is, after application of the dye (e.g. a dye composition as described herein), and a subsequent washing of the colored surface, as described herein in the context of "washable", a surface having the cationic polymer applied therein remains colored, with more than 80 %, preferably more than 90 % (e.g., from 90 % to 100 %) of the dye remaining on the surface upon washing, whereby a surface with a cationic polymer remains non-colored, with less than 20 %, preferably less than 10 % (e.g., from 1-10 %) of the dye remaining on the surface upon washing.

In exemplary embodiments, the surface to be colored is hair. Human and non- human hair may be colored using embodiments of the invention. The hair may be attached to a body or detached from a body, e.g. coloring wool, coloring hair for wigs.

As described herein, embodiments of the invention are especially useful for coloring hair attached to a body without exposing the body to harsh reagents and messy procedures.

Hence, in some embodiments, the hair is hair attached to a body of a subject. In some embodiments, the subject is human. In some embodiments, the hair is on a head of the subject.

In embodiments wherein the surface comprises hair, application of the cationic polymer onto the hair is performed so as to thereby obtain a cationic polymer coating individual hair strands. The cationic polymer may be applied to substantially all of the hair on a head, or to only a portion of the hair. The dye composition is then contacted with the coated hair strands. Such contacting may be effected by contacting the dye composition solely with hair onto which the cationic polymer has been applied, or by contacting hair including hair strands not coated by the cationic polymer.

Without being bound by any particular theory, it is believed that the two-step method described herein, comprising a priming step (by application of the cationic polymer) and a coloring step (by application of the dye composition) allows for high degree of accuracy in coloring a surface, particularly hair, because only surfaces subjected to both steps will be strongly colored. Thus, for example, a portion of the surface which has been inadvertently treated with a cationic polymer or dye composition will not be strongly colored unless inadvertently treated with both the cationic polymer and the dye composition. Consequently, a portion of a surface is considerably less likely to be inadvertently colored.

In some embodiments, the method further comprises selectively removing the dye composition from the hair onto which the cationic polymer has not been applied, for example, by washing with water. Such a procedure is facilitated by use of a composition-of-matter, e.g. as described herein, with lower adhesion to hair than to hair coated with the cationic polymer.

In some embodiments, the method further comprises removing a carrier of the dye composition from substantially all the hair, for example, by washing with water. In some embodiments, such a step is performed concomitantly with selectively removing the dye composition from the hair onto which the cationic polymer has not been applied.

For example, in some embodiments, the cationic polymer is applied to a portion of the hair, the dye composition is contacted with all of the hair, and hair then washed so as to remove the dye composition from the portions of the hair not coated by the cationic polymer. The washing may also remove a carrier of the dye composition from the hair, leaving only the composition-of-matter in the portions of the hair coated by the cationic polymer.

Application of the cationic polymer and coloring of only a portion of the hair may be useful for example, for coloring grey roots and/or for creating highlights in the hair. In some embodiments, the surface is contacted with a plurality of dye compositions. In some embodiments, each dye composition exhibits a different shade.

In some embodiments, the method is effected by contacting a portion of the surface, e.g. hair, with a first dye composition as described herein, and contacting another portion of the surface, e.g. hair, with a second dye composition as described herein (having a different shade than the first dye composition), and so forth (contacting each of a plurality of portions with a different dye composition) so as to color the surface with a plurality of shades, with different shades in different regions of the surface (e.g. hair).

In this manner a surface may be colored with any number of different shades, using different dye compositions.

The different dye compositions may comprise different compositions-of-matter, which provide different shades. In some embodiments, different compositions-of-matter may comprise the same dye molecules associated with macromolecules, but in different proportions, thereby resulting in different shades.

In any of the embodiments described herein, the method further comprises contacting hair coated with the cationic polymer with a conditioner selected to be capable of adhering to the cationic polymer. In some embodiments, contacting with a conditioner is effected after the hair has been contacted with a dye composition, such that the conditioner will not impede contact between the cationic polymer and composition-of-matter.

Any hair conditioner known in the art may be used with embodiments of the invention.

In some embodiments, the conditioner comprises a polymer.

In some embodiments, the conditioner comprises an anionic compound. In some embodiments, the conditioner comprises an anionic polymer.

In some embodiments, the conditioner comprises a lipophilic compound. In some embodiments, the conditioner comprises a lipophilic polymer.

In some embodiments, the conditioner comprises a polymer comprising both anionic groups and lipophilic moieties. In some embodiments, the polymer is a surfactant. Without being bound by any particular theory, it is believed that anionic compounds associate relatively strongly with the cationic polymer and also offset the positive charge of the cationic polymer, thereby effectively reducing the effect of the cationic polymer on surface properties of the hair strands. It is further believed that hydrophobic compounds also adhere to the cationic polymer-coated hair stands in view of their low water-solubility, and thereby reduce the cationic nature of the coated hair strands. It is further believed that a combination of lipophilic regions and moderately negatively charged regions is particularly effective for reproducing the surface properties of hair.

In any of the embodiments described herein, the applied composition-of-matter on hair primed with the cationic polymer (with or without application of a conditioner), as described herein, is resistant to shampooing.

Herein, the phrase "resistant to shampooing" means that less than 10 % of the composition-of-matter is removed by cleansing the hair once with shampoo.

In some embodiments, less than 10 % of the composition-of-matter is removed by cleansing the hair once with a standard shampoo. In some embodiments, less than 5 % of the composition-of-matter is removed by cleansing the hair once with shampoo. In some embodiments, less than 2 % of the composition-of-matter is removed by cleansing the hair once with shampoo. In some embodiments, less than 1 % of the composition- of-matter is removed by cleansing the hair once with shampoo.

In some embodiments, less than 10 % of the composition-of-matter is removed by cleansing the hair 4 times (on different days) with a standard shampoo. In some embodiments, less than 10 % of the composition-of-matter is removed by cleansing the hair 7 times (on different days) with a standard shampoo. In some embodiments, less than 10 % of the composition-of-matter is removed by cleansing the hair 10 times (on different days) with a standard shampoo.

Herein, a coloring is said to "last" so long as most (i.e., more than 50 %) of the coloring agent (e.g. composition-of-matter) remains on the surface (e.g. hair).

In some embodiments, the coloring afforded by the composition-of-matter lasts for at least 4 weeks. In some embodiments, the coloring afforded by the composition- of-matter lasts for at least 7 weeks. In some embodiments, the coloring afforded by the composition-of-matter lasts for at least 10 weeks. In some embodiments, the coloring afforded by the composition-of-matter lasts for at least 15 weeks. In some embodiments, the coloring afforded by the composition-of-matter lasts for at least 20 weeks. In some embodiments, the coloring afforded by the composition-of-matter lasts for at least 30 weeks. In some embodiments, the coloring afforded by the composition- of-matter lasts for the lifetime of the hair strands.

Priming composition:

As described herein, a cationic polymer may be used in various aspects of embodiments of the invention in combination with a dye composition described herein, in order to efficiently effect coloring by the dye composition.

In any of the embodiments described herein, the cationic polymer is used within a priming composition.

Herein, the term "priming composition" encompasses a cationic polymer described herein per se, and combinations of two or more different cationic polymers as described herein, as well as compositions comprising the cationic polymer and one or more additional ingredients, such as a carrier as described herein.

The cationic polymer is selected such that the compositions-of-matter as described herein is adhered thereto. Adherence of the composition-of-matter can be effected via, for example, physical interactions such as absorption, adsorption, swelling, entrapment, and the like.

In some embodiments, the cationic polymer is selected or designed such that the composition-of-matter is adhered thereto.

In any of the embodiments described herein, the cationic polymer is selected capable of adhering to hair (or any other surface to be colored).

In any of the embodiments described herein, the cationic polymer comprises amine groups. The amine groups may comprise primary amine groups, secondary amine groups, tertiary amine groups and/or quaternary amine groups. Quaternary amine groups are inherently cationic, and the other amine groups are usually cationic in neutral and acidic aqueous solutions.

As used herein, the term "amine" describes both a -NRxRy group and a - N⁺RxRyRz group, wherein Rx, Ry and Rz are each independently hydrogen, alkyl, cycloalkyl, heteroalicyclic, heteroaryl or aryl, as these terms are defined herein. When Rx, Ry or Rz is heteroalicyclic or heteroaryl, the neighboring (nitrogen) atoms are bound to a carbon atom in the heteroalicyclic or heteroaryl.

The amine group -NRxRy can therefore be a primary amine, where both Rx and Ry are hydrogen; a secondary amine, where Rx is hydrogen and Ry is alkyl, cycloalkyl, heteroalicyclic, heteroaryl or aryl; a tertiary amine, where each of Rx and Ry is independently alkyl, cycloalkyl, heteroalicyclic, heteroaryl or aryl.

The amine group -N⁺RxRyRz can be a quaternary amine, where each of Rx, Ry and Rz is independently alkyl, cycloalkyl, heteroalicyclic, heteroaryl or aryl; or a protonated form of a primary, secondary or tertiary amine described hereinabove, where Rz is hydrogen and Rx and Ry are as defined above.

In some embodiments, the cationic polymer comprises primary amine groups.

In some embodiments, the cationic polymer comprises primary alkylamine groups.

Herein, the phrase "primary alkylamine" refers to a group having the formula - Y-N¾, wherein Y is alkyl or cycloalkyl, and to its protonated form (-Y-NH₃ ⁺).

In some embodiments, Y is alkyl.

In some embodiments, the alkyl has from 1 to 10 carbon atoms.

In some embodiments, the primary alkylamine has at least two carbon atoms. Examples of such groups include aminoethyl, aminopropyl, aminobutyl, aminopentyl, aminohexyl, aminoheptyl, aminooctyl, aminononyl, aminodecyl, as well as longer alkylamines. In some embodiments, the primary alkylamine has from 2-10 carbon atoms. In some embodiments, the alkylamine has from 2-8 carbon atoms. In some embodiments, the alkylamine has from 2-6 carbon atoms. In some embodiments, the alkylamine has from 3-5 carbon atoms. In exemplary embodiments, the alkylamine has 4 carbon atoms.

In some embodiments, the alkylamine is non-substituted.

In some embodiments, the primary alkylamine group is a linear amino-n-alkyl group e.g. 2-aminoethyl, 3-amino-n-propyl, 4-amino-n-butyl, 5-amino-n-pentyl, 6- amino-n-hexyl, 7-amino-n-heptyl, 8-amino-n-octyl, 9-amino-n-nonyl, 10-amino-n- decyl, and so forth. 4-amino-n-butyl is an exemplary alkylamine group.

Without being bound by any particular theory, it is believed that alkylamine groups comprising such alkyl chains, and particularly unbranched alkyl chains, e.g. as in linear amino-n-alkyl groups, facilitate interaction between the amine groups and other substances, such as a surface of a hair and/or a composition-of-matter e.g. by reducing steric hindrance.

In some embodiments, the cationic polymer comprises a polyacrylamide derivative, e.g. polyacrylamide modified so as to include cationic groups. In some embodiments, the polyacrylamide comprises acrylamide residues, cross-linked acrylamide residues, e.g. N,N'-methylenebisacrylamide residues, and derivatized acrylamide residues having the general formula:

wherein:

Ri, R₂ and R₃ are each independently selected from the group consisting of hydrogen and alkyl having 1-4 carbon atoms; and

X is selected from the group consisting of alkyl, cycloalkyl, aryl, heteroalicyclic and heteroaryl. When X is heteroalicyclic or heteroaryl, the neighboring nitrogen atoms are bound to a carbon atom in the heteroalicyclic or heteroaryl.

In some embodiments, each of Ri, R₂ and R₃ is hydrogen, such that the cationic amine group is a primary amine group.

In some embodiments, X is alkyl or cycloalkyl, such that the residue comprises an alkylamine group e.g. a primary alkylamine group as described herein. In some embodiments, X is alkyl.

In some embodiments, X is an alkyl having from 1 to 10 carbon atoms. In some embodiments, the alkyl has at least two carbon atoms. In some embodiments, the alkyl has from 2-10 carbon atoms. In some embodiments, the alkyl has from 2-8 carbon atoms. In some embodiments, the alkyl has from 2-6 carbon atoms. In some embodiments, the alkyl has from 3-5 carbon atoms. In exemplary embodiments, the alkyl has 4 carbon atoms (butyl). In some embodiments, X is non-substituted. In some embodiments, X is selected from the group consisting of non-substituted alkyl, cycloalkyl and aryl. In some embodiments, X is a non-substituted alkyl.

In some embodiments, X is a linear alkyl, wherein the neighboring -NR₁R₂R₃ and -NHC(=0)- groups are attached to the termini of the linear alkyl.

In some embodiments, the cationic polymer comprises a non-ionic polymer, e.g. polyacrylamide, modified so as to include cationic groups.

In some embodiments, a percentage of cationic residues in a cationic polymer described herein, e.g. a derivatized polyacrylamide described herein, is in a range of from 1 % to 50 % of all the residues. In some embodiments, the cationic residues are in a range of from 2 % to 30 % of the residues. In some embodiments, the cationic residues are in a range of from 3 % to 20 % of the residues. In some embodiments, the cationic residues are in a range of from 4 % to 15 % of the residues. In some embodiments, the cationic residues are in a range of from 6 % to 10 % of the residues.

In some embodiments, the concentration of cationic groups, e.g. amine groups, in a cationic polymer described herein is in a range of from 0.1 to 10 milliequivalents per gram of the cationic polymer (dry weight). In some embodiments, the concentration is in a range of from 0.2 to 5 milliequivalents per gram. In some embodiments, the concentration is in a range of from 0.3 to 3 milliequivalents per gram. In some embodiments, the concentration is in a range of from 0.5 to 2 milliequivalents per gram. In some embodiments, the concentration is in a range of from 0.7 to 1.5 milliequivalents per gram. In exemplary embodiments, the concentration is about 1.0 milliequivalents per gram.

In some embodiments, a cationic polymer as described herein is designed or selected as being spatially rigid, namely, having a minimal degree of free rotation. The degree of free rotation can be determined by the nature of the polymeric backbone, the nature and degree of cross-linking, and the nature and concentration of the cationic groups.

In some embodiments, the cationic polymer is water-insoluble. In some embodiments, the water-insoluble cationic polymer maintains a solid phase when in contact with water, thereby forming a solid or semi-solid substance e.g. a gel. The solid or semi-solid substance may be, for example, in a form of a film, discrete particles, e.g. beads, and/or a continuous bulk solid or semi-solid.

In some embodiments, the cationic polymer is in a form of particles being at least 0.25 μιη in width (along the narrowest dimension of the particles). In some embodiments, the particles are at least 0.5 μιη in width. In some embodiments, the particles are at least 1 μιη in width. In some embodiments, the particles are at least 2 μηι in width. In some embodiments, the particles are at least 4 μιη in width. In some embodiments, the particles are at least 8 μιη in width. In some embodiments, the particles are at least 16 μιη in width.

In some embodiments, the cationic polymer is in a form of particles no more than 100 μιη in width (along the narrowest dimension of the particles). In some embodiments, the particles are no more than 50 μιη in width. In some embodiments, the particles are no more than 25 μιη in width. In some embodiments, the particles are no more than 15 μιη in width. In some embodiments, the particles are no more than 10 μηι in width.

In exemplary embodiments, the cationic polymer is in a form of beads i.e. roundish particles.

Polymer beads may be prepared by polymerization in an emulsion e.g. wherein droplets in the emulsion become beads upon polymerization as described, for example, in Reuveny et al. [Biotechnology and Bioengineering 1983, 25:2969-2980] and Reuveny et al. [Biotechnology and Bioengineering 1983, 25:469-480]. In addition, many types of polymer beads are commercially available, for example, for use in various chromatographic techniques. The beads may be prepared from a cationic polymer, or derivatized subsequent to formation of the bead so as to include cationic groups.

In some embodiments, the cationic polymer is cross-linked. The cross-linking of the cationic polymer may be characterized by any type of cross-links known in the art, including, for example, cross-linking by cross-linking reagents which bind to more than one polymer chain e.g. glutaraldehyde, cross-linking by inclusion of a multi- functional monomer e.g. N,N'-methylenebisacrylamide in the polymer, and cross- linking by ionizing radiation, e.g. gamma ray irradiation, X-ray irradiation, UV irradiation, electron beam exposure, or oxidizing agents e.g. hydrogen peroxide. Cross-linking reduces the water-solubility of many polymers. In some embodiments, the cross-linking of the cationic polymer is sufficient to render the cationic polymer water-insoluble, whereas the non-cross -linked cationic polymer is water-soluble.

In some embodiments, the cationic polymer, e.g. a water-insoluble cationic polymer, is characterized by a porous polymer network. In some embodiments, the porous network forms a three dimensional gel structure.

In some embodiments, a porosity of the cationic polymer is characterized by an exclusion limit of at least 100 kDa. In some embodiments, the exclusion limit is at least 150 kDa. In some embodiments, the exclusion limit is at least 200 kDa. In some embodiments, the exclusion limit is at least 300 kDa.

In some embodiments, a porosity of the cationic polymer is characterized by an exclusion limit no more than 300 kDa. In some embodiments, the exclusion limit is no more than 200 kDa. In some embodiments, the exclusion limit is no more than 150 kDa. In some embodiments, the exclusion limit is no more than 100 kDa.

In some embodiments, a porosity of the cationic polymer is characterized by an exclusion limit in a range of from 50 to 200 kDa. In some embodiments, a porosity of the cationic polymer is characterized by an exclusion limit in a range of from 75 to 150 kDa.

Herein and in the art, an "exclusion limit" represents the maximal size of molecules capable of fitting into pores of a polymer. The exclusion limit may be calculated as known in the art, for example, by plotting a partition coefficient (which indicates the ability of a molecule to enter the polymer) of various molecules as a function of a logarithm of their molecular weight, and extrapolating to obtain a molecular weight at which the partition coefficient is expected to be zero i.e. the exclusion limit.

In some embodiments, the exclusion limit of the cationic polymer is higher than the molecular weight of the composition-of-matter used in combination with the cationic polymer, so as to facilitate absorption of the composition-of-matter by the cationic polymer. In some embodiments, the exclusion limit is at least 110 % of the molecular weight of the composition-of-matter. In some embodiments, the exclusion limit is at least 125 % of the molecular weight of the composition-of-matter. In some embodiments, the exclusion limit is at least 150 % of the molecular weight of the composition-of-matter. In some embodiments, the exclusion limit is at least 200 % of the molecular weight of the composition-of-matter.

In some embodiments, the exclusion limit of the cationic polymer is selected to be only moderately higher than a molecular weight of the composition-of-matter used in combination with the cationic polymer. In some embodiments, the exclusion limit is up to 300 % of the molecular weight of the composition-of-matter. In some embodiments, the exclusion limit is up to 200 % of the molecular weight of the composition-of-matter. In some embodiments, the exclusion limit is up to 150 % of the molecular weight of the composition-of-matter. In some embodiments, the exclusion limit is up to 125 % of the molecular weight of the composition-of-matter. In some embodiments, the exclusion limit is up to 110 % of the molecular weight of the composition-of-matter.

Without being bound by any particular theory, it is believed that having an exclusion limit which is not much higher than a molecular weight of the composition- of-matter (indicating a pore size which is not much larger than that required for the composition-of-matter to fit in the pores) is advantageous because it minimizes absorption of molecules other than the composition-of-matter and/or reduces desorption of the composition-of-matter from the cationic polymer by providing a tighter fit in the pores.

In some embodiments, the cationic polymer is substantially colorless. Such a cationic polymer will generally not affect the color of the surface, e.g. hair, significantly, except by mediating binding of the composition-of-matter to the surface.

Herein, the term "colorless" encompasses white, transparent and translucent appearances, as well as faint coloring which is not visible when the cationic polymer is applied to a surface.

In some embodiments, the colorless cationic polymer is characterized by an attenuation coefficient of less than 0.1 cm^"1 for all visible wavelengths (400-750 nm) of light.

In exemplary embodiments, the cationic polymer comprises cross-linked polyacrylamide beads derivatized to generate n-butylamine side chains at a predetermined charging degree of about 1.0 milliequivalent per gram (dry weight). Such a cationic polymer is suitable for use in combination with exemplary compositions-of-matter and/or dye compositions as described herein.

In some embodiments, a pH of the priming composition is in a range of from 5 to 8. In some embodiments, a pH of the priming composition is in a range of from 5.5 to 7.5. In some embodiments, a pH of the priming composition is in a range of from 6 to 7. In exemplary embodiments, a pH of the priming composition is about 6.5.

Formulation of priming composition and dye composition:

The dye composition and/or priming composition described herein may comprise one or more additional ingredients, such as a carrier, formulated in combination with the composition-of-matter of the dye composition and/or the cationic polymer of the priming composition.

In embodiments including a carrier, the chemical composition of the carrier is typically selected according to the desired form of the priming composition or dye composition. Further, the chemical composition of the carrier is selected so as to suit the desired purpose of the composition.

For example, compositions for coloring hair as described herein can be formulated in any form that is suitable for application to hair.

In some embodiments, the carriers for use in a dye composition and/or priming composition are selected so as to be quick-drying, facilitate the spreading of the composition through the hair but minimize dripping out of the hair and/or onto the scalp, are non-harmful, and do not cause the hair to have an unappealing feel, appearance or aroma.

For example, dripping may be prevented by using a viscous carrier, e.g. cream, gel and/or foam, which is fluid enough to facilitate spreading through the hair, but sufficiently viscous so as to avoid dripping, as well as by using a quick-drying spray carrier, in which the carrier is sprayed onto the desired location, and then dries up before dripping out of the desired location.

By selecting the appropriate carrier and optionally other ingredients that can be included in the composition, as is detailed herein, compositions described herein may be formulated into any form typically employed for topical application such as application to hair. Hence, the dye composition and priming composition can be, for example, in a form of a cream, an ointment, a paste, a gel, a lotion, a milk, a suspension, an aerosol, a spray, a foam, a shampoo, a hair conditioner, a swab, a pledget, a pad, and a soap.

Ointments are semisolid preparations, typically based on vegetable oil e.g. shea butter and/or cocoa butter, petrolatum or petroleum derivatives. The specific ointment base to be used is one that provides for optimum delivery for the composition-of-matter or cationic polymer chosen for a given formulation, and, preferably, provides for other desired characteristics as well (e.g. emolliency). As with other carriers or vehicles, an ointment base should be inert, stable, non-irritating and non- sensitizing.

Lotions are preparations that may to be applied to the hair without friction. Lotions are typically liquid or semi-liquid preparations in which solid particles, including, for example, particles of a composition-of-matter or cationic polymer described herein, are present in a water or alcohol base. Lotions are typically preferred for treating large areas, due to the ease of applying a more fluid composition. Lotions are typically suspensions of solids, and oftentimes comprise a liquid oily emulsion of the oil-in-water type. It is generally necessary that the insoluble matter in a lotion be finely divided. Lotions typically contain suspending agents to produce better dispersions as well as compounds useful for localizing and holding the composition-of- matter or cationic polymer in contact with the hair.

Creams are viscous liquids or semisolid emulsions, either oil-in-water or water- in-oil. Cream bases typically contain an oil phase, an emulsifier and an aqueous phase. The oil phase, also called the "internal" phase, is generally comprised of petrolatum and/or a fatty alcohol such as cetyl or stearyl alcohol. The aqueous phase typically, although not necessarily, exceeds the oil phase in volume, and generally contains a humectant. The emulsifier in a cream formulation is generally a nonionic, anionic, cationic or amphoteric surfactant.

Pastes are semisolid dosage forms in which the composition-of-matter or cationic polymer is suspended in a suitable base. Depending on the nature of the base, pastes are divided between fatty pastes or those made from single-phase aqueous gels. The base in a fatty paste is generally petrolatum, hydrophilic petrolatum and the like. The pastes made from single -phase aqueous gels generally incorporate carboxymethylcellulose or the like as a base. Gel formulations are semisolid, suspension-type systems. Single-phase gels contain organic macromolecular gelling agents distributed substantially uniformly throughout the carrier liquid, which is typically aqueous, but may also contain a nonaqueous solvent and, optionally, an oil. Examples of suitable organic gelling agents include, without limitation, crosslinked acrylic acid polymers such as the family of carbomer polymers, e.g. carboxypolyalkylenes that may be obtained commercially under the trademark Carbopol™; hydrophilic polymers such as polyethylene oxides, polyoxyethylene-polyoxypropylene copolymers and polyvinylalcohol; cellulosic polymers such as hydroxypropyl cellulose, hydroxyethyl cellulose, hydroxypropyl methylcellulose, hydroxypropyl methylcellulose phthalate, and methyl cellulose; gums such as tragacanth and xanthan gum; sodium alginate; and gelatin. In order to prepare a uniform gel, dispersing agents such as alcohol or glycerin can be added, or the gelling agent can be dispersed by trituration, mechanical mixing or stirring, or combinations thereof.

Sprays generally provide the hair coloring agent in an aqueous and/or volatile solvent solution which can be misted onto the hair for delivery. Such sprays include those formulated to provide for concentration of the composition-of-matter or cationic polymer at the site of administration following delivery, e.g. the spray solution can be primarily composed of a volatile liquid in which the composition-of-matter or cationic polymer can be dissolved or suspended. Upon delivery to the hair, the carrier evaporates, leaving concentrated composition-of-matter or cationic polymer at the site of administration.

Foam compositions are typically formulated in a single or multiple phase liquid form and housed in a suitable container, optionally together with a propellant which facilitates the expulsion of the composition from the container, thus transforming it into foam upon application. Other foam forming techniques include, for example the "Bag- in-a-can" formulation technique. Compositions thus formulated typically contain a low- boiling hydrocarbon, e.g. isopropane. Application and agitation of such a composition at body temperature causes the isopropane to vaporize and generate the foam, in a manner similar to a pressurized aerosol foaming system. Foams can be water-based or hydro-alcoholic, but are typically formulated with high volatile solvent content which, upon application to the hair of a user, quickly evaporates, leaving concentrated composition-of-matter or cationic polymer on the hair.

Representative examples of suitable carriers according to embodiments of the present invention therefore include, without limitation, water, liquid alcohols, liquid glycols, liquid polyalkylene glycols, liquid esters, liquid amides, liquid protein hydrolysates, liquid alkylated protein hydrolysates, liquid lanolin and lanolin derivatives, and like materials commonly employed in cosmetic compositions.

In some embodiments the dye composition comprises a solution of the composition-of-matter, that is, the composition-of-matter dissolves in at least a portion of the carrier. In some embodiments, the solution is an aqueous solution.

In some embodiments, the dye composition consists essentially of a solution, e.g. aqueous solution, of the composition-of-matter as described herein. In exemplary embodiments, the carrier of the dye composition is water, the dye composition being an aqueous solution of the composition-of-matter.

In some embodiments, the solution, e.g. aqueous solution, of the composition- of-matter represents a portion of the dye composition, for example, one phase, e.g. an aqueous phase, of an emulsion, or the liquid portion of a single-phase gel formulation.

In some embodiments, the carrier of the priming composition is an aqueous carrier, the cationic polymer being dissolved or suspended in the carrier. In some embodiments, the carrier is a buffered aqueous solution. The aqueous solution may be buffered so as to have a desired pH value such as described herein.

In exemplary embodiments, the carrier of the priming composition is a phosphate buffer with a pH of 6.5.

The carrier may further comprise additional ingredients, other than those listed herein.

Additional ingredients which may be included in a priming composition and/or dye composition described herein include, without limitation, coloring agents other than the composition-of-matter described herein, such as conventional hair coloring agents and other colorants; surfactants for enhancing solubility of ingredients and/or emulsification, for enhancing foaming, and/or for cleaning hair (shampoo); conventional hair conditioners, such as polyquaternium-10; substances which provide pearlescence, such as waxes; fragrances; antioxidants; preservatives; and pH regulators. In some embodiments, the dye composition is readily removable from un- primed hair by washing with water.

In some embodiments, the dye composition is washable (as defined herein) from hair (which has not been primed as described herein), with a time period of less than 5 minutes under a water shower stream of 20 liters per minutes and a water temperature of 37 °C, wherein the hair has a length of 5 cm. In some embodiments, the dye composition is washable within a time period of less than 4 minutes, less than 3 minutes, less than 2 minutes, less than 1 minutes and even less than 30 seconds or less than 20 seconds, under the above-indicated conditions. In some embodiments, a washable dye composition comprises a washable composition-of-matter (as described herein) in combination with a washable carrier.

Kits:

According to another aspect of embodiments of the invention, there is provided a kit for coloring hair. The kit comprises a priming composition comprising a cationic polymer as described herein and at least one dye composition comprising a macromolecule having at least one dye molecule associated with the macromolecule, as described herein e.g. a dye composition comprising at least one composition-of-matter such as described herein. In some embodiments, the priming composition and each dye composition are packaged individually within the kit.

In some embodiments, the cationic polymer and composition-of-matter are such that upon application of the priming composition on hair, e.g. according to a method described herein, the dye composition selectively colors (as this phrase is defined herein) the cationic polymer. Thus, the hair becomes colored by coloring the cationic polymer applied thereon.

In some embodiments, the kit comprises a plurality of dye compositions, each dye composition comprising at least one composition-of-matter, as described herein. Each dye composition has a different shade.

In some embodiments, the kit comprises at least 3 distinct dye compositions. In some embodiments, the kit comprises at least 4 distinct dye compositions. In some embodiments, the kit comprises at least 5 distinct dye compositions. In some embodiments, the kit comprises at least 6 distinct dye compositions. In some embodiments, the kit comprises at least 8 distinct dye compositions. In some embodiments, the kit comprises at least 10 distinct dye compositions. In some embodiments, the kit comprises at least 12 distinct dye compositions. In some embodiments, the kit comprises at least 15 distinct dye compositions. In some embodiments, the kit comprises at least 20 distinct dye compositions. In some embodiments, the kit comprises at least 30 distinct dye compositions.

In some embodiments, the different dye compositions comprise compositions-of- matter which share a common macromolecule, the common macromolecule being associated with different dye molecules in the compositions-of-matter of the different dye compositions, thereby resulting in the different shades of the different dye compositions.

In some embodiments, at least a portion of the different dye compositions comprise different macro molecules.

In some embodiments, the kit further comprises a conditioner selected to be capable of adhering to the cationic polymer, as described herein.

In some embodiments, the kit further comprises instructions for applying the priming composition and at least one dye composition to the hair, for example, in accordance with a method described herein.

In some embodiments, the instructions include instructions for applying one of the dye compositions to one portion of the hair, and another of the dye compositions to another portion of the hair, so as to obtain hair colored with a plurality of shades.

In some embodiments, the instructions include instructions for applying the priming composition to hair prior to applying a dye composition to hair.

In some embodiments, the instructions include instructions for applying the priming composition to a portion of the hair at which coloring is desired.

Applicators:

According to another aspect of embodiments of the invention, there is provided an applicator configured for applying to hair a priming composition comprising the cationic polymer e.g. as described herein. In some embodiments, the applicator is configured for applying the cationic polymer onto a portion of the hair.

According to another aspect of embodiments of the invention, there is provided an applicator configured for contacting hair with at least one dye composition such as described herein. In some embodiments, the applicator is configured for contacting a portion of the hair with the dye composition(s).

In some embodiments, there is provided an applicator configured for contacting hair with at least one dye composition such as described herein and for applying to hair a priming composition.

In some embodiments, any of the abovementioned applicators is configured for being capable of applying a composition, e.g. dye composition and/or priming composition, onto a portion of hair less than 4 cm in width. In some embodiments, the portion is less than 3 cm in width. In some embodiments, the portion is less than 2 cm in width. In some embodiments, the portion is less than 1 cm in width. In some embodiments, the portion is less than 0.5 cm in width.

Without being bound by any particular theory, it is believed that such applicators with high resolution are particularly suitable for utilizing the advantageous ability of embodiments of invention to provide spatial accuracy in coloring a portion of hair (as described herein).

In some embodiments, the applicator is configured for applying at least two dye compositions. The applicator may be configured for applying the dye compositions simultaneously, e.g. so as to color hair with a mixture of dye compositions, and/or sequentially e.g. so as to color different portions of the hair with different dye compositions. In some embodiments, the applicator is configured for applying at least 3 dye compositions. In some embodiments, the applicator is configured for applying at least 4 dye compositions. In some embodiments, the applicator is configured for applying at least 5 dye compositions. In some embodiments, the applicator is configured for applying at least 6 dye compositions.

In some embodiments, the applicator is in communication with a plurality of reservoirs for comprising different dye compositions, and the applicator is configured for contacting at least a portion of hair with any dye compositions in a reservoir. In some embodiments, the applicator is further in communication with a reservoir for comprising priming composition.

In some embodiments, the reservoirs are fixed to the applicator, e.g. the applicator and reservoirs are marketed as a single object. In some embodiments, the reservoirs are detachable and may be marketed separately, for example, in a form of replaceable cartridges comprising a dye composition or priming composition.

In some embodiments the detachable reservoirs are configured so as to be refillable.

In some embodiments the detachable reservoirs are configured so as to be disposable, for being replaced when depleted.

In some embodiments, the applicator is configured for being attached to a plurality of reservoirs, the plurality of reservoirs being selected in accordance with a desired coloring task.

In some embodiments, the applicator comprises one or more knobs, buttons, switches or the like, each being configured so as to induce release of one or more identified compositions when manipulated.

The applicator is preferably configured for facilitating flow of a composition having the consistency of the composition to be applied. For example, in embodiments wherein the cationic polymer comprises solid or semisolid particles, e.g. beads, such as described herein, the applicator for applying the cationic polymer should be configured to allow flow of such particles.

Any of the applicators described herein may be utilized according to other aspects of embodiments of the invention described herein.

Thus, in some embodiments, a kit described herein further comprises at least one applicator described herein. In some embodiments, the kit further comprises a plurality of reservoirs such as described herein, which maybe empty or filled with compositions included in the kit, as described herein.

In some embodiments, a method described herein is effected by using an applicator described herein for applying the priming composition and/or the dye composition described herein.

General:

The term "alkyl", as used herein, describes a saturated aliphatic hydrocarbon including straight chain and branched chain groups. Preferably, the alkyl group has 1 to 20 carbon atoms. Whenever a numerical range; e.g. "1-20", is stated herein, it implies that the group, in this case the alkyl group, may contain 1 carbon atom, 2 carbon atoms, 3 carbon atoms, etc., up to and including 20 carbon atoms. More preferably, the alkyl is a medium size alkyl having 1 to 10 carbon atoms. Most preferably, unless otherwise indicated, the alkyl is a lower alkyl having 1 to 4 carbon atoms. The alkyl group may be substituted or non-substituted. Substituted alkyl may have one or more substituents, whereby each substituent group can independently be, for example, cycloalkyl, alkenyl, alkynyl, aryl, heteroaryl, heteroalicyclic, amine, halide, sulfonate, sulfoxide, phosphonate, hydroxy, alkoxy, aryloxy, thiohydroxy, thioalkoxy, thioaryloxy, cyano, nitro, azo, sulfonamide, carbonyl, carboxy, thiocarbamate, urea, thiourea, carbamate, amide, guanyl, guanidine and hydrazine.

The alkyl group can be an end group, as this phrase is defined herein, wherein it is attached to a single adjacent atom, or a linking group e.g. an alkylene, as this phrase is defined herein, which connects two or more moieties.

Herein throughout, the phrase "end group" describes a group (a substituent) that is attached to a single moiety in the compound via one atom thereof.

The phrase "linking group" describes a group (a substituent) that is attached to two or more moieties in the compound.

The term "cycloalkyl" describes an all-carbon monocyclic or fused ring (i.e., rings which share an adjacent pair of carbon atoms) group where one or more of the rings does not have a completely conjugated pi-electron system. The cycloalkyl group may be substituted or unsubstituted. Substituted cycloalkyl may have one or more substituents, whereby each substituent group can independently be, for example, cycloalkyl, alkenyl, alkynyl, aryl, heteroaryl, heteroalicyclic, amine, halide, sulfonate, sulfoxide, phosphonate, hydroxy, alkoxy, aryloxy, thiohydroxy, thioalkoxy, thioaryloxy, cyano, nitro, azo, sulfonamide, carbonyl, carboxy, thiocarbamate, urea, thiourea, carbamate, amide, guanyl, guanidine and hydrazine. The cycloalkyl group can be an end group, as this phrase is defined hereinabove, wherein it is attached to a single adjacent atom, or a linking group, as this phrase is defined hereinabove, connecting two or more moieties.

The term "aryl" describes an all-carbon monocyclic or fused-ring polycyclic (i.e., rings which share adjacent pairs of carbon atoms) groups having a completely conjugated pi-electron system. The aryl group may be substituted or unsubstituted. Substituted aryl may have one or more substituents, whereby each substituent group can independently be, for example, cycloalkyl, alkenyl, alkynyl, aryl, heteroaryl, heteroalicyclic, amine, halide, sulfonate, sulfoxide, phosphonate, hydroxy, alkoxy, aryloxy, thiohydroxy, thioalkoxy, thioaryloxy, cyano, nitro, azo, sulfonamide, carbonyl, carboxy, thiocarbamate, urea, thiourea, carbamate, amide, guanyl, guanidine and hydrazine. The aryl group can be an end group, as this term is defined hereinabove, wherein it is attached to a single adjacent atom, or a linking group, as this term is defined hereinabove, connecting two or more moieties.

The term "heteroaryl" describes a monocyclic or fused ring (i.e., rings which share an adjacent pair of atoms) group having in the ring(s) one or more atoms, such as, for example, nitrogen, oxygen and sulfur and, in addition, having a completely conjugated pi-electron system. Examples, without limitation, of heteroaryl groups include pyrrole, furan, thiophene, imidazole, oxazole, thiazole, pyrazole, pyridine, pyrimidine, quinoline, isoquinoline and purine. The heteroaryl group may be substituted or unsubstituted. Substituted heteroaryl may have one or more substituents, whereby each substituent group can independently be, for example, cycloalkyl, alkenyl, alkynyl, aryl, heteroaryl, heteroalicyclic, amine, halide, sulfonate, sulfoxide, phosphonate, hydroxy, alkoxy, aryloxy, thiohydroxy, thioalkoxy, thioaryloxy, cyano, nitro, azo, sulfonamide, carbonyl, carboxy, thiocarbamate, urea, thiourea, carbamate, amide, guanyl, guanidine and hydrazine. The heteroaryl group can be an end group, as this phrase is defined hereinabove, where it is attached to a single adjacent atom, or a linking group, as this phrase is defined hereinabove, connecting two or more moieties. Representative examples are pyridine, pyrrole, oxazole, indole, purine and the like.

The term "heteroalicyclic" describes a monocyclic or fused ring group having in the ring(s) one or more atoms such as nitrogen, oxygen and sulfur. The rings may also have one or more double bonds. However, the rings do not have a completely conjugated pi-electron system. The heteroalicyclic may be substituted or unsubstituted. Substituted heteroalicyclic may have one or more substituents, whereby each substituent group can independently be, for example, cycloalkyl, alkenyl, alkynyl, aryl, heteroaryl, heteroalicyclic, amine, halide, sulfonate, sulfoxide, phosphonate, hydroxy, alkoxy, aryloxy, thiohydroxy, thioalkoxy, thioaryloxy, cyano, nitro, azo, sulfonamide, carbonyl, carboxy, thiocarbamate, urea, thiourea, carbamate, amide, guanyl, guanidine and hydrazine. The heteroalicyclic group can be an end group, as this phrase is defined hereinabove, where it is attached to a single adjacent atom, or a linking group, as this phrase is defined hereinabove, connecting two or more moieties. Representative examples are piperidine, piperazine, tetrahydrofuran, tetrahydropyran, morpholine and the like.

The term "phosphonate" refers to an -0-P(=0)(ORx)- end group or a -0-P(=0)-

0-, with Rx as defined hereinabove.

The term "halide" and "halo" describes fluorine, chlorine, bromine or iodine. The term "aminoalkyl" describes an alkyl group as defined above, further substituted by one or more amines.

The term "sulfoxide" or "sulfinyl" describes a -S(=0) Rx end group or an -

S(=0)- linking group, as these phrases are defined hereinabove, where Rx is as defined hereinabove.

The terms "sulfonate" and "sulfonyl" describe a -S(=0)₂-Rx end group or an - S(=0)₂- linking group, as these phrases are defined hereinabove, where Rx is as defined herein.

The term "sulfonamide", as used herein, encompasses both S-sulfonamides and N-sulfonamides.

The term "S- sulfonamide" describes a -S(=0)₂-NRxRy end group or a -S(=0)₂- NRx- linking group, as these phrases are defined hereinabove, with Rx and Ry as defined herein.

The term "N- sulfonamide" describes an RxS(=0)₂-NRy- end group or a -S(=0)₂-NRx- linking group, as these phrases are defined hereinabove, where Rx and Ry are as defined herein.

The term "carbonyl" as used herein, describes a -C(=0)-Rx end group or a - C(=0)- linking group, as these phrases are defined hereinabove, with Rx as defined herein.

The term "aldehyde" as used herein, describes a carbonyl end group wherein Rx is hydrogen.

The terms "hydroxy" and "hydroxyl" describe a -OH group.

The term "alkoxy" describes both an -O-alkyl and an -O-cycloalkyl group, as defined herein. The term "aryloxy" describes both an -O-aryl and an -O-heteroaryl group, as defined herein.

The term "thiohydroxy" describes a -SH group.

The term "thioalkoxy" describes both a -S-alkyl group, and a -S-cycloalkyl group, as defined herein.

The term "thioaryloxy" describes both a -S-aryl and a -S-heteroaryl group, as defined herein.

The terms "cyano" and "nitrile" describe a -C≡N group.

The term "nitro" describes an -N0₂ group.

The term "azo" describes an -N=NR' end group or an -N=N- linking group, as these phrases are defined hereinabove, with R' as defined hereinabove.

The terms "carboxy" and "carboxyl", as used herein, encompasses both C- carboxy and O-carboxy groups.

The term "C-carboxy" describes a -C(=0)-ORx end group or a -C(=0)-0- linking group, as these phrases are defined hereinabove, where Rx is as defined herein.

The term "O-carboxy" describes a -OC(=0)-Rx end group or a -OC(=0)- linking group, as these phrases are defined hereinabove, where R' is as defined herein.

The term "urea" describes a -NRxC(=0)-NRyRw end group or a -NR_xC(=0)- NRy- linking group, as these phrases are defined hereinabove, where Rx and Ry are as defined herein and Rw is as defined herein for Rx and Ry.

The term "thiourea" describes a -NRx-C(=S)-NRyRw end group or a -NRx- C(=S)-NRy- linking group, with Rx, Ry and Ry as defined herein.

The term "amide", as used herein, encompasses both C-amides and N-amides.

The term "C-amide" describes a -C(=0)-NRxRy end group or a -C(=0)-NRx- linking group, as these phrases are defined hereinabove, where Rx and Ry are as defined herein.

The term "N-amide" describes an RxC(=0)-NRy- end group or an RxC(=0)-N- linking group, as these phrases are defined hereinabove, where Rx and Ry are as defined herein.

The term "carbamyl" or "carbamate", as used herein, encompasses both N- carbamates and O-carbamates. The term "N-carbamate" describes an RyOC(=0)-NRx- end group or a -OC(=0)-NRx- linking group, as these phrases are defined hereinabove, with Rx and Ry as defined herein.

The term "O-carbamate" describes an -OC(=0)-NRxRy end group or an - OC(=0)-NRx- linking group, as these phrases are defined hereinabove, with Rx and Ry as defined herein.

The term "thiocarbamyl" or "thiocarbamate", as used herein, encompasses both O-thiocarbamates and N-thiocarbamates.

The term "O-thiocarbamate" describes a -OC(=S)-NRxRy end group or a -OC(=S)-NRx- linking group, as these phrases are defined hereinabove, with Rx and Ry as defined herein.

The term "N-thiocarbamate" describes an RyOC(=S)NRx- end group or a -OC(=S)NRx- linking group, as these phrases are defined hereinabove, with Rx and Ry as defined herein.

The term "guanyl" describes an RxRyNC(=N)- end group or an -RxNC(=N)- linking group, as these phrases are defined hereinabove, where Rx and R_y are as defined herein.

The term "guanidine" describes an -RxNC(=N)-NRyRw end group or an - RxNC(=N)-NRy- linking group, as these phrases are defined hereinabove, where Rx, Ry and Rw are as defined herein.

The term "hydrazine", as used herein, describes a -NRx-NRyRw end group or a - NR_x-NRy- linking group, as these phrases are defined hereinabove, with Rx, Ry, and Rw as defined herein.

As used herein the term "about" refers to ± 10 %.

The terms "comprises", "comprising", "includes", "including", "having" and their conjugates mean "including but not limited to".

The term "consisting of means "including and limited to".

The term "consists essentially of means that the composition, method or structure may include additional ingredients, steps and/or parts, but only if the additional ingredients, steps and/or parts do not materially alter the basic and novel characteristics of the claimed composition, method or structure. As used herein, the singular form "a", "an" and "the" include plural references unless the context clearly dictates otherwise. For example, the term "a compound" or "at least one compound" may include a plurality of compounds, including mixtures thereof.

Throughout this application, various embodiments of this invention may be presented in a range format. It should be understood that the description in range format is merely for convenience and brevity and should not be construed as an inflexible limitation on the scope of the invention. Accordingly, the description of a range should be considered to have specifically disclosed all the possible subranges as well as individual numerical values within that range. For example, description of a range such as from 1 to 6 should be considered to have specifically disclosed subranges such as from 1 to 3, from 1 to 4, from 1 to 5, from 2 to 4, from 2 to 6, from 3 to 6 etc., as well as individual numbers within that range, for example, 1, 2, 3, 4, 5, and 6. This applies regardless of the breadth of the range.

Whenever a numerical range is indicated herein, it is meant to include any cited numeral (fractional or integral) within the indicated range. The phrases "ranging/ranges between" a first indicate number and a second indicate number and "ranging/ranges from" a first indicate number "to" a second indicate number are used herein interchangeably and are meant to include the first and second indicated numbers and all the fractional and integral numerals therebetween.

As used herein the term "method" refers to manners, means, techniques and procedures for accomplishing a given task including, but not limited to, those manners, means, techniques and procedures either known to, or readily developed from known manners, means, techniques and procedures by practitioners of the chemical, cosmetic, pharmacological, biological, biochemical and medical arts.

It is appreciated that certain features of the invention, which are, for clarity, described in the context of separate embodiments, may also be provided in combination in a single embodiment. Conversely, various features of the invention, which are, for brevity, described in the context of a single embodiment, may also be provided separately or in any suitable sub-combination or as suitable in any other described embodiment of the invention. Certain features described in the context of various embodiments are not to be considered essential features of those embodiments, unless the embodiment is inoperative without those elements.

Various embodiments and aspects of the present invention as delineated hereinabove and as claimed in the claims section below find experimental support in the following examples.

EXAMPLES

Reference is now made to the following examples, which together with the above descriptions illustrate some embodiments of the invention in a non limiting fashion. EXAMPLE 1

Preparation of dyed macromolecules

General procedure: A macromolecule (bovine serum albumin) is dyed by being mixed with standard reagents of an oxidative dye, namely dye precursors and oxidizing agent, e.g. 0.5 % H₂O₂, in aqueous solution, under alkaline conditions, e.g. pH 9.5.

Using this general procedure, the following dyed macromolecules were prepared:

Sample 1 - dark brown: 3.33 mM of 4-aminophenol (4 AMP), 3.33 mM of p- phenylenediamine (p-PND), 3.33 mM of 3-aminophenol (3AMP), 10 mg/ml bovine serum albumin (BSA) and 0.5 % H₂O₂ were incubated in aqueous 0.6 M K₂CO₃ (pH 9.5) for 30 minutes at room temperature.

Sample 2 - light brown: 3.33 mM of 4 AMP, 3.33 mM of p-PND, 3.33 mM of resorcinol, 10 mg/ml BSA and 0.5 % H₂O₂ were incubated in aqueous 0.6 M K₂CO₃ (pH 9.5) for 30 minutes at room temperature.

Sample 3 - medium dark brown: 5 mM of 4AMP, 5 mM of 3AMP, 10 mg/ml BSA and 0.5 % H₂O₂ were incubated in aqueous 0.6 M K₂CO₃ (pH 9.5) for 30 minutes at room temperature.

Sample 4 - dark yellow: 5 mM of p-PND, 5mM of resorcinol, 10 mg/ml BSA and 0.5 % H₂O₂ were incubated in aqueous 0.6 M K₂CO₃ (pH 9.5) for 30 minutes at room temperature.

Sample 5 - red-brown: 5 mM of p-PND, 5 mM of 3 AMP, 10 mg/ml BSA and

0.5 % H₂O₂ were incubated in aqueous 0.6 M K₂CO₃ (pH 9.5) for 30 minutes at room temperature. Sample 6 - yellow: 5 niM of 4AMP, 5 niM of resorcinol, 10 mg/ml BSA and 0.5 % H₂O₂ were incubated in aqueous 0.6 M K₂CO₃ (pH 9.5) for 30 minutes at room temperature.

Sample 7 - dark red: 10 mM of p-PND, 10 mg/ml BSA and 0.5 % H₂0₂ were incubated in aqueous 0.6 M K₂C0₃ (pH 9.5) for 30 minutes at room temperature.

In addition, bovine serum albumin was dyed by being mixed with a non- oxidative dye, as follows:

Sample 8 - light reddish brown: 5 mM of 2-hydroxy-l,4-naphthoquinone (Natural Orange 6, lawsone), the dye in henna, and 10 mg/ml BSA were incubated in aqueous 0.6 M K₂C0₃ (pH 9.5) for 30 minutes at room temperature.

For each of the abovementioned colors, the reaction was arrested by removal of unbound reagents from the macromolecules. The mixtures were centrifuged for 10 minutes at 4000 rotations per minute, using ultrafiltration Centricon tubes (cutoff: 50,000 Da), and washed three times with purified water.

In order to monitor the reaction, samples were also taken immediately after

(time = 0 minutes), 10 minutes and 25 minutes after initiation of the reaction, and the macromolecules were purified by ultrafiltration.

As shown in Figure 5, the colors of the dyed macromolecules developed gradually during the 30 minute reaction, but the reaction was mostly complete after 10 minutes.

Solutions of the purified dyed macromolecules after a 30 minute reaction are shown in Figure 6.

EXAMPLE 2

Activation of hair surface and hair coloring

In order to prime the hair surface, Bio-Gel® P60 cross-linked polyacrylamide (PAA) microbeads (diameter 45-90 μιη when hydrated) were chemically derivatized by controlled aminolysis with 1,4-diaminobutane, to generate n-butylamine side chains at a predetermined charging degree (PAA-AB) of 1.0 milliequivalent per gram (dry weight), using procedures such as described in Freeman et al. [Biotechnology and Bioengineering 2004, 86: 196-200], Reuveny et al. [Biotechnology and Bioengineering 1983, 25:2969-2980] and Reuveny et al. [Biotechnology and Bioengineering 1983, 25:469-480]. Dry polyacrylamide beads were heated in ethylene glycol with 1,4- diaminobutane. The derivatized beads were then filtered and washed with water and saline. The PAA-AB microbeads were suspended in 50 niM sodium phosphate buffer (pH 6.5), and added to samples of untreated white hair and partially bleached hair for 10 minutes incubation, followed by 3 washings with water.

As shown in Figures 7 A and 7B, the PAA-AB microbeads adhered to and thoroughly covered the hair surface.

Dyed protein samples (Sample 3 - medium dark brown, and Sample 8 - light reddish brown) prepared as described in Example 1, were added to samples of the primed hair (untreated white hair and partially bleached hair) as well as to unprimed hair samples which served as a control. Following incubation for 10 minutes, unbound dye was washed from the hair samples (3 times with water).

As shown in Figures 8 A and 8B, untreated white hair was colored medium dark brown and light reddish brown by the respective dyed proteins when primed beforehand with PAA-AB beads, but not in the absence of priming.

As shown in Figures 9 A and 9B, partially bleached hair was colored medium dark brown and light reddish brown by the respective dyed proteins when primed beforehand with PAA-AB beads, but not in the absence of priming. Significantly, in coloring the partially bleached hair (shown in Figure 9B, left) light reddish brown, a lighter shade was achieved (Figure 9B, right), as compared with the initial hair color.

As shown in Figure 10, the beads adhering to the hair surface were colored brown by the medium dark brown dyed protein.

These results indicate that a combination of dyed macromolecules as described herein and priming with adherent beads as described herein is effective for coloring hair externally, without performing chemical reactions in the hair. These results further indicate that the external dying obscures the initial hair color, such that lighter shades may be obtained without bleaching.

Although the invention has been described in conjunction with specific embodiments thereof, it is evident that many alternatives, modifications and variations will be apparent to those skilled in the art. Accordingly, it is intended to embrace all such alternatives, modifications and variations that fall within the spirit and broad scope of the appended claims. All publications, patents and patent applications mentioned in this specification are herein incorporated in their entirety by reference into the specification, to the same extent as if each individual publication, patent or patent application was specifically and individually indicated to be incorporated herein by reference. In addition, citation or identification of any reference in this application shall not be construed as an admission that such reference is available as prior art to the present invention. To the extent that section headings are used, they should not be construed as necessarily limiting.

Claims

WHAT IS CLAIMED IS:

1. A composition-of-matter comprising a macromolecule and at least one oxidative dye associated with said macromolecule, said oxidative dye being a reaction product of at least one oxidative dye precursor and an oxidizing agent.

2. The composition-of-matter of claim 1, wherein said oxidative dye is a reaction product of said at least one oxidative dye precursor and said oxidizing agent in an alkaline aqueous solution.

3. A process for preparing a composition-of-matter which comprises a macromolecule and at least one dye molecule being in association with said macromolecule, the process comprising contacting the macromolecule with a reaction mixture that comprises at least one oxidative dye precursor of the oxidative dye and an oxidizing agent.

4. The process of claim 3, wherein said reaction mixture further comprises an alkaline aqueous solution.

5. The process of claim 3, further comprising isolating said composition-of-matter by filtration.

6. The process of claim 3, wherein said macromolecule has a molecular weight of at least 10 kDa.

7. The process of claim 3, wherein said macromolecule comprises a polypeptide.

8. The process of claim 3, wherein said macromolecule has a neutral or negative net charge in aqueous solution.

9. The process of claim 3, wherein said at least one oxidative dye precursor is selected from the group consisting of 1-naphthol, l-acetoxy-2-methylnaphthalene, 2,7- naphthalenediol, 1,5-naphthalenediol, 2-aminopyridine, 2-amino-3-hydroxypyridine, 2,4,5, 6-tetraaminopyrimidine, 2,6-diaminopyridine, 2,6-dimethoxy-3,5-pyridinediamine, 2,5,6-triamino-4-pyrimidinol, l-hydroxyethyl-4,5-diaminopyrazole, hydroquinone, pyrocatechol, 4-amino-2-hydroxytoluene, 4-aminophenol, p-methylaminophenol, 3- aminophenol, 3-amino-2,4-dichlorophenol, 5-amino-6-chloro-o-cresol, 2-methyl-5- hydroxyethylaminophenol, 2-aminophenol, 2-amino-5-ethylphenol, 6-amino-m-cresol, 6-amino-2,4-dichloro-m-cresol, o-phenylenediamine, m-phenylenediamine, p- phenylenediamine, hydroxyethyl-p-phenylenediamine, N,N-bis(2-hydroxyethyl)-p- phenylenediamine, N-phenyl-p-phenylenediamine, 2-methoxy-p-phenylenediamine, 2- chloro-p-phenylenediamine, 2,6-diaminotoluene, 2,6-dihydroxyethylaminotoluene, 2,5- diaminotoluene, 2,5-diamino-hydroxyethylbenzene, 2,4-diaminoanisole, 2,4- diaminophenoxyethanol, 2-amino-4-hydroxyethylaminoanisole, benzodioxoles, 1,2,4- trihydroxybenzene, resorcinol, 4-chlororesorcinol, 2-methylresorcinol, l,3-bis(2,4- diaminophenoxy)propane, hydroxyethyl-3,4-methylenedioxyaniline, and phenyl-methyl- pyrazolone.

10. A composition-of-matter comprising a macromolecule and at least one oxidative dye associated with said macromolecule, the composition-of-matter being prepared according to the process of any one of claims 3-9.

11. The composition-of-matter of any one of claims 1 and 10, being water-soluble.

12. The composition-of-matter of claim 1, wherein said macromolecule has a molecular weight of at least 10 kDa.

13. The composition-of-matter of claim 1, wherein said macromolecule comprises a polypeptide.

14. The composition-of-matter of claim 1, wherein said macromolecule has a neutral or negative net charge in aqueous solution.

15. The composition-of-matter of claim 1, wherein said at least one oxidative dye precursor is selected from the group consisting of 1-naphthol, l-acetoxy-2- methylnaphthalene, 2,7-naphthalenediol, 1,5-naphthalenediol, 2-aminopyridine, 2- amino-3-hydroxypyridine, 2,4,5, 6-tetraaminopyrimidine, 2,6-diaminopyridine, 2,6- dimethoxy-3,5-pyridinediamine, 2,5,6-triamino-4-pyrimidinol, l-hydroxyethyl-4,5- diaminopyrazole, hydroquinone, pyrocatechol, 4-amino-2-hydroxytoluene, 4- aminophenol, p-methylaminophenol, 3-aminophenol, 3-amino-2,4-dichlorophenol, 5- amino-6-chloro-o-cresol, 2-methyl-5-hydroxyethylaminophenol, 2-aminophenol, 2- amino-5-ethylphenol, 6-amino-m-cresol, 6-amino-2,4-dichloro-m-cresol, o- phenylenediamine, m-phenylenediamine, p-phenylenediamine, hydroxyethyl-p- phenylenediamine, N,N-bis(2-hydroxyethyl)-p-phenylenediamine, N-phenyl-p- phenylenediamine, 2-methoxy-p-phenylenediamine, 2-chloro-p-phenylenediamine, 2,6- diaminotoluene, 2,6-dihydroxyethylaminotoluene, 2,5-diaminotoluene, 2,5-diamino- hydroxyethylbenzene, 2,4-diaminoanisole, 2,4-diaminophenoxyethanol, 2-amino-4- hydroxyethylaminoanisole, benzodioxoles, 1,2,4-trihydroxybenzene, resorcinol, 4- chlororesorcinol, 2-methylresorcinol, 1 ,3-bis(2,4-diaminophenoxy)propane, hydroxyethyl-3 ,4-methylenedioxyaniline, and phenyl-methyl-pyrazolone.

16. A kit for coloring hair, the kit comprising:

(i) a priming composition comprising a cationic polymer; and

(ii) at least one dye composition comprising a macromolecule having at least one dye molecule associated therewith,

said cationic polymer and said dye composition being packaged individually within the kit and being such that upon application of said priming composition on a hair of a subject, the dye composition selectively colors said cationic polymer.

17. The kit of claim 16, wherein the dye composition is washable from hair upon which said priming composition is not applied, within a time period of less than 5 minutes under a water shower stream of 20 liters per minutes and a water temperature of 37 °C, wherein said hair has a length of 5 cm.

18. The kit of claim 16, wherein said dye composition comprises a solution of said macromolecule associated with said at least one dye molecule.

19. The kit of claim 16, wherein said dye composition comprises a composition-of- matter defined in any one of claims 1-2 and 10-15.

20. The kit of claim 16, wherein said cationic polymer is colorless.

21. The kit of claim 16, wherein said cationic polymer comprises primary alkylamine groups.

22. The kit of claim 16, wherein said cationic polymer is a cross-linked polymer.

23. The kit of claim 22, wherein said cationic polymer is porous, and wherein a porosity of the cationic polymer is characterized by an exclusion limit in a range of from 50 to 200 kDa.

24. The kit of claim 16, wherein said cationic polymer comprises a polyacrylamide derivative.

25. The kit of claim 16, wherein said cationic polymer is in a form of beads.

26. The kit of any one of claims 16-25, further comprising a conditioner selected to be capable of adhering to said cationic polymer.

27. The kit of claim 16, comprising a plurality of dye compositions, each dye composition being individually packaged within the kit and comprising a macromolecule associated with at least one dye molecule, and each dye composition of said plurality of dye compositions having a different shade.

28. The kit of claim 16, further comprising instructions for applying said priming composition to hair prior to applying said dye composition to hair.

29. The kit of claim 16, further comprising instructions for applying said priming composition to a portion of the hair at which coloring is desired.

30. The kit of claim 16, further comprising an applicator configured for applying at least one of said at least one dye composition to hair.

31. The kit of claim 16, further comprising an applicator configured for applying said priming composition to hair.

32. An applicator for coloring hair, the applicator being configured for contacting a portion of the hair with at least one dye composition.

33. The applicator of claim 32, being in communication with a plurality of reservoirs for comprising different dye compositions, the applicator being configured for contacting said portion of the hair with any of said dye compositions.

34. An applicator for priming hair for coloring, the applicator being configured for applying a composition comprising a cationic polymer onto a portion of the hair.

35. The applicator of any one of claims 32-34, wherein said cationic polymer is in a form of beads.

36. A method of coloring hair, the method comprising contacting the hair with a dye composition comprising a composition-of-matter as defined in any one of claims 1-2 and 10-15, thereby coloring the hair.

37. A method of coloring a surface, the method comprising:

(i) applying a priming composition comprising a cationic polymer onto at least a portion of the surface, to thereby obtain a cationic polymer coating a surface; and (ii) subsequent to said applying, contacting the surface with a dye composition comprising a macromolecule having at least one dye molecule associated therewith, thereby coloring the surface, said cationic polymer and said dye composition being such that upon application of said cationic polymer on the surface, the dye composition selectively colors said cationic polymer.

38. A method of coloring hair, the method comprising:

(i) applying a priming composition comprising a cationic polymer onto the hair, to thereby obtain a cationic polymer coating hair strands; and

(ii) subsequent to said applying, contacting the hair with a dye composition comprising a macromolecule having at least one dye molecule associated therewith, thereby coloring the hair, said cationic polymer and said dye composition being such that upon application of said cationic polymer on a hair of a subject, the dye composition selectively colors said cationic polymer.

39. The method of claim 38, wherein the dye composition is washable from hair upon which said cationic polymer is not applied, within a time period of less than 5 minutes under a water shower stream of 20 liters per minutes and a water temperature of 37 °C, wherein said hair has a length of 5 cm.

40. The method of claim 38, wherein said cationic polymer is colorless.

41. The method of claim 38, wherein said cationic polymer comprises primary alkylamine groups.

42. The method of claim 38, wherein said cationic polymer is a cross-linked polymer.

43. The method of claim 38, wherein said cationic polymer is porous, a porosity of the cationic polymer being characterized by an exclusion limit in a range of from 50 to 200 kDa.

44. The method of claim 38, wherein said cationic polymer comprises a polyacrylamide derivative.

45. The method of claim 38, wherein said cationic polymer is in a form of beads.

46. The method of claim 38, further comprising contacting the coated hair with a conditioner selected to be capable of adhering to said cationic polymer.

47. The method of claim 38, wherein said applying a cationic polymer is effected by an applicator configured for applying to hair a composition comprising said cationic polymer.

48. The method of any one of claims 36 and 38, comprising contacting a portion of the hair with a first dye composition comprising a macromolecule having at least one dye molecule associated therewith, and contacting another portion of the hair with a second dye composition comprising a macromolecule having at least one dye molecule associated therewith, said first dye composition and said second dye composition having different shades, thereby coloring the hair with a plurality of shades.

49. The method of any one of claims 36 and 38, wherein said contacting is effected by an applicator configured for applying to hair at least one dye composition comprising a macromolecule having at least one dye molecule associated therewith.

50. The method of any one of claims 37 and 38, wherein said dye composition comprises the composition-of-matter as defined in any one of claims 1-2 and 10-15.