EP0403530A1 - Storage-stable compositions and methods for treating keratinous tissue - Google Patents

Abstract

Cette invention concerne de nouvelles compositions activées contenant des protéines, stables au stockage et comportant des agents réducteurs tels que du thioglycolate d'ammonium et, le cas échéant, des agents oxydants tels que le peroxyde d'hydrogène et/ou des antioxydants, de préférence des antioxydants volatils. Ces compositions sont utiles pour traiter des tissus kératineux cornés de mammifères tels que les cheveux et les ongles de l'homme et les sabots et la fourrure des animaux afin d'en améliorer la résistance et l'apparence. En outre, ces compositions aident à favoriser la pousse des cheveux et des ongles.This invention relates to novel activated compositions containing proteins, stable in storage and comprising reducing agents such as ammonium thioglycolate and, where appropriate, oxidizing agents such as hydrogen peroxide and / or antioxidants, preferably volatile antioxidants. These compositions are useful for treating horny keratinous tissues of mammals such as human hair and nails and animal hooves and fur in order to improve their resistance and appearance. In addition, these compositions help promote the growth of hair and nails.

Description

STORAGE—STABLE COMPOSITIONS AND METHODS FOR TREATING

KERATINOUS TISSUE

This is a continuation-in-part application of United States Patent Application Serial No. 223,295, entitled "Storage Stable Compositions and Methods for Treating Keratinous Tissue", filed July 22, 1988, which is a continuation-in-part application of United States Patent Application Serial No. 164,009, entitled "Compositions For Treating Keratinous Tissue", filed March 4, 1988. BACKGROUND OF THE INVENTION

This invention relates to novel storage stable activated protein containing compositions comprising reducing agents, oxidizing agents and/or anti-oxidants. The composi¬ tions are useful for conditioning horny keratinous tissue of mammals such as human hair and nails, and the hooves and fur of animals to improve their strength and appearance. In addi¬ tion, these compositions are useful for promoting hair growth.

United States Patent 4,438,102 describes compositions which are useful for promoting the growth of normal dermal and epidermal tissue, more specifically the compositions are use¬ ful to promote wound healing in the soft keratinous tissue of the epidermis. Several examples of wound healing are provided in the specification.

The compositions of the prior patent are described as containing defined percentages of thioglycollic acid, ammonium hydroxide, glycerine, citric acid, hydrogen peroxide, commer¬ cial gelatin, a lower alkanol, and a solvent such as acetone or diethyl ether. The commercial grade gelatins disclosed by the patent contain do not contain sufficient cysteinyl content for use in the present invention.

It is known to wave hair permanently by a first treat¬ ment with a reducing.mixture of thioglycollic acid and ammonium hydroxide. The mixture may also contain a protein. The hair is subsequently treated in a second step with an oxidizing agent such as hydrogen peroxide or sodium bromate. The procedures are described, for example, in U.S. Patents 4,158,704; 3,957,605; 3,842,848; and several others.

The reducing step is believed to break the disulfide bonds of cystine disulfide bridges in the hair to form sulfh dryl groups. The hair is shaped under tension or pres¬ sure in the presence of the reducing agent. In a subsequent step, the oxidizing agent is contacted with the hair while it is still under tension to oxidize the sulfhydryl groups and reform disulfide bonds with the hair in the new shape.

U.S. Patent 3,842,848 describes a method of bonding especially prepared hydrolyzed peptide products of keratinaceous materials to human hair. The process is effected by conducting the reducing step of permanent waving in the presence of the peptide products and, thereafter, in a second step, oxidizing.

It has now been discovered that activated protein com¬ positions which are used to chemically bond to hair and other horny keratinous tissue may be formulated with reducing agents, oxidizing agents and/or anti-oxidant components to produce compositions for treating horny keratinous tissue. Especially advantageous characteristics of these compounds include storage stability and the ability to effectuate a reduction and oxidation process to form covalent disulfide linkages without having to resort to two separate solutions. Thus, the essential ingredients of the compositions of this invention are the reducing agent, the oxidizing agent, the protein and/or the antioxidant„ Numerous additional com¬ ponents may also be included, for example water, bases, acids, buffering agents, emulsifying agents or surfactants, thick¬ eners, preservatives, coloring agents and perfuming agents.

It is a surprising result that compositions of the present invention may be formulated in such a way that activated proteinaceous material in combination with reducing agent and oxidizing agent may produce a composition which can activate natural keratinous tissue and bind activated protein to the activated natural keratinous tissue.

It is an object of the present invention to provide novel^' compositions containing at least one activated thiol- containing protein in combination with a reducing agent and an oxidizing agent. It is an additional object of the present invention to provide storage stable compositions for treating horny keratinous tissue of mammals at least in part by for- mulating the activated thiol-containing protein in combination with an antioxidant.

It is a further object of the present invention to provide storage stable compositions for treating horny keratinous tissue which are capable of maintaining the activity of an activated protein in the presence of an oxidiz¬ ing agent to promote the shelf-life of the composition, and which can promote the formation of disulfide linkages between the activated protein and the treated keratinous tissue in one composition.

It is an additional object of the present invention to provide a new method for conditioning horny keratinous tissue which includes reducing the horny keratinous tissue and oxidizing said tissue without having to apply two separate solutions to the treated tissue to perform the reducing and oxidizing steps. BRIEF DESCRIPTION OF THE INVENTION

The novel compositions of the present invention are aqueous compositions having a pH of from about 4.0 to about 9, preferably a pH of about 7 to about 8, and most preferably a pH of about 7.6 (physiological pH) . Compositions of the pres¬ ent invention may comprise a reducing agent, an activated protein, and an oxidizing agent, in addition to other less essential components. Other embodiments of the present inven¬ tion may comprise a reducing agent, an activated protein, and an antioxidant. The compositions containing an antioxidant may also contain an oxidizing agent. In those compositions where an anti-oxidant is used in combination with an oxidizing agent, it is preferred that the antioxidant used is a volatile antioxidant and the oxidizing agent used is a non-volatile oxidizing agent.

The protein used is an activated protein. An activated protein is a protein which has been subjected to a reducing agent, for example a thiol-containing compound, which results in the breaking of disulfid bonds of cystine residues within the protein structure to produce free thiol or mercap- tide groups on cysteine residues. The ability of a protein to bind to native keratin is believed to be related in part to the number of thiol or mercaptide groups on the protein which are free to bind to mercaptide or thiol groups on the native keratin. Also, it is believed that the ability of the com¬ position to oxidize the keratin mercaptide and a proximal protein mercaptide enhances the probability of disulfide covalent bond formation. Production of disulfide covalent bonds may produce the strongest interaction between activated protein and keratinous tissue. Any promotion in growth related to the binding of protein to keratinous tissue would be expected to be of greatest duration where covalent bonding as opposed to electrostatic or ionic binding occurs.

The protein in the compositions of the present inven¬ tion may react with and form chemical bonds with the hard keratin of human and animal hair, nails and skin, thus effect¬ ing decreased hair breakage, increased hair thickness, increased nail hardness, decreased nail splitting and delamination and attachment of moist hydrated proteins to skin. The compositions are therefore useful for treating human hair, nails and skin to chemically bond the activated protein. In addition, compositions of the present invention are useful to promote wound healing in mammals. The composi¬ tions may also function as topical pharmaceutical carriers and vehicles»

The amount of activated protein that bonds to the keratinous tissue will vary with the concentration of reducing agents in the composition and the number of activated thiol or mercapto groups in the activated protein and the keratinous tissue. The time that the reducing agent is in contact with the keratinous tissue is also important; the longer the keratinous tissue is in contact with the reducing agent, the greater will be likelihood of a protein-keratinous tissue covalent bond formation.

The treatment using compositions of the present inven¬ tion is preferably initially conducted at ambient tempera¬ tures, i.e., about 20'C to about 35°C; however, higher temperatures may be used. The keratinous tissue may be treated over a period of time ranging from about 10 minutes to about 6 hours. The compositions of the present invention may also be formulated as sustained or controlled release formula¬ tions for prolonging treatment beyond 6 hours. Reaction" is effected by bringing the compositions of the present invention into contact with the keratinous substrate to be treated and allowing the treated tissue to dry. The time of contact may be varied at will.

In certain embodiments of the present invention employing a volatile antioxidant, it may be preferred to sub¬ ject the treated hair or tissue to heat under a hair dryer or similar heat source to volatilize and remove the antioxidant. By removing the antioxidant in this way, oxidation to promote covalent disulfide formation by the oxygen in ambient air may be promoted. Preferred compositions may employ non-volatile oxidizing agents which may promote oxidation after the volatile antioxidants are removed from the formulations.

In a first aspect of the present invention, exemplary embodiments comprise about 0.01 to about 12.0% by weight of an activated protein component, about 0.1 to about 15% by weight of a compatible reducing agent, about 0.001 to about 4.0% by weight of an oxidizing agent, and at least one component selected from the group consisting of water, acids, bases, buffering agents, emulsifying agents or surfactants, thick¬ eners, preservatives, organic solvents, coloring agents and perfuming agents.

Alternative embodiments of the present invention may include an anti-oxidant instead of an oxidizing agent. Exemplary compositions comprise about 0.01 to about 12.0% by weight of an activated protein component, about 0.1 to about 15% by weight of a compatible reducing agent, about 0.01 to about 2.0% by weight of an antioxidant and at least one com¬ ponent selected from the group consisting of water, acids, bases, buffering agents, emulsifying agents or surfactants, thickeners, preservatives, organic solvents, coloring agents, preservatives and perfume agents.

Additional embodiments of the present invention com¬ prise about 0.01% to about 12.0% by weight of an activated protein component, about 0.1 to about 15% of a compatible reducing agent, about 0.001 to about 4.0% by weight of an oxidizing agent and about 0.01 to about 4.0% by weight of antioxidant, preferably a volatile antioxidant and at least one component selected from the group consisting of water, acids, bases, buffering agents, solvents, emulsifying agents or surfactants, thickeners, coloring agents, preservatives and perfume agents.

The compositions of the present invention are for¬ mulated to enhance the formation of free mercaptide or thiol groups in the protein and the keratinous tissue to maximize the probability that a free thiol in the protein and a free thiol in the keratinous tissue will form a covalent disulfide bond. The inclusion of an oxidizing agent in the same com¬ position as the reducing agent and activated protein is designed to maximize covalent disulfide formation without using a second oxidizing solution.

The compositions of the present invention may be for¬ mulated as gels, creams, lotions, sprays or liquids of varying viscosities. DETAILED DESCRIPTION OF THE INVENTION

In a first aspect of the present invention, exemplary compositions are comprised of an activated protein, a com¬ patible reducing agent, an oxidizing agent and at least one component selected from the group consisting of water, bases, acids, buffering agents, emulsifying agents or surfactants, thickeners, preservatives, organic solvents, coloring agents and perfuming agents.

In this first aspect of the invention, the activated protein comprises about 0.01 to about 12% by weight of the composition, preferably about 1.0 to about 5.0% by weight for certain formulations and about 6 to 10% by weight for other applications where more concentrated formulations are found to be advantageous.

Activated proteins for use in compositions of the present invention are preferably exemplified by proteins which have sufficient cysteinyl content, i.e., at least about 1 cysteine amino acid for every 200 amino acids in a peptide chain (approximately, at least about 0.5% by weight cysteine, preferably, at least about 1.0% by weight cysteine and most preferably, at least about 5% by weight cysteine) to covalently bind to the keratinous tissue of hair, skin and nails to produce a durable permanent bond to keratinous tis¬ sue. By permanent bond we mean that the protein is not easily washed or rubbed off from the keratinous tissue and becomes as permanent as normal hair and nails. A large number of exemplary proteins may be used in the present invention and include keratin, food proteins, for example, casein, alpha and beta-lactalbumin, seed proteins, for example, soybean proteins, linseed protein, cotton seed protein, corn protein and peanut protein, among others, hemogrlobin, insulin, myosin, zein, ovalbumin, hemoglobin, trypsin, chymotrypsin, chymotrypsinogen, elastases, thrombins, plasminogen, fibrinogen/fibrin, lysozyme, papain, human serum albumin, heat coagulable mucoproteins isolated from cartilage, bones and skin, gamma globulin blood proteins, and a number of the blood factor proteins, including, for example, factor VIII, XII, IXa and Xa, among others. Of course, proteins which contain large numbers of cysteinyl residues are preferred, because these proteins would form the greatest number of covalent bonds with the keratinous tissue and thus, produce the greatest durability. It is to be noted that the commercial gelatins disclosed in U.S. Patent No. 4,438,102 contain at most trace cysteinyl residues and most commercial grade gelatins, includ¬ ing Grade A edible gelatin, do contain undetectable amounts of cysteine. Such gelatin proteins, which do not contain suffi¬ cient cysteinyl content to covalently bind to keratinous tis¬ sue in any significant way, i.e., to produce a permanent attachment of the protein to the keratinous tissue, are there¬ fore not contemplated for use in the present invention.

Preferred proteins for use in the present invention include proteins containing high percentages by weight of cysteine, for example, ribonuclease Tl, human serum albumin and gamma globulins. An especially preferred protein for use in the present invention is keratin, because of its particularly high cysteine content (about 12% to about 17% by weight of cysteine) . Proteins are activated by being sub¬ jected to a reducing agent at a pH of about 9.0 or above for a time sufficient to produce free thiol group. This period is generally about 5 minutes up to about one hour. Activation periods of greater than one hour are less preferred because although such activation periods may marginally increase the amount of activated thiol groups in the protein, such periods may also result in hydrolysis of the protein into shorter, less advantageous peptide units. A number of globular proteins contain cysteinyl residues within hydrophobic pock¬ ets. To activate these proteins and expose cysteinyl groups which exist in hydrophobic pockets to the keratinous tissue, it may be necessary to subject the proteins to denaturation and activation so that an activated cysteinyl residue of the denatured protein may be placed in proximity to the cysteinyl residues in the keratinous tissue to promote covalent binding.

The proteins of the present invention are preferably activated in the presence of reducing agent separately before they are formulated with the other components, because the addition of components other than a reducing agent at a pH above about 9.0 may adversely affect the rate at which cystinyl disulfide bonds (-S-S-) in the protein are converted to cysteinyl mercaptide groups (S-H) . This may lower the overall activity of the protein. However, although less preferred, it is possible to generate activated protein after formulation by simply exposing unactivated protein to reducing agents during storage below pH 7.0, provided that the cysteinyl content of the protein is sufficiently high to activate sufficient cysteinyl residues to promote covalent binding to keratinous tissue.

The activated protein is preferably a keratin, but any protein which contains sufficient cysteinyl content to promote covalent binding to activated keratinous tissue is con¬ templated for use in the present invention. A particularly preferred keratin is Kerasol^^m from Croda Chemicals Interna¬ tional, Chesire, England. The molecular weight of proteins useful in the present invention preferably varies between about 5,000 and 500,000 Daltons, and most preferably varies between about 120,000 and 130,0-00 Daltons.

Reducing agents which are useful to activate protein in the present invention include sulfides, thiol-containing compositions including dithiothreitol, trithiohexitol, glutathione, cysteine, mercaptoethanol, thioglycerol, thioalkanoic acid and mercaptocarboxylic acid, for example, mercaptosuccinic acid, thiolactic acid and their pharmaceuti¬ cally acceptable salts, among others, including thioglycollic acid and salts of thioglycollic acid. Preferred reducing agents for activating the protein are thioglycerol, cysteine, thiolactic acid and thioglycollic acid, and their pharmaceuti¬ cally acceptable salts. An especially preferred reducing agent for use in the present invention is ammonium thioglycol¬ late. It is preferred that the reducing agent for activating the protein should be the same as the biologically compatible reducing agent which is used in the final formulation of the invention. The use of strong biologically incompatible reduc¬ ing agents to activate the protein are less preferred and may make the use of the protein more difficult because the reduc¬ ing agent may have to be removed from the activated protein before formulation.

Compositions of the present invention also contain a compatible reducing agent in an amount equal to about 0.1 to about 15% by weight of the formulation. Preferred composi¬ tions contain about 3.0 to about 10% by weight of a compatible reducing agent. The amount of compatible reducing agent varies according to the therapeutic use for which the composi¬ tions are intended, but generally falls within the range of about 3.0 to about 10% by weight. A compatible reducing agent is an agent which reduces cystinyl disulfide linkages in keratinous tissue to produce free thiol or mercaptide groups and is compatible with biological and/or pharmaceutical systems. Compatible reducing agents which are contemplated for use in the present invention include mercaptoethanol, dithiothreitol, glutathione, cysteine and salts of thioglycol¬ lic acid. An especially preferred reducing agent is ammonium thioglycollate.

Compositions of the present invention may additionally comprise about 0.001 to about 4.0% by weight of an oxidizing agent. Preferred embodiments comprise about 0.1 to about 1.0% of the oxidizing agent and most preferably comprise about 0.5% to about 1.0% of the oxidizing agent. The oxidizing agent is included in compositions of the present invention to enhance oxidation and promote the formation of covalent disulfide bonds between activated protein and keratinous tissue. Exemplary oxidizing agents include hydrogen peroxide (which may or may not be stabilized with known stabilizers, for exam¬ ple urea) and its salts including ammonium sulfate peroxide, urea peroxide, pyrophosphate peroxide, carbonate peroxide, organic peroxides including acetyl peroxide, benzoyl peroxide, among others, alkali metal perborates including sodium per¬ borate,, the alkali metal bromates including sodium and potas¬ sium bromate and sodium and potassium iodate. In embodiments of the present invention which do not include an antioxidant, hydrogen peroxide is the preferred oxidizing agent. In the embodiments which include hydrogen peroxide, it is preferred that the amount of hydrogen peroxide should be in an amount equal, to about 0.001 to about 1.5% by weight of the composi¬ tion and most preferably about 0.05 to about 1.0%. Where oxidizing agents other than hydrogen peroxide are used, a higher percentage by weight is usually used compared to hydrogen peroxide which has a high oxidation equivalent per unit weight.

In addition to the above-described ingredients, addi¬ tional components may be added to the formulation to enhance the effects of the compositions. In addition to water, addi¬ tional components may include bases, acids, buffering agents, solvents, emulsifying agents or surfactants, thickeners, pre¬ servatives, organic solvents, coloring agents and perfuming agents.

Exemplary acids and bases are added to adjust the pH of the formulation to desired levels. Preferred acids include organic acids for example acetic acid, citric acid and tartaric acid, among others, and inorganic phosphoric acid including its salts such as the salts of mono- and di- hydrogen phosphoric acid. The inorganic phosphoric acid salts may also be included in the formulations as buffering agents. Preferred bases include organic amines, for example. monoethanolamine, triethanolamine, trimethylamine and triethylamine. Most preferred bases include ammonium hydroxide.

Buffering agents, for example the inorganic phosphoric acid salts indicated above as well as other buffering agents, for example the salts of organic acids such as acetic acid and citric acid may be included in the formulations of the present invention in amounts effective to maintain the pH of the for¬ mulation over time. Preferably, the amount of buffering agent is no more than about 1.5% by weight of the formulation and most preferably is less than 0.75% by weight. The pH of the formulation may be a factor in determining its stability and in maintaining the activity of certain components in the for¬ mulation, especially the activated protein and the compatible reducing agent. Thus, a buffering agent may be included within the formulation to maintain the pH at a relatively con¬ stant level over time.

To add homogeneity to and promote the solubility of the formulation, certain organic solvents may be included. Among the solvents that may be useful in certain embodiments of the present formulation include water, soluble polar organic solvents for example, alkanols such as methanol, ethanol, propanol, butanol and carbonyl containing solvents for example acetone, butanone and the like, among others.

Additional solvents include ethers and amines, for example diethyl or dipropyl ether and trimethyl or triethyl amine. Trimethylamine and triethylamine may also be added as bases.

The solvent added to the formulation may enhance the solubility of certain components. Where liquid formulations are contemplated, it is sometimes advisable to add an organic solvent to promote the solubility of certain less polar com¬ ponents, which without the added organic solvent may be only marginally soluble in water resulting in formulations having more than one phase. The addition of the organic solvent may produce a uniform, homogeneous single phase.

Emollients may also be included, especially in lotions to produce a uniform, homgeneous single phase and provide other favorable characteristics. An especially preferred emollient for use in formulations of the present invention is PPG 15-sterol ether, which also may be added to the formula¬ tions of the present invention for its emulsifying character¬ istics.

An emulsifying agent or surfactant is often added to embodiments of the present invention to enhance the character¬ istics of the formulation, to promote the solubility of the protein and other components and the phase stability of the formulation. Such agents also provide detergent-like qualities to the formulations. Suitable surfactants or emul¬ sifying agents may be nonionic, anionic or amphoteric. Non- ionic emulsifying compositions include, for example the lower alkylene oxide condensation products of hydrophobic compounds, for example ethylene oxide condensation products with higher fatty acids, higher fatty alcohols or alkylated aromatic hydrocarbons, higher molecular weight polypropylene glycols, amide and amine condensation products of which N-bis (2- hydroxyethyl)-lauramide is exemplary. Preferred nonionic emulsifying compositions include polyoxyethylene ethers including polyoxyethyleneisohexadecyl ether, for example Arlasolve 200^'tm (available from ICI Americas, Wilmington Delaware) , polyoxyethylenelauryl ether, for example Brij 35^tm (ICI) , polyoxyethylenestearyl ether, for example Brij 72^tm, and Brij 78^tm (ICI) and polyoxypropylenestearyl ether, for example PPG-15 stearyl ether (Arlamol E, ICI) . Other exemplary emulsifiers include ethoxylated lanolin, for exam¬ ple, Lanogel 41 (Amerchol, Inc., Edison, N.J.). Exemplary anionic surfactants include sulfuric acid esters of polyhydric alcohols, e.g. lauryl sulfate, cetyl sulfate, etc., higher fatty alcohol sulfates derived from coconut oil, hydroxy sul- fonated higher fatty acid esters such as, e.g., higher fatty acid esters of 2,3-dihydroxy-propane sulfonic acid, higher fatty acid esters of low molecular weight alkylol sulfonic acids, e.g., oleic acid ester of isethionic acid, sulfated higher fatty acid alkylolamides such as e.g., ethanolamide sulfates, higher fatty acid amides of amine alkyl sulfonic acids, e.g., lauric amide of taurine, among others, and aromatic containing anionic synthetic surfactants. Exemplary amphoteric surfactants include the salts of N-alkyl compounds of betaamino propionic acid wherein the alkyl group is derived from a fatty acid such as a mixture of coconut oil fatty acids, among others.

It may be preferable to add an anti-foaming agent to certain compositions to promote homogeneity and prevent foam¬ ing from surfactant action. A preferred anti-foaming agent for use in embodiments of the present invention includes, for example, Dimethicone^tm, available from Dow Chemical Corp., Midland, Michigan.

Thickeners or gelling agents may be added to provide additional weight and a more viscous feel to the formulations. Suitable thickening agents include polyvinyl pyrollidone, for example PVP K30 (GAF Charllotte, N.C.) polyacrylates, car- bomers, for example carboxyvinyl polymer such as Carbapol 940 (available from B.F. Goodrich, Cleveland, Ohio) polyoxyethylene stearyl ethers, for example, polyoxyethylene-2 stearyl ether such as Steareth 2^tm (ICI) and polyoxyethylene- 20 stearyl ether such as Steareth 20^tm (ICI) , sodium alginate, carageenan, agar, ethoxylated polyvinyl alcohol, gums, for example methylcellulose, hydroxymethylcellulose, car- boxymethylcellulose, propylcellulose and hydroxypropylcel- lulose, acacia, tragacanth, guar, and quince, among others. In compositions which are contemplated to be formulated as a gel or lotion, Isoseteth 20^tm (polyoxyethyleneisohexadecyl ether, ICI) , and Steareth 2^tm and 20^t:m are preferred for use as thickening agents. In compositions which are contemplated to be formulated as creams, preferred thickeners include Steareth 2^tm and Steareth 20^tm and the carbomer polymers, for example Carbopol 940^tm.

Preservatives are added for preventing microbial growth in the presence of protein nutrients. Exemplary pre¬ servatives include benzoic acid analogs including, among others, sodium benzoate. Other presevatives include propyl and methyl paraben, Dowicil^tιn (Dow Chemical Corp., Midland, Mi.) and formaldehyde solution. An especially preferred pre¬ servative is Germaben II^^m, available from Sutton Laboratories, New Jersey.

Coloring agents and perfume agents may also be added to enhance the characteristics of the formulations.

In another aspect of the present invention, exemplary compositions include an antioxidant instead of an oxidizing agent. These compositions comprise about 0.01 to about 10% by weight of an activated protein component, about 0.1 to about 15% by weight of a compatible reducing agent, about 0.001 to about 2.0% by weight of an antioxidant, and at least one com¬ ponent selected from the group consisting of water, acids, bases, buffering agents, emulsifying agents or surfactants, thickeners, coloring agents and perfume agents.

In compositions comprising an antioxidant, the antioxidant is included to promote the storage stability of the formulations. Exemplary antioxidants may include alpha- tocopherol, hydroxyquinone, unipherol, tocopherol ascorbate, lecithin, chlorophyll, ascorbylpalmitate, linseed oil, tongue oil, other natural product antioxidants such as the steam dis¬ tillation extract of rosemary as disclosed in U.S. Patent No. 4,450,097, thiazoline carboxylate, dihydroquinolines, methyl gallate, propyl gallate, alkylaryl and diarylamines.

Certain chelating agents, for example, EDTA, may be employed to enhance the antioxidant effect of the above agents. The chelating agent may function to chelate any dis- solved metals which may be responsible for the in situ gener¬ ation of oxygen. Preferably, the chelating agent comprises between about 0.001 to about 0.5% of the formulation and most preferably the chelating agent comprises no more than about 0.1% of the formulation.

Preferably, in compositions which employ an antioxidant, a volatile antioxidant is used. Volatile antioxidants provide the advantage of protecting the activated protein and reducing agent from oxidation during storage. In addition, because the antioxidants are volatile, after the compositions are placed on the treatment area and exposed to air or a heat source, the antioxidant will evaporate from the treatment area leaving the remaining protein and activated keratinous tissue to be air oxidized. Volatile antioxidants include voltile carbonyl containing compounds, hindered phenolic compounds, for example 2,4,6-trialkyl phenols, buty- lated hydroxyanisole (BHA) , butylated hydroxytoluene (BHT) , p- hydroxytoluene and p-hydroxyanisole.

A volatile antioxidant as used in preferred embodi¬ ments of the present invention is an antioxidant that volatil¬ izes or evaporates from the treatment area under normal drying conditions. Non-volatile antioxidants, although useful in certain aspects of the present invention, are less preferred than are volatile antioxidants, which are added to formula¬ tions for their ability to stabilize the active ingredients over time while in storage and their ability to be removed from the treatment area under normal drying conditions. Preferred volatile antioxidants include those that are more easily volatilized, i.e., will evaporate more quickly from the treatment surface.

Certain antioxidants may be formulated in combination with solvents including water. This may promote azeotrope formation and volatility of the antioxidant. Azeotrope forma¬ tion with water or with other solvents may result in the antioxidant volatilizing at a temperature lower than normal. Thus, by formulating the compositions with, for example, an alcoholic or other solvent, the volatilization of the antioxidant may be enhanced, resulting in an- enhanced rate of oxidation of the treated keratinous tissue.

In preferred embodiments of the present invention, when an oxidizing agent is not included in the formulation, about 0.01% to about 2.0% of antioxidant is included in the formulations. Without the additional oxidizing agent, the antioxidant is included to prevent atmospheric oxygen or oxygen dissolved in the solution from deactivating the protein during storage. In compositions in which oxidizing agents are employed to promote the oxidation of free thiols or mercap- tides to covalent disulfide bonds, the oxidizing agent com¬ prises about 0.001% to about 4.0% by weight of an oxidizing agent and the antioxidant comprises about 0.01% to about 4.0% of the formulation.

In formulations of the present invention comprising an antioxidant or an antioxidant and oxidizing agent, the for¬ mulations may additionally comprise acids, bases, buffering agents, emulsifying agents or surfactants, thickeners, pre¬ servatives, organic solvents, coloring agents and perfume agents as described for other embodiments of the present invention. In formulations comprising an antioxidant, a volatile antioxidant is preferred. In such formulations addi¬ tional organic solvent may be added to promote the volatiliza¬ tion of the antioxidant. It is recognized that the choice of additives is made to avoid any interactions that may affect the activity of the the activated protein, reducing agent, oxidizing agent or antioxidant.

The compositions of the present invention are applied to the treatment area as a liquid, cream, gel or lotion by rubbing the compositions into the hair or other tissue to be treated. After the compositions have been in contact with the treated tissue for a time period of about 20 minutes to six hours, the treated area is then dried at ambient air tempera¬ ture or preferably, at elevated temperatures under a hair dryer or other heat source.

In a method for treating the nails of humans and the hooves of animals, compositions of the present invention which do not contain an oxidizing agent or an antioxidant may be used. Thus, the treatment of the nails of humans and the hooves of animals comprises exposing the nails or hooves to a composition comprising about 0.01 to about 12% by weight of an activated protein as described hereinabove, preferably keratin, about 0.1 to about 15% by weight of a pharmaceuti¬ cally compatible reducing agent as described hereinabove, preferably thioglycollic acid or ammonium thioglycollate and the remainder.of the composition comprises at least one com¬ ponent selected from the group consisting of water, acids, bases, buffering agents, emulsifying agents or surfactants, thickeners, preservatives, organic solvents, coloring agents and perfuming agents.

The following examples are provided to illustrate the present invention and should not be construed to limit the scope of the invention of the present application in any way.

EXAMPLE 1 Dilute Liquid

Component Weight Percent

Ammonium Thio¬ glycollate Ph 9 3.27 A Kerasol^tm 0.95

Propylene Glycol 0.05

Lanogel 41^tm 0„05 B

Brij 35^tm 0.13

PVP-K30^tm25% Sol. 0.22

Glycerine 0.16

Citric Acid Sol (5.88%) 0.04

Hyd. Peroxide (3%) 0.53 C

Acetone 0.13

Isopropyl Alcohol 70% 0.53 Kerasol^tm 1.90

Water - Distilled 85.40 D

Germaben II^tm 0.93 E

Fragrance 1.90

Arlasolve^tm 3.79 F

Procedure: Add a mixture of Component B to 1/2 of Component D at 100° F and mix for 5 minutes in a high speed blender. After thoroughly mixed, add a mixture of Component C and mix for 5 minutes. Let the mixture cool and add the balance of component D to the mixture at room temperature and mix for 2 minutes. Prepare A by adding 9.64% of a 28% ammonia solution to 90.64% of a 60% ammonium thioglycollate water solution. pH should read 9; if not, add more ammonia solution. Add kerasol^tm to ammonium thioglycollate solution and mix until well homogenized. Let stand for at least five minutes. This produces mixture A. Add A to the BDC mixture and then mix in a high speed blender until thoroughly mixed. Next, add com¬ ponent E to mixture ABCD until thoroughly mixed. Finally add components F to mixture ABCDE and mix at high speed until homogeneous. EXAMPLE II Concentrated Liquid

Component Weight Percent

Ammmonium Thio¬ glycollate pH 9 9.75 A Kerasol^tm 2.82

Propylene Glycol 0.13

Lanagel 41^tm 0.13 B

Brij 35^tm 0.39

PVP-K30^tm 25% 0.64

Glycerine 0.47

Citric Acid (5.88%) 0.12

Hydrogen Peroxide (3%) 1.55 C

Acetone 0.39

Isopropyl Alcohol 70% 1.68

Kerasol^tm 5.65

Water 67.81 D

Germaben II^tm 2.82 E

Fragrance and 5.65 F

Solubilizer (1:2)

Procedure: Add a mixture of Components B to 1/2 of Component D at 100° F and mix for 5 minutes in a high speed blender. After thoroughly mixed, add a mixture of Components C and mix for 5 minutes. Let mixture cool and add the balance of com¬ ponent D to the mixture at room temperature and mix for 2 minutes. Prepare A by adding 9.64% of a 28% ammonia solution, to 90.64% of a 60% ammonium thioglycollate water solution. pH should read 9; if not, add more ammonia solution. Add kerasol^^m to ammonium thioglycollate solution and mix until well homogenized. Let stand for at least five minutes. This produces mixture A. Add A to the BDC mixture and then mix in a high speed blender until thoroughly mixed. Next, add com¬ ponents E to mixture ABCD until thoroughly mixed. Finally add components F to mixture ABCDE and mix at high speed until homogeneous. Example III Lotion

Component Weight Percent

Example II Concentrated 32.19 A liquid

Non-Perfumed Moisturizing 45.85 B

Lotion

Carbopol 940 - 2% Sol 19.51 C

Triethylamine 1.225

H₂0 1.225 D

Procedure: Add Component A to Component B (see below) and thoroughly mix. Then add Component C until a homogeneous mix¬ ture is made. Finally, adjust the pH of mixture ABC with the triethylamine/H2θ mixture to about 7.0.

Non-Perfumed Moisturizing Lotion

Component Weight Percent

Arlamol E^tm 1.36

Brij 72^tm 5.23

Brij 78^tm 1.32

Mineral Oil 11.64

Propyl Paraben 0.18

Water 77.71

Sodium EDTA 0.09

Anti-Foam Dimethicone^tm 0.09 B

Methyl Paraben 0.36

Propylene Glycol 1.36

Dowicil 200^tm 0.45 C

Formaldehyde (37%) 0.21 D

Procedure forNon-perfumed Moisturizing Lotion Component: Add A to B at 160°F mixing thoroughly. Mix and cool to 100"F. Add Components C and D, mixing thoroughly. Allow to cool to 80^βF. Use at 80^βF. EXAMPLE IV Thin Cream

Component Weight Percent

Non-perfumed Moisturizing 8.70

Lotion Component

Carbopol 940^tm 58.26 A

Triethano1amine 2.61

Concentrated Liquid 28.70 B

Example II

Fragrance 1.74 C

Procedure: Mix Components of A together and adjust pH with triethanolamine. Add Component B and thoroughly mix until homogeneous. Mix in fragrance until homogeneous.

EXAMPLE V-THICK CREAM

Component Weight Percent

Example II Concentrated 30.22 A liquid

Concentrated Non-Perfumed 9.16 B

Moisturizing Lotion

Carbapol 940 - 4% Sol 58.50. C

Fragrance 2.14% D

Procedure: Add Component A to component B (see below) and thoroughly,mix. Then add Component C until a homogeneous mix¬ ture is made. Add Component D and mixture until a homogeneous thick cream is formed. Concentrated Non-Perfumed Moisturizing Lotion Component Weight Percent

Arlamol E^tm 2.33

Brij 72j^m 8.95

Brij 78^tm 2.26

Mineral Oil 19.94

Propyl Paraben 0.31

Water 63.15

Sodium EDTA 0.016

Anti-Foam Dimethicone^tm 0.016 B

Methyl Paraben 0.7

Propylene Glycol 2.33

Procedure for Concentrated Non-Perfumed Moisturizing Lotion Component: Add A to B at 160^βF mixing thoroughly. Mix and cool to 80°F. Use at 80°F.

EXAMPLE VI

Component Weight Percent

Ammonium Thioglycollate 0.1 to 4.0%

Ammonium Hydroxide 0.005 to 2.0%

Keratin 0.01 to 10.0%

Hydrogen Peroxide 0.001 to 1.0 9.

Water . 83.0 to 99.884%

Procedure: Mix the ammonium thioglycollate, ammonium hydroxide and a small amount of water to form a mixture with a pH of about 9 to 11. About half of the protein is added to this mixture which may be agitated or stirred for about 15 minutes followed by the addition of water, hydrogen peroxide and the remaining protein. Alternatively, after the first half of the protein is added, the water, hydrogen peroxide and remaining protein may be added. EXAMPLE VII

Component Percent by Weight

Ammonium Thioglycollate 0.1 to 4.0%

Ammonium hydroxide 0.05 to 2.0%

Hydrogen Peroxide 0.001 to 1.0%

Keratin 0.01 to 10.0%

Glycerine 0.15 to 0.25%

Citric Acid 0.095 to.0.29%

Solvent 0.16 to 1.28%

Water 81.18 to 99.434%

Procedure: Mix the ammonium thioglycollate, ammonium hydroxide and a small amount of water to form a mixture with a pH of about 9 to 11. About half of the protein is added to this mixture which may be agitated or stirred for about 15 minutes followed by the addition of water, hydrogen peroxide and the remaining protein. Alternatively, after the first half of the protein is added, the water, hydrogen peroxide and remaining protein may be added. The other ingredients are then added after the hydrogen peroxide and stirred or agitated sufficiently to produce a homogeneous composition.

EXAMPLE VIII Concentrated Stock Solution

Component Weight Percent

Purified Water 32.55

Propylene Glycol 0.14

Lanogel 41 0.14

Brij 35 0.40

Purified Water 0.487 B

PVP-K30^tιn 25% 0.163

Glycerine 99% 0.48

Citric Acid 5.88% 0.12

Solution

Hydrogen Peroxide (3%) 1.55

Acetone 0.40

Isopropyl Alcohol 99% 0.91 Kerasol^tm 5.65

Water 41.62

Germaben II^tm 2.82

Ammonium pH 9.0 8.81

Thioglycollate (60%) E

Ammonia Sol'n (28%) 0.94%

Kerasol^tm 2.82- ^•δ Procedure: Step 1: In a separate tank agitate B (water) very strongly and sprinkle B (PVP K-30) onto the Vortex. Mix until PVP K-30 solution is complete. Step 2: Charge a mixing tank with water at 35-40"C. Add the A phase ingredient and mix thoroughly. Add the PVP K-30 solution and mix in well. Step 3: Mix C phase together in a plastic container. Warm to 35- 40^βC. Add to step 2 and add the D phase. Step 4: In a plastic container, add ammonium thioglycollate and then add ammonia solution slowly to bring the pH to 9.0. Add the Kerasol^ia. Mix this solution well and add it to the batch.

EXAMPLE IX

Component Weight Perdent

Purified Water 30.53

PVP K-30 0.163

Lanogel 41 0.14

Propylene Glycol 0.14

Brij 35 0,40

Essence of Pellitory 0.50%

Essence of Elder 0.50%

Glycerine 0.48%

Citric Acid 5.88% 0.12

Hydrogen Peroxide 3% 1.55 B

Acetone 0.40

Isopropanol 99% 0.91

Kerasol^tm 5.65

Germaben II^tm 2.82

Purified Water 36.24

Dehyquart A 1.00

Ammonium pH 9.0 8.81

Thioglycollate (( 50%)

Ammonia (28%) 0.94%

Kerasol^tm 2.82

Arlasolve 200^tm 4.00%

Fragrance 1.88

Procedure: Agitate purified water (A) rapidly with "lightnin" mixer and sprinkle PVP K-30 slowly onto the surface. Allow PVP to go into solution. Add remainder of A ingredients and mix in well. Add B ingredients individually and mix well after each addition. Add C ingredients individually and mix in well after each addition. In a separate container add ammonium thioglycollate (D) and use ammonia solution to bring the pH to 9.0. Add kerasoltm and mix in very well. Add this D phase to the batch and blend it in very well. In a separate container heat Arlasolve 200-tm very gently to liquify. Add remaining E ingredients separately and mix very well. Add this to the batch and mix until the product is uniform.

EXAMPLE X

Component Weight

Purified Water 41.91

PVP K-30 0.22

Lanogel 41 0.05

Propylene Glycol 0.05

Brij 35 0.13

Essence of Rosemary 0.33%

Essence of Pimpernil 0.33%

Allantoin 0.34%

Glycerine 0.16%

Citric Acid 5.88% 0.04

Hydrogen Peroxide 3% 0.52 B

Acetone 0.13

Isopropanol 99% 0.30

Kerasol^tm 1.88

Germaben 11^"tin 0.94

Purified Water 42.81

Ammonium pH 9.0 2.95

Thioglycollate (60%)

Ammonia (28%) 0.09%

Kerasol^tm 0.94

Arlasolve 200^tm 4.00%

Fragrance 1.88 E

Procedure: Agitate purified water (A) rapidly with "lightnin" mixer and sprinkle PVP K-30 slowly onto the surface. Allow PVP to go into solution. Add remainder of A ingredients and mix in well. Add B ingredients individually.and mix well after each addition. Add C ingredients individually and mix in well after each addition. In a separate container add ammonium thioglycollate and use ammonia solution to bring the pH to 9.0. Add kerasol^tm and mix in very well. Add this D phase to the batch and blend it in very well. In a separate container heat Arlasolve 200-fc_m very gently to liquify. Add remaining E ingredients separately and mix very well. Add this to the batch and mix until the product is uniform.

EXAMPLE XI

Component Weight Percent

Purified Water 41.57

PVP K-30 .... 0.22

Lanogel 41 0.05 A

Propylene Glycol 0.05

Brij 35 0.13

Biotin 0.001

Elastin 0.001

Glycerine 0.16%

Citric Acid 5.88% 0.04

Hydrogen Peroxide 3% 0.52 B

Acetone 0.13

Isopropanol 99% 0.30

Kerasol^tm 1.88

Germaben II^tm 0.94

Purified Water 44.28 C

Ammonium pH 9.0 2.95

Thioglycollate (60%) D

Ammonia (28%) 0.09%

Kerasol^tm 0.94

Arlasolve 200^tm 4.00%

Fragrance 1.75 E

Procedure: Agitate purified water (A) rapidly with "lightnin" mixer and sprinkle PVP K-30 slowly onto the surface. Allow PVP to go into solution. Add remainder of A ingredients and mix in well. Add B ingredients individually and mix well after each addition. Add C ingredients individually and mix in well after each addition. In a separate container premix ammonium thioglycollate (D) into purified water (D) and use ammonia solution to bring the pH to 9.0. Add kerasol^tm and this to the batch and mix until the product is uniform.

EXAMPLE XI

Component Weight Percent

Purified Water 41.57 PVP K-30 0.22 Lanogel 41 0.05 Propylene Glycol 0.05 Brij 35 0.13 Biotin 0.001 Elastin 0.001

Glycerine 0.16%

Citric Acid 5.88% 0.04

Hydrogen Peroxide 3% 0.52 B

Acetone 0.13

Isopropanol 99% 0.30

Kerasol^tm 1.88

Germaben Il^m 0.94 Purified Water 44.28

Ammonium pH 9.0 2.95

Thioglycollate (60%) D Ammonia (28%) 0.09% Kerasol^tm 0.94

Arlasolve 200^tm 4.00%

Fragrance 1.75 E

Procedure: Agitate purified water (A) rapidly with "lightnin" mixer and sprinkle PVP K-30 slowly onto the surface. Allow PVP to go into solution. Add remainder of A ingredients and mix in well. Add B ingredients individually and mix well after each addition. Add C ingredients individually and mix in well after each addition. In a separate container premix ammonium thioglycollate (D) into purified water (D) and use ammonia solution to bring the pH to 9.0. Add kerasol^m and mix in very well. Add this D phase to the batch and blend it in very well. In a separate container heat Arlasolve 200^_m very gently to liquify. Add remaining E ingredients sepa¬ rately and mix very well. Add this to the batch and mix until the product is uniform. EXAMPLE XII Concentrated Non-Perfumed Moisturizing Lotion

Component Weight Percent

Arlamol E 2.33

Brij 72 8.93

Brij 78 2.25

Mineral Oil 70 19.89

Propylparaben 0.31

Purified Water 62.15

Disodiu EDTA 0.16

Dimethicone 0.16 B

Methylparaben 0.70

Propylene Glycol 2.33

Germaben II 0.79 C

Procedure: Charge main mixing kettle with B ingredients and heat while mixing to 80-85"C. In a separate container heat A ingredients to 80-85^βC and mix until uniform. At 80-85"C add mixed A ingredients to mixed B ingredients while thoroughly mixing. Cool to 50-55°C. At 50-55"C add Germaben and blend in very well. Continue to cool to 30°C and use at this temperature.

EXAMPLE XIII Component Weight Percent

Concentrated Non-Perfumed 8.85 A

Moisturizing Lotion (Ex. XII)

Purified Water 56.94

Elastin 0.001 B

Biotin 0.001

Carbopol 940 2.36

Concentrated Stock Sol'n 29.23 C

Example VIII

Fragrance 1.75 D

Coloring Agent 0.87 E Procedure: This is a 4% Carbopol dispersion. Measure water and agitate at high speed. Add elastin and then biotin and allow them to disperse. Add Carbopol 940 to the lip of the vortex and mix well until the dispersion is complete. Add component A to the mixing kettel at 25-30°C and add B phase from above and mix until uniform. Add component C to mixture of A and B and mix until uniform. Add fragrance and coloring agent.

EXAMPLE XIV Non-Perfumed Moisturizing Lotion Component Weight Percent

Arlamol E 1. . 36 Brij 72 5 . B -ώ .

Mineral Oil 70 11. . 60 Propylparaben 0. . 18

Purified Water 77 . . 4

Disodium EDTA 0. . 10

Dimethicone 0 0.. .0099 B

Methylparaben 0 . . 41

Propylene Glycol 1. . 36

Dowicil 200 0 . . 05 Purified Water 0. . 50

Formaldehyde 37% 0 0..2 2 D

Germaben II 0 0..2 233 E

Procedure: Heat A phase components to 70-75°C and mix until uniform. Charge main kettle with water and begin heating to 70-75°C. Add the remainder of phase B components and mix to dissolve the solids. Add, at 70-75°C, A phase to B phase while mixing. Blend well and cool to 35-40"C. Premix C phase and add to the batch when the solution is clear. Add D and E phases one at a time and mix in well. Cool to 25-30°C and use at this temperature. EXAMPLE XV Component Weight Percent

Non-Perfumed Moisturizing 8.70 A-

Lotion (Ex. XIV)

Purified Water 56.11

Carbopol 940 1.15

Essence of Rosemary 0.20 B

Essence of Althea 0.20

Essence of Bilberry 0.20

Essence of Jaborand. 0.20

Essence of Verbena 0.20

Triethanolamine 99% 2.60 C

Concentrated Stock Sol'n 28.70 D

Example VIII

Fragrance 1.74 E

Procedure: In a separate mixing tank agitate water at high speed and spring Carbopol 940 onto the vortex. Disperse Car¬ bopol and add remaining B phase components. Mix until car¬ bopol dispersion is complete. Charge mixing tank with A com¬ ponent and hold at 25-30°C. Add B mixture and blend until homogeneous. Add triethanolamine and mix in well until homogeneous. Add component D until homogeneous. Add com¬ ponent E and blend in well.

EXAMPLE XVI STORAGE STABLE COMPOSITION

Component Weight Percent

Activated Protein 0.1 to 12.0%

Reducing Agent 0.1 to 15.0%

Antioxidant 0.01 to 4.0%

Water, acids, bases 79.0 to 99.79% buffering agents, emulsifying agents, thickeners, preservatives, organic solvents coloring agents, fragrance Procedure: Form activated protein separately with solution containing reducing agent at a pH above about 9. Mix in remaining components until final mixture is homogeneous. EXAMPLE XVII STORAGE STABLE COMPOSITION INCLUDING OXIDIZING AGENT

Component Weight Percent

Activated Protein 0.1 to 12.0%

Reducing Agent 0.1 to 15.0%

Oxidizing Agent 0.001 to 4.0%

Antioxidant 0.01 to 4.0%

Water, acids, bases 75.0 to 99.789% buffering agents, emulsifying agents, thickeners, preservatives, organic solvents coloring agents, fragrance

Procedure: Form activated protein separately with solution containing reducing agent at a pH above about 9. Mix in remaining components until final mixture is homogeneous.

Examples XVIII and XIX Hair Growth

A 32 year old New York City man with a history of hair loss, who had tried many formulations and had been using minoxidil four times a day for two years as prescribed by a physician with no results. Two weeks after beginning the for¬ mulation of Example I daily he noticed that his hair was thicker. Within a month he noticed new growth which became profound within 3 months.

A 63 year old Florida man with a long history of hair loss skeptically used the formulation of Example I on the top and front of his head, up to his greatly receded hairline. Within 3 weeks there was visible new growth in areas that had not grown visible hair in 20 years.

Examples XX and XXI Hair Thickening and Strengthening

A 27 year old male hairdresser from New York who noticed considerable hair loss due to breakage on his pillow, in his comb, in the drains, etc. began using the formulation of Example I daily. Within 3 weeks his estimates of the hair on his comb in the morning decreased from over 100 hairs to under 10 hairs. He no longer noticed hair on his pillow in the morning, and only a normal amount of hair in his drains. A 31 year old New Jersey man with very thin fly-away hair began using the formulation of Example I daily, and within one month found his hair to be thicker, with more body, and stayed where he wanted it with much less fly-away.

Example XXII Nail Hardening

A 42 year old New York man with chronically thin brittle nails began using the formulation of Example III on his nails daily. Within 1 month he reported his nails were considerably harder, less prone to breaking, and he could grow them longer; especially one nail which had been injured years ago and had developed a chronic thin spot. This thin spot persisted relative to the rest of his nails, but since all of his nails were harder and thicker it was no longer a problem. In addition, the rate of nail growth was markedly faster, based upon the frequency with which he had to file his nails before and after treatment.

Examples XXIII-XXVIII Animal Coat Restorationof Acute and Chronic Conditions

A thoroughbred used as a carriage horse developed back rubs on the withers and back from the tack. Daily application of the formulation of Example IV resulted in 80% restoration of the acute condition within 8 days despite continued appli¬ cation of the tack and carriage driving. The attending veterinarian estimated that this level of fur restoration should have taken 6-8 weeks in the absence of driving and the application of tack.

The formulation from Example IV was used on over 8 horses at a major show barn for coat deficits due to wounds, abrasions and back rubs. In all instances, the deficits resolved at a vastly accelerated rate in the estimate of a highly experienced groom who travels with the U.S. Olympic Equestrian Team.

A horse with a chronic hairless spot on the head which was present when the horse was purchased over 2 years ago experienced coat restoration in that area following 10 days of daily application of the formulation of example IV.

A horse with chronic coat loss on the lower legs due to bandages experienced coat restoration following 3 weeks of daily application of the formulation from Example I.

A cat with a paralyzed forelimb which for years was continually dragging on the ground experienced coat restora¬ tion within 1 month after beginning daily applications of the formulation of Example I.

Example XXVIII Coat Restoration of Chronic Condition

A dog had a severe pyoderma with hair loss, oozing and bleeding, and itching resulting in self-inflicted lacerations of over 1 year duration. Suspected IgA deficiency. Two weeks of lincomycin almost a year earlier provided a slight positive effect but was discontinued due to toxicity. In late, 1987 thyroid supplementation was tried without results. Euthanasia was considered. In the middle of 1988, experimental treatment with the composition from Example X was initiated. Within 4 days all open sores scabbed over. Within 8 days all lesions healed with appreciable hair growth. Within 21 days the dog was approximately normal in appearance. Complete hair regrowth over the entire body has begun. After 3 months, the dog is completely restored. No more deficits, self- mutilation, etc. The treatment was stopped and the problem seems to be completely and permanently resolved. There has been no loss of hair since ceasing treatment.

Example XXIX Hoof Growth

A mare with seedy toe, an anaerobic infection of the lamina was treated by excising the tissue over the infection and the infection was treated with the composition of Example XIII. After only one week of use the hoof wall appeared appreciably healthier and of better texture. The coronary band looked much better. She started using this composition on all of the hooves of her animals.

Example XXX Coat Restoration of Chronic Condition

A horse had hip rubs of over 1 year duration which refused to heal. After using 1 ounce of the composition of Example XV the open wounds healed and grew hair. Example XXXI Grooming Daily misting gets dog coats into show condition in one to two weeks which would otherwise have taken four to six months. Beskie terriers looked "fantastic" after one month using the composition of Example X. The coats became silky and shiny and the dogs pads were strengthened with no crack¬ ing.

Examples XXXII and XXXIII Grooming and Hoof Conditioning Use of the composition of Example X greatly facili¬ tated the transition from winter to summer coats of horses following a gloomy spring. After two applications the coat transition was greatly improved. Use of the composition of Example XI for hoof conditioning resulted in a nice texture, not soft and spongey yet moist and resilient, yet not dry and brittle.

Example XXXIV Hoof Conditioning In a number of cases, the composition of Example XIII resulted in hoof reconditioning, and the strengthening of "shelly" hooves, and dry, brittle and/or cracking hooves. Example XXXV Restoration of Chronic Coat Condition

Photographer for a New Jersey newspaper used the com¬ position of Example XV for treating hair loss on a biopsy for her Burmese mountain dog which had not grown hair at the site of biopsy for over a year and a half. After 3 days of treat¬ ment, the photographer reported preliminary regrowth of hair.

This invention has been described in terms of specific embodiments set forth in detail herein, but it should be understood that these are by way of illustration and the invention is not necesarily limited thereto. Modifications and variations will be apparent from the disclosure and may be resorted to without departing from the spirit of the inven¬ tions those of skill in the art will readily understand. Accordingly, such variations and modifications are considered to be within the purview and scope of the invention and the following claims.

Claims

WHAT IS CLAIMED IS:

1. A composition for treating keratinous tissue in mam¬ mals comprising: a), about 0.01 to about 12% by weight of an activated protein component; b) . about 0.1 to about 15% by weight of a compatible reducing agent; c) . about 0.001 to about 4.0% by weight of an oxidiz¬ ing agent; and d) . about 81.0% to about 99;889% by weight of at least one component selected from the group consisting of water, acids, bases, buffering agents, emulsifying agents, thick¬ eners, solvents, preservatives, coloring agents and perfuming agents.

2. The composition according to claim 1 wherein said activated protein component is an activated keratin protein.

3. The composition according to claim 1 wherein said com¬ patible reducing agent is a salt of thioglycollic acid.

4. The composition according to claim 3 wherein said com¬ patible reducing agent is ammonium thioglycollate.

5. The composition according to claim 2 wherein said oxidizing agent is selected from the group consisting of sodium perborate and hydrogen peroxide.

6. The composition according to claim 5 wherein said oxidizing agent comprises about 0.001 to about 1.5% by weight of said composition.

7. The composition according to claim 6 wherein said oxidizing agent is hydrogen peroxide.

8. The composition according to claim 1 wherein said activated protein component is an activated keratin protein, said compatible reducing agent is ammonium thioglycollate and said oxidizing agent is hydrogen peroxide.

9. A composition for treating keratinous tissue in mam¬ mals comprising: a), about 0.01 to about 12% by weight of an activated protein component; b) . about 0.1 to about 15% by weight of a compatible reducing agent: c) . about 0.001 to about 2.0% by weight of an antioxidant; and d) . about 81.0% to about 99.889% by weight of at least one component selected from the group consisting of water, acids, bases, buffering agents, emulsifying agents, thick¬ eners, solvents, preservatives, coloring agents and perfuming agents.

10. The composition according to claim 9 wherein said activated protein component is a keratin protein.

11. The composition according to claim 9 wherein said compatible reducing agent is a salt of thioglycollic acid.

12. The composition according to claim 9 wherein said compatible reducing agent is ammonium thioglycollate.

13. The composition according to claim 9 wherein said antioxidant is a volatile antioxidant.

14. The composition according to claim 9 wherein said activated protein component is a keratin protein, said com¬ patible reducing agent is ammonium thioglycollate, and said antioxidant is a volatile antioxidant.

15. A composition for treating keratinous tissue in mam¬ mals comprising: a), about 0.01 to about 12% by weight of an activated protein component; b) . about 0.1 to about 15% by weight of a compatible reducing agent: c) . about 0.001 to about 4.0% by weight of an antioxidant; d) . about 0.001 to about 4.0% by weight of an oxidiz¬ ing agent; and e) . about 81.0% to about 99.889% by weight of at least one component selected from the group consisting of water, acids, bases, buffering agents, emulsifying agents, thick¬ eners, solvents, preservatives, coloring agents and perfuming agents.

16. The composition according to claim 15 wherein said activated protein component is a keratin protein.

17. The composition according to claim 15 wherein said compatible reducing agent is a salt of thioglycollic acid.

18. The composition according to claim 15 wherein said compatible reducing agent is ammonium thioglycollate.

19. The composition according to claim 15 wherein said antioxidant is a volatile antioxidant.

20. The composition according to claim 19 wherein said oxidizing agent is a nonvolatile oxidizing agent.

21. The composition according to claim 15 wherein said activated protein component is a keratin protein, said com¬ patible' reducing agent is ammonium thioglycollate, said antioxidant is a volatile antioxidant and said oxidizing agent is a nonvolatile oxidizing agent.

22. A method of treating keratinous tissue to improve its appearance and strength by contacting the tissue with a com¬ position comprising:

(a) about 0.01 to about 12% by weight of an activated protein;

(b) about 0.1 to about 15% by weight of a compatible reducing agent;

(c) about 0.001 to about 4.0% by weight of an oxidiz¬ ing agent; and

(d) about 61% to about 99.899% by weight of at least one component selected from the group consisting of water, bases, acids, buffering agents, emulsifying agents, thick¬ eners, coloring agents and perfuming agents.

23. The method according to claim 22 wherein said activated protein is an activated keratin protein, said com¬ patible reducing agent is ammonium thioglycollate and said oxidizing agent is hydrogen peroxide.

24. The method according to claim 23 additionally com¬ prising drying said keratinous tissue after said contacting step.

25. A method of treating keratinous tissue to improve its appearance and strength by contacting the tissue with a com¬ position comprising: a), about 0.01 to about 12% by weight of an activated protein; b) . about 0.1 to about 15% by weight of a compatible reducing agent; c) . about 0.001 to about 2.0% by weight of an antioxidant; and d) . about 81.0% to about 99.889% by weight of at least one component selected from the group consisting of water, acids, bases, buffering agents, emulsifying agents, thick¬ eners, solvents, preservatives, coloring agents and perfuming agents.

26. The method according to claim 25 Wherein said activated protein component is a keratin protein, said com¬ patible reducing agent is ammonium thioglycollate, and said antioxidant is a volatile antioxidant.

27. The method according to claim 26 additionally com¬ prising drying said keratinous tissue after said contacting step.

28. A method of treating keratinous tissue to improve its appearance and strength and promote its growth by contacting the tissue with a composition comprising: a), about 0.01 to about 12% by weight of an activated protein component; b) . about 0.1 to about 15% by weight of a compatible reducing agent: c) . about 0.001 to about 4.0% by weight of an antioxidant; d) . about 0.001 to about 4.0% by weight of an oxidiz¬ ing agent; e) . about 81.0% to about 99.889% by weight of at least one component selected from the group consisting of water, acids, bases, buffering agents, emulsifying agents, thick¬ eners, solvents, preservatives, coloring agents and perfuming agents.

29. The method according to claim 28 wherein said activated protein component is a keratin protein, said com¬ patible reducing agent is ammonium thioglycollate, said antioxidant is a volatile antioxidant and said oxidizing agent is a nonvolatile oxidizing agent.

30. The method according to claim 29 additionally com¬ prising drying said keratinous tissue after said contacting step.

31. A method of treating human nails and animal hooves to improve their appearance and strength and promote their growth by contacting the tissue with a composition comprising: a), about 0.01 to about 12% by weight of an activated protein component; b) . about 0.1 to about 15% by weight of a compatible reducing agent; and c) . about 75.0% to about 99.89% by weight of at least one component selected from the group consisting of water, acids, bases, buffering agents, emulsifying agents, thick¬ eners, solvents, preservatives, coloring agents and perfuming agents.

32. The method according to claim 30 wherein said activated protein is keratin.

33. The method according to claim 32 wherein said com¬ patible reducing agent is selected from the group consisting of thioglycollic acid and ammonium thioglycollate.