Compositions and methods for treating skin conditions and promoting wound healing

This invention relates to activated protein-containing compositions comprising reducing agents, oxidizing agents and/or anti-oxidants and methods of use. The therapeutic compositions and methods of the present invention are particularly effective in promoting wound healing, and in inhibiting certain skin disorders, including eczema and seborrhea, sclerodema and acne. The therapeutic compositions and methods of the present invention have also shown enhanced effect as veterinary tools in reducing the debilitation associated with certain skin conditions in mammals including eczematoid dermatitis, chronic dermatitis, equine exuberant granuloma ("proud flesh"), decubitis ulcers, and canine cutaneous granulomas ("lick" granuloma).

This invention relates to therapeutic compositions using novel activated 
protein-containing ingredients and includes reducing agents and 
optionally, oxidizing agents and/or anti-oxidants. The therapeutic 
compositions of the present invention are particularly effective in 
promoting wound healing, and in inhibiting certain skin disorders, 
including eczema and seborrhea, dandruff, psoriases and other rash-like 
indications, scleroderma and acne. The therapeutic methods of the present 
invention have also shown enhanced effect as veterinary tools in reducing 
the debilitation associated with certain skin conditions in mammals 
including eczematoid dermatitis, chronic dermatitis, equine exuberant 
granuloma ("proud flesh"), decubitis ulcers, and canine cutaneous 
granulomas ("lick" granuloma). 
U.S. Pat. No. 4,438,102 describes compositions which are useful for 
promoting the growth of normal dermal and epidermal tissue, and described 
as being useful to promote wound healing in the soft keratin tissue of the 
epidermis. The compositions of the patent are described as containing 
defined percentages of thioglycollic acid, ammonium hydroxide, glycerine, 
citric acid, hydrogen peroxide, gelatin, a lower alkanol, and a solvent 
such as acetone or diethyl ether. Several examples of wound healing are 
provided in the specification. The commercial gelatins described in the 
patent do not contain sufficient cysteinyl residues to covalently bind to 
keratinous tissue, especially skin and the patent does not disclose that 
the gelatin covalently binds to skin. 
U.S. Pat. No. 3,954,974 describes disinfectant emulsions comprised of 
hydrogen peroxide in an oil-in-water phase. Exemplary compositions 
comprise hydrogen peroxide, an emulsifier or surfactant, an oil or gel and 
water. Certain embodiments may additionally include solvents. The 
compositions of the patent may be used for disinfecting hands and for 
treating skin diseases, irritations and injuries. 
U.S. Pat. No. 4,195,095 describes the use of certain formulations 
comprising thioglycollic acid for use in the treatment of fatty cysts, 
dandruff, scleroderma and other dermatological disorders, including acne 
vulgaris. Exemplary compositions comprise thioglycollic acid, 
hexachlorophene, sodium hydroxide, water and other ingredients, including 
bulking or gelling polymers and preservatives, among others. 
It has now been discovered that activated protein compositions which are 
formulated with reducing agents, and with or without oxidizing agents 
and/or anti-oxidant compositions exhibit surprising activity for treating 
wounds, pyoderma, sebborhea, psoriasis, acne, itching, callouses, corns, 
burns, eczematoid dermatitis, chronic dermatitis, decubitis ulcers, 
miscellaneous rashes, non-specific dermatitis and certain veterinary 
conditions including equine exuberant granuloma ("proud flesh") and canine 
cutaneous granulomas ("lick" granuloma). 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 and the activated protein, and optionally an oxidizing 
agent and/or an antioxidant may be added. Numerous additional components 
may also be included, for example water, bases, acids, buffering agents, 
emulsifying agents or surfactants, thickeners, preservatives, coloring 
agents and perfuming agents. 
It is an object of the present invention to provide novel therapeutic 
compositions and methods for treating certain skin conditions for example, 
wounds, eczematoid dermatitis, chronic dermatitis, decubitis ulcers, 
sebborhea, psoriasis, pyoderma, dandruff, itching, allergic skin 
reactions, miscellaneous rashes, acne and certain veterinary conditions 
including equine exuberant granuloma, and canine cutaneous granulomas 
("lick" granuloma) utilizing compositions containing at least one 
activated thiol-containing protein in combination with a reducing agent. 
It is a further object of the present invention to provide therapeutic 
compositions and methods for treating the aboveidentified skin conditions 
with storage stable compositions which are capable of maintaining the 
activity of an activated protein in the presence of an oxidizing agent to 
promote the shelf-life of the composition. 
It is a further object of the present invention to provide compositions for 
treating chronic veterinary skin conditions which do not respond to 
traditional therapeutic methods, for example, equine exuberant granuloma 
("proud flesh") and canine cutaneous granuloma ("lick" granuloma). 
It is an additional object of the present invention to provide new 
compositions and methods for conditioning keratinous tissue including 
skin, by exposing the tissue to activated protein and reducing agent and 
oxidizing said tissue without having to apply two separate solutions to 
the treated tissue to perform the reducing and oxidizing steps. This 
approach has shown surprising efficacy in treating the above-mentioned 
skin conditions. 
BRIEF DESCRIPTION OF THE INVENTION 
The thereapeutic compositions and methods of the present invention are 
useful for treating certain skin conditions for example, wounds, 
sebborhea, psoriasis, dandruff, allergic skin reactions, acne, itching, 
callouses, corns, burns, miscellaneous rashes, non-specific dermatitis and 
certain veterinary conditions including eczematoid dermatitis, chronic 
dermatitis, equine exuberant granuloma ("proud flesh"), decubitis ulcers, 
and canine cutaneous granuloma ("lick" granuloma). In using the present 
invention a composition comprising an activated protein, a compatible 
reducing agent, and optionally an oxidizing agent and/or an antioxidant is 
contacted with an area of keratinous tissue affected with one of the above 
conditions. The therapeutic compositions and methods of the present 
invention have shown especially surprising activity against non-healing 
skin conditions, especially canine cutaneous granulomas ("lick" granuloma) 
and equine exuberant granuloma ("proud flesh"). 
Compositions useful in the therapeutic methods of the present invention 
include 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 present invention may comprise 
a reducing agent, and an activated protein, in addition to other less 
essential components, and optionally an oxidizing agent and/or an 
antioxidant. Additional embodiments of the present invention 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 antioxidant 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 disulfide 
bonds of cystine residues within the protein structure to produce free 
thiol or mercaptide groups on cysteine residues. The ability of a protein 
to bind to keratinous tissue 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 keratinous tissue. Also, it is 
believed that the ability to oxidize the keratin mercaptide and a proximal 
protein mercaptide aids the formation of disulfide covalent bonds. 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. 
It is preferred that the activated protein should be hydrated. A hydrated 
protein may be beneficial to certain types of skin conditions and wounds, 
especially burns, because the hydrated protein may be expected to provide 
additional moisture to the skin. The protein in the compositions of the 
present invention may react with and form chemical bonds with the keratin 
of human and animal skin, thus effecting an attachment of moist hydrated 
proteins to skin. The compositions are therefore useful for treating human 
and animal skin to chemically bond the activated protein to the skin, 
moisturize dry skin and provide a moisturizing vehicle to carry other 
agents into dehydrated skin. 
The percent 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 
protein-keratinous tissue covalent bond formation. 
The therapeutic compositions of the present invention are preferably 
utilized at ambient temperatures, i.e., about 20.degree. C. to about 
35.degree. C.; however, higher temperatures may be used. Obviously, when 
treating a wound, especially a burn, treatment is kept to a lower 
temperature to avoid exacerbating the wound condition. Where the 
application of heat is viewed as therapeutic, conducting the treatment at 
higher temperatures is recommended. The keratinous tissue may be treated 
over a period of time ranging from about 10 minutes to about 6 hours. The 
keratinous tissue may be treated acutely or chronically, with or without a 
dressing as needed. In certain embodiments, compositions useful in 
practicing the therapeutic methods of the present invention may be 
formulated with sustained or controlled release polymers to produce 
formulations capable of delivering active agent for extended periods of 
time. 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. 
Depending upon the anti-oxidant used the reaction may be facilitated by 
heat. 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 one aspect of the present invention, compositions of the present 
invention 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, 
thickeners, preservatives, organic solvents, coloring agents and perfuming 
agents. 
Compositions for use in the therapeutic method of the present invention may 
include an antioxidant instead of an oxidizing agent, and in certain 
embodiments no oxidizing agent or antioxidant is used. 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, at least one component 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, and optionally about 0.001 to about 4.0% 
of an antioxidant and/or an oxidizing agent. 
Additional embodiments of the method of the present invention utilize a 
composition comprising 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 utilized in the methods of the present invention are 
formulated 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 
interact to form a covalent disulfide bond. The inclusion of an oxidizing 
agent in the same composition as the reducing agent and activated protein 
is designed to maximize covalent disulfide formation without having to 
resort to a second oxidizing solution. 
In addition to covalent disulfide bond formation, a number of other 
mechanisms may be responsible for the enhanced activity displayed by the 
therapeutic methods of the present invention. 
The present invention utilizes compositions formulated as gels, creams, 
lotions, sprays or liquids of varying viscosities. 
DETAILED DESCRIPTION OF THE INVENTION 
The present invention employs compositions comprising an activated protein, 
a compatible reducing agent, optionally an oxidizing agent and/or an 
antioxidant, 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 these compositions, 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 used in compositions employed in the present invention 
are exemplified by keratins or certain types of gelatins containing 
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 tissue. 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, hemoglobin, 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. Pat. No. 4,438,102 
contain at most trace cysteinyl residues and most commercial grade 
gelatins, including Grade A edible gelatin, contain undetectable amounts 
of cysteine. Such gelatin proteins, which do not contain sufficient 
cysteinyl content to covalently bind to keratinous tissue in any 
significant way, i.e., to produce a permanent attachment of the protein to 
the keratinous tissue, are therefore 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 T1, 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). In compositions to be used to treat certain wounds, 
exemplary compositions utilize keratin either alone or in combination with 
fibrinogen/fibrin or modified fibrinogen. Other compositions which may be 
used to treat wounds may comprise fibrinogen/fibrin or modified fibrinogen 
alone. 
The proteins used in the present invention are preferably activated in the 
presence of 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 
also result in hydrolysis of the protein into shorter, less advantageous 
peptide units. A number of globular proteins contain cysteinyl residues 
within hydrophobic pockets. 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 of 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 may adversely affect the rate 
at which cystinyl disulfide bonds in the protein are converted to 
cysteinyl mercaptide groups. 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 enough 
cysteinyl residues to promote covalent binding to the keratinous tissue. 
The activated protein is preferably a keratin, but any protein which 
contains sufficient cysteinyl content to promote binding to activated 
keratinous tissue is contemplated for use in the present invention. 
Preferred proteins are keratins because of their high cysteinyl content 
(generally, greater than 10% by weight of the protein and often as high as 
15-17% by weight of the protein) which may be obtained by hydrolysis of 
skin, feathers, wool and hair. A particularly preferred keratin is 
Kerasol.TM. from Croda Chemicals International, 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 130,000 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 analogues, for 
example, mercaptosuccinic acid, thiolactic acid and their pharmaceutically 
acceptable salts, among others, 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 pharmaceutically acceptable salts. Especially preferred 
reducing agents for use in the present invention include thioglycerol and 
salts of thioglycollic acid, especially ammonium thioglycollate. It is 
preferred that the reducing agent for activating the protein should be the 
same as the pharmaceutically compatible reducing agent which is used in 
the final formulation of the invention. The use of strong pharmaceutically 
incompatible reducing agents to activate the protein are less preferred 
and may make the use of the protein more difficult because the reducing 
agent may have to be removed from the activated protein before 
formulation. 
Compositions for use in the method of the present invention also contain a 
pharmaceutically compatible reducing agent in an amount equal to about o.1 
to about 15% by weight of the formulation. Preferred compositions contain 
about 3.0 to about 10% by weight of a pharmaceutically compatible reducing 
agent. The amount of pharmaceutically compatible reducing agent varies 
according to the therapeutic use for which the compositions are intended, 
but generally falls within the range of about 3.0 to about 10% by weight. 
A pharmaceutically 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, especially for use on the skin of humans and/or animals. 
Pharmaceutically compatible reducing agents which are contemplated for use 
in the present invention include mercaptoethanol, dithiothreitol, 
thioglycerol, thiolactic acid, glutathione, cysteine and thioglycollic 
acid and its salts. An especially preferred reducing agent is ammonium 
thioglycollate. 
Compositions used in 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 also to promote the formation of covalent disulfide 
bonds between activated protein and keratinous tissue. Additionally, the 
oxidizing agent may function as a disinfecting agent to clean the 
keratinous tissue and enhance healing. Exemplary oxidizing agents include 
hydrogen peroxide (which may or may not be stabilized with, for example, 
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 perborate, the alkali metal bromates including sodium and 
potassium 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 
composition 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. 
Of course, many of the therapeutic compositions and methods, especially 
those which are used to treat the veterinary conditions of equine 
exuberant granuloma ("proud flesh") and canine cutaneous granuloma ("lick" 
granuloma) do not require the inclusion of an oxidizing agent, but it is 
often advantageous to include the oxidizing agent. Thus, where oxidizing 
agent is not included to treat keratinous tissue conditions, the 
compositions comprise no greater than about 12.0% of an activated protein 
component and no greater than about 15.0% of a reducing component, the 
remainder of the composition comprising 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 addition to the above-described ingredients, additional components may 
be added to the formulation to enhance the effects of the compositions. In 
addition to water, additional components may include bases, acids, 
buffering agents, solvents, emulsifying agents or surfactants, thickeners, 
preservatives, 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 formulation 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 formulation, 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 constant 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 components, 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, homogeneous 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 formulations of the present invention for its emulsifying 
characteristics. 
An emulsifying agent or surfactant is often added to embodiments of the 
present invention to enhance the characteristics 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 emulsifying agents 
may be nonionic, anionic or amphoteric. Nonionic emulsifying compositions 
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 Del.), 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 example, 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 sulfonated higher fatty acid esters such as, e.g., higher fatty 
acid esters of 2,3-dihydroxypropane 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 cocoanut 
oil fatty acids, among others. 
It may be preferable to add an anti-foaming agent to certain compositions 
to promote homogeneity and prevent foaming 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, Mich. 
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, carbomers, 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, carboxymethylcellulose, propylcellulose and 
hydroxypropylcellulose, 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.TM. 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 Carbapol 940.TM.. 
Preservatives are added for preventing microbial growth in the presence of 
protein nutrients. Exemplary preservatives include benzoic acid analogs 
including, among others, sodium benzoate. Other presevatives include 
propyl and methyl paraben, Dowicil.TM. (Dow Chemical Corp., Midland, Mi.) 
and formaldehyde solution. An especially preferred preservative is 
Germaben II.TM., available from Sutton Laboratories, N.J. 
Coloring agents and perfume agents may also be added to enhance the 
characteristics of the formulations. 
In another aspect of the present invention, the compositions include an 
antioxidant instead of an oxidizing agent. These compositions comprise 
about 0.01 to about 12% 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 component 
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 alphatocopherol, hydroxyquinone, unipherol, tocopherol 
ascorbate, lecithin, chlorophyll, ascorbylpalmitate, linseed oil, tongue 
oil, other natural antioxidants such as the steam distillation extract of 
rosemary as disclosed in U.S. Pat. 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 dissolved metals which may be responsible for the in situ 
generation of oxygen. Preferably, the chelating agent comprises between 
about 0.001 to about 0.5% of the formulation and most preferably 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, butylated 
hydroxyanisole (BHA), butylated hydroxytoluene (BHT), p-hydroxyanisole and 
p-hydroxytoluene. 
A volatile antioxidant as used in preferred embodiments of the present 
invention is an antioxidant that volatilizes 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 formulations 
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. Formulations including solvents may promote 
solvent/antioxidant azeotrope formation and volatility of the antioxidant. 
Azeotrope formation 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. Where 
solvents are used, it is preferred that a pharmaceutically compatible 
solvent is used. 
In preferred embodiments of the present invention, when an oxidizing agent 
is not included in the formulations utilized, 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 mercaptides to covalent disulfide bonds, 
the oxidizing agent comprises 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 useful in the present invention comprising an antioxidant 
or an antioxidant and oxidizing agent, the formulations may additionally 
comprise acids, bases, buffering agents, emulsifying agents or 
surfactants, thickeners, preservatives, 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, additional organic solvent 
may be added to promote the volatilization of the antioxidant. It is 
recognized that the choice of additives is made to avoid any interactions 
that may affect the activity of the activated protein, reducing agent, 
oxidizing agent or antioxidant. 
Compositions of the present invention may be used to treat acne. Preferred 
compositions for treating acne include the compositions of the present 
invention in combination with pharmaceutically effective amounts of 
benzoyl peroxide, tetracycline and other anti-acne agents. The 
compositions of the present invention may also be used to resolve pock 
marks and acne scars in individuals who have not had acne for years. 
Compositions of the present invention may also be used to treat dandruff. 
Especially preferred compositions for treating dandruff include the 
compositions of the present invention in combination with an effective 
amount of a traditional anti-dandruff agent, for example, zinc pyrithione. 
The compositions useful in present invention are applied to the treatment 
area as a liquid, cream, gel or lotion by rubbing the compositions into 
the keratinous tissue to be treated. Following application, the compounds 
are allowed to dry. This may be accelerated by the use of heat or 
circulating air. Subsequent to drying, formulations may be washed off all 
together, although this is not preferred when the compositions are used to 
treat wounds. When treating dandruff or sebborhea, the compositions may be 
washed off before drying. 
The methods of the present invention comprise exposing the area of 
keratinous tissue to be treated to any one of the compositions previously 
described hereinabove. Depending on the type of skin condition to be 
treated, the therapeutic compositions are applied to the affected area at 
least once a day and as many times a day as are necessary to produce an 
improved condition. Generally, the therapeutic compositions are applied to 
the effected area between one and four times a day. Applications of fewer 
than once a day may be performed depending upon the indication and 
severity. For example, in the treatment of dandruff, sebborhea and skin 
conditions other than wounds, the formulation need only be applied after 
or during shampooing, which may only be once or twice a week. The duration 
of therapy will depend on the condition treated and on the response of the 
condition to the thereapy. Thus, when chronic conditions that do no 
respond to traditional therapy are being treated, it is to be expected 
that the duration of therapy will be longer than when less severe 
conditions are treated. 
Although the present invention is not bound by theory, it is believed that 
the surprising therapeutic activity exhibited by compositions useful in 
the present invention may be the result of a combination of 
pharmacological effects. It is believed that the activity of these 
compositions may be the result of the combined effects of many possible 
mechanisms of action, including; moisturization of the affected area by 
hydrated protein, film formation, cytoskeletal interaction and mediation 
of the clotting process, disulfide covalent bond formation, the release of 
endogenous mediators including fibronectin, receptor modification and 
vasodilation.

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 Thioglycollate 
3.27 A 
pH 9 
Kerasol 0.95 
Propylene Glycol 0.05 B 
Lanogel 41 0.05 
Brij 35 0.13 
PVP-K30-25% Sol. 0.22 
Glycerine 0.16 C 
Citric Sol (2%) 0.04 
Hyd. Peroxide (3%) 
0.53 
Acetone 0.13 
Isopropyl Alcohol 70% 
0.53 
Kerasol 1.90 
Water - Distilled 
85.42 D 
Germaben II 0.93 E 
Fragrance 1.90 F 
Arlasolve 3.79 
______________________________________ 
Procedure: Add a mixture of Components B to 1/2 of Component D at 
100.degree. 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 
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 procedures mixture 
A. Add A to the BDC mixture and then mix in a high speed blender until 
thoroughly mixed. Next, add component 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 Thioglycollate 
9.75 A 
pH 9 
Kerasol .TM. 2.82 
Propylene Glycol 0.13 B 
Lanagel 41 .TM. 0.13 
Brij 35 .TM. 0.39 
PVP-K30 .TM. 25% 0.64 
Glycerine 0.47 C 
Citric Acid Solution 
0.12 
Hydrogen Peroxide (3%) 
1.55 
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.degree. 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 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 components 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 
Carbapol 940 - 2% Sol 
19.51 C 
Triethylamine 1.225 D 
H.sub.2 O 1.225 
______________________________________ 
Procedure: Add Component A to Component B (see below) and thoroughly mix. 
Then add Component C until a homogeneous mixture is made. Finally, adjust 
the pH of mixture ABC with the triethylamine/H.sub.2 O mixture to about 
7.0. 
______________________________________ 
Non-Perfumed Moisturizing Lotion 
Component Weight Percent 
______________________________________ 
Arlamed E .TM. 1.36 A 
Brij 72 .TM. 5.23 
Brij 78 .TM. 1.32 
Mineral Oil 11.64 
Propyl Paraben 0.18 
Water 77.71 B 
Sodium EDTA 0.09 
Anti-Foam Dimethicone .TM. 
0.09 
Methyl Paraben 0.36 
Propylene Glycol 1.36 
Dowicil 200 .TM. 0.45 C 
Formaldehyde (37%) 
0.21 D 
______________________________________ 
Procedure for Non-Perfumed Moisturizing Lotion Component: Add A to B at 
160.degree. F. mixing thoroughly. Mix and cool to 100.degree. F. Add 
Components C and D, mixing thoroughly. Allow to cool to 80.degree. F. Use 
at 80 F. 
______________________________________ 
EXAMPLE IV Thin Cream 
Component Weight Percent 
______________________________________ 
Non-perfumed Moisturizing 
8.70 A 
Lotion 
Carbopol 940 .TM. 58.26 
Triethanolamine 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 
4% Solol 940 .TM. 58.50 C 
Fragrance 2.14% 
______________________________________ 
Procedure: Add Component A to component B (see below) and thoroughly mix. 
Then add Component C until a homogeneous mixture is made. Add Component D 
to produce a homogeneous thick cream. 
______________________________________ 
Concentrated Non-Perfumed Moisturizing Lotion 
Component Weight Percent 
______________________________________ 
Arlamol E .TM. 2.33 A 
Brij 72 .TM. 8.95 
Brij 78 .TM. 2.26 
Mineral Oil 19.94 
Propyl Paraben 0.31 
Water 63.15 B 
Sodium EDTA 0.016 
Anti-Foam Dimethicone .TM. 
0.016 
Methyl Paraben 0.7 
Propylene Glycol 2.33 
______________________________________ 
Procedure for Concentrated Non-Perfumed Moisturizing Lotion Component: Add 
A to B at 160.degree. F. mixing thoroughly. Mix and cool to 80.degree. F. 
Use at 80.degree. F. 
______________________________________ 
EXAMPLE VI Concentrated Stock Solution 
Component Weight Percent 
______________________________________ 
Purified Water 32.55 A 
Propylene Glycol 0.14 
Lanogel 41 0.14 
Brij 35 0.40 
Purified Water 0.487 B 
PVP-K30 .TM. 25% 0.163 
Glycerine 99% 0.48 C 
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 D 
Germaben II .TM. 2.82 
Ammonium pH 9.0 8.81 E 
Thioglycollate (60%) 
Ammonium Sol'n (28%) 
0.94% 
Kerasol .TM. 2.82% 
______________________________________ 
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.degree.-40.degree. 
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.degree.-40.degree. C. Add to step 2 and add the D 
phase. Step 4: In a plastic container, add ammonium thioglycollate and 
then ammonia solution slowly to bring the pH to 9.0. Add the Kerasol.TM.. 
Mix this solution well and add it to the batch. 
______________________________________ 
EXAMPLE VII 
Component Weight Percent 
______________________________________ 
Purified Water 30.53 A 
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% B 
Citric Acid 5.88% 
0.12 
Hydrogen Peroxide 3% 
1.55 
Acetone 0.40 
Isopropanol 99% 0.91 
Kerasol .TM. 5.65 
Germaben II .TM. 
2.82 C 
Purified Water 36.24 
Dehyquart A 1.00 
Ammonium pH 9.0 8.81 D 
Thioglycollate (60%) 
Ammonia (28%) 0.94% 
Kerasol .TM. 2.82 
Arlasolve 200 .TM. 
4.00% E 
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 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.TM. 
very gently to liquify. Add remaining E ingredients separately and mix 
very well. Add this to the batch and mix until the produce is uniform. 
______________________________________ 
EXAMPLE VIII 
Component Weight Percent 
______________________________________ 
Purified Water 41.91 A 
PVK-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% B 
Citric Acid 5.88% 
0.04 
Hydrogen Peroxide 3% 
0.52 
Acetone 0.13 
Isopropanol 99% 0.30 
Kerasol .TM. 1.88 
Germaben II .TM. 
0.94 C 
Purified Water 42.81 
Ammonium pH 9.0 2.95 D 
Thioglycollate (60%) 
Ammonia (28%) 0.09% 
Kerasol .TM. 0.94 
Arlasolve 200 .TM. 
4.00% E 
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 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.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 IX 
Component Weight Percent 
______________________________________ 
Purified Water 41.57 A 
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% B 
Citric Acid 5.88% 
0.04 
Hydrogen Peroxide 3% 
0.52 
Acetone 0.13 
Isopropanol 99% 0.30 
Kerasol .TM. 1.88 
Germaben II .TM. 
0.94 C 
Purified Water 44.28 
Ammonium pH 9.0 2.95 D 
Thioglycollate (60%) 
Ammonia (28%) 0.09% 
Kerasol .TM. 0.94 
Arlasolve 200 .TM. 
4.00% E 
Fragrance 1.75 
______________________________________ 
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.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 Concentrated Non-Perfumed Moisturizing Lotion 
Component Weight Percent 
______________________________________ 
Arlamol E 2.33 A 
Brij 72 8.93 
Brij 78 2.25 
Mineral Oil 70 
19.89 
Propylparaben 0.31 
Purified Water 
62.15 B 
Disodium EDTA 0.16 
Dimethicone 0.16 
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.degree.-85.degree. C. In a separate container heat A 
ingredients to 80.degree.-85.degree. C. and mix until uniform. At 
80.degree.-85.degree. C. add mixed A ingredients to mixed B ingredients 
while thoroughly mixing. Cool to 50.degree.-55.degree. C. At 
50.degree.-55.degree. C. add Germaben and blend in very well. Continue to 
cool to 30.degree. C. and use at this temperature. 
______________________________________ 
EXAMPLE XI 
Component Weight Percent 
______________________________________ 
Concentrated Non-Perfumed 
8.85 A 
Moisturizing Lotion 
(Ex. XII) 
Purified Water 56.94 B 
Elastin 0.001 
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.degree.-30.degree. 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 XII Non-Perfumed Moisturizing Lotion 
Component Weight Percent 
______________________________________ 
Arlamol E 1.36 A 
Brij 72 5.21 
Mineral Oil 70 11.60 
Propylparaben 0.18 
Purified Water 77.4 B 
Disodium EDTA 0.10 
Dimethicone 0.09 
Methylparaben 0.41 
Propylene Glycol 
1.36 
Dowicil 200 0.05 C 
Purified Water 0.50 
Formaldehyde 37% 
0.2 D 
Germaben II 0.23 E 
______________________________________ 
Procedure: Heat A phase components to 70.degree.-75.degree. C. and mix 
until uniform. Charge main kettle with water and begin heating to 
70.degree.-75.degree. C. Add the remainder of phase B components and mix 
to dissolve the solids. Add, at 70.degree.-75.degree. C., A phase to B 
phase while mixing. Blend well and cool to 35.degree.-40.degree. 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.degree.-30.degree. C. and 
use at this temperature. 
______________________________________ 
EXAMPLE XIII 
Component Weight Percent 
______________________________________ 
Non-Perfumed Moisturizing 
8.70 A 
Lotion (Ex. XIV) 
Purified Water 56.11 B 
Carbopol 940 1.15 
Essence of Rosemary 
0.20 
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 Carbopol and add remaining B phase 
components. Mix until carbopol dispersion is complete. Charge mixing tank 
with A component and hold at 25.degree.-30.degree. C. Add B mixture and 
blend until homogeneous. Add triethanolamine and mix in well until 
homogeneous. Add component D until homogeneous. Add component E and blend 
in well. 
______________________________________ 
EXAMPLE XIV 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: From activated protein separately with solution containing 
reducing agent at a pH above about 9. Mix in remaining components until 
final mixture is homogeneous. 
______________________________________ 
EXAMPLE XV 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 XVI-XXII (Wound Healing) 
A 35 year old man who had a match stick to his finger upon lighting it 
applied the composition from Example I immediately to the wound. It left a 
black mark where the matchhead stuck to the skin. Within 10 minutes the 
pain disappeared. Within 18 hours it was as if there was no wound except 
for a small, dark skin discoloration. There was no blistering, no 
tightening of the skin at the site, no pain and no loss or compromise in 
the use of the digit. Wound healing was greatly accelerated by the 
classical measures of "rubor, dolor and calor." 
A 35 year old New York man cut the temporal aspect of the second phalange 
of his left index finger and applied the formulation of Example III 
immediately. The bleeding wound closed within 15 minutes and the bleeding 
stopped while several people observed it. They tried to separate the edges 
of the wound with their fingers at this time and could not. Clot 
retraction and reepithelialization were complete within 2 days with no 
sclerification. This healing process would have been expected to take 7 to 
10 days otherwise. 
A horse with a 3 year chronic history of a lower leg proud flesh wound 
(equine exuberant granulation) which was treated with many medications 
including systemic antibiotics, laser debulking and skin grafts to no 
avail, responded to the formulation of Examples I and IV. The initial 
lesion was about 3 inches in diamter. Following 3 months of therapy the 
granulation bed reepithelialized and the coat regrew. Skin function and 
appearance is completely restored. This horse is now back in jumping 
competition for the first time in 3 years. Three veterinarians who treated 
this horse did not expect the wound to ever resolve. 
A dog was treated with the formulation of Example IV at the Oradell (N.J.) 
Animal Clinic with a Lick Granuloma of 2 years in duration. The condition 
had been treated by six vets previously to no avail. After thousands of 
dollars of diagnostics and treatment the formulation of Example IV was 
used, and the granuloma resolved. It is now covered with a thick skin, no 
longer weeps or bleeds, and is covered with a healthy coat. The animal no 
longer licks it. After a long and chronic course, the attending 
veterinarian did not expect this granulation to resolve. When it did 
resolve, both he and the owner of the dog call it "miraculous." 
A St. Bernard with bilateral lick granulomas on its forepaws of 6 months 
duration had the formulation of Example II applied daily to one paw. 
Within 4 days the treated wound closed and stopped weeping and bleeding, 
while the untreated wound is unresolved. The owner of this dog, an 
experienced breeder and kennel owner said the response of this wound to 
the formulation was the most dramatic wound healing she had ever observed. 
Two horses owned by different owners (one, a veterinarian) had hock 
injuries that were sutured dehised and reopened due to the continual 
flexion and tension at the hock. Both injuries were subacute and of 6 
weeks and 9 weeks duration. Upon the initial application of the compound 
of Example II a dramatic improvement in both horses was evident within 12 
hours. A clear acceleration of granulation bed formation, 
re-epithelialization and coat restoration was observed over what was 
expected. Both wounds closed and resolved within 14 days of daily therapy 
after not resolving for months previously despite being treated with 
standard therapy. 
Veterinarians at the Meadowlands racetrack (N.J., USA) used the formulation 
of Example II for lower leg injuries in race horses and report greatly 
accelerated time to healing. Specifically, heal cracks and coronary band 
overreach injuries which are common in race horses responded by filling, 
re-epithelialization and regaining a normal appearance and function which 
does not limit their performance. This occurred within 5-15 days versus 
the customary three to eight weeks. 
Examples XXIII-XXV (Hoof Wounds) 
A New Jersey girl found her horse had a split hoof in January of 1988 which 
was not responsive to any therapy. This wound consisted of a split in the 
coronary band and the upper part of the hoof wall which exposed the 
sensitive tissues underneath. In May of 1988 the formulation of Example IV 
was applied daily. Within 2 weeks the wound was 90% healed as determined 
by hoof regrowth filling in the crack. Complete resolution took 4 weeks. 
A horse at the U.S. Equestrian Team barns had kicked its foreleg with a 
rear leg and experienced an overrach injury to the cornary band which was 
expected to heal without treatment in 10-12 weeks. Using the formulation 
of Example IV daily initial healing as defined as a cessation of weeping 
and bleeding and a closing of the wound was observed in 3 days, and 
complete healing occurred within 10 days. 
A horse with a hoof wound resulting in an evulsion which severed the front 
of the hoof wall completely as well as some of the underlying sensitive 
tissue wept and bled, and treatment with traditional therapies for 2 
months was not adequate to stem the weeping and bleeding from the wound. 
Following initiation of daily therapy with the formulation of Example IV 
the wound stopped weeping and bleeding within 2 days. Healing progressed 
at a greatly accelerated rate, and at 2 weeks the sensitive underlying 
tissues had stabilized. At the end of a 3 month period of daily 
application this hoof is completely healed and indistinguishable from the 
other hooves on the horse. 
Examples XXVI-XXVIII (Itching and Rashes) 
A New York dermatologist with a chronic history of a full scalp cap of 
sebborhea characterized by itching and dandruff, which was not responsive 
to any therapy completely responded to the forrmulation of Example I 
within 1 week of daily applications. Itching ceased within 2 days after 
initiating therapy. The characteristic exudate stopped as did flaking of 
the scalp. Continued daily applications of this formulation has completely 
resolved his condition. 
A dog with a history of ideopathic chronic itching which was not resolved 
by any medication or biweekly bathing was treated with a single 
application of this formulation and the itching resolved. It is speculated 
that subsequent applications may be necessary, but have not been needed at 
the time of this writing, which is 3 weeks after the initial application. 
A horse with a history of tail rubbing (resulting in hair loss) which 
began, presumably, as a result of worms but persisted despite effective 
deworming was treated with a single application of this compound following 
a 2 week episode of tail rubbing subsequent to effective deworming. The 
tail rubbing immediately stopped, no further treatment was required, and 
the tail hair grew back in. Tail rubbing is not uncommon in horses. No one 
knows why they do it, and heretofore there was no effective therapy to 
treat it. 
Example XXIX (Rashes) 
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 VIII 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 XXX (Wound Healing) 
A cat had a radial nerve paralysis resulting in a forelimb dragging which 
caused a chronic hair loss and abrasion. The underlying wound was several 
years old. Treatment with the composition of Example VIII resulted in the 
wound closing within days and the growing of fur over a 10 day period 
Example XXXI (Flea Bites) 
Flea bite allergic reactions in dogs cease immediately upon application. 
Usually a Shar-pei gets 1" bald, raised inflamed area when bit. Dramatic 
decrease in flea allergy response in Shar-pei, St. Bernard and Seskie 
Terrier using the composition of Example VIII. 
Example XXXII (Wound Healing) 
The composition of Example VIII sealed a hole in two-three weeks in the 
cheek of a foal which could not be sutured. 
Example XXXIII (Wound Healing) 
A horse stepped upon a stub of a metal fence post and had a 3" wide 
puncture going down to the tendons. After application of the composition 
of Example VIII, the wound healed quickly with no "proud flesh" occurring. 
Example XXXIV (Lick Granuloma) 
A 14 year old doberman had a 7 year history of "lick granuloma." 3 small 
animal vets recommended amputation. Dog got progressively worse until the 
point was reached where the dog could not even walk. Euthanasia was 
considered. The composition of Example VIII was applied to the condition. 
After two months of treatment the dog walked. After two and one half 
months the dog was running, which had not occurred in years. 
Examples XXXV and XXXVI (Resolution of Existing Acne Scars) 
A 29 year old female stock broker used a daily application of the 
composition of Example V and within two months noticed her acne scars had 
significantly decreased in size. 
A 36 year old housewife utilizing the composition of Example V noticed an 
improvement in existing acne pitting within a month of daily application. 
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 necessarily limited thereto. 
Modifications and variations will be apparent from the disclosure and may 
be resorted to without departing from the spirit of the inventions 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.