WO1996025943A1 - Compositions et procedes pour ameliorer la croissance des cheveux et pour leur restituer leur couleur d'origine - Google Patents

Compositions et procedes pour ameliorer la croissance des cheveux et pour leur restituer leur couleur d'origine Download PDF

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Publication number
WO1996025943A1
WO1996025943A1 PCT/US1996/002341 US9602341W WO9625943A1 WO 1996025943 A1 WO1996025943 A1 WO 1996025943A1 US 9602341 W US9602341 W US 9602341W WO 9625943 A1 WO9625943 A1 WO 9625943A1
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Prior art keywords
hair
composition according
growth
insulin
medium
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PCT/US1996/002341
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English (en)
Inventor
Ella Lindenbaum
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Life Medical Sciences, Inc.
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Publication date
Application filed by Life Medical Sciences, Inc. filed Critical Life Medical Sciences, Inc.
Priority to EP96906564A priority Critical patent/EP0812210A4/fr
Priority to AU49904/96A priority patent/AU4990496A/en
Publication of WO1996025943A1 publication Critical patent/WO1996025943A1/fr

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
    • A61Q19/00—Preparations for care of the skin
    • A61Q19/08—Anti-ageing preparations
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • A61K38/22—Hormones
    • A61K38/28—Insulins
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00—Cosmetics or similar toiletry preparations
    • A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
    • A61K8/40—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing nitrogen
    • A61K8/44—Aminocarboxylic acids or derivatives thereof, e.g. aminocarboxylic acids containing sulfur; Salts; Esters or N-acylated derivatives thereof
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00—Cosmetics or similar toiletry preparations
    • A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
    • A61K8/64—Proteins; Peptides; Derivatives or degradation products thereof
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
    • A61Q19/00—Preparations for care of the skin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
    • A61Q3/00—Manicure or pedicure preparations
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
    • A61Q5/00—Preparations for care of the hair
    • A61Q5/06—Preparations for styling the hair, e.g. by temporary shaping or colouring
    • A61Q5/065—Preparations for temporary colouring the hair, e.g. direct dyes
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
    • A61Q7/00—Preparations for affecting hair growth
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
    • C12N5/06—Animal cells or tissues; Human cells or tissues
    • C12N5/0602—Vertebrate cells
    • C12N5/0625—Epidermal cells, skin cells; Cells of the oral mucosa
    • C12N5/0627—Hair cells
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2500/00—Specific components of cell culture medium
    • C12N2500/05—Inorganic components
    • C12N2500/10—Metals; Metal chelators
    • C12N2500/20—Transition metals
    • C12N2500/24—Iron; Fe chelators; Transferrin
    • C12N2500/25—Insulin-transferrin; Insulin-transferrin-selenium
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2500/00—Specific components of cell culture medium
    • C12N2500/90—Serum-free medium, which may still contain naturally-sourced components
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2501/00—Active agents used in cell culture processes, e.g. differentation
    • C12N2501/30—Hormones
    • C12N2501/305—Growth hormone [GH], aka. somatotropin
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2501/00—Active agents used in cell culture processes, e.g. differentation
    • C12N2501/30—Hormones
    • C12N2501/38—Hormones with nuclear receptors
    • C12N2501/395—Thyroid hormones

Definitions

  • the present invention relates to methods for stimulat ⁇ ing hair follicles and promoting the growth of hair in intact skin, especially the scalp. More particularly, the invention relates to a method for inducing angiogenesis and causing increased vascularization and increased circulation (vasodila- tion) of intact skin tissue and stimulating dormant, dying or weak hair follicles to produce hair or promote the growth of hair in such skin and reduce hair loss.
  • the present invention also relates to a method for stimulating nail growth and producing stronger nail (ungual) tissue.
  • the present invention also relates to a method for promoting melanogenesis and restoring the natural color to hair in which the color is diminished or diminishing, such as in gray or graying hair.
  • compositions of the present invention which are used to enhance hair and nail growth and promote melanogenesis and restore natural hair color are compositions which are based on a minimum essential medium in combination with preferably at least two non-steroidal anabolic hormones one of which hormones is insulin and the other which is selected from triiodothyronine, thyroxine and growth hormone.
  • a minimum essential medium in combination with preferably at least two non-steroidal anabolic hormones one of which hormones is insulin and the other which is selected from triiodothyronine, thyroxine and growth hormone.
  • Three anabolic hormones, including insulin, triiodothyronine or thyroxine and growth hormone in combination with minimum essential media may be preferably used.
  • an enriched growth media such as MCDB 153 or related enriched growth media is used in combination with insulin, triiodothyronine or thyroxine and growth hormone.
  • Composi ⁇ tions for promoting hair and nail growth may also contain an effective amount of a penetration enhancement agent for promoting the penetration of the active constituents of the compositions through the surface of the skin where angiogenesis, melanogenesis and vascularization may be promoted.
  • Hair follicles produce different types of hair at dif ⁇ ferent stages in an animal's life.
  • the hairs formed may change quite dramatically in thickness, color, and length.
  • hair follicles have a transforming ability which allows animals to adjust to seasonal changes and to life cycle changes (i.e., hormonal or metabolic changes) over the course of an animal's life. This allows the rapid distinction between young animals and adults and between the adult sexes.
  • hair growth plays a major role in social and sexual com ⁇ munication in mammals and explains why hair disorders such as hirsutism, androgenetic alopecia and alopecia areata often create psychological problems.
  • hair fol ⁇ licles pass through regular cycles of hair development and growth (anagen) followed by periods of resting (telegen) .
  • the hair produced after a resting period may be very similar to the previous one produced, as seen in many hair follicle cycles on the eyelids and the young human scalp; or, it may be slightly or even markedly different.
  • the hair may actually regress in qual ⁇ ity and cease growing.
  • the precise mechanism of how altera ⁇ tions occur in the type of hair produced by a hair follicle are unknown. Methods which are shown to stimulate hair fol ⁇ licles and enhance the growth of hair may be used to regulate hair follicle disorders and increase hair production by domesticated animals such as sheep.
  • the hair follicle is composed of epithelial components (the matrix and outer root sheath) and dermal components (the dermal papilla and connective tissue sheath) .
  • Hair growth is effected by the division of the hair follicle matrix cells under the control of the dermal papilla.
  • Three distinct stages of hair growth can be identified, an active phase (anagen) during which hair growth occurs, an intermediate regressive phase (catagen) during which hair actually ceases growing or even regresses and a resting phase (telogen) during which no cell proliferation occurs.
  • the factors that regulate cell division are poorly understood, although growth factors, steroid hormones, dermoepithelial interactions and the immune system have been implicated. Philpott, et al., J.
  • EGF epidermal growth factor
  • TGF transforming growth factor
  • Minoxidil tln has been shown to be efficacious in enhancing hair growth. Its mechanism of action is based upon its vasodilatory effect which causes shifting of the fluid volume from the vascular compartment into the extracellular compartment, especially in the dermal area. This, in turn, may cause an increase in blood circulation in the dermis and increased supply of nutrients to the dermis affecting the hair follicles.
  • MCDB 153 supplemented with non-steroidal anabolic hormones was used as treatment on surgical wounds.
  • the skin on the dorsum of all the animals was depilated prior to the extirpation of the skin patches.
  • the application of MCDB 153 including anabolic hormones induced vascularized granulation tissue formation as well as epithelialization.
  • the barrier for penetration through the skin, the keratinized layer was overcome by the opening in the wound area. This condition allows for unhindered contact between the medium and the deeper layers of the skin including the hair follicle cells and the capillary endothelial cells.
  • the present invention relates to formulations and methods for promoting hair or nail (ungual tissue) growth in intact skin of animals, especially including the scalp of humans.
  • the formulations according to the present invention are also useful for promoting angiogenesis in the dermis and for stimulating dormant, dying or weak hair follicles to pro ⁇ quiz hair or promote the growth of hair in such skin.
  • the natural hair color of the growing hair is restored in hair diminished in its natural color. This is an unexpected result.
  • Compositions according to the present invention are also useful for enhancing the growth and increasing the strength of ungual tissue as well as correcting irregularities in such tissue.
  • compositions according to the present invention are useful for enhancing hair growth by causing increased vascularization of intact skin tissue and stimulating dormant, dying or weak hair follicles to produce hair or promote the growth of hair in such skin.
  • the formula ⁇ tions according to the present invention are also useful for promoting melanogenesis in hair follicles and for restoring the hair color in hair which exhibits diminished hair color such as in gray, graying and damaged hair.
  • the present formulations comprise a hair or nail growth enhancing effective amount of a non-steroidal anabolic hormone selected from insulin, growth hormone, triiodothyronine and thyroxine (T 3 or T4) , and mixtures thereof, most preferably a mixture of all three hormones, in combination with a hair or nail growth enhancing effective amount of minimum essential medium, preferably a supplemented medium such as MCDB 153.
  • the formulations may be adjusted to enhance penetration of the individual components through intact skin in order to promote angiogenesis of the underlying dermal layers and consequently, to stimulate hair follicles and ungual tissue and promote hair or ungual tissue growth. All of the components used in the compositions according to the present invention are included in amounts effective for an intended use generally- enhancing or stimulating hair and/or nail growth or restoring hair color in hair exhibiting diminished color.
  • the non-steroidal anabolic hormone is a mixture of insulin and at least one or more anabolic hormones such as triiodothyronine, thyroxine and growth hormone. It has been unexpectedly discovered that a mixture of the anabolic hormones insulin and triiodothyronine, thyroxine or growth hormone produces a synergistic enhancement in hair growth when combined with a minimum essential medium.
  • the anabolic hormone comprises a mixture of an amount of insulin, growth hormone and triiodothyronine or thyroxine in amounts effective to substantially enhance the growth of hair or ungual tissue, also synergistically.
  • Embodiments in which the anabolic hormone is a mixture of effective amounts of triiodothyronine or thyroxine and growth hormone or insulin and growth hormone are also contemplated by the present invention.
  • hair color restoration occurs in hair which is diminished in hair color.
  • Composi- tions which restore hair color are generally made of the same componentry and such components are included in the same gen ⁇ eral amounts which are useful for promoting hair growth.
  • insulin is included in compositions according to the present invention at concentrations ranging from about 5 ng/ml (nanograms/ml) to about 100 ug/ml (micro- grams/ml) (preferably, at least about 50 ng/ml within this range), more preferably about 500 ng/ml to about 20 ug/ml.
  • compositions according to the present invention when a non-steroidal anabolic hormone other than insulin is included in compositions according to the present invention, for example, triiodothyronine, thyroxine or human growth hormone, among others, each of these other hormones is included in an amount effective to enhance the growth of hair or ungual tissue, i.e., in an amount of at least about 0.05 ng/ml of the formulation, with a preferred range of about 0.5 ng/ml to about 100 ng/ml.
  • compositions which are delivered in solid or concentrated form i.e.
  • the anabolic hormone is included in concentrations similar to those contained in the solutions (based upon the general assumption that 1 ml of solution is approximately equal to about 1 gram in weight of the final composition) . Percent weights may fall outside of these ranges, depending upon the ability to deliver the individual components of the formulations through the skin, the level of stability of the hormone and other factors, as well recognized by one of ordinary skill in the art.
  • compositions according to the present invention preferably include at least one anabolic hormone selected from triiodothyronine, thyroxine, and growth hormone and most preferably both growth hormone and triidothyronine or thyroxine, in combination with at least a minimum essential medium, preferably a supplemented minimum essential medium such as MCDB 153.
  • the preferred growth hormone is human growth hormone, preferably in combination with triiodothyronine (T 3 ) , thyroxine (T4) or insulin and more preferably in combination with both insulin and triiodothyronine (T 3 ) or thyroxine (T 4 ) .
  • anabolic hormone other than insulin used will generally depend on the extent and rate of hair or nail growth desired or damage to hair, nail or skin tissue which is to be corrected, but in most of the cases the amount of hormone will fall within a preferred range of about 0.5 ng/ml and about 100 ng/ml by weight or more of the com ⁇ position.
  • growth hormone preferably human growth hormone
  • Triiodothyronine (T 3 ) or thyroxine (T 4 ) is preferably included in amounts ranging from about 0.5 ng/ml to about 100 ng/ml or more. Triiodothyronine (T 3 ) may be preferred over thyroxine (T4) because it has greater potency and the same general activity as thyroxine. Thyroxine (T4) , however, is more storage stable than is triiodothyronine (T 3 ) and thyroxine's stability should be taken into account and is preferred when formulating compositions which are to be stored for at least several weeks or more.
  • Triiodothyronine (T 3 ) and thyroxine (T4) may be readily substituted for each other, however, with the general rule that at least about three to five times the amount of thyroxine (T4) is substituted for triiodothyronine (T 3 ) .
  • Figures 1-2 represent the results of the experiments performed and described in Example 3. These graphs show the relative degree of increase in blood flow measured by Laser- Doppler Flow changes on burn wounds treated with a composition of the present invention and conventional therapy (Silverol) .
  • angiogenesis is used throughout the speci ⁇ fication to describe processes which result in the development of new blood vessels and the vasculature (neovasculature) .
  • vasculature neovasculature
  • melanogenesis is used throughout the speci ⁇ fication to describe processes which result in the development or restoration of pigment or color in the hair as it grows.
  • rejuvenated i.e., smoother, more moist, softer, oily/waxy, and more cosmetically pleasing to the vision and touch.
  • delivery polymer or “gelling agent” is used throughout the specification to describe a polymer or other gelling agent which can be used in combination with minimum essential medium and non-steroidal anabolic hormones selected from insulin, triiodothyronine or thyroxine and growth hormone and mixtures thereof, and optionally, a penetration enhance ⁇ ment agent, the amount of delivery polymer included in an amount effective to gel the composition and hold it in place on the intact skin to be treated without running off.
  • delivery polymer and gelling agent are synonymous where the amount of delivery polymer or gelling agent included is effective to gel the composition.
  • These delivery polymers include, for.example, numerous hydrogels in hydrated or unhydrated form, such as those derived from hydroxyethylmethacrylate (HEMA) , glycerolmethacrylate (GMA) and polyvinylpyrrolidone (PVP) , polyethylene glycol (PEG) , various carbohydrates, cellulose, cellulose ethers, including methyl cellulose, hydroxyethyl cellulose and hydroxypropyl cellulose, dextran, polythyleneoxide, dextran-polyethylene, acrylamide, polyacrylamide, amylose, collagen, gelatin, sepharose, agarose (for example, as an agarose saturated gel) , related polymers and mixtures thereof, among numerous others.
  • HEMA hydroxyethylmethacrylate
  • GMA glycerolmethacrylate
  • PVP polyvinylpyrrolidone
  • PEG polyethylene glycol
  • various carbohydrates cellulose, cellulose ethers, including methyl
  • the cellulose ethers especially including methyl cel ⁇ lulose, hydroxymethyl cellulose, hydroxyethylcellulose and hydroxypropylcellulose are preferred and are preferably included at a weight ratio of about 0.1% to about 20% by weight, more preferably about 0.5% to about 10% by weight of the hair growth compositions.
  • Methylcellulose and hydroxyethyl cellulose are preferred cellulose ethers for use in the hair gel compositions according to the present inven ⁇ tion, with hydroxyethylcelllulose being more preferred.
  • delivery polymer is also used to describe polymers which instill slow-release or sustained release characteristics to the hair and nail growth formulations of the invention.
  • minimum essential medium is used throughout the specification to describe a medium or mixture which con ⁇ tains no serum, and in combination with anabolic hormone and optionally, a penetration enhancement agent, comprises the compositions according to the present invention.
  • minimum essential medium is readily understood by those in the art to comprise a nutrient media which supports cellular growth.
  • the minimum essential medium preferably comprises the following elements: (a) essential amino acids; (b) non-essential amino acids; (c) vitamins selected from the group consisting of biotin, folate, lipoate, niacinamide, pantothenate, pyridoxine, riboflavin, thiamin and vitamin B 12 an ⁇ mixtures thereof, preferably a vitamin mixture comprising folate, niacinamide, pantothenate, pyridoxine, riboflavin and thiamin; (d) glucose; and (e) a mixture of inorganic ions selected from the group consisting of calcium, sodium, potassium, magnesium, chloride and mix ⁇ tures thereof, preferably a mixture comprising calcium, sodium, potassium, magnesium and chloride.
  • vitamin C may be included as one of the preferred vitamins in the present formulations.
  • a minimum essen ⁇ tial medium for use in the present invention may exclude non- essential amino acids (b) , but preferably, non-essential amino acids are included in combination with essential amino acids.
  • Especially preferred amino acids include glutamine, serine and cysteine.
  • All of the above-described elements (a) , (b) , (c) , (d) and (e) are included with the anabolic hormone mixture in con- centrations and/or amounts effective for enhancing or stimulating the growth of hair and/or nails or revitalizing skin tissue. It is noted that amounts of these components which stimulate or enhance the growth of hair also quite unex ⁇ pectedly restore the natural hair color to the growing hair in many instances where the hair color is diminished and is char ⁇ acterized as being gray or graying.
  • the preferred concentra ⁇ tion of essential and optional non-essential amino acids used in the present invention ranges from about 5.0 urn (10 ⁇ 6 mole) to about 50 mmol. (10 ⁇ 3 mole) .
  • the preferred concentrations of vitamins used in the present invention ranges from about 1 nanomole (10 ⁇ 9 mol.) to about 10 um.
  • the preferred concentra ⁇ tions of glucose used in the invention ranges from about 1 umol. to about 10 or more mmol.
  • these inorganic ions are preferably included in the present compositions at a concentration range of about 1 umol to about 50 mmol.
  • a penetration enhancing agent is also included in the formulations.
  • the nutrient medium according to the present invention may also contain any one or more of the following elements: (f) purines and pyrimidines; (g) other organic com ⁇ pounds; (h) other inorganic ions; (i) trace elements; and (j) buffers and indicators.
  • All of these optional elements (f) , (9) i (h) , (i) and (j), when they are included in the nutrient medium according to the present invention, are included in amounts effective, in combination with the anabolic hormone(s), for promoting angiogenesis, enhancing vasculariza ⁇ tion and/or stimulating dormant, dying or weak hair follicles to promote hair growth and, in certain instances, to restore the growing hair to its natural pigment or color.
  • these components are added to stimulate or enhance the growth of ungual tissue, including nail tissue of humans.
  • these components are added in amounts effective to stimulate, enhance and revitalize the growth and appearance of intact skin tissue of animals, including humans.
  • components (f) , (g) , (j) and (k) range in concentration from about 1 nmol. to about 10 mmol. In the case of components (h) and (j), the concentration preferably ranges from about 1 umol. to about 50 mmol.
  • concentration preferably ranges from about 1 umol. to about 50 mmol.
  • the present invention may also make use of medium containing serum, although the use of a serum containing cellular nutrient medium is generally less preferred than is serum free medium.
  • examples of such nutrient medium include, among numerous others, DMEM, HAM F12 and HAM F10, all containing serum.
  • minimum essential medium is used throughout the specification to describe all types of nutrient medium contemplated for use in the present invention which contain at least the basic elements as described hereinabove, as well as supplemental components.
  • MCDB 153 is a preferred supplemented minimum essential medium for use in the present invention. MCDB 153 is believed to contain the components which are associated with an enhanced rate of hair growth as well as other components which enhance the rate of angiogenesis and vascularization of tissue surrounding the hair follicle and consequently, the growth of hair from the follicle and, in many instances, the restoration of natural hair color in growing hair.
  • the minimum essential medium according to the present invention may include one or more commercially available media in solution or lyophilate (solid) form.
  • the cellular nutrient medium used may be in the form of a lyophilate which may be reconstituted with water, preferably sterilized, distilled water and then supplemented with an anabolic hormone such as insulin, triiodothyronine, thyroxine, growth hormone or mix ⁇ tures thereof, and optionally, certain penetration enhancing agents or other additives. Lyophilized forms of insulin, growth hormone and triiodothyronine or thyroxine may also be used in the present compositions.
  • the medium may be used directly in formulations according to the present invention in the form of a lyophilate, or related solid-type aterial, rather than a solution, especially when a gel is to be used for delivery. It is clearly preferred when utilizing solid-type materials for delivering the wound healing composi ⁇ tions according to the present invention that the delivery system in the form of a gel or other form contain moistening quantities of water.
  • non-steroidal anabolic hormone is used throughout the specification to describe the primary hormones which are included in the instant invention in combination with minimum essential media to enhance or stimulate hair or nail growth.
  • These primary hormones include insulin, triiodothyronine, thyroxine, and growth hormone, among others.
  • growth hormone it is preferred to use it in com ⁇ bination with triiodothyronine or thyroxine or insulin or most preferably with a mixture of triiodothyronine or thyroxine and insulin.
  • non-steroidal anabolic hormone includes naturally isolated (preferably, human) or synthetically produced versions of these hormones which are known to function substantially the same as the naturally occuring hormones and includes, where relevant, compounds pro ⁇ substituted by genetic engineering processes and recombinant techni ⁇ ques. While not being limited by way of theory, it is believed that the inclusion of at least one non-steroidal anabolic steroid selected from insulin and triiodothyronine or thyroxine (preferably, insulin) serves to enhance the effect of the minimum essential media in unexpectedly increasing the rate and quality of hair (particularly, with respect to restoration of hair color) and nail growth.
  • a combination of at least two anabolic hormones e.g., insulin and growth hormone, insulin and triiodothyronine or thyroxine, or growth hormone and triiodothyronine or thyroxine is preferably used because a combination of non-steroidal anabolic hormones and minimum essential medium creates an enhancement of tissue growth (hair, ungual tissue, skin revitalization) which is greater than the sum of the individual parts.
  • Three anabolic hormones (insulin, triiodothyronine or thyroxine and growth hormone) in effective amounts in combination with minimum essential medium acts synergistically to promote angiogenesis, vascularization and consequently, hair growth and in many instances, melanogenesis and hair color restoration.
  • the non-steroidal anabolic hormone actually enables the cells to utilize or process the nutrients in the media, which action results in angiogenesis, and an enhance ⁇ ment of vascularization and rate of hair and nail growth and skin revitalization. Restoration of hair color is an auxiliary beneficial result which occurs along with enhanced hair growth in many instances.
  • compositions according to the present invention may be used with or without the inclusion of a penetration enhancement agent. It is an unexpected result that compositions which exclude a penetration enhancement agent will be absorbed into intact skin and evidence a sur ⁇ prising degree of activity (i.e., hair or nail growth enhance ⁇ ment or skin revitalization) . While not being limited by way of theory, it is believed that the components used in the present composition are absorbed through the skin by way of hair roots and pores in the skin in a manner sufficient to evidence significant activity.
  • Exemplary penetration enhancement agents which may be used to treat the skin before applying the present formula- -lo ⁇
  • tions include, for example, soaps and detergents (solvents for removing natural skin oils) , depilatories, epilatories such as trichloroacetic acid and phenol, enzymes which detach an epithelial layer from the dermal substratum including trypsin, collagenase, hyaluronidase, elastase and dispase and keratin solvents such as urea and DMSO.
  • water may be used as a penetration enhancement agent, as hydration of the skin is associated with better pen ⁇ etration. All of these agents are used in concentrations and amounts which are effective for treating the skin in order to enhance the penetration of the individual components in the hair and nail growth compositions according to the present invention through intact skin.
  • these enzymes trypsin, collagenase, dis ⁇ pase, hyaluronidase and elastase, these enzymes are used to treat intact skin in an amount ranging from about 0.1 to about 10 mg/ml., more preferably about 1 mg/ml.
  • this epilatory should be used in dilute concentrations in water- generally about 1% by weight or less.
  • urea this is generally used as a mixture of 20% by weight urea in lanolin (water may be sub ⁇ stituted for the lanolin) .
  • DMSO may also be used, generally as a dilute solution in water (0.25% to about 5% by weight, preferably about 1% to about 2% by weight of the solution) .
  • Water may also be used preferably as a penetration enhancer in certain instances. Where water is used, the amount used is that which will wet or hydrate the surface of the skin or scalp to which the compositions of the present invention are applied.
  • penetration enhancement agents which may be used in the present compositions include, for example, ionophoresis agents such as the mucopolysaccharides, for example chondroitin, chondroitin-6-sulphate and dermatan sulphate, among others. These compounds are generally included in com ⁇ positions according to the present invention in effective amounts to enhance penetration of the individual components through intact skin.
  • a negatively charged synthetic membrane (which generates a static charge) may also be used for enhancing the individual components in composi ⁇ tions according to the present invention.
  • each penetration enhancement agent which is used in the formulations according to the present invention will depend upon the requirement for hair or nail growth or skin revitalization, but each component is included in an amount effective for producing the intended results, for exam ⁇ ple, substantially enhancing the rate of penetration of the formulations through the skin. This amount will vary accord ⁇ ing to the type of agent used.
  • One of ordinary skill in the art will know to vary the amount and type of agent within the weight ranges defined above to enhance penetration through intact skin and promote the efficacy of compositions according to the present invention.
  • the formulations include an anabolic hormone other than insulin at a concentration of at least about 0.05 ng/ml, preferably about 0.5 ng/ml to about 100 ng/ml or more.
  • the amount of insulin generally falls outside of this range.
  • the anabolic hormone comprises a mixture of insulin, triiodothyronine or thyroxine and growth hormone because of the known benefits these hormones have in promoting the growth and elaboration of cells and their general absence of toxicity.
  • it is this combination of anabolic hormones which evidences unexpected synergistic activity in promoting angiogenesis, vascularization and hair or nail growth in the instant invention.
  • the preferred insulin is human insulin (more preferably human recombinant or genetically engineered insulin) , which is a well-known protein which is readily available commercially from a number of sources (for example, Novo Nordisk, Copenhagen, Denmark, among others) . It is con ⁇ stituted from a number of amino acids (approximately 51) with a total molecular weight of about 5,500.
  • Human insulin for use in the present invention is generally prepared using genetic engineering techniques. Depending upon the manufac ⁇ turer, the insulin may have slightly different activity based upon weight, however the activity of insulin defined in units is, of course, standard. While not being limited by way of theory, in the present invention, it is believed that the insulin promotes hair or nail growth at least in part by enhancing and stimulating the transport and utilization of glucose as an energy source by the hair or nail growing cells.
  • Growth hormone may also be used in the present inven ⁇ tion, preferably in combination with insulin or triiodothyronine or thyroxine and most preferably in combina ⁇ tion with both triiodothyronine or thyroxine and insulin.
  • the preferred human growth hormone is a well-known defined protein which is readily available and results from a pituitary secre ⁇ tion into the blood system. It is constituted from a number of amino acids with a total molecular weight of about 193,000.
  • the human growth hormone which may be used in the present invention can be obtained from a variety of sources, including genetic engineering processes and techniques.
  • the growth hormone serves to stimulate and enhance angiogenesis, i.e., the elaboration and development of the vascular system which is believed to also enhance the blood supply and the delivery of nutrients to hair follices and nail tissue so as to produce an enhancement or stimulation in hair or nail growth and, in certain instances, restoration of natural color.
  • the action of growth hormone is believed to be synergistic with insulin and/or triiodothyronine or thyroxine.
  • the present invention also contemplates the inclusion of effective amounts of triiodothyronine or thyroxine either alone, but preferably in combination with other non-steroidal anabolic hormones.
  • the preferred triiodothyronine is human triiodothyronine, which is a well-known defined hormone and readily available commercially.
  • Triiodothyronine and thyroxine are naturally occurring amino acids of the thyroid gland which exert a stimulating effect on metabolism. Although virtually identical in metabolic effects, triiodothyronine is more potent than is thyroxine but is less stable.
  • thyroxine is substituted for triiodothyronine, to obtain the same effect as triiodothyronine, thyroxine is added in an amount between about three and five times that of triiodothyronine.
  • Thyroxine being more stable, is preferred for use in the present invention in those compositions which advantageously have storage stable characteristics. While not being limited by way of theory, it is believed that the triiodothyronine or thyroxine utilized in the present inven ⁇ tion stimulates vascularization and facilitates the resupply of blood borne components in cells responsible for the growth of hair.
  • a particularly preferred composition according to the present invention comprises a mixture of an effective amount of human growth hormone in the presence of an effective amount of insulin and triiodothyronine (T 3 ) or thyroxine (T4) , preferably in a serum free cellular nutrient medium, most preferably MCDB 153.
  • the anabolic hormones other than insulin i.e., growth hormone or triiodothyronine
  • the anabolic hormones other than insulin are generally included in the final composition in a concentration range of about 0.05 ng/ml to about 100 ng/ml, preferably about 0.5 ng/ml to about 20 ng/ml or more and most preferably about 1 ng/ml to about 20 ng/ml.
  • thyroxine is generally included in an amount ranging from about 0.5 ng/ml to about 100 ng/ml or more, generally at a concentration of at least about three-five times that of triiodothyronine.
  • insulin is also included in an effective amount, generally an amount which is substantially greater than the other anabolic hormones.
  • the amount of insulin is preferably included in amounts ranging from about 5ng/ml to about 100 ug/ml more preferably about 50 ng/ml to about 20 ug/ml and even more preferably about 500 ng/ml to about 20 ug/ml.
  • One of ordinary skill in the art will know to vary the amount of anabolic hormones within effective ranges based upon the type and potency of the preparation of the compound in order to enhance or stimulate the growth of hair or nail tis ⁇ sue according to the present invention.
  • compositions according to the present invention preferably comprise effective amounts of minimum essential medium in combination with effective amounts of insulin, triiodothyronine or thyroxine and growth hormone. It is a combination of these three anabolic hormones in combination with minimum essential medium which has exhibited the greatest synergistic activity in promoting angiogenesis, vasculariza ⁇ tion, increased blood flow and hair or nail growth.
  • Suppl- mented minimum essential medium is preferably used and MCDB 153 is the most preferred media to be used in the instant invention.
  • the inclusion of effective concentrations of selenide are optional.
  • the inclusion of effective amounts of vitamin C (ascorbic acid) is preferred.
  • the compositions are preferably transferrin-free, although transferrin may be added.
  • the minimum essential medium which is used in the present invention is any medium having the effect of promoting hair growth when used in effective amounts in combination with the non-steroidal anabolic hormones.
  • the nutrient media, com ⁇ prised of the componentry set forth hereinabove is mixed with an effective amount of the non-steroidal anabolic hormone(s) to form the compositions according to the present invention.
  • Minimum essential medium is readily recog ⁇ nized by those of ordinary skill in the art.
  • Minimum essen ⁇ tial medium is known to preferably comprise effective amounts of the following constituents: (a) essential amino acids; (b) non-essential amino acids; (c) vitamins as previously described; (d) inorganic ions as previously described and (e) glucose; and optionally, (f) purines and pyrimidines; (g) other organic compounds; (h) other inorganic ions; (i) trace elements; (j) buffers and indicators and (k) other supple ⁇ ments.
  • the medium used herein also contains effective amounts of elements (f) through (k) . Serum free nutrient medium is preferred.
  • non-essential amino acids (b) are preferably added, but are not required.
  • the preferred serum free nutrient medium is modified MCDB 153, a well-known medium. Mixtures of standard commercial nutrient media may also be used with favorable results in the instant invention.
  • the mechanism which might be assumed is that new capillaries appear in the tissue surround ⁇ ing the hair follicles and related nail tissue from the first day on and reach their maximum levels after one week or so.
  • the new vessels in granulation tissue originate as budlike structures on nearby vessels, enhance vascularization, become canalized and ramify throughout the dermis in proximity to the hair follicles and nail growth tissue.
  • the resulting increase in vascularization provides more nutrition and stimulatory factors for the hair follicle and nail and skin tissue, the consequence of which is the stimulation of the hair follicle and related nail and skin tissue and the enhancement of pro ⁇ duction of hair, nail and skin tissue.
  • the function of the medium is to provide nutrients to normal, distressed and injured fol ⁇ licles of the dermis in order to stimulate the follicle and enhance the growth of hair.
  • the medium functions with the non-steroidal anabolic hormone to promote the normal processes of stimulation of hair follicles and the enhanced growth of hair.
  • the color of the hair returns to its natural color.
  • a number of nutrient media may be used in the present invention, including commercially available media or other media well known in the art.
  • examples of such media include ADC-1, LPM (Bovine Serum Albumin-free) , F10 (HAM), F12 (HAM), DCCM1, DCCM2, RPMI 1640, MCDB 105, MCDB 110, MCDB 202, MCDB 402, MCDB 153, BGJ Medium (with or without Fitton-Jackson Modification) , Minimum Essential Medium Eagle, Basal Medium Eagle (BME-with the addi ⁇ tion of Earle's salt base) , Dulbecco's Modified Eagle Medium (DMEM-without serum) , Glasgow Modification Eagle Medium (GMEM) , Leibovitz L-15 Medium, McCoy's 5A Medium, Medium M199 (M199E- with Earle's salt base), Medium M199 (M199H- with Hank's salt
  • serum-containing nutrient media may also be used in compositions according to the present invention, but the use of serum-containing media is less preferred because of the possibility that the serum may be contaminated with microbial agents and because the patient may develop immunological reactions to certain antigenic components con- tained in the serum.
  • a preferred nutrient media for use in the present invention is modified MCDB 153.
  • Vitamin B12 3.0 x 10-7
  • Weights of each of the above components in the medium may be varied within the concentrations described hereinabove (in hair or nail growth enhancing effective amounts) to pro ⁇ vide formulations workable within the description of the pres ⁇ ent invention.
  • the non-steroidal anabolic hormone to be incorporated into the modified MCDB 153 composition is a mixture of at least two hormones selected from insulin, triiodothyronine/thyronine and growth hormone at hair growth enhancing effective concentra ⁇ tions.
  • the anabolic hormone includes a mix ⁇ ture of human growth hormone, insulin (containing transferrin or transferrin-free) and triiodothyronine (T 3 ) or thyroxin (T 4 ) , each hormone included in a hair growth enhancing effec ⁇ tive amount.
  • the three hormone combination exhibits an unex ⁇ pected synergistic effect in promoting hair growth.
  • Hormones other than insulin are included in an amount ranging from at least about 0.05ng/ml, preferably at least about 0.5ng/ml to about 100 ng/ml, and more preferably about 1 ng/ml to about 100 ng/ml.
  • thyroxine it is generally sub ⁇ stituted for triiodothyronine at a concentration of at least about three to five times the concentration of triiodothyronine used.
  • the effective amount of insulin generally ranges from about 5ng/ml to about lOOug/ml and more preferably about 50 ng/ml to about 20ug/ml, even more preferably about 500 ng/ml to about 20 ug/ml within this range.
  • Insulin is a desirable constituent anabolic hormone, found to impart a maturing stimulus of the growing culture. Insulin may be commercially obtained and is generally provided in mU quantities (about 41 ng of insulin) .
  • the Standard Preparation is a quantity of purified Zinc Insulin crystals extracted 52% from Bovine and 48% from Porcine pancreas (See, Martindale Pharmacopoeia, 26th Ed.).
  • formulations according to the present invention may also contain transfer- rin (which is believed to improve iron transport) in amounts which preferably range from about 500 ng/ml to about 50 ug/ml, more preferably about 5-10 ug/ml and selenite (preferably, in the form of sodium selenite) in amounts preferably ranging from about 0.5 to about 50 ng/ml, more preferably about 5-10 ng/ml.
  • transferrin is not critical to the use of the instant invention and in certain cases may be less preferred.
  • the formulations according to the present invention may also include an effective amount of an anti-dandruff agent or other ingredients which are commonly applied to the scalp or hair, including antimicrobial agents, where desirable, generally in amounts found useful in topical applications.
  • an anti-fungal agent for use in preventing the outgrowth or fur ⁇ ther growth of nail fungus infections is also optional.
  • One of ordinary skill in the art can easily determine the type and amount of anti-dandruff or antimicrobial agent chosen for use in formulations according to the present invention.
  • the formulations as described herein are further for ⁇ mulated with gelation agents or related delivery polymers for gelling the formulations according to the present invention for delivery to the area of intact skin to be treated.
  • the formulations comprising effective amounts of anabolic hormone(s) and nutrient media, either alone or in addition to other optional components, especially including an effective amount of a penetration enhancement agent, are admixed with amounts of a delivery polymer effec ⁇ tive for producing a gel, for example a hydrogel polymer derived from HEMA (hydroxyethylmethacrylate) or NVP (N- vinylpyrrolidone) , polyethylene glycol (PEG) , polyethylene, gelatin, various carbohydrates, sepharose, agarose, methylcel- lulose and related hydrophilic cellulose ethers including cel ⁇ lulose, hydroxymethyl cellulose, hydroxyethyl cellulose and hydroxypropylcellulose, dextran, polyethyleneoxid
  • the amount of delivery polymer which is added to the formulations to produce a gel is that amount which solidifies the composition to a point where the composition does not easily flow off of the intact skin to be treated and generally ranges from about 0.1% by weight to about 20% by weight, preferably about 0.5% to about 10% by weight, depending upon the type of delivery polymer used.
  • the gel compositions according to the present invention preferably contain sufficient water or moisture to maintain the area to be treated in a relatively moist state- a condition shown to be beneficial for penetration enhancement of the individual components of the formulations through the skin.
  • compositions according to the present invention may have to be varied in order to maintain effective concentrations.
  • these compositions may also contain an amount of a pharmaceutically acceptable excipient and, in addition, other additives such as diluents, compound ⁇ ing agents, bulking agents, surfactants and the like.
  • the formulations as described hereinabove are topically applied to the skin tissue to be treated as a liquid or gel at least once a day and preferably at least twice a day and up to six or more times a day. In many instances it may be advisable to apply the compositions to the scalp or other area of the skin to be treated at least once a day after showering or bathing. In the case of formulations containing a delivery polymer, preferably a moistened delivery polymer, the formulations may be administered less frequently than when the formulations are applied as a liquid.
  • a delivery polymer preferably a moistened delivery polymer
  • One possible regimen for utilizing the compositions according to the instant invention is to first treat the skin with a penetration enhancement agent in an amount (i.e., at a concentration) and for a time effective for enhancing pene ⁇ tration of the components of the hair or nail growth composi ⁇ tion.
  • the penetration enhancement agent is generally applied to the intact skin to be treated with the hair growth composition for 30 seconds up to one half hour or more (preferably from about 1 to about 5 minutes) to condition the skin for penetration. It is noted that the inclusion of a penetration enhancement agent -is unexpectedly not required in order to deliver effective amounts of the present compositions to promote hair growth, nail growth and skin revitalization.
  • the skin to which the composition is to be applied is first wet with water, after which time, the composition is applied.
  • a cover ⁇ ing such as a polyethylene shower cap, may be used to keep the composition in contact with the area to be treated.
  • the scalp is simply washed with soap or detergents to remove all natural oils from the scalp before application of the hair growth enhancing composition.
  • the treated skin is then exposed to the hair growth composition (preferably, after wetting with water to promote penetration) and massaged into the scalp.
  • the hair growth composition is a gel which is rubbed onto the area of the skin to be treated and is left on the skin for a convenient period, generally at least 1-3 hours and preferably for an eight hour period such as overnight.
  • a covering is preferably placed over the gel in order to keep the gel in contact with the treated skin area. To remove the composition, it can be washed off with clear water.
  • the amount of material which is to be spread on the skin or massaged into the scalp to be treated will be readily apparent.
  • the formulations should be applied in a manner which is cosmetically appealing. In general, in solution or gel form, about 0.5-2.0 cc of formulation is applied per 5-10 cm 2 to the area of the scalp. Depending upon the requirement of hair growth and the extent of baldness, an amount greater or less than 0.5-1 cc of formulation per 5-10 cm 2 of the wound surface may be utilized. In many instances, the depth of the formulation on the skin should be at least about 0.05-0.1 mm. Greater depths may also be used, but generally at the expense of the cosmetic appeal of the product.
  • more material may have to be added to the hair in order to assure the indi ⁇ vidual that a sufficient amount of the composition will come into contact with the underlying skin or scalp area.
  • the treatment regimen and compositions used are virtually identical to the method used for enhancing the growth of hair.
  • Restoration of hair color generally will occur as an auxiliary result of enhanced growth of hair.
  • the hair color is diminished in color, for example, in gray, graying or even white hair
  • the methods of use will be the same as if the desired result is enhanced hair growth, but in such a case, the more clearly desired result will be restoration of hair color.
  • Enhanced hair growth most likely will also occur.
  • treatment may be at night or during the day.
  • the tissue of the patient is treated by applying the compositions according to the present invention directly on the tissue after first wetting the tis ⁇ sue and, in the case of ungual tissue, to the cuticle area of the nail or tissue where the nail or ungual tissue is attached to the nail or tissue bed.
  • this night treatment about 1 ml per cm 2 is placed on the tissue as described above and the composition and tissue is covered with gauze and adhesive.
  • a cover may be advantageously applied to keep the composition in contact with the treated tissue.
  • a preferred treat ⁇ ment includes washing off the remainder of the gel under run ⁇ ning water and then drying the tissue.
  • the compositions according to the invention may be applied as for the night treatment, as above, or alternatively, the com ⁇ positions are applied to the tissue liberally and is allowed to be absorbed into the nail and surrounding skin.
  • the skin is first moistened with water before applying the composition.
  • a protective covering may be used in order to keep the composition in contact with the treated scalp.
  • Methyl cellulose Methyl cellulose (Methocel MC 4000 cp, Fluka AG) in 90 ml of double distilled water was autoclaved.
  • the preferred media contained essential and non- essential amino acids, vitamins, other organic constituents, major inorganic salts, trace elements and buffers and was sup ⁇ plemented with CaCl and L-glutamine and with the non-steroidal anabolic hormones, insulin, thyroxin, growth hormone and insulin-like growth factor (IGF) at the concentrations as indicated below.
  • IGF insulin-like growth factor
  • Insulin 5-10 ug/ml (Novo Nordisk) (about 143 mU/ml)
  • Thyroxine(T4) 1.0 X 10" 7 M (Sigma) (77.69 ng/ml)
  • Methylcellulose (Methocel 4000 cp, Fluka AG) 4% or Hydroxyethylcellulose 6%
  • methyl cellulose is autoclaved in 90 ml of double distilled water.
  • the water/methyl cellulose mixture is cooled to 4*C and stirred until the gel dissolves and the solution clears (overnight with a magnetic stirrer in the cold room) .
  • To the water/methyl cellulose mixture is added 10 ml of the concentrated MCDB 153 solution (X 10 con ⁇ centrated media available from Biological Industries, Beth Haemek, Israel) containing the supplements as described above (adjusted to a final volume of 100 ml of formulation) .
  • the solution is then mixed well and then decanted into 50 ml sterile test tubes.
  • the formulation is stored at 4 ⁇ C.
  • the scalp is treated once a day, preferably before going to sleep. If the scalp is oily or if other hair groom ⁇ ing products are in use, the hair is hydrated (in certain cases, also shampooed) to remove the oil or other deposits and then towel dried. Massage the gel into the scalp and leave on overnight. Preferably, the scalp is first moistened before applying the composition. In the morning, wash out the remains of the gel with clear water.
  • the individuals treated in example 1 were studied for a further period to determine the effects of the instant invention on hair growth and cessation of hair loss. All of the individuals, prior to the study suffered from alopecia androgenetica or traumatic alopecia of variable severity.
  • the individuals were instructed to use the hair growth formulation of the instant invention as described in example l (except that transferrin was excluded from the formulation) and this use continued for more than six months.
  • the volunteers were instructed to apply the hair growth formula ⁇ tion to the scalp once a day, before retiring, by massaging it into the scalp area. The gel was left on during the night, and washing of the remains of the gel was performed the fol ⁇ lowing morning.
  • Tables 2 and 3 The results of the study are presented in Tables 2 and 3, below.
  • Example 1 the reaction to the treatment was seen 1-3 weeks after the start of the treatment, when cessation of hair loss was reported by all participants. In addition, generally within 4-6 weeks, the increase in the rate of existing hair growth was reported as well as the start of new hair growth. In the further results, after four months of treatment, all of the participants in the study evidenced moderate or dense hair growth, with most show moderate hair growth. After more than 6 months of treatment several individuals within the treatment group evidenced dense hair growth and two individuals exhibited a complete cover of the deficit area.
  • mice which were depilated manually on Day 0 of the experiment.
  • anagen ensued (as judged by gray coloration produced by melanogenesis) .
  • an injection of the cytoxic agent cytophosphan (0.5 ml per gram) was made ip.
  • catagen resulted form the administration of the cytophosphan.
  • anagen was seen and on that day blood samples were taken for CBC analy ⁇ sis, as well as punch biopsies.
  • the second cytophosphan injection was given.
  • catagen occurred and on Day 28, anagen was present. Second blood samples were taken on Day 30, at which time punch biopsies were also taken.
  • Group No. 2 Treatment with Hair Growth Composition starting on Day 10 (three times daily) after cytophosphan injection and continued for 30 days.
  • mice which were depilated manually on Day 0 of the experiment.
  • anagen occurred (Day 7 - 30%, Day 8 - 60%, Day 9 - 70%).
  • cytophosphan 0.5 ml per gram
  • all animals were in anagen (100%) at that time.
  • catagen occurred and on Days 16-17, anagen was seen and on Day 17 blood samples were taken for analysis and biopsies were also taken.
  • the second cytophosphan injection was given.
  • catagen occurred and on Day 28-29, anagen occurred.
  • Second blood samples were taken on Day 30, at which time punch biopsies were also taken.
  • Group No. 3 Treatment with vehicle placebo starting on Day 0 (three times daily) and continued for 28 days.
  • mice which were depilated manually on Day 0 of the experiment. On Days 7-8, anagen occurred. On Day 9, cytophosphan (0.5 ml per gram) was administered and on Day 15, catagen occurred. On Days 16-17, anagen was seen and on Day 20 vellus hair was seen. On Day 24, terminal hair was observed and on Day 28, the number of animals that had terminal hair was 4 (mild) , 10 (moderate) and 6 (extensive terminal growth) .
  • Group No. 4 Treatment with vehicle placebo starting on Day 9 following cytophosphan injections and continued for 20 days.
  • mice which were depilated on Day 0 and angagen was observed on Day 9, when the mice were injected with cytophosphan.
  • Catagen was observed on Day 15, and anagen followed on Day 17-18.
  • Vellus hairs were seen on Day 20 and Day 24.
  • terminal hair growth was mild in five naimals, moderate in three animals and extensive in two animals.
  • Group No. 5 No Treatment given to this control group- used as a baseline for the other studies.
  • mice This group consisted of 16 mice, which were depilated on Day 0 and anagen ensued on Days 9-10. On Day 10, cytophosphan was injected and catagen occurred on Days 15-17. On Days 18-21, anagen occurred and on Day 21, punch biopsies were taken.
  • results In the experimental mice, the use of the present invention produced significant enhancement of the growth of hair in experimental animals under conditions in which the hair growth composition was used to treat the animals prior to injection with cytophosphan and after cytophosphan. (Table 4, below) . Although the standard deviation of the results of the test animals was rather large, nonetheless, the overall results evidenced significant enhancement of hair growth in the test animals which had been treated with the present invention compared to untreated control animals. In addition, the use of the present invention did not produce mortality in any of the test animals (total of 234 animals) .
  • Case #1 A man in his 40's, who had a diffuse vertex hair loss of 5 years duration has been using the present composition (composi ⁇ tion lc.2) for 5 and 1/2 months. His natural hair coloring is light-reddish brown, but in the vertex area, where hair loss appeared, the color had faded to yellowish-white. After using the present composition, the natural hair coloring of hair in the vertex area was restored. The hair has the same reddish-brown coloring as the res of the hair on the head.
  • a 54 year old Caucasian male who has a beard in which the original dark coloration became streaked with white used the present composition as recommended for 1 month (nighttime appli ⁇ cation sufficient to allow contact of the formulation with the underlying skin, washing off the composition in the morning) .
  • the treatment has induced pigment change in which the original coloration has returned in the newly growth hair, and in the white hair, the roots have become black, evidencing restoration of hair color in the newly grown portion of the hair.
  • the material as defined in the materials and methods sec ⁇ tion, above, in liquid or gel form was used (with the exception that triiodothyronine at 13.02 ng/ml was substituted for the thyroxine, the amount of CaCl 2 used was 4.0 X 10" 5 and no Phenol Red was included in the formulation- in liquid form, no thickener was used- in gel form, 4 g of methyl cellulose was used) to treat corneal ulcers in Hartley derived female guinea pigs.
  • Hartley derived female guinea pigs weighing 350-400g were housed in individual cages and fed guinea pig chaw and vitamin C enriched water ad libitum.
  • the right eyes of 7 animals were treated with the formulation as described above in liquid form using two drops every two hours and in liquid form in the eve ⁇ ning. After 24 hours and 48 hours, the eyes were examined under the microscope and photographed.
  • a total number of 19 guinea pig eyes were treated with Cariel while a total number of 16 eyes were used as a control (untreated) .
  • Hartley-derived albino female guinea-pigs weighing 250 g were used in this study.
  • the animals were housed in individual cages and fed regular guinea-pig chow and water enriched with Vitamin C ad libitum. All surgical procedures to impart burn wounds to the animals were performed under general anaesthesia of Katamin HC1 150 mg/kg i.m./d,l-2-(chlorophenyl)-2- (methylamino)cyclohexanone hydrochloride, Parke Davis).
  • the animals were divided into two groups. In the first group, eight guinea pigs were treated with Silverol, which is currently used as a preparation for burn wounds in all military trauma units in Israel. The second group of eight animals was treated essentially with composition lcl. of the materials and methods section (5 ug/ml insulin, methyl cellulose as gelling agent) . The experiment was repeated on three different occa ⁇ sions.

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Abstract

La présente invention concerne un procédé pour stimuler les follicules pileux dormants présents normalement dans la peau intacte, en particulier sur le cuir chevelu intact de l'homme, améliorer la croissance des cheveux et leur rendre leur couleur d'origine lorsque celle-ci s'est détériorée. La formulation selon la présente invention est utile pour favoriser la croissance des cheveux en stimulant les follicules dormants, faibles ou mourants, pour produire des cheveux. Un effet secondaire inattendu de cette croissance des cheveux est que la couleur naturelle des cheveux est restituée lorsque cette couleur s'est détériorée, par exemple dans les cheveux grisonnants et/ou gris, à mesure que les cheveux poussent. En plus, les formulations ont un effet angiogénique sur le tissu de l'épiderme entourant les follicules pileux, et elles favorisent la croissance du tissu ungéal ou revitalisent la peau chez les animaux. Ces formulations contiennent généralement une quantité suffisante pour avoir l'effet recherché d'un mélange d'hormones anabolisantes non stéroïdiennes choisies parmi l'insuline, l'hormone de croissance, la tri-iodothyronine ou la thyroxine, en combinaison avec un milieu essentiel minimum et de préférence enrichi, tel que le milieu MCDB 153.
PCT/US1996/002341 1995-02-23 1996-02-21 Compositions et procedes pour ameliorer la croissance des cheveux et pour leur restituer leur couleur d'origine WO1996025943A1 (fr)

Priority Applications (2)

Application Number Priority Date Filing Date Title
EP96906564A EP0812210A4 (fr) 1995-02-23 1996-02-21 Compositions et procedes pour ameliorer la croissance des cheveux et pour leur restituer leur couleur d'origine
AU49904/96A AU4990496A (en) 1995-02-23 1996-02-21 Compositions and methods for enhancing the growth of hair and restoring hair color

Applications Claiming Priority (4)

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US39400595A 1995-02-23 1995-02-23
US08/394,005 1995-02-23
US53751595A 1995-10-02 1995-10-02
US08/537,515 1995-10-02

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WO1996025943A1 true WO1996025943A1 (fr) 1996-08-29

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PCT/US1996/002341 WO1996025943A1 (fr) 1995-02-23 1996-02-21 Compositions et procedes pour ameliorer la croissance des cheveux et pour leur restituer leur couleur d'origine

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EP (1) EP0812210A4 (fr)
AU (1) AU4990496A (fr)
CA (1) CA2213481A1 (fr)
IL (1) IL117236A0 (fr)
WO (1) WO1996025943A1 (fr)

Cited By (21)

* Cited by examiner, † Cited by third party
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EP0991319A2 (fr) * 1997-04-18 2000-04-12 Hadasit Medical Research Services & Development Co., Ltd. Traitement du diabete
WO2000072920A1 (fr) * 1999-06-01 2000-12-07 The University Of Texas Southwestern Medical Center Composes de diarylether substitues
WO2000073292A1 (fr) * 1999-06-01 2000-12-07 The University Of Texas Southwestern Medical Center Composes biaryles
WO2000073265A1 (fr) * 1999-06-01 2000-12-07 The University Of Texas Southwestern Medical Centre Derives de thyroxane contenant du soufre et leur utilisation en tant que promoteurs de croissance capillaire
WO2000072810A1 (fr) * 1999-06-01 2000-12-07 The University Of Texas Southwestern Medical Center Methode de traitement de la perte des cheveux a l'aide de composes de sulfonyle thyromimetiques
US6183761B1 (en) 1998-03-16 2001-02-06 The Procter & Gamble Company Compositions for regulating skin appearance
GB2369572A (en) * 2000-11-29 2002-06-05 Raft Trustees Ltd Wound treatment composition comprising insulin
US6525094B1 (en) 1999-06-01 2003-02-25 The University Of Texas Southwestern Medical Center Method of treating hair loss using diphenylether derivatives
US6680344B1 (en) 1999-06-01 2004-01-20 The University Of Texas Southwestern Medical Center Method of treating hair loss using diphenylmethane derivatives
US6723717B1 (en) * 1999-06-01 2004-04-20 The University Of Texas Southwestern Medical Center Sulfur-containing thyroxane derivatives and their use as hair growth promotors
FR2850864A1 (fr) * 2003-02-12 2004-08-13 Oreal Utilisation d'un inhibiteur de 15-hydroxy prostaglandine deshydrogenase pour favoriser la pigmentation de la peau ou des phaneres
WO2004073594A2 (fr) * 2003-02-12 2004-09-02 L'oreal Utilisation d’un inhibiteur de 15-hydroxy prostaglandine deshydrogenase pour favoriser la pigmentation de la peau ou des phaneres
WO2005034890A2 (fr) * 2003-09-15 2005-04-21 Allergan, Inc. Procedes de traitement de cheveux gris a l'aide d'amides d'acide cyclopentane(ene) heptan(en)oique
US7198648B2 (en) * 2000-06-20 2007-04-03 Henkel Kommanditgesellschaft Auf Aktien (Henkel Kgaa) Coupling component for oxidative hair dyes
EP1818061A1 (fr) * 2005-12-02 2007-08-15 Charite-Universitätsmedizin Berlin Utilisation de la ghréline pour stimuler la pousse des cheveux
WO2008067833A1 (fr) * 2006-12-05 2008-06-12 Charité Universitätsmedizin - Berlin Utilisation de la ghréline pour inverser la chute des poils/cheveux
US7622103B1 (en) * 2003-03-27 2009-11-24 Cielo Jill M Hair and scalp toxin remover composition and method of its making
EP2465581A1 (fr) * 2010-12-15 2012-06-20 Han-Min Chen Composition permettant de favoriser la croissance des cheveux
WO2013081722A1 (fr) * 2011-11-28 2013-06-06 A66, Incorporated Poly-haptène ayant un système topique de repousse de cheveux d'alopécie hormonodépendante
EP2695611A1 (fr) 2012-08-06 2014-02-12 Dr. August Wolff GmbH & Co. KG Arzneimittel Eprotirome pour l'utilisation dans la prévention et/ou le traitement de troubles de cheveux et compositions correspondantes
US20150150771A1 (en) * 2012-06-19 2015-06-04 Debraj Shome Composition and method for an intradermal hair growth solution

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Cited By (34)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0991319A4 (fr) * 1997-04-18 2000-10-18 Hadasit Med Res Service Traitement du diabete
EP0991319A2 (fr) * 1997-04-18 2000-04-12 Hadasit Medical Research Services & Development Co., Ltd. Traitement du diabete
US6183761B1 (en) 1998-03-16 2001-02-06 The Procter & Gamble Company Compositions for regulating skin appearance
US6680344B1 (en) 1999-06-01 2004-01-20 The University Of Texas Southwestern Medical Center Method of treating hair loss using diphenylmethane derivatives
WO2000072920A1 (fr) * 1999-06-01 2000-12-07 The University Of Texas Southwestern Medical Center Composes de diarylether substitues
WO2000072810A1 (fr) * 1999-06-01 2000-12-07 The University Of Texas Southwestern Medical Center Methode de traitement de la perte des cheveux a l'aide de composes de sulfonyle thyromimetiques
WO2000073292A1 (fr) * 1999-06-01 2000-12-07 The University Of Texas Southwestern Medical Center Composes biaryles
WO2000073265A1 (fr) * 1999-06-01 2000-12-07 The University Of Texas Southwestern Medical Centre Derives de thyroxane contenant du soufre et leur utilisation en tant que promoteurs de croissance capillaire
US6525094B1 (en) 1999-06-01 2003-02-25 The University Of Texas Southwestern Medical Center Method of treating hair loss using diphenylether derivatives
US6646005B1 (en) 1999-06-01 2003-11-11 The University Of Texas Southwestern Medical Center Method of treating hair loss using sulfonyl thyromimetic compounds
US6723717B1 (en) * 1999-06-01 2004-04-20 The University Of Texas Southwestern Medical Center Sulfur-containing thyroxane derivatives and their use as hair growth promotors
US7198648B2 (en) * 2000-06-20 2007-04-03 Henkel Kommanditgesellschaft Auf Aktien (Henkel Kgaa) Coupling component for oxidative hair dyes
US9308240B2 (en) 2000-11-29 2016-04-12 Pharmecosse Limited Method of preventing or reducing scarring of human skin
GB2369572A (en) * 2000-11-29 2002-06-05 Raft Trustees Ltd Wound treatment composition comprising insulin
US8557770B2 (en) 2000-11-29 2013-10-15 Pharmecosse Ltd. Method of preventing or reducing scarring of human skin
US20110014250A1 (en) * 2003-02-12 2011-01-20 L'oreal Inhibitors of 15-hydroxyprostaglandin dehydrogenase for stimulating pigmentation of the skin or skin appendages
FR2850864A1 (fr) * 2003-02-12 2004-08-13 Oreal Utilisation d'un inhibiteur de 15-hydroxy prostaglandine deshydrogenase pour favoriser la pigmentation de la peau ou des phaneres
WO2004073594A2 (fr) * 2003-02-12 2004-09-02 L'oreal Utilisation d’un inhibiteur de 15-hydroxy prostaglandine deshydrogenase pour favoriser la pigmentation de la peau ou des phaneres
WO2004073594A3 (fr) * 2003-02-12 2004-10-07 Oreal Utilisation d’un inhibiteur de 15-hydroxy prostaglandine deshydrogenase pour favoriser la pigmentation de la peau ou des phaneres
US8735445B2 (en) 2003-02-12 2014-05-27 L'oreal Inhibitors of 15-hydroxyprostaglandin dehydrogenase for stimulating pigmentation of the skin or skin appendages
US7705041B2 (en) 2003-02-12 2010-04-27 L'oreal Inhibitors of 15-hydroxyprostaglandin dehydrogenase for stimulating pigmentation of the skin or skin appendages
US7622103B1 (en) * 2003-03-27 2009-11-24 Cielo Jill M Hair and scalp toxin remover composition and method of its making
WO2005034890A3 (fr) * 2003-09-15 2005-06-09 Allergan Inc Procedes de traitement de cheveux gris a l'aide d'amides d'acide cyclopentane(ene) heptan(en)oique
WO2005034890A2 (fr) * 2003-09-15 2005-04-21 Allergan, Inc. Procedes de traitement de cheveux gris a l'aide d'amides d'acide cyclopentane(ene) heptan(en)oique
EP1818061A1 (fr) * 2005-12-02 2007-08-15 Charite-Universitätsmedizin Berlin Utilisation de la ghréline pour stimuler la pousse des cheveux
WO2008067833A1 (fr) * 2006-12-05 2008-06-12 Charité Universitätsmedizin - Berlin Utilisation de la ghréline pour inverser la chute des poils/cheveux
EP2465581A1 (fr) * 2010-12-15 2012-06-20 Han-Min Chen Composition permettant de favoriser la croissance des cheveux
WO2013081722A1 (fr) * 2011-11-28 2013-06-06 A66, Incorporated Poly-haptène ayant un système topique de repousse de cheveux d'alopécie hormonodépendante
US8697146B2 (en) * 2011-11-28 2014-04-15 A66 Incorporated Poly-hapten with topical hormone alopecia hair regrowth system
US20140314681A1 (en) * 2011-11-28 2014-10-23 George Davey Poly-hapten with topical hormone alopecia hair regrowth system
US20150150771A1 (en) * 2012-06-19 2015-06-04 Debraj Shome Composition and method for an intradermal hair growth solution
US9700504B2 (en) * 2012-06-19 2017-07-11 Debraj Shome Composition and method for an intradermal hair growth solution
EP2695611A1 (fr) 2012-08-06 2014-02-12 Dr. August Wolff GmbH & Co. KG Arzneimittel Eprotirome pour l'utilisation dans la prévention et/ou le traitement de troubles de cheveux et compositions correspondantes
WO2014023698A1 (fr) 2012-08-06 2014-02-13 Dr. August Wolff Gmbh & Co. Kg Arzneimitttel Éprotirome destiné à être utilisé dans la prévention et/ou le traitement de troubles capillaires et compositions correspondantes

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AU4990496A (en) 1996-09-11
EP0812210A4 (fr) 1998-08-19
CA2213481A1 (fr) 1996-08-29
EP0812210A1 (fr) 1997-12-17

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