WO2013129723A1 - Composition for improving skin conditions comprising hordenine - Google Patents

Composition for improving skin conditions comprising hordenine Download PDF

Info

Publication number
WO2013129723A1
WO2013129723A1 PCT/KR2012/001994 KR2012001994W WO2013129723A1 WO 2013129723 A1 WO2013129723 A1 WO 2013129723A1 KR 2012001994 W KR2012001994 W KR 2012001994W WO 2013129723 A1 WO2013129723 A1 WO 2013129723A1
Authority
WO
WIPO (PCT)
Prior art keywords
hordenine
composition
skin
propafenone
present
Prior art date
Application number
PCT/KR2012/001994
Other languages
French (fr)
Inventor
Sang Cheol Kim
Seoung Woo SHIN
Seung Beom Kim
Eun Sun Jung
Jong Sung Lee
Deok Hoon Park
Original Assignee
Bio Spectrum, Inc.
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Bio Spectrum, Inc. filed Critical Bio Spectrum, Inc.
Publication of WO2013129723A1 publication Critical patent/WO2013129723A1/en

Links

Images

Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/13—Amines
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/18—Magnoliophyta (angiosperms)
    • A61K36/88—Liliopsida (monocotyledons)
    • A61K36/899—Poaceae or Gramineae (Grass family), e.g. bamboo, corn or sugar cane
    • A61K36/8998—Hordeum (barley)
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/13—Amines
    • A61K31/135—Amines having aromatic rings, e.g. ketamine, nortriptyline
    • A61K31/137—Arylalkylamines, e.g. amphetamine, epinephrine, salbutamol, ephedrine or methadone
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/13—Amines
    • A61K31/135—Amines having aromatic rings, e.g. ketamine, nortriptyline
    • A61K31/138—Aryloxyalkylamines, e.g. propranolol, tamoxifen, phenoxybenzamine
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/16—Amides, e.g. hydroxamic acids
    • A61K31/165—Amides, e.g. hydroxamic acids having aromatic rings, e.g. colchicine, atenolol, progabide
    • A61K31/166—Amides, e.g. hydroxamic acids having aromatic rings, e.g. colchicine, atenolol, progabide having the carbon of a carboxamide group directly attached to the aromatic ring, e.g. procainamide, procarbazine, metoclopramide, labetalol
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/18—Magnoliophyta (angiosperms)
    • A61K36/88—Liliopsida (monocotyledons)
    • A61K36/89—Cyperaceae (Sedge family)
    • 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
    • A61K8/41—Amines
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/0012—Galenical forms characterised by the site of application
    • A61K9/0014—Skin, i.e. galenical aspects of topical compositions
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/06—Ointments; Bases therefor; Other semi-solid forms, e.g. creams, sticks, gels
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/20—Pills, tablets, discs, rods
    • A61K9/2004—Excipients; Inactive ingredients
    • A61K9/2013—Organic compounds, e.g. phospholipids, fats
    • A61K9/2018—Sugars, or sugar alcohols, e.g. lactose, mannitol; Derivatives thereof, e.g. polysorbates
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/20—Pills, tablets, discs, rods
    • A61K9/2004—Excipients; Inactive ingredients
    • A61K9/2022—Organic macromolecular compounds
    • A61K9/205—Polysaccharides, e.g. alginate, gums; Cyclodextrin
    • A61K9/2059—Starch, including chemically or physically modified derivatives; Amylose; Amylopectin; Dextrin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P17/00—Drugs for dermatological disorders
    • 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
    • A61Q19/02—Preparations for care of the skin for chemically bleaching or whitening the skin
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L29/00—Foods or foodstuffs containing additives; Preparation or treatment thereof
    • A23L29/03—Organic compounds
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/14—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles
    • A61K9/141—Intimate drug-carrier mixtures characterised by the carrier, e.g. ordered mixtures, adsorbates, solid solutions, eutectica, co-dried, co-solubilised, co-kneaded, co-milled, co-ground products, co-precipitates, co-evaporates, co-extrudates, co-melts; Drug nanoparticles with adsorbed surface modifiers
    • A61K9/145—Intimate drug-carrier mixtures characterised by the carrier, e.g. ordered mixtures, adsorbates, solid solutions, eutectica, co-dried, co-solubilised, co-kneaded, co-milled, co-ground products, co-precipitates, co-evaporates, co-extrudates, co-melts; Drug nanoparticles with adsorbed surface modifiers with organic compounds
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/14—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles
    • A61K9/141—Intimate drug-carrier mixtures characterised by the carrier, e.g. ordered mixtures, adsorbates, solid solutions, eutectica, co-dried, co-solubilised, co-kneaded, co-milled, co-ground products, co-precipitates, co-evaporates, co-extrudates, co-melts; Drug nanoparticles with adsorbed surface modifiers
    • A61K9/146—Intimate drug-carrier mixtures characterised by the carrier, e.g. ordered mixtures, adsorbates, solid solutions, eutectica, co-dried, co-solubilised, co-kneaded, co-milled, co-ground products, co-precipitates, co-evaporates, co-extrudates, co-melts; Drug nanoparticles with adsorbed surface modifiers with organic macromolecular compounds
    • 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

Definitions

  • the present invention relates to a composition comprising hordenine as an active ingredient, which has excellent stability, can be used safely without adversely affecting the skin, and has excellent effects on wrinkle reduction, skin whitening, anti-inflammation, antioxidant activity, hair loss prevention and hair growth.
  • Wrinkles are caused by aging of the skin, and skin aging results from the natural changes associated with the aging process.
  • Skin aging is broadly classified into two types: physiological aging showing age-related changes in the skin function, structure or shape throughout the skin surface; and photo-aging caused by UV light.
  • Changes in the dermis result from changes in substances having low molecular weight in the extracellular matrix due to decreases in the number of fibroblasts and the ability to form fibroblasts. Specific examples of these changes include separation of collagen bundles, a decrease in mucopolysaccharide synthesis, decreases in the number and diameter of collagen and elastin, decomposition of collagen and elastin, and blood vessel expansion.
  • the human skin color is determined according to the concentration and distribution of melanin in the skin.
  • the melanin pigment that is produced in the melanocytes of the human skin is a phenolic polymer consisting of a complex of a black pigment and a protein and plays an important role in preventing skin damage caused by UV light.
  • tyrosinase present in melanocytes was reported to be the most important factor in melanin biosynthesis, and tyrosinase plays an important role in the skin darkening process by converting tyrosine into DOPA and DOPA-quinone, which are intermediate products of melanin polymer production.
  • the present inventors have made extensive efforts to develop novel natural substances, which have excellent effects on wrinkle reduction, skin whitening, hair loss prevention, hair growth promotion, antioxidant activity and anti-inflammatory activity, are highly safe and stable, and do not adversely affect the skin.
  • the present inventors have searched for plant-derived compounds, and as a result, have found that hordenine increases collagen synthesis and inhibits collagenase and that a composition containing hordenine as an active ingredient has high efficacy and safety, improves skin conditions and shows excellent anti-inflammatory, antioxidant and anti-obesity effects, thereby completing the present invention.
  • Another object of the present invention is to provide an anti-inflammatory composition.
  • Still another object of the present invention is to provide an antioxidant composition.
  • Yet another object of the present invention is to provide a method of improving skin conditions, inhibiting inflammatory responses, inhibiting oxidative stress, and ameliorating, treating or preventing the disorders, diseases or symptoms associated therewith, by administering said composition to a subject.
  • composition for improving skin conditions comprising hordenine as an active ingredient.
  • an anti-inflammatory composition comprising hordenine as an active ingredient.
  • an antioxidant composition comprising hordenine as an active ingredient.
  • a method of improving the skin conditions of a subject by administering said skin condition-improving composition to the subject in a dermatologically acceptable way.
  • Hordenine exhibits an excellent effect of reducing wrinkles through molecular mechanisms such as the promotion of collagen synthesis and the protection of fibroblasts and has the effects of promoting hair growth and preventing hair loss. Also, it shows an excellent skin-whitening effect, and particularly, the skin-whitening effect thereof is synergistically increased when hordenine is used together with propafenone at a suitable ratio to prepare a composition. Furthermore, hordenine has an excellent effect of inhibiting inflammation-associated immune responses and an antioxidant effect of protecting mitochondria from reactive oxygen species. In addition, it has excellent safety, because it is not cytotoxic and does not adversely affect the skin.
  • the inventive composition comprising hordenine can be used as cosmetic, pharmaceutical and food compositions which have exhibit excellent effects of improving skin conditions, including the effects of reducing wrinkles, promoting skin whitening, promoting hair growth and preventing hair loss, and, at the same time, has ensured safety.
  • FIG. 1 is a graph showing the effect of hordenine on the promotion of collagen (type I collagen) biosynthesis.
  • FIG. 2 is a graph showing the effects of compositions, comprising hordenine and/or propafenone, on the reduction of melanin synthesis in human melanocytes.
  • FIG. 3 is a graph showing the effect of hordenine on the proliferation of hair follicle dermal papilla cells.
  • Hordenine which is used as an active ingredient in the composition according to the present invention is a phenethylamine alkaloid present in plants of the family Cyperaceae, sprouted barley (Hordeum vulgare), Cactus, Citrus aurantium and the like and is typically known to have antibacterial effects. Also, it functions to enhance the sympathetic nervous system together with synephrine that is an adrenergic alkaloid, and thus it is used as a diet food that induces weight loss.
  • US Patent Publication No. 2002/0058075 discloses that a citrus-derived compound comprising hordenine has the effect of reducing body weight by increasing motor ability.
  • Hordenine is known as N,N-dimethyltyramine and has an IUPAC name of 4-(2-dimethylaminoethyl)phenol. Also, it has a structure of the following formula 1:
  • This hordenine can be extracted from natural sources, including plants of the family Cyperaceae, sprouted barley (Hordeum vulgare), Cactus, Citrus aurantium and the like. Specifically, hordenine can be obtained from the above natural sources using various extraction methods known in the art.
  • an extraction solvent selected from among (a) water, (b) an anhydrous or hydrated lower alcohol containing 1 to 4 carbon atoms (methanol, ethanol, propanol, butanol, etc.), (c) a mixed solvent of the lower alcohol with water, (d) acetone, (e) ethyl acetate, (f) chloroform, (g) 1,3-butylene glycol, or a mixture of two or more thereof. More preferably, it can be extracted using an aqueous solution of methanol, ethanol and butanol, and most preferably, it can be extracted using an aqueous solution of butanol.
  • an extraction solvent selected from among (a) water, (b) an anhydrous or hydrated lower alcohol containing 1 to 4 carbon atoms (methanol, ethanol, propanol, butanol, etc.), (c) a mixed solvent of the lower alcohol with water, (d) acetone, (e) ethyl acetate, (f)
  • the extracts of the present invention include, in addition to the extracts obtained using the above-described extraction solvents, extracts obtained through conventional purification processes. For example, fractions obtained through various additional purification methods, such as separation with an ultra filtration membrane having a given molecular weight cut-off or separation by various chromatography systems (manufactured for separation according to size, charge, hydrophobicity or affinity), are also included in the scope of the extract of the present invention.
  • hordenine that is used in the present invention may be prepared by chemical synthesis or may be a commercially available product (for example, hordenine available from Chromadex).
  • the term “improving skin conditions” means improvement and betterment of all skin conditions. Preferably, it means a reduction in skin wrinkles, an improvement in skin whitening, promotion of hair growth, or prevention of hair loss.
  • the inventive composition for improving skin conditions serves to reduce wrinkles.
  • the composition of the present invention has low cytotoxicity and exhibits an excellent effect of reducing skin wrinkles through molecular mechanisms such as the protection of fibroblasts and the promotion of collagen synthesis, as clearly demonstrated in examples below. It is understood that the reduction in skin wrinkles includes skin protection (e.g., prevention of skin wrinkles, removal of skin wrinkles, and prevention of skin aging).
  • the inventive composition for improving skin conditions serves to whiten the skin.
  • whitening refers to the effect of reducing skin troubles by preventing skin darkening caused by melanin pigmentation.
  • Hordenine in the skin whitening composition of the present invention inhibits the expression of tyrosinase in cells to inhibit melanin production, thus exhibiting a skin whitening effect. Also, it is highly stable and there is little or no change of the content of hordenine with the passage of time. In addition, it has little or no adverse effects, such as skin irritation. Particularly, the skin whitening effect of hordenine is synergistically increased when it is used together with propafenone, known to having a skin shortening effect, at a suitable ratio, to prepare a composition.
  • Propafenone which is used together with hordenine in a skin whitening composition was approved for use as an anti-arrhythmic drug in the year 1980 and is used mainly for supraventricular arrhythmias.
  • Na ions are rapidly introduced into cells by propafenone while the Vmax value of action potential is lowered, thus inhibiting the generation of stimulation and the conduction of excitation between cells.
  • Propafenone is structurally similar to propranolol and has weak beta-blocking activity.
  • Japanese Patent Laid-Open Publication No. 2003-397665 discloses that propafenone has the effect of inhibiting neuropathic pain.
  • Propafenone which is used in the present invention may be prepared by chemical synthesis or may be a commercially available product (for example, propafenone available from Sigma).
  • Hordenine and propafenone may be present at a weight ratio ranging from 1:1 to 1:5.
  • hordenine and propafenone are present at a weight ratio of 1:1.5 to 1:2.5, and in this case, the most excellent whitening effect is exhibited.
  • the content of hordenine and propafenone in the composition of the present invention is 0.0001-10 wt%, and preferably 0.001-1.0 wt%, based on the total weight of the composition.
  • the inventive composition for improving skin conditions has excellent effects on the promotion of hair growth and the prevention of hair loss.
  • prevention of hair loss and “promotion of hair growth” are used in the same sense.
  • the composition of the present invention comprises hordenine as an active ingredient and thus exhibits the effect of inhibiting inflammatory responses.
  • the active ingredient hordenine generally acts on COX-2 which causes pain in inflammatory responses.
  • arachidonic acid that is a precursor of prostaglandin (PGE2) is present in the cell membrane and is converted into prostaglandin through a cyclooxygenase pathway.
  • PGE2 arachidonic acid that is a precursor of prostaglandin
  • the produced prostaglandin acts as a chemical mediator in allergic and inflammatory processes and irritates a nociceptive nerve or sustains inflammatory responses or influences carcinogenesis.
  • a substance that inhibits COX-2 activity is highly likely to inhibit inflammatory responses.
  • the anti-inflammatory composition of the present invention can be applied to various disorders, diseases or abnormal conditions, which can be prevented or treated by inhibiting inflammatory responses.
  • inflammation-related diseases which can be prevented or treated by the composition of the present invention include, but are not limited to, bullous pemphigoid, cicatricial pemphigoid, discoid lupus erythematosis, lupus erythematosus, systemic lupus erythematosus, allergy and atopy.
  • the anti-inflammatory composition of the present invention is preferably used for the prevention or treatment of atopy.
  • Hordenine that is the active ingredient of the inventive composition exhibits an antioxidant effect by protecting mitochondria from oxidative damage caused by reactive oxygen species (ROS) which occur due to various irritations.
  • ROS reactive oxygen species
  • the antioxidant composition of the present invention can be applied to various disorders, diseases or abnormal conditions, which can be prevented or treated by inhibiting or removing oxidative stress.
  • oxidative stress-related diseases which can be prevented or treated by the composition of the present invention include, but are not limited to, atherosclerosis, coronary artery disease, cardiac artery restenosis, reperfusion injury, neurodegenerative diseases such as Parkinson's disease or Alzheimer's disease, stroke, cancer, aging, cardiovascular disease, osteoporosis, disorders of the central nerve system, peripheral vascular disease, and respiratory distress syndrome.
  • the antioxidant composition of the present invention is used for the prevention or treatment of aging.
  • the antioxidant composition of the present invention exhibits the effect of protecting intracellular mitochondria from oxidative injury caused by the stimulation of rotenone, which induces the production of reactive oxygen species, and reactive oxygen species induced thereby.
  • examples of hordenine that may be used in the present invention include, in addition to the hordenine compound of formula 1, hordenine derivatives, which show the effects of promoting collagen synthesis, reducing wrinkles, whitening the skin, promoting hair growth, preventing hair loss, or exhibiting anti-inflammatory and antioxidant effects, among derivatives obtained by adding substituents to hordenine or substituting hordenine according to a conventional method known in the art. More specifically, examples of hordenine that may be used in the present invention include, in addition to the compound of formula 1, derivatives obtained by linking various substituents to the structure of formula 1 as a nucleus according to a method known in the art.
  • hordenine may be contained in the composition in an amount of 0.00001-15.0 wt%, more preferably 0.0001-10 wt%, and more preferably 0.0001-5 wt%, based on the total weight of the composition.
  • the composition of the present invention may be used as a cosmetic composition.
  • the cosmetic composition of the present invention may contain, in addition to the active ingredient hordenine, components that are conventionally used in cosmetic compositions, for examples, conventional auxiliaries, such as antioxidants, stabilizers, solubilizers, vitamins, pigments and fragrances, and carriers.
  • conventional auxiliaries such as antioxidants, stabilizers, solubilizers, vitamins, pigments and fragrances, and carriers.
  • the cosmetic composition of the present invention may be formulated into any form known in the art.
  • the formulation include, but are not limited to, a solution, a suspension, an emulsion, a paste, a gel, a cream, a lotion, a powder, a soap, a surfactant-containing cleanser, an oil, a powder foundation, an emulsion foundation, a wax foundation and a spray.
  • the cosmetic composition of the present invention can be formulated in the form of nourishing cream, astringent lotion, skin softener, lotion, essence, nourishing gel, or massage cream.
  • the formulation of the present invention is paste, cream or gel, it may contain, as carrier components, animal oil, vegetable oil, wax, paraffin, starch, tragacanth gum, cellulose derivatives, polyethylene glycol, silicone, bentonite, silica, talc or zinc oxide.
  • the formulation of the present invention is a powder or spray formulation, it may contain, as carrier components, lactose, talc, silica, aluminum hydroxide, calcium silicate or polyamide powder. Particularly, if it is spray, it may additionally contain a propellant, such as chlorofluorohydrocarbon, propane/butane or dimethyl ether.
  • a propellant such as chlorofluorohydrocarbon, propane/butane or dimethyl ether.
  • the formulation of the present invention is a solution or an emulsion, it may contain, as carrier components, a solvent, a solubilizing agent or an emulsifying agent, for example, water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butyl glycol oil, glycerol fatty ester, polyethylene glycol or sorbitan fatty acid ester.
  • a solvent for example, water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butyl glycol oil, glycerol fatty ester, polyethylene glycol or sorbitan fatty acid ester.
  • the formulation of the present invention is a suspension
  • it may contain, as carrier components, a liquid diluent, such as water, ethanol or propylene glycol, and a suspending agent, such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester, polyoxyethylene sorbitan ester, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar-agar or tragacanth gum.
  • a liquid diluent such as water, ethanol or propylene glycol
  • a suspending agent such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester, polyoxyethylene sorbitan ester, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar-agar or tragacanth gum.
  • the formulation of the present invention is a surfactant-containing cleansing, it may contain, as carrier components, aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic monoester, isethionate, imidazolium derivatives, methyl taurate, sarcosinate, fatty acid amide ether sulfate, alkyl amido betaine, aliphatic alcohol, fatty acid glyceride, fatty acid diethanolamide, vegetable oil, lanoline derivatives, or ethoxylated glycerol fatty acid ester.
  • carrier components aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic monoester, isethionate, imidazolium derivatives, methyl taurate, sarcosinate, fatty acid amide ether sulfate, alkyl amido betaine, aliphatic alcohol, fatty
  • the composition of the present invention may be used as a pharmaceutical composition.
  • the pharmaceutical composition of the present invention contains a pharmaceutically acceptable carrier.
  • the pharmaceutically acceptable carrier contained in the pharmaceutical composition of the present invention is commonly used in pharmaceutical formulations, and examples thereof include, but are not limited to, lactose, dextrose, sucrose, sorbitol, mannitol, starch, rubber acacia, potassium phosphate, alginate, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrups, methylcellulose, methylhydroxy benzoate, propylhydroxy benzoate, talc, magnesium stearate, and mineral oils.
  • the pharmaceutical composition according to the present invention may contain, in addition to these components, a lubricant, a wetting agent, a sweetener, a flavoring agent, an emulsifier, a suspending agent, and a preservative.
  • a lubricant for lubricating a lubricant for lubricating a lubricating a glycerol, a glycerol, a g., g., g., g., g., g., g., g., glycerin, sorbitol, sorbitol, sorbitol, sorbitol, sorbitol, sorbitol, sorbitol, sorbitol, sorbitol, sorbitol, sorbitol, sorbitol, sorbitol, sorbitol, sorbitol, sorbitol, sorbitol, sorbito
  • the pharmaceutical composition of this invention may be administered orally or parenterally. Preferably, it is administered parenterally, and most preferably, it is administered by topical application.
  • the suitable dose of the pharmaceutical composition of the present invention may vary depending on various factors, such as formulation methods, administration methods, the patient's age, body weight, sex, pathogenic state, diet, administration time, administration route, excretion rate and reaction sensitivity.
  • the suitable dose of the pharmaceutical composition of the present invention is in the range of 0.001-100 mg/kg for an adult.
  • the composition is a preparation for external use, it is preferably applied 1-5 times a day in amount of 1.0-3.0 ml for 1 month or more for an adult.
  • the above-described dose of the composition does not limit the scope of the present invention.
  • the pharmaceutical composition of the present invention may be formulated with the above-described pharmaceutically acceptable carrier and/or vehicle, thus providing several forms, including a unit dosage form and a multi-dosage form.
  • the formulations include, but are not limited to, a solution, suspension, syrup or emulsion in oil or aqueous medium, an elixir, a powder, a granule, a tablet and a capsule, and may additionally contain a dispersion agent or a stabilizer.
  • composition of the present invention may be used as a food composition.
  • the food composition of the present invention may contain, in addition to the active ingredient hordenine, conventional additives for preparing food compositions, for example, proteins, carbohydrates, fats, nutrients, seasonings and flavors.
  • carbohydrates include conventional sugars, such as monosaccharides (e.g., glucose and fructose), disaccharides (e.g., maltose, sucrose and oligosaccharide), and polysaccharides (e.g., dextrin and cyclodextrin); and sugar alcohols (e.g., xylitol, sorbitol and erithritol).
  • flavors include natural flavors [thaumatin and extract of stevia (e.g., rebaudioside A and glycyrrhizin)] and synthetic flavors (e.g., saccharin and aspartame).
  • the food composition of the present invention may comprise, in addition to the active ingredient hordenine, citric acid, liquid fructose, sugar, glucose, acetic acid, malic acid, fruit juice, an extract of Eucommia ulmoides oliv, a jujube extract or an extract of glycyrrhiza uralensis.
  • hordenine of the present invention may be used with confidence for a long period, because it has little or no toxicity and adverse effect. Particularly, it may be used safely in the above-described cosmetic, pharmaceutical and food compositions.
  • the present invention also provides a method of improving the skin conditions of a subject, inhibiting inflammatory responses in a subject or inhibiting oxidative stress in a subject by administering said composition to the subject.
  • the term “subject” is meant to include humans and mammals, for example, dogs, pigs, horses and cattle, in which poor skin conditions, for example, wrinkles, skin darkening and hair loss, are to be improved or prevented, or inflammation or inflammation-related diseases are to be treated or prevented, or oxidative stress-related disorders, diseases or abnormal conditions are to be treated or prevented, and these purposes can be achieved by administering the composition of the present invention.
  • administering means introducing a given substance into a subject according to any suitable method.
  • the composition of the present invention may be administered orally or parenterally through any general route, so long as it can reach a target tissue.
  • the composition of the present invention may be administered by any device which delivers the active ingredient into a target, for example, a cell.
  • composition of the present invention comprises hordenine as an active ingredient.
  • the active ingredient hordenine has excellent effects of improving skin conditions, including the effects of reducing wrinkles, improving skin whitening, promoting hair growth and preventing hair loss.
  • Hordenine exhibits an excellent effect of reducing wrinkles through molecular mechanisms, including the promotion of collagen synthesis and the protection of fibroblasts and has the effects of promoting hair growth and preventing hair loss.
  • it has an excellent effect of whitening the skin, and particularly, the skin whitening effect thereof is synergistically increased when hordenine is used together with propafenone at a suitable ratio to prepare a composition.
  • the active ingredient hordenine has excellent effects of inhibiting inflammation-related immune responses and protecting mitochondria from reactive oxygen species, thus exhibiting antioxidant effects.
  • the active ingredient hordenine can be used safely in cosmetic, pharmaceutical and food compositions, because it has no cytotoxicity and does not adversely affect the skin.
  • Example 1 Measurement of wrinkle-reducing effect of hordenine
  • Wrinkle-reducing effects can be generally measured by carrying out clinical testing for the ability to biosynthesize collagen in humans.
  • human normal fibroblasts (Department of Dermatology, Ajou University) were inoculated into a 24-well micro plate containing DMEM medium at a density of 2 x 10 5 cells/well and were cultured in a 5% CO 2 incubator at 37 °C for 24 hours. Then, the medium was removed from each well, and the cells were treated with various concentrations of a sample, after which the cells were cultured for 24 hours. Then, the cell medium was collected, thereby preparing samples.
  • FIG. 1 shows the results of observing collagen biosynthesis. As shown in FIG. 1, hordenine concentration-dependently increased the production of collagen in the human fibroblasts.
  • Example 2 Measurement of wrinkle-reducing effect of cosmetic composition containing hordenine
  • the wrinkle-reducing effects of the creams were evaluated by measuring a change in skin elasticity.
  • each of three kinds of nourishing creams described in Preparation Example 1 and the nourishing cream of the Comparative Preparation Example was applied to the face of 30 healthy women (25 to 35 years old) twice a day for 3 months under the conditions of a temperature of 24 ⁇ 26 °C and a humidity of 38-40%, and then the skin elasticity of the face of each subject was measured using Cutometer SEM 474 (Courage+Khazaka, Cologne, Germany).
  • the criteria of evaluation were as follows: 0 indicating no improvement in skin elasticity; and 1-5 indicating improvements in skin elasticity.
  • the results of the test are shown in Table 1 below.
  • the creams of Preparation Example 1 showed significantly excellent effects on wrinkle reduction compared to the cream of the Comparative Preparation Example. Also, the skin elasticity increased in a concentration-dependent manner as the hordenine concentration of the creams increased.
  • compositions of nourishing creams containing hordenine are shown in Table 2 below.
  • An aqueous phase comprising purified water, triethanolamine and propylene glycol was dissolved by heating to 70 °C, and an oil phase comprising fatty acid, oily components, an emulsifier and a preservative was dissolved by heating to 70 °C and added to the aqueous phase, thus preparing an emulsion.
  • the emulsion was cooled to 45 °C, and hordenine and a fragrance were added to and dispersed in the cooled emulsion, followed by cooling to 30 °C.
  • Table 2 Components and contents of nourishing creams containing hordenine Component Content (wt%) Hordenine 0.01, 0.05 or 1.00 Jojoba oil 5.0 Liquid paraffin 7.0 Cetylaryl alcohol 2.0 Polyglyceryl-3-methyl glucose distearate 2.0 Glyceryl stearate 0.5 Squalane 3.0 Propylene glycol 4.0 Glycerine 5.0 Triethanolamine 0.3 Carboxyvinyl polymer 0.3 Tocopheryl acetate 0.2 Preservative and fragrance Trace Purified water Balance Sum 100
  • Aging is a process occurring in all living organisms over time in every tissues and organs.
  • the animal skin is an organ having the largest surface area and forming the outer layer of the individual, and thus the degree of skin wrinkles is a convenient indicator of aging.
  • UV irradiation can artificially induce an aging process, and thus oral administration or transdermal administration of a compound into animals after UV irradiation can be used to evaluate the effect of the compound on anti-aging.
  • hairless mice were selected as the experimental animal model. 6-7-week-old hairless mice (Skh:HR-1) were divided into following groups each consisting of 8 mice: a normal group, an UV control group, a UV/hordenine treatment group. The mice were raised during the experimental period. For the normal group and the UV control group, 0.5 ml of saline solution was orally administered to the mice.
  • UV/hordenine treatment group 100 mg/Kg of body weight of hordenine (a solid basis) or a culture medium of stem cells (human cord blood-derived mesenchymal stem cells) was mixed with 0.5 ml of saline solution and administered orally to the mice using a syringe for liquid administration.
  • the animals were administered at the same time for 5 days a week for a total of 5 weeks. Two weeks to five weeks after the oral administration, the UV control group and the UV/hordenine treatment group were radiated with UV light similar as sunlight 3 times a week. The total UV irradiation was 600 mJ/cm 2 over the period of the experiment.
  • skin replicas were collected from the back side of the hairless mice using a silicon polymer before biopsy. After 5 weeks, skin replicas were collected from the sample treatment group and were used to assess the anti-aging effect, that is, the wrinkle-reducing effect.
  • R1 value skin roughness
  • R2 value maximum roughness
  • R3 value average roughness
  • R4 value smoothness depth
  • R5 value Arithmetic average roughness (International Journal of Cosmetric Science, 2005 Jun;27(3):155-60).
  • Example 4 Measurement of effect of hordenine on the inhibition of melanin production in human melanocytes
  • the effect of a test substance on the inhibition of melanin production was measured using human melanocytes.
  • human epidermal melanocytes (derived from neonates) purchased from Cascade Biologics were cultured in Medium 254 containing a human melanocyte growth supplement purchased from Cascade Biologics. Specifically, human epidermal melanocytes were inoculated into a 6-well plate at a density of 1 ⁇ 10 5 cells/well and cultured in a 5 % CO 2 incubator at 37 °C under a confluence of about 80% was reached.
  • the medium was removed, and the cells were treated with each of propafenone/hordenine mixtures having propafenone and hordenine contents of 15 ug/ml: 15 ug/ml (1:1), 7.5 ug/ml: 22.5 ug/ml (1:3), 10 ug/ml: 20 ug/ml (1:2), 20 ug/ml: 10 ug/ml (2:1), and 22.5 ug/ml: 7.5 ug/ml (3:1), propafenone (30 ug/ml), hordenine (30 ug/ml), and the positive control kojic acid (30 ug/ml), and were then cultured for 5 days under conditions of 5% CO 2 and 37 °C while replacing medium at two-day intervals.
  • the medium was removed, and the cells were washed with PBS (phosphated buffer saline) and treated with trypsin, followed by collecting the cells.
  • the collected cells were counted using a hematocytometer, and then the cells of the treatment groups were adjusted to the same number, and the cells of each group were placed in a 2 mL tube. Then, the cells were centrifuged at 5,000-10,000 rpm for 10 minutes, and the supernatant was removed to obtain pellets.
  • the cell pellets were dried at 60 °C, and then 100 ⁇ l of 1M sodium hydroxide solution containing 10% DMSO was added to the cell pellets in a 60 °C incubator, thereby obtaining intracellular melanin.
  • the resulting solution was measured for absorbance at 490 nm using a microplate reader, thus determining the amount of melanin per cell count (FIG. 2). As shown in FIG. 2, when the ratio of propafenone and hordenine was 1: 2, the highest ability to inhibit the production of melanin was observed.
  • the inhibition of melanin synthesis compared to the negative control group was 44% for propafenone alone, 26% for hordenine alone, 49% for the 1:1(w/w) mixture of propafenone and hordenine, 52% for the 1:3 (w/w) mixture of propafenone and hordenine, 55% for the 2:1 (w/w) mixture of propafenone and hordenine, and 54% for the 3:1 (w/w) mixture of propafenone and hordenine, whereas it was 76% for the 1:2 (w/w) mixture of propafenone and hordenine according to the present invention, suggesting that the 1:2 (w/w) mixture of propafenone and hordenine showed a melanin synthesis inhibition which was significantly higher than those shown by propafenone alone, hordenine alone and the propafenone/hordenine mixtures having other mixing weight ratios.
  • nourishing creams A to H prepared in Formulation Example 1-4 were used.
  • Artificial skin purchased from MEL-300-B (MaTek. USA) was treated with a sample for 2 weeks while replacing the sample at 3-day intervals. After completion of the treatment, the tissue was fixed in 10% formaldehyde and embedded in paraffin, after it was sectioned and subjected to Fontana-Masson staining of melanin. Then, the amount of melanin per area was determined using Image-pro plus program (MediaCybernetics, U.S.A).
  • Example 5 Safety test of hordenine on human skin
  • a formulation comprising each of 0.1% hordenine, 0.5% hordenine and 1% hordenine added to squalane serving as a base was prepared. Using each of the prepared formulations, a 24-hour cumulative patch test was performed on the upper arm of 30 healthy adults a total of 9 times at 2-day intervals, thereby determining whether hordenine irritated the skin.
  • the Finn chamber (Epitest Ltd, Finland) was chosen as the patching method. 15 ⁇ l of each of the above formulations for skin application was dropped into each chamber, and a patch was applied. The level of a reaction shown on the skin for each test was scored using the following equation 1 below, and the results are shown in Table 5 below.
  • Average reaction level [ ⁇ (Reaction index x reaction level)/No. of test subjects x maximum score (4 points) ⁇ 100]/number of tests (9 times)
  • Points were marked in accordance with reaction level, for example, ⁇ for 1 point, + for 2 points, and ++ for 4 points.
  • the composition is considered safe if the average reaction level is below 3.
  • hordenine has an anti-inflammatory effect
  • a test for the ability to inhibit COX (cyclooxygenase) activity was performed in human monocyte RAW 264.7 cells (Korean Cell Line Bank) according to a conventional method.
  • the measurement of COX activity was carried out according to the method described in Wadleigh DJ, J Biol Chem. 2000. 3;275(9):6259-66. Specifically, RAW 264.7 cells were cultured and then dispensed in a 24-well plate.
  • a COX-2 luciferase reporter vector was transfected into the cells using Superfect (Qiagen) reagent.
  • the cells were treated with 100 ⁇ g/ml of LPS (lipopolysacchride) and 100 nM of PMA (phorbol myristate acetate) to activate the COX-2 promoter, while these cells were treated with a test sample diluted at various concentrations.
  • the cells were cultured for 16 hours under the same conditions as described above, after the cells were collected to obtain a cell lysis.
  • the cell lysis was centrifuged to remove the cell pellets, and a specific amount of the supernatant was allowed to react with a luciferase substrate, after which the OD value at 450 nm was measured using a luminometer. The results of the measurement are shown in Table 6 below.
  • Example 7 Effect of the protection of fibroblasts from reactive oxygen species
  • Human normal fibroblasts (Department of Dermatology, Ajou University) were inoculated into a 24-well plate containing DMEM medium (2 x 10 5 cells/well) and then were cultured in a 5% CO 2 incubator at 37 °C for 24 hours. Then, the medium was replaced with serum-free DMEM medium, and the cells were treated with various concentrations of a sample and then treated with 1 uM of rotenone known to induce mitochondrial reactive oxygen species. 3 days after treatment with the test sample, cell viability was observed using the MTT method.
  • MTT 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide
  • a lemon-yellowish substrate was cleaved to formazan by the mitochondrial respiratory chain enzyme of living cells. Because no reaction occurs in dead cells, the amount of formazan produced is used for counting of viable cells. The results of the measurement are shown in Table 7 below.
  • the cell regeneration rate is calculated according to the following equation 2:
  • hordenine reduced the fibroblast cytotoxicity induced by rotenone. This indicates that hordenine protects fibroblasts through a mechanism of protecting mitochondria from being injured by reactive oxygen species.
  • Example 8 Effect on the proliferation of hair follicle dermal papilla cells
  • Hair follicle dermal papilla cells were cultured in a serum-free medium containing hordenine, and the effect of hordenine on the viability of these cells was observed.
  • the hair follicle dermal papilla cells were purchased from Cell Applications, Inc. and cultured in hair follicle dermal papilla cell growth medium (Cell Applications, Inc., USA).
  • the hair follicle dermal papilla cells cultured in the hair follicle dermal papilla cell growth medium were inoculated into a 6-well plate at a density of 3 x 10 5 cells per well, and the next day, the cells were washed once with serum-free medium. Then, the cells were cultured in serum-free medium containing hordenine for 72 hours, after which the cells were washed once with serum-free medium. Then, the medium was replaced with serum-free medium containing 10% MTT, and after 3 hours, the O.D. value was measured and calculated as a percentage of control. As a result, as shown in FIG. 3, hordenine increased the proliferation of hair follicle dermal papilla cells in a concentration-dependent manner.
  • a composition for hair growth containing hordenine was prepared as a hydrogel base having the components and contents shown in Table 8 below.
  • test groups 1 and 2 are hair growth compositions containing hordenine.
  • Test group 2 Hordenine 0.1 1.0 Preservative (Gramben) 0.8 0.1 Thickener (Xanthan-gum) 0.3 0.1 Total weight 100 100
  • hair loss patients (age: 40s to late 60s) were divided into the following four groups each consisting of 10 persons: alopecia patients having follicular atrophy and baldness; typical male-pattern alopecia patients; and acute alopecia areata patients.
  • Each of the hair loss compositions prepared in Example 9-1 was applied to the hair loss portion of each alopecia patient twice a day in an amount of 3 cc each time for 6 months. The conditions of hair loss and hair growth were observed at 1-month intervals.
  • Moxidil® Hami Pharm Co., Ltd., Korea
  • 50% ethanol alone was used.
  • bristle hairs including downy hairs started to appear from 1 or 2 months after treatment with the compositions. 6 months after treatment with the compositions, the hair growth effect was shown 80% of the patients. In addition, it was observed that new hairs grew continuously and no hair loss phenomenon was found.
  • Carboxyvinyl polymer 0.1
  • Triethanolamine 0.1
  • Sorbitan sesquioleate 0.5
  • Liquid paraffin 5.0
  • Carboxyvinyl polymer 0.1
  • Sorbitan sesquioleate 0.5
  • Nourishing creams compositions containing propafenone and hordenine
  • Nourishing creams containing propafenone and hordenine were prepared using the compositions shown in Table 10 below according to a conventional method.
  • Sorbitan sesquioleate 0.8
  • Nonylphenylether 0.3
  • Lactose 1 g
  • the above components were mixed with each other and filled into a gelatin capsule according to a conventional method, thereby preparing a capsule formulation.

Abstract

Disclosed is a composition containing hordenine as an active ingredient. The active ingredient hordenine has the effects of improving skin conditions, including the effects of reducing wrinkles, improving skin whitening, promoting hair growth or preventing hair loss. Hordenine exhibits an excellent effect of reducing wrinkles through molecular mechanisms such as the promotion of collagen synthesis and the protection of fibroblasts and has the effects of promoting hair growth and preventing hair loss. Also, it has an excellent skin-whitening effect which is synergistically increased when it is present together with propafenone at a suitable ratio in the composition. Hordenine has the effects of inhibiting inflammatory responses and exhibits an antioxidant effect by a mechanism of protecting mitochondria from oxidative damage caused by reactive oxygen species. Also, it may be applied safely to cosmetic, pharmaceutical and food compositions, because it has no cytotoxicity and does not adversely affect the skin.

Description

COMPOSITION FOR IMPROVING SKIN CONDITIONS COMPRISING HORDENINE
The present invention relates to a composition comprising hordenine as an active ingredient, which has excellent stability, can be used safely without adversely affecting the skin, and has excellent effects on wrinkle reduction, skin whitening, anti-inflammation, antioxidant activity, hair loss prevention and hair growth.
Wrinkles are caused by aging of the skin, and skin aging results from the natural changes associated with the aging process. Skin aging is broadly classified into two types: physiological aging showing age-related changes in the skin function, structure or shape throughout the skin surface; and photo-aging caused by UV light.
As skin aging progresses, changes in the dermis appear, and dermal atrophy appearing in people 70 years or older is a typical aging phenomenon. Changes in the dermis result from changes in substances having low molecular weight in the extracellular matrix due to decreases in the number of fibroblasts and the ability to form fibroblasts. Specific examples of these changes include separation of collagen bundles, a decrease in mucopolysaccharide synthesis, decreases in the number and diameter of collagen and elastin, decomposition of collagen and elastin, and blood vessel expansion. Generally, among various factors, including the skin’s moisture content, collagen content and immune responses to external environments, the expression level and activity of collagenase, a collagen-degrading enzyme that reduces the production and content of collagen, has the greatest effect on the formation of wrinkles. The effect of hordenine on collagen and collagenase has not yet been found.
The human skin color is determined according to the concentration and distribution of melanin in the skin. The melanin pigment that is produced in the melanocytes of the human skin is a phenolic polymer consisting of a complex of a black pigment and a protein and plays an important role in preventing skin damage caused by UV light.
The activity of tyrosinase present in melanocytes was reported to be the most important factor in melanin biosynthesis, and tyrosinase plays an important role in the skin darkening process by converting tyrosine into DOPA and DOPA-quinone, which are intermediate products of melanin polymer production.
Meanwhile, the imbalance of hormones is becoming more severe due to environmental pollution, automobile exhaust gases and the like. For this reason, the rate of hair loss increases, the age of people having hair loss decreases, and the skin immune system is affected, thus causing various skin diseases such as atopy or psoriasis. In addition, a change in eating habits toward instant foods and meats, young people suffering from obesity increase rapidly. Thus, there have been many studies on the development of substances which can improve beauty and can effectively solve various pathological phenomena, for example, formation of wrinkles by intrinsic aging and UV light, discolorations or freckles, obesity, immune imbalance, and hair loss.
Under such circumstances, the present inventors have made extensive efforts to develop novel natural substances, which have excellent effects on wrinkle reduction, skin whitening, hair loss prevention, hair growth promotion, antioxidant activity and anti-inflammatory activity, are highly safe and stable, and do not adversely affect the skin. During such studies, the present inventors have searched for plant-derived compounds, and as a result, have found that hordenine increases collagen synthesis and inhibits collagenase and that a composition containing hordenine as an active ingredient has high efficacy and safety, improves skin conditions and shows excellent anti-inflammatory, antioxidant and anti-obesity effects, thereby completing the present invention.
It is, therefore, an object of the present invention to provide a composition for improving skin conditions.
Another object of the present invention is to provide an anti-inflammatory composition.
Still another object of the present invention is to provide an antioxidant composition.
Yet another object of the present invention is to provide a method of improving skin conditions, inhibiting inflammatory responses, inhibiting oxidative stress, and ameliorating, treating or preventing the disorders, diseases or symptoms associated therewith, by administering said composition to a subject.
These objects, other objects and advantages of the present invention will be more apparent from the following detailed description, the appended claims and the accompanying drawings.
To achieve the above objects, in accordance with one aspect of the present invention, there is provided a composition for improving skin conditions comprising hordenine as an active ingredient.
In accordance with another aspect of the present invention, there is provided an anti-inflammatory composition comprising hordenine as an active ingredient.
In accordance with still another aspect of the present invention, there is provided an antioxidant composition comprising hordenine as an active ingredient.
In accordance with still another aspect of the present invention, there is provided a method of improving the skin conditions of a subject by administering said skin condition-improving composition to the subject in a dermatologically acceptable way.
In accordance with still another aspect of the present invention, there is provided a method of inhibiting inflammation in a subject by administering said anti-inflammatory composition to the subject.
In accordance with yet another aspect of the present invention, there is provided a method of inhibiting oxidative stress by administering said antioxidant composition to a subject.
Hordenine exhibits an excellent effect of reducing wrinkles through molecular mechanisms such as the promotion of collagen synthesis and the protection of fibroblasts and has the effects of promoting hair growth and preventing hair loss. Also, it shows an excellent skin-whitening effect, and particularly, the skin-whitening effect thereof is synergistically increased when hordenine is used together with propafenone at a suitable ratio to prepare a composition. Furthermore, hordenine has an excellent effect of inhibiting inflammation-associated immune responses and an antioxidant effect of protecting mitochondria from reactive oxygen species. In addition, it has excellent safety, because it is not cytotoxic and does not adversely affect the skin. Thus, the inventive composition comprising hordenine can be used as cosmetic, pharmaceutical and food compositions which have exhibit excellent effects of improving skin conditions, including the effects of reducing wrinkles, promoting skin whitening, promoting hair growth and preventing hair loss, and, at the same time, has ensured safety.
FIG. 1 is a graph showing the effect of hordenine on the promotion of collagen (type I collagen) biosynthesis.
FIG. 2 is a graph showing the effects of compositions, comprising hordenine and/or propafenone, on the reduction of melanin synthesis in human melanocytes.
FIG. 3 is a graph showing the effect of hordenine on the proliferation of hair follicle dermal papilla cells.
Hordenine which is used as an active ingredient in the composition according to the present invention is a phenethylamine alkaloid present in plants of the family Cyperaceae, sprouted barley (Hordeum vulgare), Cactus, Citrus aurantium and the like and is typically known to have antibacterial effects. Also, it functions to enhance the sympathetic nervous system together with synephrine that is an adrenergic alkaloid, and thus it is used as a diet food that induces weight loss. In connection with this, US Patent Publication No. 2002/0058075 discloses that a citrus-derived compound comprising hordenine has the effect of reducing body weight by increasing motor ability. Hordenine is known as N,N-dimethyltyramine and has an IUPAC name of 4-(2-dimethylaminoethyl)phenol. Also, it has a structure of the following formula 1:
[Formula 1]
Figure PCTKR2012001994-appb-I000001
This hordenine can be extracted from natural sources, including plants of the family Cyperaceae, sprouted barley (Hordeum vulgare), Cactus, Citrus aurantium and the like. Specifically, hordenine can be obtained from the above natural sources using various extraction methods known in the art. Preferably, it can be extracted using an extraction solvent selected from among (a) water, (b) an anhydrous or hydrated lower alcohol containing 1 to 4 carbon atoms (methanol, ethanol, propanol, butanol, etc.), (c) a mixed solvent of the lower alcohol with water, (d) acetone, (e) ethyl acetate, (f) chloroform, (g) 1,3-butylene glycol, or a mixture of two or more thereof. More preferably, it can be extracted using an aqueous solution of methanol, ethanol and butanol, and most preferably, it can be extracted using an aqueous solution of butanol. It will be obvious to a person skilled in the art that, even when extraction solvents other than the above-mentioned extraction solvents are used, extracts that exhibit substantially the same effects as the above extraction solvents can be obtained. In addition to the extraction solvent, specific extraction conditions (temperature, time, etc.) can be suitably selected by a person skilled in the art depending on the properties of the extraction solvent.
The extracts of the present invention include, in addition to the extracts obtained using the above-described extraction solvents, extracts obtained through conventional purification processes. For example, fractions obtained through various additional purification methods, such as separation with an ultra filtration membrane having a given molecular weight cut-off or separation by various chromatography systems (manufactured for separation according to size, charge, hydrophobicity or affinity), are also included in the scope of the extract of the present invention. In addition, hordenine that is used in the present invention may be prepared by chemical synthesis or may be a commercially available product (for example, hordenine available from Chromadex).
As used herein, the term “improving skin conditions” means improvement and betterment of all skin conditions. Preferably, it means a reduction in skin wrinkles, an improvement in skin whitening, promotion of hair growth, or prevention of hair loss.
The inventive composition for improving skin conditions serves to reduce wrinkles. The composition of the present invention has low cytotoxicity and exhibits an excellent effect of reducing skin wrinkles through molecular mechanisms such as the protection of fibroblasts and the promotion of collagen synthesis, as clearly demonstrated in examples below. It is understood that the reduction in skin wrinkles includes skin protection (e.g., prevention of skin wrinkles, removal of skin wrinkles, and prevention of skin aging).
The inventive composition for improving skin conditions serves to whiten the skin. As used herein, the term “whitening” refers to the effect of reducing skin troubles by preventing skin darkening caused by melanin pigmentation.
Hordenine in the skin whitening composition of the present invention inhibits the expression of tyrosinase in cells to inhibit melanin production, thus exhibiting a skin whitening effect. Also, it is highly stable and there is little or no change of the content of hordenine with the passage of time. In addition, it has little or no adverse effects, such as skin irritation. Particularly, the skin whitening effect of hordenine is synergistically increased when it is used together with propafenone, known to having a skin shortening effect, at a suitable ratio, to prepare a composition.
Propafenone which is used together with hordenine in a skin whitening composition was approved for use as an anti-arrhythmic drug in the year 1980 and is used mainly for supraventricular arrhythmias. In the anti-arrhythmic action of propafenone, Na ions are rapidly introduced into cells by propafenone while the Vmax value of action potential is lowered, thus inhibiting the generation of stimulation and the conduction of excitation between cells. Propafenone is structurally similar to propranolol and has weak beta-blocking activity. Japanese Patent Laid-Open Publication No. 2003-397665 discloses that propafenone has the effect of inhibiting neuropathic pain. Also, a paper prepared by Huh S., et al reported that propafenone has the effect of whitening the skin by inhibiting the expression tyrosinases, TRP-1 and TRP-2 (Arch Dermatol Res. 2010; 302(7): 561-5). The IUPAC name of propafenone is 1-{2-[2-hydroxy-3-(propylamino) propoxy]phenyl}-3-phenylpropan-1-one and has a structure of the following formula 2:
[Formula 2]
Figure PCTKR2012001994-appb-I000002
Propafenone which is used in the present invention may be prepared by chemical synthesis or may be a commercially available product (for example, propafenone available from Sigma). In one preferred embodiment, Hordenine and propafenone may be present at a weight ratio ranging from 1:1 to 1:5. Preferably, hordenine and propafenone are present at a weight ratio of 1:1.5 to 1:2.5, and in this case, the most excellent whitening effect is exhibited. The content of hordenine and propafenone in the composition of the present invention is 0.0001-10 wt%, and preferably 0.001-1.0 wt%, based on the total weight of the composition.
Also, the inventive composition for improving skin conditions has excellent effects on the promotion of hair growth and the prevention of hair loss. As used herein, the terms “prevention of hair loss” and “promotion of hair growth” are used in the same sense.
The composition of the present invention comprises hordenine as an active ingredient and thus exhibits the effect of inhibiting inflammatory responses. Specifically, the active ingredient hordenine generally acts on COX-2 which causes pain in inflammatory responses. Meanwhile, arachidonic acid that is a precursor of prostaglandin (PGE2) is present in the cell membrane and is converted into prostaglandin through a cyclooxygenase pathway. The produced prostaglandin acts as a chemical mediator in allergic and inflammatory processes and irritates a nociceptive nerve or sustains inflammatory responses or influences carcinogenesis. Thus, a substance that inhibits COX-2 activity is highly likely to inhibit inflammatory responses.
Thus, the anti-inflammatory composition of the present invention can be applied to various disorders, diseases or abnormal conditions, which can be prevented or treated by inhibiting inflammatory responses. Examples of inflammation-related diseases which can be prevented or treated by the composition of the present invention include, but are not limited to, bullous pemphigoid, cicatricial pemphigoid, discoid lupus erythematosis, lupus erythematosus, systemic lupus erythematosus, allergy and atopy. The anti-inflammatory composition of the present invention is preferably used for the prevention or treatment of atopy.
Hordenine that is the active ingredient of the inventive composition exhibits an antioxidant effect by protecting mitochondria from oxidative damage caused by reactive oxygen species (ROS) which occur due to various irritations.
The antioxidant composition of the present invention can be applied to various disorders, diseases or abnormal conditions, which can be prevented or treated by inhibiting or removing oxidative stress. Examples of oxidative stress-related diseases which can be prevented or treated by the composition of the present invention include, but are not limited to, atherosclerosis, coronary artery disease, cardiac artery restenosis, reperfusion injury, neurodegenerative diseases such as Parkinson's disease or Alzheimer's disease, stroke, cancer, aging, cardiovascular disease, osteoporosis, disorders of the central nerve system, peripheral vascular disease, and respiratory distress syndrome. Most preferably, the antioxidant composition of the present invention is used for the prevention or treatment of aging.
The correlation between various disorders and free radical damage has been reported generally and the component of various cells comprising enzyme, ion channel, structural protein and membrane lipid is a potential target against reactive free radical species (Rice-Evans C, Mol Aspects of Med 13(1):1-111(1992)). The reaction of the potential target and free radicals impairs cell function of specific ranges, induces pathological changes and allows for cell death ultimately. A variety of diseases caused by physiological oxidation are disclosed in US Patent Nos. 5,750,351; 5,773,209; 5,773,231 and 5,846,959.
In one specific embodiment, the antioxidant composition of the present invention exhibits the effect of protecting intracellular mitochondria from oxidative injury caused by the stimulation of rotenone, which induces the production of reactive oxygen species, and reactive oxygen species induced thereby.
It will be obvious to a person skilled in the art that examples of hordenine that may be used in the present invention include, in addition to the hordenine compound of formula 1, hordenine derivatives, which show the effects of promoting collagen synthesis, reducing wrinkles, whitening the skin, promoting hair growth, preventing hair loss, or exhibiting anti-inflammatory and antioxidant effects, among derivatives obtained by adding substituents to hordenine or substituting hordenine according to a conventional method known in the art. More specifically, examples of hordenine that may be used in the present invention include, in addition to the compound of formula 1, derivatives obtained by linking various substituents to the structure of formula 1 as a nucleus according to a method known in the art. For example, it is believed that derivatives obtained by linking a acetyl group, a hydroxyl group, a halo group, a nitro group or a C1-3 alkyl group to the compound of formula 1 are likely to exhibit effects identical or similar to the compound of formula 1. Thus, these derivatives also fall within the scope of the present invention.
In a preferred embodiment of the present invention, hordenine may be contained in the composition in an amount of 0.00001-15.0 wt%, more preferably 0.0001-10 wt%, and more preferably 0.0001-5 wt%, based on the total weight of the composition.
In a preferred embodiment of the present invention, the composition of the present invention may be used as a cosmetic composition.
The cosmetic composition of the present invention may contain, in addition to the active ingredient hordenine, components that are conventionally used in cosmetic compositions, for examples, conventional auxiliaries, such as antioxidants, stabilizers, solubilizers, vitamins, pigments and fragrances, and carriers.
The cosmetic composition of the present invention may be formulated into any form known in the art. Examples of the formulation include, but are not limited to, a solution, a suspension, an emulsion, a paste, a gel, a cream, a lotion, a powder, a soap, a surfactant-containing cleanser, an oil, a powder foundation, an emulsion foundation, a wax foundation and a spray. More specifically, the cosmetic composition of the present invention can be formulated in the form of nourishing cream, astringent lotion, skin softener, lotion, essence, nourishing gel, or massage cream.
If the formulation of the present invention is paste, cream or gel, it may contain, as carrier components, animal oil, vegetable oil, wax, paraffin, starch, tragacanth gum, cellulose derivatives, polyethylene glycol, silicone, bentonite, silica, talc or zinc oxide.
If the formulation of the present invention is a powder or spray formulation, it may contain, as carrier components, lactose, talc, silica, aluminum hydroxide, calcium silicate or polyamide powder. Particularly, if it is spray, it may additionally contain a propellant, such as chlorofluorohydrocarbon, propane/butane or dimethyl ether.
If the formulation of the present invention is a solution or an emulsion, it may contain, as carrier components, a solvent, a solubilizing agent or an emulsifying agent, for example, water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butyl glycol oil, glycerol fatty ester, polyethylene glycol or sorbitan fatty acid ester.
If the formulation of the present invention is a suspension, it may contain, as carrier components, a liquid diluent, such as water, ethanol or propylene glycol, and a suspending agent, such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester, polyoxyethylene sorbitan ester, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar-agar or tragacanth gum.
If the formulation of the present invention is a surfactant-containing cleansing, it may contain, as carrier components, aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic monoester, isethionate, imidazolium derivatives, methyl taurate, sarcosinate, fatty acid amide ether sulfate, alkyl amido betaine, aliphatic alcohol, fatty acid glyceride, fatty acid diethanolamide, vegetable oil, lanoline derivatives, or ethoxylated glycerol fatty acid ester.
In another preferred embodiment of the present invention, the composition of the present invention may be used as a pharmaceutical composition. The pharmaceutical composition of the present invention contains a pharmaceutically acceptable carrier. The pharmaceutically acceptable carrier contained in the pharmaceutical composition of the present invention is commonly used in pharmaceutical formulations, and examples thereof include, but are not limited to, lactose, dextrose, sucrose, sorbitol, mannitol, starch, rubber acacia, potassium phosphate, alginate, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrups, methylcellulose, methylhydroxy benzoate, propylhydroxy benzoate, talc, magnesium stearate, and mineral oils. The pharmaceutical composition according to the present invention may contain, in addition to these components, a lubricant, a wetting agent, a sweetener, a flavoring agent, an emulsifier, a suspending agent, and a preservative. Details of suitable pharmaceutically acceptable carriers and formulations can be found in Remington's Pharmaceutical Sciences (19th ed., 1995), which is incorporated herein by reference.
The pharmaceutical composition of this invention may be administered orally or parenterally. Preferably, it is administered parenterally, and most preferably, it is administered by topical application.
The suitable dose of the pharmaceutical composition of the present invention may vary depending on various factors, such as formulation methods, administration methods, the patient's age, body weight, sex, pathogenic state, diet, administration time, administration route, excretion rate and reaction sensitivity. The suitable dose of the pharmaceutical composition of the present invention is in the range of 0.001-100 mg/kg for an adult. When the composition is a preparation for external use, it is preferably applied 1-5 times a day in amount of 1.0-3.0 ml for 1 month or more for an adult. However, the above-described dose of the composition does not limit the scope of the present invention.
According to the conventional techniques known to those skilled in the art, the pharmaceutical composition of the present invention may be formulated with the above-described pharmaceutically acceptable carrier and/or vehicle, thus providing several forms, including a unit dosage form and a multi-dosage form. Examples of the formulations include, but are not limited to, a solution, suspension, syrup or emulsion in oil or aqueous medium, an elixir, a powder, a granule, a tablet and a capsule, and may additionally contain a dispersion agent or a stabilizer.
In addition, the composition of the present invention may be used as a food composition. The food composition of the present invention may contain, in addition to the active ingredient hordenine, conventional additives for preparing food compositions, for example, proteins, carbohydrates, fats, nutrients, seasonings and flavors. Examples of the above-described carbohydrates include conventional sugars, such as monosaccharides (e.g., glucose and fructose), disaccharides (e.g., maltose, sucrose and oligosaccharide), and polysaccharides (e.g., dextrin and cyclodextrin); and sugar alcohols (e.g., xylitol, sorbitol and erithritol). Examples of flavors include natural flavors [thaumatin and extract of stevia (e.g., rebaudioside A and glycyrrhizin)] and synthetic flavors (e.g., saccharin and aspartame).
For example, where the food composition of the present invention is provided as a drink, it may comprise, in addition to the active ingredient hordenine, citric acid, liquid fructose, sugar, glucose, acetic acid, malic acid, fruit juice, an extract of Eucommia ulmoides oliv, a jujube extract or an extract of glycyrrhiza uralensis.
Meanwhile, in the example of the present invention, a cumulative skin irritation test for hordenine was carried out, and as a result, it was found that the natural substance hordenine was harmless to the human body. Therefore, hordenine of the present invention may be used with confidence for a long period, because it has little or no toxicity and adverse effect. Particularly, it may be used safely in the above-described cosmetic, pharmaceutical and food compositions.
The present invention also provides a method of improving the skin conditions of a subject, inhibiting inflammatory responses in a subject or inhibiting oxidative stress in a subject by administering said composition to the subject.
As used herein, the term “subject” is meant to include humans and mammals, for example, dogs, pigs, horses and cattle, in which poor skin conditions, for example, wrinkles, skin darkening and hair loss, are to be improved or prevented, or inflammation or inflammation-related diseases are to be treated or prevented, or oxidative stress-related disorders, diseases or abnormal conditions are to be treated or prevented, and these purposes can be achieved by administering the composition of the present invention.
As used herein, the term "administering" means introducing a given substance into a subject according to any suitable method. The composition of the present invention may be administered orally or parenterally through any general route, so long as it can reach a target tissue. In addition, the composition of the present invention may be administered by any device which delivers the active ingredient into a target, for example, a cell.
Those skilled in the art will understand from the above description and the following examples that skin conditions, inflammation, oxidative stress, and disorders, diseases or abnormal conditions resulting therefrom can be improved, ameliorated, prevented or treated by administering the composition of the present invention to a subject using the above-described methods.
The features and advantages of the present invention will be summarized as follows:
(i) The composition of the present invention comprises hordenine as an active ingredient.
(ii) The active ingredient hordenine has excellent effects of improving skin conditions, including the effects of reducing wrinkles, improving skin whitening, promoting hair growth and preventing hair loss. Hordenine exhibits an excellent effect of reducing wrinkles through molecular mechanisms, including the promotion of collagen synthesis and the protection of fibroblasts and has the effects of promoting hair growth and preventing hair loss. In addition, it has an excellent effect of whitening the skin, and particularly, the skin whitening effect thereof is synergistically increased when hordenine is used together with propafenone at a suitable ratio to prepare a composition.
(iii) The active ingredient hordenine has excellent effects of inhibiting inflammation-related immune responses and protecting mitochondria from reactive oxygen species, thus exhibiting antioxidant effects.
(iv) In addition, the active ingredient hordenine can be used safely in cosmetic, pharmaceutical and food compositions, because it has no cytotoxicity and does not adversely affect the skin.
Hereinafter, the present invention will be described in further detail with reference to examples. It is to be understood, however, that these examples are for illustrative purposes and are not intended to limit the scope of the present invention.
Example 1: Measurement of wrinkle-reducing effect of hordenine
Wrinkle-reducing effects can be generally measured by carrying out clinical testing for the ability to biosynthesize collagen in humans. In testing for the ability to biosynthesize collagen, human normal fibroblasts (Department of Dermatology, Ajou University) were inoculated into a 24-well micro plate containing DMEM medium at a density of 2 x 105 cells/well and were cultured in a 5% CO2 incubator at 37 ℃ for 24 hours. Then, the medium was removed from each well, and the cells were treated with various concentrations of a sample, after which the cells were cultured for 24 hours. Then, the cell medium was collected, thereby preparing samples.
The synthesis of collagen was determined by measuring the amount of procollagen type I C-peptide (PICP) using a Procollagen Type I C-peptide EIA kit (MK101; Takara, Kyoto, Japan). The measurement was carried out according to the manufacture’s instruction of the kit (Takara). FIG. 1 shows the results of observing collagen biosynthesis. As shown in FIG. 1, hordenine concentration-dependently increased the production of collagen in the human fibroblasts.
Example 2: Measurement of wrinkle-reducing effect of cosmetic composition containing hordenine
The wrinkle-reducing effect of a cosmetic composition containing hordenine was measured by clinical testing. Nourishing creams prepared in Preparation Example A (nourishing creams containing 0.01%, 0.05% and 1% hordenine) and Comparative Preparation Example (nourishing cream containing purified water) were used.
The wrinkle-reducing effects of the creams were evaluated by measuring a change in skin elasticity. In the measurement of skin elasticity, each of three kinds of nourishing creams described in Preparation Example 1 and the nourishing cream of the Comparative Preparation Example was applied to the face of 30 healthy women (25 to 35 years old) twice a day for 3 months under the conditions of a temperature of 24~26 ℃ and a humidity of 38-40%, and then the skin elasticity of the face of each subject was measured using Cutometer SEM 474 (Courage+Khazaka, Cologne, Germany). The criteria of evaluation were as follows: 0 indicating no improvement in skin elasticity; and 1-5 indicating improvements in skin elasticity. The results of the test are shown in Table 1 below.
Table 1
Test item Preparation Example 1 (Cream Containing 0.01% hordenine) Preparation Example 1 (Cream Containing 0.05% hordenine) Preparation Example 1 (Cream Containing 1% hordenine) Comparative Preparation Example (Cream Containing 0% hordenine)
Skin elasticity 2.85 3.09 3.19 2.72
As can be seen in Table 1 above, the creams of Preparation Example 1 showed significantly excellent effects on wrinkle reduction compared to the cream of the Comparative Preparation Example. Also, the skin elasticity increased in a concentration-dependent manner as the hordenine concentration of the creams increased.
Preparation Example 1: Preparation of creams containing hordenine extract
The compositions of nourishing creams containing hordenine are shown in Table 2 below. An aqueous phase comprising purified water, triethanolamine and propylene glycol was dissolved by heating to 70 ℃, and an oil phase comprising fatty acid, oily components, an emulsifier and a preservative was dissolved by heating to 70 ℃ and added to the aqueous phase, thus preparing an emulsion. Then, the emulsion was cooled to 45 ℃, and hordenine and a fragrance were added to and dispersed in the cooled emulsion, followed by cooling to 30 ℃.
Table 2
Components and contents of nourishing creams containing hordenine
Component Content (wt%)
Hordenine 0.01, 0.05 or 1.00
Jojoba oil 5.0
Liquid paraffin 7.0
Cetylaryl alcohol 2.0
Polyglyceryl-3-methyl glucose distearate 2.0
Glyceryl stearate 0.5
Squalane 3.0
Propylene glycol 4.0
Glycerine 5.0
Triethanolamine 0.3
Carboxyvinyl polymer 0.3
Tocopheryl acetate 0.2
Preservative and fragrance Trace
Purified water Balance
Sum
100
Example 3: Anti-aging effect by oral administration
Aging is a process occurring in all living organisms over time in every tissues and organs. The animal skin is an organ having the largest surface area and forming the outer layer of the individual, and thus the degree of skin wrinkles is a convenient indicator of aging. Especially, UV irradiation can artificially induce an aging process, and thus oral administration or transdermal administration of a compound into animals after UV irradiation can be used to evaluate the effect of the compound on anti-aging.
To examine the effect of the composition on photoaging symptoms, hairless mice were selected as the experimental animal model. 6-7-week-old hairless mice (Skh:HR-1) were divided into following groups each consisting of 8 mice: a normal group, an UV control group, a UV/hordenine treatment group. The mice were raised during the experimental period. For the normal group and the UV control group, 0.5 ml of saline solution was orally administered to the mice. For the UV/hordenine treatment group, 100 mg/Kg of body weight of hordenine (a solid basis) or a culture medium of stem cells (human cord blood-derived mesenchymal stem cells) was mixed with 0.5 ml of saline solution and administered orally to the mice using a syringe for liquid administration.
The animals were administered at the same time for 5 days a week for a total of 5 weeks. Two weeks to five weeks after the oral administration, the UV control group and the UV/hordenine treatment group were radiated with UV light similar as sunlight 3 times a week. The total UV irradiation was 600 mJ/cm2 over the period of the experiment. For an objective evaluation of the effect on improving skin wrinkles, which is an important phenomenon of anti-aging, skin replicas were collected from the back side of the hairless mice using a silicon polymer before biopsy. After 5 weeks, skin replicas were collected from the sample treatment group and were used to assess the anti-aging effect, that is, the wrinkle-reducing effect. As shown in Table 3 below, R1, R2, R3, R4 and R5, which indicate the extent of wrinkle reduction, were lower in the hordenine-administered hairless mice than in the UV/control groups. These results suggest that the composition of the present invention has the effect of reducing skin wrinkles. In other words, the oral administration of hordenine showed the effects of reducing skin wrinkles, suggesting that hordenine has an anti-aging effect.
Table 3
R-parameter UV/Control group UV/hordenine
R1 value 0.57±0.014 0.51±0.008
R2 value 0.48±0.009 0.36±0.010
R3 value 0.27±0.018 0.22±0.015
R4 value 0.15±0.007 0.08±0.009
R5 value 0.31±0.011 0.24±0.013
R1 value: skin roughness, R2 value: maximum roughness, R3 value: average roughness, R4 value: smoothness depth, R5 value: Arithmetic average roughness (International Journal of Cosmetric Science, 2005 Jun;27(3):155-60).
Example 4: Measurement of effect of hordenine on the inhibition of melanin production in human melanocytes
The effect of a test substance on the inhibition of melanin production was measured using human melanocytes.
In the experiments, human epidermal melanocytes (derived from neonates) purchased from Cascade Biologics were cultured in Medium 254 containing a human melanocyte growth supplement purchased from Cascade Biologics. Specifically, human epidermal melanocytes were inoculated into a 6-well plate at a density of 1× 105 cells/well and cultured in a 5 % CO2 incubator at 37 ℃ under a confluence of about 80% was reached. After the culture, the medium was removed, and the cells were treated with each of propafenone/hordenine mixtures having propafenone and hordenine contents of 15 ug/ml: 15 ug/ml (1:1), 7.5 ug/ml: 22.5 ug/ml (1:3), 10 ug/ml: 20 ug/ml (1:2), 20 ug/ml: 10 ug/ml (2:1), and 22.5 ug/ml: 7.5 ug/ml (3:1), propafenone (30 ug/ml), hordenine (30 ug/ml), and the positive control kojic acid (30 ug/ml), and were then cultured for 5 days under conditions of 5% CO2 and 37 ℃ while replacing medium at two-day intervals. After completion of the treatment, the medium was removed, and the cells were washed with PBS (phosphated buffer saline) and treated with trypsin, followed by collecting the cells. The collected cells were counted using a hematocytometer, and then the cells of the treatment groups were adjusted to the same number, and the cells of each group were placed in a 2 mL tube. Then, the cells were centrifuged at 5,000-10,000 rpm for 10 minutes, and the supernatant was removed to obtain pellets. The cell pellets were dried at 60 ℃, and then 100 ㎕ of 1M sodium hydroxide solution containing 10% DMSO was added to the cell pellets in a 60 ℃ incubator, thereby obtaining intracellular melanin. The resulting solution was measured for absorbance at 490 nm using a microplate reader, thus determining the amount of melanin per cell count (FIG. 2). As shown in FIG. 2, when the ratio of propafenone and hordenine was 1: 2, the highest ability to inhibit the production of melanin was observed. Specifically, the inhibition of melanin synthesis compared to the negative control group was 44% for propafenone alone, 26% for hordenine alone, 49% for the 1:1(w/w) mixture of propafenone and hordenine, 52% for the 1:3 (w/w) mixture of propafenone and hordenine, 55% for the 2:1 (w/w) mixture of propafenone and hordenine, and 54% for the 3:1 (w/w) mixture of propafenone and hordenine, whereas it was 76% for the 1:2 (w/w) mixture of propafenone and hordenine according to the present invention, suggesting that the 1:2 (w/w) mixture of propafenone and hordenine showed a melanin synthesis inhibition which was significantly higher than those shown by propafenone alone, hordenine alone and the propafenone/hordenine mixtures having other mixing weight ratios. These results suggest that the effect of propafenone and hordenine on the inhibition of melanin production synergistically increased to the highest level at a specific mixing ratio.
Example 5: Measurement of whitening effect
In this Example, nourishing creams A to H prepared in Formulation Example 1-4 were used. Artificial skin purchased from MEL-300-B (MaTek. USA) was treated with a sample for 2 weeks while replacing the sample at 3-day intervals. After completion of the treatment, the tissue was fixed in 10% formaldehyde and embedded in paraffin, after it was sectioned and subjected to Fontana-Masson staining of melanin. Then, the amount of melanin per area was determined using Image-pro plus program (MediaCybernetics, U.S.A).
Table 4
Skin-whitening effects of nourishing creams containing propafenone, hordenine and a mixture of propafenone and hordenine on artificial skin
Test item Preparation Example A Peparation Example B Peparation Example C mparative Preparation Example (Containing no propafenone and hordenine) Peparation Example D Peparation Example E Peparation Example F Peparation Example G
elanine density
1 6 2 8 3 1 3 4
As can be seen in Table 4 above, in Preparation Example D in which propafenone and hordenine were mixed at a ratio of 1:2, propafenone and hordenine had significantly excellent synergistic effects on skin whitening compared to the effects of the Comparative Preparation Example containing no propafenone and hordenine and the effects of the mixtures having other propafenone/hordenine mixing ratios.
Example 5: Safety test of hordenine on human skin
In order to examine whether the hordenine extract is safe for the human skin, a skin safety test was performed. For this purpose, a cumulative skin irritation test was carried out.
A formulation comprising each of 0.1% hordenine, 0.5% hordenine and 1% hordenine added to squalane serving as a base was prepared. Using each of the prepared formulations, a 24-hour cumulative patch test was performed on the upper arm of 30 healthy adults a total of 9 times at 2-day intervals, thereby determining whether hordenine irritated the skin. The Finn chamber (Epitest Ltd, Finland) was chosen as the patching method. 15 ㎕ of each of the above formulations for skin application was dropped into each chamber, and a patch was applied. The level of a reaction shown on the skin for each test was scored using the following equation 1 below, and the results are shown in Table 5 below.
[Equation 1]
Average reaction level=[{(Reaction index x reaction level)/No. of test subjects x maximum score (4 points)}×100]/number of tests (9 times)
Points were marked in accordance with reaction level, for example, ± for 1 point, + for 2 points, and ++ for 4 points. The composition is considered safe if the average reaction level is below 3.
Table 5
Figure PCTKR2012001994-appb-T000001
As can be seen in Table 5 above, the numbers of subjects corresponding to ± , + and ++ in for test groups 1, 2 and 3 were 0, 0 and 1, respectively, and the average reactive levels in test groups 1, 2 and 3 were 0.00, 0.00 and 0.09, respectively. Because the average reaction level was below 3, hordenine was proven to be safe for use on the human skin without showing any significant cumulative irritation.
Example 6: Evaluation of anti-inflammatory effect
In order to determine whether hordenine has an anti-inflammatory effect, a test for the ability to inhibit COX (cyclooxygenase) activity was performed in human monocyte RAW 264.7 cells (Korean Cell Line Bank) according to a conventional method. The measurement of COX activity was carried out according to the method described in Wadleigh DJ, J Biol Chem. 2000. 3;275(9):6259-66. Specifically, RAW 264.7 cells were cultured and then dispensed in a 24-well plate. A COX-2 luciferase reporter vector was transfected into the cells using Superfect (Qiagen) reagent. The cells were treated with 100 μg/ml of LPS (lipopolysacchride) and 100 nM of PMA (phorbol myristate acetate) to activate the COX-2 promoter, while these cells were treated with a test sample diluted at various concentrations. The cells were cultured for 16 hours under the same conditions as described above, after the cells were collected to obtain a cell lysis. The cell lysis was centrifuged to remove the cell pellets, and a specific amount of the supernatant was allowed to react with a luciferase substrate, after which the OD value at 450 nm was measured using a luminometer. The results of the measurement are shown in Table 6 below.
Table 6
Sample COX activity(%)
LPS (1㎍/㎖) 100
LPS (1㎍/㎖) + hordenine (1㎍) 98
LPS (1㎍/㎖) + hordenine (10㎍) 79
LPS (1㎍/㎖) + hordenine (100㎍) 71
As can be seen in Tabel 6 above, hordenine concentration-dependently inhibited the activity of COX. This suggests that hordenine can exhibit an anti-inflammatory effect.
Example 7: Effect of the protection of fibroblasts from reactive oxygen species
Human normal fibroblasts (Department of Dermatology, Ajou University) were inoculated into a 24-well plate containing DMEM medium (2 x 105 cells/well) and then were cultured in a 5% CO2 incubator at 37 ℃ for 24 hours. Then, the medium was replaced with serum-free DMEM medium, and the cells were treated with various concentrations of a sample and then treated with 1 uM of rotenone known to induce mitochondrial reactive oxygen species. 3 days after treatment with the test sample, cell viability was observed using the MTT method. MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide), a lemon-yellowish substrate, was cleaved to formazan by the mitochondrial respiratory chain enzyme of living cells. Because no reaction occurs in dead cells, the amount of formazan produced is used for counting of viable cells. The results of the measurement are shown in Table 7 below.
The cell regeneration rate is calculated according to the following equation 2:
[Equation 2]
Cell regeneration rate = {(sample-treated group ? rotenone-treated group)/ rotenone-treated group} x 100
Table 7
Test item Hordenine (10μM) Hordenine (50μM) Hordenine (100μM)
Cell regeneration rate 12 31 49
As can be seen in Table 7 above, hordenine reduced the fibroblast cytotoxicity induced by rotenone. This indicates that hordenine protects fibroblasts through a mechanism of protecting mitochondria from being injured by reactive oxygen species.
Example 8: Effect on the proliferation of hair follicle dermal papilla cells
In order to examine the effects of hordenine of the present invention on hair loss prevention and hair growth under in vitro conditions, the following test was performed.
Hair follicle dermal papilla cells were cultured in a serum-free medium containing hordenine, and the effect of hordenine on the viability of these cells was observed. The hair follicle dermal papilla cells were purchased from Cell Applications, Inc. and cultured in hair follicle dermal papilla cell growth medium (Cell Applications, Inc., USA).
The hair follicle dermal papilla cells cultured in the hair follicle dermal papilla cell growth medium were inoculated into a 6-well plate at a density of 3 x 105 cells per well, and the next day, the cells were washed once with serum-free medium. Then, the cells were cultured in serum-free medium containing hordenine for 72 hours, after which the cells were washed once with serum-free medium. Then, the medium was replaced with serum-free medium containing 10% MTT, and after 3 hours, the O.D. value was measured and calculated as a percentage of control. As a result, as shown in FIG. 3, hordenine increased the proliferation of hair follicle dermal papilla cells in a concentration-dependent manner.
Example 9: Effects on hair loss prevention and hair growth
In order to examine the effects of hordenine of the present invention on hair loss prevention and hair growth, the following test was performed.
9-1: Preparation of composition for hair growth
A composition for hair growth containing hordenine was prepared as a hydrogel base having the components and contents shown in Table 8 below. In Table 8, test groups 1 and 2 are hair growth compositions containing hordenine.
Table 8
Componet Content(%)
Test group 1 Test group 2
Hordenine 0.1 1.0
Preservative (Gramben) 0.8 0.1
Thickener (Xanthan-gum) 0.3 0.1
Total weight 100 100
9-2. Measurement of effects on hair loss preservation and hair growth
40 hair loss patients (age: 40s to late 60s) were divided into the following four groups each consisting of 10 persons: alopecia patients having follicular atrophy and baldness; typical male-pattern alopecia patients; and acute alopecia areata patients. Each of the hair loss compositions prepared in Example 9-1 was applied to the hair loss portion of each alopecia patient twice a day in an amount of 3 cc each time for 6 months. The conditions of hair loss and hair growth were observed at 1-month intervals. In a comparative group, Moxidil® (Hanmi Pharm Co., Ltd., Korea) was used, and in a control group, 50% ethanol alone was used.
The criteria of evaluation were as follows:
4: having high effect = new hairs appear
3: having moderate effect = new hairs (downy hairs) appear
2: having slight effect = degree of hair loss decreases
1: having no effect = no change appears
The results of the test are shown in Table 9 below
Table 9
- Control group Comparative group Test group 1 Test group 2
Efficacy/effect 1.5 3.8 2.4 3.2
As shown in Table 9 above, in the alopecia patients treated with the hair loss compositions containing hordenine of the present invention, bristle hairs including downy hairs started to appear from 1 or 2 months after treatment with the compositions. 6 months after treatment with the compositions, the hair growth effect was shown 80% of the patients. In addition, it was observed that new hairs grew continuously and no hair loss phenomenon was found.
Hereinafter, formulation examples for the composition of the present invention will be described.
Formulation Example 1: Cosmetic formulations
1-1: Skin lotion (skin softener) (content: wt%)
Hordenine: 0.01
Glycerin: 3.0
Butylene glycol: 2.0
Propylene glycol: 2.0
Carboxyvinyl polymer: 0.1
Ethanol 10.0:
Triethanolamine: 0.1
Preservative, trace pigment, trace fragrance and trace purified water: balance
Sum: 100.0
1-2: Milk lotion (content: wt%)
Hordenine: 0.01
Beeswax: 4.0
Polysorbate 60: 1.5
Sorbitan sesquioleate: 0.5
Liquid paraffin: 5.0
Squalane: 5.0
Caprylic/capric triglyceride: 5.0
Glycerin: 3.0
Butylene glycol 3.0
Propylene glycol: 3.0
Carboxyvinyl polymer: 0.1
Triethanolamine: 0.2
Preservative, trace pigment, trace fragrance and trace purified water: balance
Sum: 100.0
1-3: Nourishing cream (content: wt%)
Hordenine: 0.01
Beeswax: 10.0
Polysorbate: 60 1.5
Sorbitan sesquioleate: 0.5
Liquid paraffin: 10.0
Squalane: 5.0
Caprylic/capric triglyceride: 5.0
Glycerin: 5.0
Butylene glycol 3.0
Propylene glycol: 3.0
Triethanolamine: 0.2
Preservative, trace pigment, trace fragrance and trace purified water: balance
Sum: 100
1-4: Nourishing creams (compositions containing propafenone and hordenine)
Nourishing creams containing propafenone and hordenine were prepared using the compositions shown in Table 10 below according to a conventional method.
Table 10
Figure PCTKR2012001994-appb-T000002
1-5: Massage cream (content: wt%)
Hordenine: 0.01
Beeswax: 10.0
Polysorbate 60: 1.5
Sorbitan sesquioleate: 0.8
Liquid paraffin: 40.0
Squalane: 5.0
Caprylic/capric triglyceride: 4.0
Glycerin: 5.0
Butylene glycol: 3.0
Propylene glycol: 3.0
Triethanolamine: 0.2
Preservative, trace pigment, trace fragrance and trace purified water: balance
Sum: 100
1-6: Pack (content: wt%)
Hordenine: 0.01
Polyvinyl alcohol: 13.0
Sodium carboxymethylcellulose: 0.2
Alantoin: 0.1
Ethanol: 5.0
Nonylphenylether: 0.3
Preservative, trace pigment, trace fragrance and trace purified water: balance
Sum: 100
Formulation Example 2: Pharmaceutical formulations
2-1: Preparation of powder formulation
Hordenine: 2 g
Lactose: 1 g
The above components were mixed with each other and placed in an airtight bag, thereby preparing a powder formulation.
2-2: Preparation of tablet formulation
Hordenine: 100 ㎎
Maize starch: 100 ㎎
Lactose: 100 ㎎
Magnesium stearate: 2 ㎎
The above components were mixed with each other and compressed into a tablet according to a conventional method, thereby preparing a tablet formulation.
2-3: Preparation of capsule formation
Hordenine: 100 ㎎
Maize starch: 100 ㎎
Lactose: 100 ㎎
Magnesium stearate: 2 ㎎
The above components were mixed with each other and filled into a gelatin capsule according to a conventional method, thereby preparing a capsule formulation.

Claims (13)

  1. A composition for improvement of skin conditions, comprising hordenine as an active ingredient.
  2. An anti-inflammatory composition comprising hordenine as an active ingredient.
  3. An antioxidant composition comprising hordenine as an active ingredient.
  4. The composition of claim 1, wherein the improvement of skin conditions is reduction of wrinkles, promotion of skin whitening, promotion of hair growth, or prevention of hair loss.
  5. The composition of claim 4, wherein the improvement of skin conditions is promotion of skin whitening.
  6. The composition of claim 5, further comprising propafenone.
  7. The composition of claim 6, wherein propafenone and hordenine are contained at a weight ratio 1: 1.5 to 1:2.5.
  8. The composition of claim 6, wherein propafenone and hordenine are contained in an amount of 0.0001-10 wt% based on the total weight of the composition.
  9. The composition of any one of claims 1 to 8, wherein hordenine is contained in an amount of 0.0001-10.0 based on the total weight of the composition.
  10. The composition of any one of claims 1 to 8, wherein the composition is a cosmetic composition.
  11. The composition of claim 10, wherein the composition has a formulation selected from the group consisting of a solution, a suspension, an emulsion, a paste, a gel, a cream, a lotion, a powder, a soap, a surfactant-containing cleanser, an oil, a powder foundation, an emulsion foundation, a wax foundation and a spray.
  12. The composition of any one of claims 1 to 8, wherein the composition is a pharmaceutical composition.
  13. The composition of any one of claims 1 to 8, wherein the composition is a food composition.
PCT/KR2012/001994 2012-02-29 2012-03-20 Composition for improving skin conditions comprising hordenine WO2013129723A1 (en)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
KR1020120021394A KR101436199B1 (en) 2012-02-29 2012-02-29 Composition for Improving Skin Conditions Comprising Hordenine
KR10-2012-0021394 2012-02-29

Publications (1)

Publication Number Publication Date
WO2013129723A1 true WO2013129723A1 (en) 2013-09-06

Family

ID=49082898

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/KR2012/001994 WO2013129723A1 (en) 2012-02-29 2012-03-20 Composition for improving skin conditions comprising hordenine

Country Status (2)

Country Link
KR (1) KR101436199B1 (en)
WO (1) WO2013129723A1 (en)

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN110151742A (en) * 2019-05-22 2019-08-23 王雄 Purposes of the hordenine in preparation treatment hypophysis tumor medicine
EP3148333B1 (en) 2014-05-26 2021-03-03 Maltaflor International GmbH Use of a fertilizer comprising the constituent hordenine
WO2022194755A1 (en) 2021-03-16 2022-09-22 Unilever Ip Holdings B.V. Use of hordenine for treating cellular senescence

Families Citing this family (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR102493627B1 (en) * 2015-11-26 2023-01-31 코웨이 주식회사 Pharmaceutical Composition Comprising Hordenine as Active Ingredient for Preventing or Treating Skin Cancer
KR20220071315A (en) 2020-11-24 2022-05-31 주식회사 에이엠메딕스 Composition for skin whitening
KR102487742B1 (en) 2020-11-24 2023-01-12 주식회사 에이엠메딕스 Composition for skin whitening

Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20020058075A1 (en) * 1998-08-31 2002-05-16 Dennis Jones Methods for inducing weight loss in a human with materials derived from citrus varieties
WO2003097077A1 (en) * 2002-05-15 2003-11-27 Zhishin Inc. Novel dosage forms for materials derived from citrus varieties
US20040146539A1 (en) * 2003-01-24 2004-07-29 Gupta Shyam K. Topical Nutraceutical Compositions with Selective Body Slimming and Tone Firming Antiaging Benefits
US20040208902A1 (en) * 2003-04-18 2004-10-21 Gupta Shyam K. Controlled-release nano-diffusion delivery systems for cosmetic and pharmaceutical compositions
US20060039887A1 (en) * 2004-08-20 2006-02-23 Infinity2 Health Sciences, Inc. Cosmetic or pharmaceutical composition for skin care

Family Cites Families (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR100919897B1 (en) * 2007-11-05 2009-09-30 바이오스펙트럼 주식회사 Compositions for Improving Skin Conditions Comprising Isomenthone as an Active Ingredient
KR101096206B1 (en) * 2008-11-20 2011-12-22 스킨큐어(주) Compositions for Improving Skin Conditions Comprising Pterocladiella capillacea

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20020058075A1 (en) * 1998-08-31 2002-05-16 Dennis Jones Methods for inducing weight loss in a human with materials derived from citrus varieties
WO2003097077A1 (en) * 2002-05-15 2003-11-27 Zhishin Inc. Novel dosage forms for materials derived from citrus varieties
US20040146539A1 (en) * 2003-01-24 2004-07-29 Gupta Shyam K. Topical Nutraceutical Compositions with Selective Body Slimming and Tone Firming Antiaging Benefits
US20040208902A1 (en) * 2003-04-18 2004-10-21 Gupta Shyam K. Controlled-release nano-diffusion delivery systems for cosmetic and pharmaceutical compositions
US20060039887A1 (en) * 2004-08-20 2006-02-23 Infinity2 Health Sciences, Inc. Cosmetic or pharmaceutical composition for skin care

Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP3148333B1 (en) 2014-05-26 2021-03-03 Maltaflor International GmbH Use of a fertilizer comprising the constituent hordenine
CN110151742A (en) * 2019-05-22 2019-08-23 王雄 Purposes of the hordenine in preparation treatment hypophysis tumor medicine
CN110151742B (en) * 2019-05-22 2021-08-24 王雄 Application of hordenine in preparation of medicine for treating pituitary tumor
WO2022194755A1 (en) 2021-03-16 2022-09-22 Unilever Ip Holdings B.V. Use of hordenine for treating cellular senescence

Also Published As

Publication number Publication date
KR20130099694A (en) 2013-09-06
KR101436199B1 (en) 2014-09-01

Similar Documents

Publication Publication Date Title
KR100823533B1 (en) COMPOSITIONS FOR IMPROVING SKIN CONDITIONS COMPRISING alpha;-BISABOLOL AS AN ACTIVE INGREDIENT
KR100862968B1 (en) Agents for Improving Wrinkles on Skin Comprising Matrine or Its Oxidized Derivatives
WO2013129723A1 (en) Composition for improving skin conditions comprising hordenine
WO2019045259A2 (en) Cosmetic composition including dendrobium candidum wallich ex lindley flower extract
WO2015156439A1 (en) Composition for improving skin condition, comprising extract of andrographis paniculata, andrographolide or salt thereof
WO2012002784A2 (en) Composition containing paper mulberry extracts
WO2019098468A1 (en) Food or cosmetic composition for skin whitening, skin moisturizing, or skin wrinkle improvement, comprising composite of sheep placenta powder and plants
WO2017146414A1 (en) Composition for skin moisturization and skin wrinkle alleviation, containing α-terpineol as active ingredient
WO2024049120A1 (en) Cosmetic composition containing extract extracted by means of eco-friendly natural eutectic solvent
KR20140012456A (en) Composition for improving skin conditions comprising akebia saponin d
WO2022085858A1 (en) Composition for whitening skin or ameliorating wrinkles comprising dendropanax morbiferus extract
WO2011096688A2 (en) Composition for improving scalp and hair health comprising a dibenzo-p-dioxin derivative
KR100848800B1 (en) Skincare composition containing brazilian pepper oil
WO2018080039A1 (en) Composition for preventing hair loss or improving hair growth, containing yellow-colored soybean leaf extract
WO2018097388A1 (en) Composition for skin whitening, wrinkle alleviation, antioxidation, and ultraviolet light blocking, containing jujube seed extract as active ingredient
WO2017119535A1 (en) Anti-aging composition comprising carnosine, soy peptide, and andrographis paniculata extract
WO2014119826A1 (en) Cosmetic composition for whitening or wrinkle improvement which includes seaweed extract as active ingredient
WO2014003224A1 (en) Compositions for whitening skin comprising madecassoside
WO2009151212A2 (en) External preparation composition for skin comprising ginseng flower or ginseng seed extracts
WO2012173383A2 (en) Skin composition for external use containing cryptotanshinone as active ingredient
WO2014088258A1 (en) Composition for improving skin conditions comprising veratric acid or acceptable salt thereof as an active ingredient
KR100863616B1 (en) Skincare composition containing copaiba balsam oil
WO2017003190A1 (en) Whitening cosmetic composition comprising caragana sinica root extract
WO2018151563A1 (en) Composition for preventing photoaging and alleviating skin wrinkles, containing cosmos bipinnatus extract
WO2013070018A1 (en) Composition comprising carnosic acid or derivative thereof

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 12869645

Country of ref document: EP

Kind code of ref document: A1

NENP Non-entry into the national phase

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 12869645

Country of ref document: EP

Kind code of ref document: A1