WO2018004252A1 - Composition for promoting hair growth or inhibiting hair loss using rosmarinus officinalis (rosemary) leaf extract - Google Patents

Composition for promoting hair growth or inhibiting hair loss using rosmarinus officinalis (rosemary) leaf extract Download PDF

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WO2018004252A1
WO2018004252A1 PCT/KR2017/006833 KR2017006833W WO2018004252A1 WO 2018004252 A1 WO2018004252 A1 WO 2018004252A1 KR 2017006833 W KR2017006833 W KR 2017006833W WO 2018004252 A1 WO2018004252 A1 WO 2018004252A1
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hair
composition
extract
essential oil
rosemary
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PCT/KR2017/006833
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French (fr)
Korean (ko)
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최한준
박상용
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최한준
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    • 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
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A23L33/105—Plant extracts, their artificial duplicates or their derivatives
    • 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)
    • A61K36/185—Magnoliopsida (dicotyledons)
    • A61K36/53—Lamiaceae or Labiatae (Mint family), e.g. thyme, rosemary or lavender
    • 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
    • A61K8/33—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing oxygen
    • A61K8/36—Carboxylic acids; Salts or anhydrides thereof
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00—Cosmetics or similar toiletry preparations
    • A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
    • A61K8/97—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from algae, fungi, lichens or plants; from derivatives thereof
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00—Cosmetics or similar toiletry preparations
    • A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
    • A61K8/97—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from algae, fungi, lichens or plants; from derivatives thereof
    • A61K8/9783—Angiosperms [Magnoliophyta]
    • A61K8/9789—Magnoliopsida [dicotyledons]
    • 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
    • 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
    • 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
    • A23V2200/00—Function of food ingredients
    • A23V2200/30—Foods, ingredients or supplements having a functional effect on health
    • A23V2200/318—Foods, ingredients or supplements having a functional effect on health having an effect on skin health and hair or coat
    • 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
    • A23V2250/00—Food ingredients
    • A23V2250/20—Natural extracts
    • A23V2250/21—Plant extracts

Definitions

  • the present invention is rosemary ( Rosmarinus officinalis ) relates to a composition for promoting hair growth or inhibiting hair loss using a leaf extract.
  • Hair is formed from hair follicles, with about 5 million adult hair follicles distributed throughout the body, except for palms and seagrass, with 1 million heads, of which 100,000 are present in the scalp. (Bulletin of Food Technology 2009, 22 (4): 651-663). Scalp hair accounts for about 20% of human hair, and has the function of protecting the head from shocks by buffering the head along with the physiological functions of the hair such as sweat evaporation and temperature control. It also has important social functions for personality and image production (Clin Dermatol 2001, 19 (2): 161-166).
  • hair follicle which is a very complex organ consisting of internal hair root, external hair root, hair shaft, and matrix cells, and hair dermal cells among various types of hair follicles.
  • papilla cells play an important role in hair growth.
  • Dermal papilla cells a type of dermal fibroblast specialized in hair bulbs, are not only normal hair cycles but also hair growth through the growth of dermal papilla cells in symptoms such as androgen alopecia (male hair loss). It plays an important role in controlling growth. Thus, factors affecting the proliferation of dermal papilla cells in hair loss are important targets for improving hair loss.
  • alopecia refers to an increase in the number of hairs that fall abnormally due to a shorter growth rate and a longer degenerative or resting period during these cycles (Differentiation, 2004, 72 (9-10): 489; Dev Cell. , 2001, 1 (1): 13; J. Invest.Dermatol., 2005, 125 (1): 42).
  • dihydrotestosterone is another testosterone produced by the testosterone called testosterone by type 2 5 ⁇ -reductase.
  • Minoxidil preparations see US Pat. Nos. 4,139,619 and 4,596,812) and finasteride (US patents). 5555457, 5760046 or Korean Patent No. 375083).
  • Minoxidil preparations are known to reduce hair loss by up to 10% in men with alopecia, but have the disadvantage of not having a very good effect on hair loss outside the kiln and can cause depression. There is a problem in that they have side effects such as decreased libido, erectile dysfunction. For this reason, efforts have recently been made to find substances having a hair growth promoting or hair loss inhibiting activity in natural products with few side effects.
  • the present invention discloses hair growth promoting activity and the like of the rosemary extract.
  • the mixture of essential oil extract of rosemary leaf by carnosic acid and hexane extraction method is rosema by carnosic acid or hexane extraction method. It was confirmed that the activity of the essential oil extract of the leaves of Lee, and especially the activity of 1: 2 mixture of essential oil extract of rosemary leaves by carnosic acid and hexane extraction method. Furthermore, the inventors of the present invention showed that when the animal experiment was conducted using a mouse depilated using a mixture of 1: 2 of the essential oil extract of rosemary leaf by carnosic acid and hexane extraction method, which was the most active in the cell test results, It was confirmed that the hair density and thickness were increased along with the promotion.
  • Carnosic acid (CAS # 3650-09-7) is a benzenediol abietane diterpene found in rosemary ( Rosmarinus officinalis ), Salvia ( Salvia officinalis ), and has a strong antioxidant activity. , Anti-cancer activity by inducing apoptosis in breast cancer cells, ovarian cancer cells, prostate cancer cells and the like (International Journal of Molecular Sciences. 2014; 15 (7) 12698-12713).
  • composition for promoting hair growth inhibition or hair loss of the present invention comprises (i) a rosemary leaf extract or (ii) a mixture of carnosic acid and rosemary leaf essential oil as an active ingredient. It is done.
  • active ingredient alone refers to a component that can exhibit the desired activity or exhibit itself with a carrier which is inactive.
  • hair loss should be understood as including all the symptoms classified as hair loss in the art regardless of the direct and indirect cause of the occurrence. Specifically, it includes all of the symptoms of hair loss due to poor blood circulation, excessive sebum secretion due to excessive production of dihydrotestosterone, deterioration of the scalp function by peroxides or bacteria, aging, genetic factors, stress, and their complex actions. .
  • extract refers to water, methanol, ethanol, butanol lower alcohol having 1 to 4 carbon atoms, acetone, methylene chloride, ethylene, acetone, hexane, ether, chloroform, ethyl acetate, butyl acetate, Supercritical extraction solvents such as extracts obtained by leaching with N, N-dimethylformamide (DMF), dimethylsulfoxide (DMSO), 1,3-butylene glycol, propylene glycol or a mixed solvent thereof, carbon dioxide, pentane, etc.
  • DMF N-dimethylformamide
  • DMSO dimethylsulfoxide
  • the extraction method is applied to any method, such as cold needle, reflux, heating, ultrasonic radiation, supercritical extraction in consideration of the polarity, degree of extraction, the degree of preservation of the active substance can do.
  • the fractionated extract includes a fraction obtained by suspending the crude extract in a specific solvent and mixing and standing with a solvent having a different polarity.
  • the crude extract is sequentially fractionated using the solvents in increasing or decreasing polarity.
  • the meaning of the extract includes a concentrated liquid or solid extract from which the extraction solvent is removed by lyophilization, vacuum drying, hot air drying, spray drying, or the like.
  • it preferably means an extract obtained by leaching using water, acetone or a mixed solvent thereof as an extraction solvent (in particular, an extract obtained using a mixed solvent of water and acetone).
  • rosemary leaf essential oil extract means the extract of the oil phase obtained by condensing the volatile component of the rosemary leaf to be extracted.
  • methods known in the art such as steam distillation, solvent extraction using volatile solvents (benzene, hexane, etc.), supercritical extraction, can be used.
  • rosemary leaf essential oil extract refers to an extract obtained by using hexane as a volatile solvent, specifically, an extract obtained by dehydration after reflux cooling with hexane.
  • the effective amount is 0.0001% by weight to 15% by weight based on the total weight of the composition. Will be determined within the% range.
  • the term "effective amount" herein refers to the present invention, which may exhibit an intended hair growth promoting effect or hair loss inhibiting effect when the composition of the present invention is administered to a mammal, preferably a human, during the administration period according to the suggestion of a medical professional or the like. Refers to the amount of the active ingredient included in the composition of. Such effective amounts can be determined experimentally within the range of ordinary skill in the art.
  • the food composition of the present invention may be prepared in any form, for example, beverages such as tea, juice, carbonated beverages, ionic beverages, processed oils such as milk, yogurt, gums, rice cakes, sweets, bread, sweets, noodles, and the like. It can be prepared as a dietary supplement, such as foods, tablets, capsules, pills, granules, liquid, powder, flaky, paste, syrup, gel, jelly, bar.
  • the food composition of the present invention can distinguish any product as long as it conforms to the enforcement regulations at the time of manufacture and distribution in the legal and functional divisions.
  • it may be a health functional food according to the Act on Health Functional Food, or confectionary, bean, tea, beverage, special purpose food, etc. according to each food type in the Food Code of the Food Sanitation Act (KFDA notification, standards and standards of food). .
  • the food composition of the present invention may include food additives in addition to the active ingredient.
  • Food additives can generally be understood as substances that are added to, mixed with, or infiltrated in the manufacture, processing, or preservation of foods, and because they are taken daily with food and for a long time, their safety must be ensured.
  • the Food Additives Code of the Food Sanitation Act (KFDA Notification, Food Additives Standards and Standards) defines the food additives that are guaranteed to be safe by dividing them into chemical synthetics, natural additives, and mixed preparations. In sweeteners, flavors, preservatives, emulsifiers, acidulants, thickeners and the like can be classified.
  • Sweeteners are used to impart a proper sweetness to foods, and may be natural or synthesized.
  • a natural sweetener is used.
  • sugar sweeteners such as corn syrup solids, honey, sucrose, fructose, lactose and maltose.
  • Flavoring agents can be used to enhance the taste or aroma, both natural and synthetic. It is the case of using a natural thing preferably.
  • the natural flavor may be obtained from apples, lemons, citrus fruits, grapes, strawberries, peaches, and the like, or may be obtained from green tea leaves, round leaves, jujube leaves, cinnamon, chrysanthemum leaves, jasmine and the like.
  • ginseng red ginseng
  • bamboo shoots aloe vera, ginkgo and the like
  • Natural flavors can be liquid concentrates or solid extracts.
  • a synthetic flavor may be used, and as the synthetic flavor, esters, alcohols, aldehydes, terpenes, and the like may be used.
  • calcium sorbate, sodium sorbate, potassium sorbate, calcium benzoate, sodium benzoate, potassium benzoate, EDTA (ethylenediaminetetraacetic acid) and the like can be used, and as an emulsifier, acacia gum, carboxymethylcellulose, xanthan gum, pectin Etc. may be used, and acidic acid, malic acid, fumaric acid, adipic acid, phosphoric acid, gluconic acid, tartaric acid, ascorbic acid, acetic acid, phosphoric acid, and the like may be used.
  • the acidulant may be added so that the food composition is at an appropriate acidity for the purpose of inhibiting the growth of microorganisms in addition to the purpose of enhancing the taste.
  • suspending implementers As the thickener, suspending implementers, sedimenting agents, gel formers, swelling agents and the like can be used.
  • the food composition of the present invention may include a bioactive substance or minerals known in the art for the purpose of supplementing and reinforcing the functionality and nutritional properties and ensuring the stability as a food additive.
  • physiologically active substances include catechins, vitamin B1, vitamin C, vitamin E, vitamin B12 and the like, tocopherol, dibenzoylthiamine, and the like contained in green tea.
  • minerals include calcium preparations such as calcium citrate and magnesium stearate.
  • Magnesium preparations, iron preparations such as iron citrate, chromium chloride, potassium iodine, selenium, germanium, vanadium, zinc and the like.
  • the food additive as described above may be included in an amount that can achieve the purpose of addition according to the product type.
  • composition of the present invention may be regarded as a pharmaceutical composition in another specific embodiment.
  • the pharmaceutical composition of the present invention can be classified into any product as long as it conforms to the relevant enforcement regulations at the time of manufacture and distribution in the legal and functional divisions.
  • it may be a quasi-drug or drug under the Pharmaceutical Affairs Act or, if it is a drug, it may be a generic or specialized drug.
  • compositions of the present invention may be prepared in oral or parenteral formulations according to the route of administration by conventional methods known in the art, including pharmaceutically acceptable carriers in addition to the active ingredients.
  • pharmaceutically acceptable here means that the subject of application (prescription) is not toxic as far as adaptable without inhibiting the activity of the active ingredient.
  • composition of the present invention is prepared in an oral dosage form, powders, granules, tablets, pills, dragees, capsules, solutions, gels, syrups, suspensions, wafers according to methods known in the art with suitable carriers It may be prepared in a formulation such as.
  • suitable pharmaceutically acceptable carriers include lactose, glucose, sucrose, dextrose, sugars such as sorbitol, mannitol, xylitol, starch such as corn starch, potato starch, wheat starch, cellulose, methylcellulose, ethylcellulose, Celluloses such as sodium carboxymethylcellulose, hydroxypropylmethylcellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, magnesium stearate, mineral oil, malt, gelatin, talc, polyol, vegetable Yu etc. can be mentioned. If formulated, it may be formulated to include diluents and / or excipients, such as fillers, extenders, binders, wetting agents, disintegrants, surfactants, if necessary.
  • diluents and / or excipients such as fillers, extenders, binders, wetting agents, disintegrants, surfactants, if necessary.
  • compositions of the present invention When the pharmaceutical compositions of the present invention are prepared in parenteral formulations, they may be formulated in the form of injections, transdermal administrations, nasal inhalants and suppositories with suitable carriers according to methods known in the art.
  • suitable carriers may be sterile water, ethanol, polyols such as glycerol or propylene glycol, or mixtures thereof, preferably PBS (phosphate buffered saline) containing Ringer's solution, triethanol amine or sterilized for injection. Water, isotonic solutions such as 5% dextrose, and the like.
  • transdermal administration When formulated as a transdermal administration, it may be formulated in the form of an ointment, cream, lotion, gel, external solution, pasta, linen, aerosol, and the like.
  • Nasal inhalants can be formulated in the form of aerosol sprays using suitable propellants, such as dichlorofluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide, etc. witepsol), tween 61, polyethylene glycols, cacao butter, laurin paper, polyoxyethylene sorbitan fatty acid esters, polyoxyethylene stearate, sorbitan fatty acid esters and the like.
  • suitable propellants such as dichlorofluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide, etc. witepsol
  • tween 61 polyethylene glycols, cacao butter, laurin paper, polyoxyethylene
  • Preferred dosages of the pharmaceutical compositions of the present invention range from 0.001 mg / kg to 10 g / kg per day, preferably 0.001 mg / kg to 1 g, depending on the condition, body weight, sex, age, severity of the patient and route of administration. It can range from / kg. Administration can be done once a day or divided into several times. Such dosage should not be construed as limiting the scope of the invention in any aspect.
  • composition of the present invention can be understood as a hair cosmetic composition for external application to hair or scalp.
  • the hair cosmetic composition is a solution, suspension, emulsion, paste, gel, cream, lotion, powder, shampoo, soap, surfactant-containing cleansing, oil, powder foundation
  • Product forms include emulsion foundations, wax foundations, and sprays.
  • it may be a softening lotion, a nourishing lotion, a nourishing cream, a massage cream, an essence, an eye cream, a cleansing cream, a cleansing foam, a cleansing water, a pack, a spray or a powder formulation.
  • the cosmetic composition of the present invention may include, in addition to the active ingredient, components conventionally used in the cosmetic composition, for example, conventional adjuvants such as stabilizers, solubilizers, surfactants, vitamins, pigments and pharmaceuticals, and carriers.
  • conventional adjuvants such as stabilizers, solubilizers, surfactants, vitamins, pigments and pharmaceuticals, and carriers.
  • the formulation of the present invention is a paste, cream or gel, animal oils, vegetable oils, waxes, paraffins, starches, trachants, cellulose derivatives, polyethylene glycols, silicones, bentonites, silicas, talc or zinc oxide may be used as carrier components.
  • animal oils, vegetable oils, waxes, paraffins, starches, trachants, cellulose derivatives, polyethylene glycols, silicones, bentonites, silicas, talc or zinc oxide may be used as carrier components.
  • animal oils vegetable oils, waxes, paraffins, starches, trachants, cellulose derivatives, polyethylene glycols, silicones, bentonites, silicas, talc or zinc oxide
  • cellulose derivatives polyethylene glycols
  • silicones bentonites
  • silicas talc or zinc oxide
  • lactose, talc, silica, aluminum hydroxide, calcium silicate or polyamide powder may be used, in particular in the case of a spray, additionally chlorofluorohydrocarbon, propane Propellant such as butane or dimethyl ether.
  • a solvent, solubilizing agent or emulsifying agent is used as the carrier component, specifically water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylglycol oil, glycerol aliphatic ester, polyethylene glycol, fatty acid ester of sorbitan and the like can be used.
  • a liquid diluent such as water, ethanol or propylene glycol
  • a suspending agent such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester, polyoxyethylene sorbitan ester, and microcrystals
  • carrier components such as water, ethanol or propylene glycol
  • ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester, polyoxyethylene sorbitan ester, and microcrystals are used as carrier components.
  • Castle cellulose, aluminum metahydroxy, bentonite, agar and the like can be used.
  • the carrier component is aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, isethionate, imidazolinium derivative, methyltaurate, sarcosinate, fatty acid amide.
  • Ether sulfates, alkylamidobetaines, aliphatic alcohols, fatty acid glycerides, fatty acid diethanolamides, vegetable oils, lanolin derivatives or ethoxylated glycerol fatty acid esters and the like can be used.
  • the hair cosmetic composition of the present invention can be prepared according to the manufacturing method of the cosmetic composition usually carried out in the art, except that the active ingredient is included.
  • according to the present invention can provide a composition for promoting hair growth or inhibiting hair loss using the rosemary extract.
  • composition of the present invention can be commercialized into food, drugs, hair cosmetics and the like.
  • Figure 2 is a result of measuring the cell growth promoting effect when treated with rosemary leaf extract and the like in human keratinocytes (HaCaT cells).
  • Figure 3 is a result of measuring the hair growth promoting effect when the rosemary leaf extract and the like treated to C57BL / 6 mice.
  • Figure 4 is the result of measuring the hair density and thickness when the rosemary leaf extract and the like treated with C57BL / 6 mice.
  • Figure 5 is the result of observing histological changes when treated with rosemary leaf extract, etc. in C57BL / 6 mice.
  • Figure 6 is the result of measuring the expression level of Bcl-2 and Bax when treated with rosemary leaf extract, etc. in C57BL / 6 mice.
  • Samples were (i) essential oil extract of rosemary leaf by hexane extraction (A), (ii) essential oil extract of rosemary leaf by steam distillation (B), (iii) carnoic acid (C), (iv) carnoic acid and hexane Mixture of 1: 5 (carnoic acid: essential oil, weight ratio) of essential oil extract of rosemary leaf by extraction method (D), (v) 1: 2 (carnoic acid: essential oil) of essential oil extract of rosemary leaf by carnosic acid and hexane extraction method , A total of five were used as a sample (E).
  • the essential oil extract of rosemary leaf by hexane extraction was added 5 times weight of hexane to the rosemary leaf powder, extracted by reflux at 70-80 °C for 2 hours, washed with sodium hydroxide, and after obtaining a nucleic acid layer, the hexane layer was dried with anhydrous sodium sulfate. Dehydrated and concentrated under reduced pressure,
  • the essential oil extract of the rosemary leaf by steam distillation is put into the distillation apparatus using rosemary leaf, and steamed at 100 ° C for 1 hour to extract the essential oil.
  • the extracted essential oil is recovered in a cooling tube and dehydrated with anhydrous sodium sulfate. It was prepared by.
  • Carnosic acid was purchased from Sigma (St. Louis, MO, USA) and used.
  • the results of three replicates are shown in FIG. 1 as a percentage compared to the negative control group, which is the sample untreated group.
  • the essential oil extract (A) by hexane extraction method is better than the essential oil extract (B) by steam distillation method of human dermal papilla cells proliferation effect, a mixture of essential oil extract by carnosic acid and hexane extraction method (D and It can be seen that E) is significantly superior to carnoic acid (C) or its essential oil extract (A). In particular, the mixture (E) of 1: 2 was the best.
  • FIG. 2 shows that the essential oil extract (A) by the hexane extraction method is superior to the essential oil extract (B) by the steam distillation method, and the growth effect of human dermal papilla cells is superior to that of the experiment example 1, and the carnosic acid and hexane extraction method.
  • Mixtures of essential oil extracts (D and E), especially mixtures (E) of 1: 2 show the best.
  • mice The back hairs of the 7-week-old C57BL / 6 mice were removed, and five mice with clear back skin were divided into 4 groups. Hair on the back of the mouse was removed by destroying hair follicles using a Wax-Rosin mixture.
  • the first group was applied with 70% ethanol for 13 days (negative control)
  • the second group was applied with Hyundai drug minoxidil for 13 days (positive control)
  • the third group was Experimental Examples 1 and 2 1: 2 mixture of carnosic acid and essential oil extract by hexane extraction (E, R3) was dissolved in DMSO and diluted to 5% (v / v) in 70% ethanol. Was applied.
  • FIG. 3 shows the results of measuring the growth and distribution of hair at 3 days, 7 days, 9 days, 11 days and 13 days after depilation.
  • the results of measuring the density and thickness of the hair by performing close-up photography of the hair on the 3rd and 11th day after depilation are shown in FIG. 4.
  • mice were killed on the 13th day after hair removal, and tissues of the hairy areas of the dorsal skin were collected. Fixed with% formaldehyde solution. After fixation, washed with water, dehydrated, embedded in paraffin, 7 ⁇ m sections were prepared, and stained with Hematoxylin & Eosin (H & E). The results of the histological changes are shown in FIG. 5.
  • mice of each group were lethal on the 13th day after hair removal, and the tissues of the hair growth areas of the dorsal skin were collected and put into liquid nitrogen. After freezing, the protein was separated from the frozen tissue, and the expression levels of Bcl-2 and Bax were confirmed by Western Blot method.

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Abstract

Disclosed is a composition for promoting hair growth or inhibiting hair loss using a Rosmarinus Officinalis (rosemary) leaf extract having the activities of increasing hair density and thickness while promoting proliferation of hair dermal papilla cells and human keratinocytes and promoting hair growth when treated on an epilated mouse.

Description

로즈마리 잎 추출물을 이용한 발모 촉진 또는 탈모 억제용 조성물Composition for promoting hair growth or inhibiting hair loss using rosemary leaf extract
본 발명은 로즈마리(Rosmarinus officinalis) 잎 추출물을 이용한 발모 촉진 또는 탈모 억제용 조성물에 관한 것이다.The present invention is rosemary ( Rosmarinus officinalis ) relates to a composition for promoting hair growth or inhibiting hair loss using a leaf extract.
모발은 모낭(hair follicles)에서 형성되는데 성인의 모낭은 약 5백만 개로 사람의 피부 중 손바닥과 바다박을 제외한 전신에 걸쳐 분포하여 있고, 두부에 1백만 개가 존재하며, 이 중 십만 개가 두피에 존재한다(Bulletin of Food Technology 2009, 22(4):651-663). 두피 모발은 인체 모발의 약 20%를 차지하는데, 땀의 증발과 체온 조절 등의 모발의 생리적 기능과 함께 두부의 완충작용으로 두부를 충격으로부터 보호하는 기능을 가지며, 외모를 중시하는 현대인의 경향에 의하여 개인의 개성과 이미지 연출에 중요한 사회적 기능을 아울러 가진다(Clin Dermatol 2001, 19(2):161-166).Hair is formed from hair follicles, with about 5 million adult hair follicles distributed throughout the body, except for palms and seagrass, with 1 million heads, of which 100,000 are present in the scalp. (Bulletin of Food Technology 2009, 22 (4): 651-663). Scalp hair accounts for about 20% of human hair, and has the function of protecting the head from shocks by buffering the head along with the physiological functions of the hair such as sweat evaporation and temperature control. It also has important social functions for personality and image production (Clin Dermatol 2001, 19 (2): 161-166).
개개의 모발은 모낭에 기원을 두어 성장하는데, 모낭은 매우 복잡한 기관으로 내모근초, 외모근초, 모간, 모기질세포 등으로 구성되며, 모낭을 형성하는 다양한 종류의 세포들 중 모유두세포(Hair dermal papilla cell)는 모발 성장에서 아주 중요한 역할을 한다.Each hair grows with its origin in the hair follicle, which is a very complex organ consisting of internal hair root, external hair root, hair shaft, and matrix cells, and hair dermal cells among various types of hair follicles. papilla cells play an important role in hair growth.
모구(Hair bulb) 내에서 특화된 섬유아세포(Dermal fibroblast)의 일종인 모유두세포(Dermal papilla cell)는 정상 모발 주기뿐만 아니라 남성형 탈모인 안드로겐(Androgen)성 탈모와 같은 증상에서 모유두세포의 증식을 통하여 모발 성장을 조절하는 중요한 역할을 하고 있다. 따라서 모발 탈락에서 모유두세포의 증식에 영향을 미치는 인자는 탈모를 개선시키기 위한 중요한 표적이 된다. Dermal papilla cells, a type of dermal fibroblast specialized in hair bulbs, are not only normal hair cycles but also hair growth through the growth of dermal papilla cells in symptoms such as androgen alopecia (male hair loss). It plays an important role in controlling growth. Thus, factors affecting the proliferation of dermal papilla cells in hair loss are important targets for improving hair loss.
모발은 생장기, 퇴행기 및 휴지기의 성장 주기를 반복하는데, 성인의 경우 두피 모발의 생장기는 약 2 ∼ 6년, 퇴행기는 2~3주, 휴지기는 2~3개월이다. 정상적인 두피에서 90~95%는 생장기에 있고 1% 미만은 퇴화기에 있으며 5~10%는 휴지기에 있는데, 매일 100개 이상의 모발이 휴지기에 접어들고 같은 수의 모낭은 다시 생장기로 들어간다(N Engl J Med 1999, 341(13):964-973). 일반적으로 탈모증이라 함은 이러한 주기 중에서 성장기의 모발 비율이 짧아지고 퇴행기 또는 휴지기가 길어져 비정상적으로 모발이 탈락하는 숫자가 많아지는 것을 일컫는다(Differentiation, 2004, 72(9-10):489; Dev Cell., 2001, 1(1):13; J. Invest. Dermatol., 2005, 125(1):42).Hair repeats the growth, degenerative, and resting phases of growth. In adults, scalp hair grows for about 2 to 6 years, for 2 to 3 weeks, and for 2 to 3 months. In normal scalp, 90-95% are in the growing season, less than 1% are in the degenerative phase, and 5-10% are in the resting phase, with more than 100 hairs entering the resting phase every day and the same number of hair follicles entering the growing season again (N Engl J). Med 1999, 341 (13): 964-973). In general, alopecia refers to an increase in the number of hairs that fall abnormally due to a shorter growth rate and a longer degenerative or resting period during these cycles (Differentiation, 2004, 72 (9-10): 489; Dev Cell. , 2001, 1 (1): 13; J. Invest.Dermatol., 2005, 125 (1): 42).
발모 및 탈모의 정확한 기전은 현재까지 완전하게 밝혀져 있지 않으나, 탈모유발 원인으로 남성호르몬 관여에 의한 모포 기능의 저하, 모포 및 모구부의 신진대사기능의 저하, 두피 긴장에 의한 국소 혈류 장해, 영양 불량, 스트레스, 약물에 의한 부작용, 유전적 요인, 자가 면역, 국소감염, 화학물질, 백혈병, 결핵 등의 질병과 모발 제품의 남용 등 매우 다양하다. 탈모의 가장 전형적인 형태인 남성형 탈모증(androgen alopecia)의 경우 유전적으로 안드로겐(androgen) 호르몬에 의존적으로 발생하는데 포유류의 대표적인 안드로겐 호르몬에는 테스토스테론(testosterone)과 디하이드로테스토스테론(dihydrotestosterone)이 있는데 탈모 증상을 지니는 사람에게서 디하이드로테스토스테론이 생체내에서 높은 수준으로 유지되고 있음이 보고 되어 있고 디하이드로테스토스테론의 과다 생성이 탈모의 주요 원인으로 주목받고 있다(Biol. Pharm. Bull., 2006 29(2):392). 디하이드로테스토스테론은 제2형 5α-리덕타아제에 의해 테스토스테론이라는 남성 호르몬으로부터 생겨나는 또 다른 남성 호르몬이다.The exact mechanism of hair growth and hair loss has not been fully understood to date, but causes of hair loss are caused by male hormone involvement, deterioration of hair follicle function, deterioration of metabolic function of hair follicles and hair follicles, local blood disorder due to scalp tension, malnutrition, Stress, drug side effects, genetic factors, autoimmune, topical infections, chemicals, leukemia, tuberculosis and other diseases, such as abuse of hair products. Androgen alopecia, the most typical form of hair loss, is genetically dependent on androgen hormones.Typical androgen hormones in mammals include testosterone and dihydrotestosterone. It has been reported that dihydrotestosterone is maintained at high levels in vivo and overproduction of dihydrotestosterone has been noted as a major cause of hair loss (Biol. Pharm. Bull., 2006 29 (2): 392). Dihydrotestosterone is another testosterone produced by the testosterone called testosterone by type 2 5α-reductase.
종래의 탈모증 치료방법으로는, 이러한 호르몬설과 관련하여 여성 호르몬을 주재로 한 제제가 있었으나 효과뿐 아니라 피부 염증 발생, 호르몬 투여에 의한 부작용 발생 등의 보고가 있어 현재는 사용이 중단되어 있다. 현재 탈모 치료에 가장 많이 사용되는 약물로는 제2형 5α-리덕타제의 활성을 억제시키는 물질로 미녹시딜(Minoxidil) 제제(미국특허 제4,139,619호 및 제4,596,812호 참조)과 피나스테라이드(finasteride)(미국 특허 제5547957호, 제5760046호 또는 대한민국 특허 제375083호)를 들 수 있다. 미녹시딜 제제는 탈모증이 있는 남성에게서 10%까지 탈모를 감소시키는 효과가 있다고 알려져 있지만 가마 부위 이외 부위의 탈모에 대해서는 그다지 좋은 효과를 발휘하지 못한다는 단점이 있고 또 우울증을 유발할 수 있으며, 피나스테라이드 역시 일부 환자들에 대해서는 성욕 감퇴, 발기 부전 등의 부작용을 나타낸다는 문제점이 있다. 이러한 이유에서 최근 부작용이 적은 천연물에서 발모 촉진 또는 탈모 억제 활성을 갖는 물질을 찾는 노력이 이루어지고 있다.Conventional methods for treating alopecia have been formulated based on female hormones in connection with such hormonal theory, but there have been reports of skin irritation, side effects caused by hormonal administration as well as effects. Currently, the most widely used drugs for the treatment of hair loss include substances that inhibit the activity of Type 2 5α-reductase, see Minoxidil preparations (see US Pat. Nos. 4,139,619 and 4,596,812) and finasteride (US patents). 5555457, 5760046 or Korean Patent No. 375083). Minoxidil preparations are known to reduce hair loss by up to 10% in men with alopecia, but have the disadvantage of not having a very good effect on hair loss outside the kiln and can cause depression. There is a problem in that they have side effects such as decreased libido, erectile dysfunction. For this reason, efforts have recently been made to find substances having a hair growth promoting or hair loss inhibiting activity in natural products with few side effects.
본 발명은 로즈마리 추출물의 발모 촉진 활성 등을 개시한다.The present invention discloses hair growth promoting activity and the like of the rosemary extract.
본 발명의 목적은 로즈마리 추출물을 이용한 발모 촉진 또는 탈모 억제용 조성물을 제공하는 데 있다.It is an object of the present invention to provide a composition for promoting hair growth or inhibiting hair loss using a rosemary extract.
본 발명의 다른 목적이나 구체적인 목적은 이하에서 제시될 것이다.Other and specific objects of the present invention will be presented below.
본 발명자(들)은 아래의 실시예 및 실험예에서 확인되는 바와 같이, (i) 헥산 추출법에 의한 로즈마리 잎의 정유 추출물, (ii) 수증기 증류법에 의한 로즈마리 잎의 정유 추출물, (iii) 카르노스산, (iv) 카르노스산과 헥산 추출법에 의한 로즈마리 잎의 정유 추출물의 1:5(카르노스산:정유, 중량비)의 혼합물, (v) 카르노스산과 헥산 추출법에 의한 로즈마리 잎의 정유 추출물의 1:2의 혼합물(카르노스산:정유, 중량비)을 제조하고 이들의 사람 모유두세포 증식 촉진 활성과 사람 각질형성세포 증식 촉진 활성을 봤을 때, 헥산 추출법에 의한 로즈마리 잎의 정유 추출물이 수증기 증류법에 의한 로즈마리 잎의 정유 추출물보다 활성이 우수하고 또 카르노스산과 헥산 추출법에 의한 로즈마리 잎의 정유 추출물의 혼합물이 카르노스산이나 헥산 추출법에 의한 로즈마리 잎의 정유 추출물보다 활성이 우수하며, 특히 카르노스산과 헥산 추출법에 의한 로즈마리 잎의 정유 추출물의 1:2의 혼합물의 활성이 우수함을 확인하였다. 나아가 본 발명자들은 이러한 세포실험 결과에 있어서 가장 활성이 우수한 카르노스산과 헥산 추출법에 의한 로즈마리 잎의 정유 추출물의 1:2의 혼합물을 사용하여 제모된 마우스를 이용하여 동물실험을 수행하였을 때, 발모를 촉진시킴과 함께 모발의 밀도와 두께를 증가시킴을 확인할 수 있었다.The inventor (s) is confirmed in the following Examples and Experimental Examples, (i) essential oil extract of rosemary leaf by hexane extraction, (ii) essential oil extract of rosemary leaf by steam distillation, (iii) carnosic acid (iv) a mixture of 1: 5 (carnoic acid: essential oil, weight ratio) of essential oil extract of rosemary leaf by carnoic acid and hexane extraction method, (v) 1: 2 of essential oil extract of rosemary leaf by carnoic acid and hexane extraction method When the mixture of (carnoic acid: essential oil, weight ratio) was prepared and their human dermal papilla cell growth promoting activity and human keratinocyte growth promoting activity were observed, essential oil extract of rosemary leaf by hexane extraction was performed on the rosemary leaf by steam distillation. It is more active than essential oil extract and the mixture of essential oil extract of rosemary leaf by carnosic acid and hexane extraction method is rosema by carnosic acid or hexane extraction method. It was confirmed that the activity of the essential oil extract of the leaves of Lee, and especially the activity of 1: 2 mixture of essential oil extract of rosemary leaves by carnosic acid and hexane extraction method. Furthermore, the inventors of the present invention showed that when the animal experiment was conducted using a mouse depilated using a mixture of 1: 2 of the essential oil extract of rosemary leaf by carnosic acid and hexane extraction method, which was the most active in the cell test results, It was confirmed that the hair density and thickness were increased along with the promotion.
상기 카르노스산(Carnosic acid, CAS #3650-09-7)은 로즈마리(Rosmarinus officinalis), 샐비어(Salvia officinalis) 등에서 발견되는 벤젠다이올 아비에탄 디테르펜(benzenediol abietane diterpene)으로서, 강력한 항산화능을 가지고 있으며, 유방암 세포, 난소암 세포, 전립선암 세포 등에서 세포사멸을 유발함으로써 항암 활성을 가진다고 보고되어 있는 화합물이다(International Journal of Molecular Sciences. 2014;15(7)12698-12713). Carnosic acid (CAS # 3650-09-7) is a benzenediol abietane diterpene found in rosemary ( Rosmarinus officinalis ), Salvia ( Salvia officinalis ), and has a strong antioxidant activity. , Anti-cancer activity by inducing apoptosis in breast cancer cells, ovarian cancer cells, prostate cancer cells and the like (International Journal of Molecular Sciences. 2014; 15 (7) 12698-12713).
본 발명은 이러한 실험 결과에 기초하여 제공되는 것으로, 본 발명의 발모 촉진 또는 탈모 억제용 조성물은 (i) 로즈마리 잎 추출물 또는 (ii) 카르노스산과 로즈마리 잎 정유의 혼합물을 유효성분으로 포함함을 특징으로 한다. The present invention is provided on the basis of the experimental results, wherein the composition for promoting hair growth inhibition or hair loss of the present invention comprises (i) a rosemary leaf extract or (ii) a mixture of carnosic acid and rosemary leaf essential oil as an active ingredient. It is done.
본 명세서에서 "유효성분"이란 단독으로 목적하는 활성을 나타내거나 또는 그 자체는 활성이 없는 담체와 함께 활성을 나타낼 수 있는 성분을 의미한다.As used herein, the term "active ingredient" alone refers to a component that can exhibit the desired activity or exhibit itself with a carrier which is inactive.
또 본 명세서에서, 상기 "탈모"란 그 발생의 직·간접적인 원인을 불문하고 당업계에서 탈모로서 분류되는 모든 증상을 포함하는 의미로서 이해되어야 한다. 구체적으로 혈액 순환 불량, 디하이드로테스토스테론의 과다 생성 등에 따른 피지 과잉 분비, 과산화물 또는 세균 등에 의한 두피 기능 저하, 노화, 유전적 요인, 스트레스, 이들의 복합적인 작용 등에 의한 탈모 증상을 모두 포함하는 의미이다.In addition, in this specification, "hair loss" should be understood as including all the symptoms classified as hair loss in the art regardless of the direct and indirect cause of the occurrence. Specifically, it includes all of the symptoms of hair loss due to poor blood circulation, excessive sebum secretion due to excessive production of dihydrotestosterone, deterioration of the scalp function by peroxides or bacteria, aging, genetic factors, stress, and their complex actions. .
또 본 명세서에서, "추출물"이란 추출 대상을 물, 메탄올, 에탄올, 부탄올 등의 탄소수 1 내지 4의 저급 알콜, 아세톤, 메틸렌클로라이드, 에틸렌, 아세톤, 헥산, 에테르, 클로로포름, 에틸아세테이트, 부틸아세테이트, N, N-디메틸포름아미드(DMF), 디메틸설폭사이드(DMSO), 1,3-부틸렌글리콜, 프로필렌글리콜 또는 이들의 혼합 용매를 사용하여 침출하여 얻어진 추출물, 이산화탄소, 펜탄 등 초임계 추출 용매를 사용하여 얻어진 추출물 또는 그 추출물을 분획하여 얻어진 분획물을 의미하며, 추출 방법은 활성물질의 극성, 추출 정도, 보존 정도를 고려하여 냉침, 환류, 가온, 초음파 방사, 초임계 추출 등 임의의 방식을 적용할 수 있다. 분획된 추출물의 경우 상기 조추출물을 특정 용매에 현탁시킨 후 극성이 다른 용매와 혼합·정치시켜 얻은 분획물을 포함하고, 상기 조추출물을 상기 용매들을 극성이 증가 또는 감소하는 순으로 사용하여 순차적으로 분획하여 얻어진 분획물을 포함하며, 나아가 크기, 전하, 소수성, 친화성 등의 성질을 이용한 크로마토그래피에 의하여 얻어진 분획물을 포함한다. 또한 상기 추출물의 의미에는 동결건조, 진공건조, 열풍건조, 분무건조 등의 방식으로 추출 용매가 제거된 농축된 액상 또는 고형상의 추출물이 포함된다. 아래의 실시예를 참조할 때, 바람직하게는 추출용매로서 물, 아세톤 또는 이들의 혼합 용매를 사용하여 침출하여 얻어진 추출물(특히 물과 아세톤의 혼합 용매를 사용하여 얻어진 추출물)을 의미한다. In the present specification, the term "extract" refers to water, methanol, ethanol, butanol lower alcohol having 1 to 4 carbon atoms, acetone, methylene chloride, ethylene, acetone, hexane, ether, chloroform, ethyl acetate, butyl acetate, Supercritical extraction solvents such as extracts obtained by leaching with N, N-dimethylformamide (DMF), dimethylsulfoxide (DMSO), 1,3-butylene glycol, propylene glycol or a mixed solvent thereof, carbon dioxide, pentane, etc. It means the extract obtained by using or fractions obtained by fractionating the extract, the extraction method is applied to any method, such as cold needle, reflux, heating, ultrasonic radiation, supercritical extraction in consideration of the polarity, degree of extraction, the degree of preservation of the active substance can do. The fractionated extract includes a fraction obtained by suspending the crude extract in a specific solvent and mixing and standing with a solvent having a different polarity. The crude extract is sequentially fractionated using the solvents in increasing or decreasing polarity. And fractions obtained by chromatography using properties such as size, charge, hydrophobicity, affinity, and the like. In addition, the meaning of the extract includes a concentrated liquid or solid extract from which the extraction solvent is removed by lyophilization, vacuum drying, hot air drying, spray drying, or the like. When referring to the examples below, it preferably means an extract obtained by leaching using water, acetone or a mixed solvent thereof as an extraction solvent (in particular, an extract obtained using a mixed solvent of water and acetone).
또 본 명세서에서, "로즈마리 잎 정유 추출물"은 추출 대상인 로즈마리 잎의 휘발 성분을 응측시켜 얻어진 오일 상의 추출물을 말한다. 정유 추출물의 제조에는 수증기 증류법, 휘발성 용제(벤젠, 헥산 등)를 이용한 용제 추출법, 초임계추출법 등 당업계에 공지된 방법을 이용할 수 있다. 특히 로즈마리 잎 정유 추출물은 휘발성 용제로서 헥산을 사용하여 얻어진 추출물, 구체적으로 헥산으로 환류 냉각 추출 후 탈수시켜 얻어진 추출물을 의미한다.In addition, in this specification, "rosemary leaf essential oil extract" means the extract of the oil phase obtained by condensing the volatile component of the rosemary leaf to be extracted. For the preparation of the essential oil extract, methods known in the art, such as steam distillation, solvent extraction using volatile solvents (benzene, hexane, etc.), supercritical extraction, can be used. In particular, rosemary leaf essential oil extract refers to an extract obtained by using hexane as a volatile solvent, specifically, an extract obtained by dehydration after reflux cooling with hexane.
본 발명의 조성물은 그 유효성분을 성상, 용도, 제형, 배합 목적 등에 따라 발모 촉진 활성, 탈모 방지 활성 등을 나타낼 수 있는 한 통상적인 유효량은 조성물 전체 중량을 기준으로 할 때 0.0001 중량 % 내지 15 중량 % 범위 내에서 결정될 것이다. 여기서 "유효량"이란 그 적용 대상인 포유동물 바람직하게는 사람에게 의료 전문가 등의 제언에 의한 투여 기간 동안 본 발명의 조성물이 투여될 때, 의도한 발모 촉진 효과나 탈모 억제 효과를 나타낼 수 있는, 본 발명의 조성물에 포함되는 유효성분의 양을 말한다. 이러한 유효량은 당업자의 통상의 능력 범위 내에서 실험적으로 결정될 수 있다.As long as the composition of the present invention can exhibit hair growth promoting activity, anti-hair loss activity, etc. according to properties, uses, formulations, formulation purposes, etc., the effective amount is 0.0001% by weight to 15% by weight based on the total weight of the composition. Will be determined within the% range. The term "effective amount" herein refers to the present invention, which may exhibit an intended hair growth promoting effect or hair loss inhibiting effect when the composition of the present invention is administered to a mammal, preferably a human, during the administration period according to the suggestion of a medical professional or the like. Refers to the amount of the active ingredient included in the composition of. Such effective amounts can be determined experimentally within the range of ordinary skill in the art.
본 발명의 식품 조성물은 어떠한 형태로도 제조될 수 있으며, 예컨대 차, 쥬스, 탄산음료, 이온음료 등의 음료류, 우유, 요구루트 등의 가공 유류, 껌류, 떡, 한과, 빵, 과자, 면 등의 식품류, 정제, 캡슐, 환, 과립, 액상, 분말, 편상, 페이스트상, 시럽, 겔, 젤리, 바 등의 건강기능식품 제제류 등으로 제조될 수 있다. 또 본 발명의 식품 조성물은 법률상·기능상의 구분에 있어서 제조·유통 시점의 시행 법규에 부합하는 한 임의의 제품 구분을 띨 수 있다. 예컨대 건강기능식품에관한법률에 따른 건강기능식품이거나, 식품위생법의 식품공전(식약처 고시, 식품의 기준 및 규격)상 각 식품유형에 따른 과자류, 두류, 다류, 음료류, 특수용도식품 등일 수 있다.The food composition of the present invention may be prepared in any form, for example, beverages such as tea, juice, carbonated beverages, ionic beverages, processed oils such as milk, yogurt, gums, rice cakes, sweets, bread, sweets, noodles, and the like. It can be prepared as a dietary supplement, such as foods, tablets, capsules, pills, granules, liquid, powder, flaky, paste, syrup, gel, jelly, bar. In addition, the food composition of the present invention can distinguish any product as long as it conforms to the enforcement regulations at the time of manufacture and distribution in the legal and functional divisions. For example, it may be a health functional food according to the Act on Health Functional Food, or confectionary, bean, tea, beverage, special purpose food, etc. according to each food type in the Food Code of the Food Sanitation Act (KFDA notification, standards and standards of food). .
본 발명의 식품 조성물에는 그 유효성분 이외에 식품첨가물이 포함될 수 있다. 식품첨가물은 일반적으로 식품을 제조, 가공 또는 보존함에 있어 식품에 첨가되어 혼합되거나 침윤되는 물질로서 이해될 수 있는데, 식품과 함께 매일 그리고 장기간 복용되므로 그 안전성이 보장되어야 한다. 식품위생법에 따른 식품첨가물공전(식약처 고시, 식품첨가물 기준 및 규격)에는 안전성이 보장된 식품첨가물이 화학적 합성품, 천연 첨가물, 혼합 제제류로 구분하여 한정적으로 규정되어 있는데, 이러한 식품첨가물은 기능적 측면에 있어서는 감미제, 풍미제, 보존제, 유화제, 산미료, 점증제 등으로 구분될 수 있다. The food composition of the present invention may include food additives in addition to the active ingredient. Food additives can generally be understood as substances that are added to, mixed with, or infiltrated in the manufacture, processing, or preservation of foods, and because they are taken daily with food and for a long time, their safety must be ensured. The Food Additives Code of the Food Sanitation Act (KFDA Notification, Food Additives Standards and Standards) defines the food additives that are guaranteed to be safe by dividing them into chemical synthetics, natural additives, and mixed preparations. In sweeteners, flavors, preservatives, emulsifiers, acidulants, thickeners and the like can be classified.
감미제는 식품에 적당한 단맛을 부여하기 위하여 사용되는 것으로, 천연의 것이거나 합성된 것을 사용할 수 있다. 바람직하게는 천연 감미제를 사용하는 경우인데, 천연 감미제로서는 옥수수 시럽 고형물, 꿀, 수크로오스, 프룩토오스, 락토오스, 말토오스 등의 당 감미제를 들 수 있다. Sweeteners are used to impart a proper sweetness to foods, and may be natural or synthesized. Preferably, a natural sweetener is used. Examples of the natural sweetener include sugar sweeteners such as corn syrup solids, honey, sucrose, fructose, lactose and maltose.
풍미제는 맛이나 향을 좋게 하기 위하여 사용될 수 있는데, 천연의 것과 합성된 것 모두 사용될 수 있다. 바람직하게는 천연의 것을 사용하는 경우이다. 천연의 것을 사용할 경우에 풍미 이외에 영양 강화의 목적도 병행할 수 있다. 천연 풍미제로서는 사과, 레몬, 감귤, 포도, 딸기, 복숭아 등에서 얻어진 것이거나 녹차잎, 둥굴레, 대잎, 계피, 국화 잎, 자스민 등에서 얻어진 것일 수 있다. 또 인삼(홍삼), 죽순, 알로에 베라, 은행 등에서 얻어진 것을 사용할 수 있다. 천연 풍미제는 액상의 농축액이나 고형상의 추출물일 수 있다. 경우에 따라서 합성 풍미제가 사용될 수 있는데, 합성 풍미제로서는 에스테르, 알콜, 알데하이드, 테르펜 등이 이용될 수 있다. Flavoring agents can be used to enhance the taste or aroma, both natural and synthetic. It is the case of using a natural thing preferably. In addition to flavors, the use of natural ones can be combined with nutritional purposes. The natural flavor may be obtained from apples, lemons, citrus fruits, grapes, strawberries, peaches, and the like, or may be obtained from green tea leaves, round leaves, jujube leaves, cinnamon, chrysanthemum leaves, jasmine and the like. In addition, ginseng (red ginseng), bamboo shoots, aloe vera, ginkgo and the like can be used. Natural flavors can be liquid concentrates or solid extracts. In some cases, a synthetic flavor may be used, and as the synthetic flavor, esters, alcohols, aldehydes, terpenes, and the like may be used.
보존제로서는 소르브산칼슘, 소르브산나트륨, 소르브산칼륨, 벤조산칼슘, 벤조산나트륨, 벤조산칼륨, EDTA(에틸렌디아민테트라아세트산) 등이 사용될 수 있고, 또 유화제로서는 아카시아검, 카르복시메틸셀룰로스, 잔탄검, 펙틴 등이 사용될 수 있으며, 산미료로서는 연산, 말산, 푸마르산, 아디프산, 인산, 글루콘산, 타르타르산, 아스코르브산, 아세트산, 인산 등이 사용될 수 있다. 산미료는 맛을 증진시키는 목적 이외에 미생물의 증식을 억제할 목적으로 식품 조성물이 적정 산도로 되도록 첨가될 수 있다.As the preservative, calcium sorbate, sodium sorbate, potassium sorbate, calcium benzoate, sodium benzoate, potassium benzoate, EDTA (ethylenediaminetetraacetic acid) and the like can be used, and as an emulsifier, acacia gum, carboxymethylcellulose, xanthan gum, pectin Etc. may be used, and acidic acid, malic acid, fumaric acid, adipic acid, phosphoric acid, gluconic acid, tartaric acid, ascorbic acid, acetic acid, phosphoric acid, and the like may be used. The acidulant may be added so that the food composition is at an appropriate acidity for the purpose of inhibiting the growth of microorganisms in addition to the purpose of enhancing the taste.
점증제로서는 현탁화 구현제, 침강제, 겔형성제, 팽화제 등이 사용될 수 있다.As the thickener, suspending implementers, sedimenting agents, gel formers, swelling agents and the like can be used.
본 발명의 식품 조성물은 전술한 바의 식품첨가물 이외에, 기능성과 영양성을 보충, 보강할 목적으로 당업계에 공지되고 식품첨가물로서 안정성이 보장된 생리활성 물질이나 미네랄류를 포함할 수 있다.In addition to the food additives described above, the food composition of the present invention may include a bioactive substance or minerals known in the art for the purpose of supplementing and reinforcing the functionality and nutritional properties and ensuring the stability as a food additive.
그러한 생리활성 물질로서는 녹차 등에 포함된 카테킨류, 비타민 B1, 비타민 C, 비타민 E, 비타민 B12 등의 비타민류, 토코페롤, 디벤조일티아민 등을 들 수 있으며, 미네랄류로서는 구연산칼슘 등의 칼슘 제제, 스테아린산마그네슘 등의 마그네슘 제제, 구연산철 등의 철 제제, 염화크롬, 요오드칼륨, 셀레늄, 게르마늄, 바나듐, 아연 등을 들 수 있다. Examples of such physiologically active substances include catechins, vitamin B1, vitamin C, vitamin E, vitamin B12 and the like, tocopherol, dibenzoylthiamine, and the like contained in green tea. Examples of the minerals include calcium preparations such as calcium citrate and magnesium stearate. Magnesium preparations, iron preparations such as iron citrate, chromium chloride, potassium iodine, selenium, germanium, vanadium, zinc and the like.
본 발명의 식품 조성물에는 전술한 바의 식품첨가물이 제품 유형에 따라 그 첨가 목적을 달성할 수 있는 적량으로 포함될 수 있다.In the food composition of the present invention, the food additive as described above may be included in an amount that can achieve the purpose of addition according to the product type.
본 발명의 식품 조성물에 포함될 수 있는 기타의 식품첨가물과 관련하여서는 식품위생법에 따른 식품공전이나 식품첨가물 공전을 참조할 수 있다.Regarding other food additives that may be included in the food composition of the present invention, reference may be made to food or food additives according to the Food Sanitation Act.
본 발명의 조성물은 다른 구체적인 양태에 있어서는 약제학적 조성물로 파악될 수 있다.The composition of the present invention may be regarded as a pharmaceutical composition in another specific embodiment.
본 발명의 약제학적 조성물은 법률상·기능상의 구분에 있어서 제조·유통 시점의 관련 시행 법규에 부합하는 한 임의의 제품 구분을 띨 수 있다. 예컨대 약사법에 따른 의약외품 또는 의약품이거나 의약품일 경우 일반의약품 또는 전문의약품일 수 있다. The pharmaceutical composition of the present invention can be classified into any product as long as it conforms to the relevant enforcement regulations at the time of manufacture and distribution in the legal and functional divisions. For example, it may be a quasi-drug or drug under the Pharmaceutical Affairs Act or, if it is a drug, it may be a generic or specialized drug.
본 발명의 약제학적 조성물은 그 유효성분 이외에 약제학적으로 허용되는 담체를 포함하여 당업계에 공지된 통상의 방법으로 투여 경로에 따라 경구용 제형 또는 비경구용 제형으로 제조될 수 있다. 여기서 "약제학적으로 허용되는" 의미는 유효성분의 활성을 억제하지 않으면서 적용(처방) 대상이 적응 가능한 이상의 독성을 지니지 않는다는 의미이다.The pharmaceutical compositions of the present invention may be prepared in oral or parenteral formulations according to the route of administration by conventional methods known in the art, including pharmaceutically acceptable carriers in addition to the active ingredients. "Pharmaceutically acceptable" here means that the subject of application (prescription) is not toxic as far as adaptable without inhibiting the activity of the active ingredient.
본 발명의 약제학적 조성물이 경구용 제형으로 제조될 경우, 적합한 담체와 함께 당업계에 공지된 방법에 따라 분말, 과립, 정제, 환제, 당의정제, 캡슐제, 액제, 겔제, 시럽제, 현탁액, 웨이퍼 등의 제형으로 제조될 수 있다. 이때 약제학적으로 허용되는 적합한 담체의 예로서는 락토스, 글루코스, 슈크로스, 덱스트로스, 솔비톨, 만니톨, 자일리톨 등의 당류, 옥수수 전분, 감자 전분, 밀 전분 등의 전분류, 셀룰로오스, 메틸셀룰로오스, 에틸셀룰로오스, 나트륨 카르복시메틸셀룰로오스, 하이드록시프로필메틸셀룰로오스 등의 셀룰로오스류, 폴리비닐 피롤리돈, 물, 메틸히드록시벤조에이트, 프로필히드록시벤조에이트, 마그네슘 스테아레이트, 광물유, 맥아, 젤라틴, 탈크, 폴리올, 식물성유 등을 들 수 있다. 제제화활 경우 필요에 따라 충진제, 증량제, 결합제, 습윤제, 붕해제, 계면활성제 등의 희석제 및/또는 부형제를 포함하여 제제화할 수 있다.When the pharmaceutical composition of the present invention is prepared in an oral dosage form, powders, granules, tablets, pills, dragees, capsules, solutions, gels, syrups, suspensions, wafers according to methods known in the art with suitable carriers It may be prepared in a formulation such as. Examples of suitable pharmaceutically acceptable carriers include lactose, glucose, sucrose, dextrose, sugars such as sorbitol, mannitol, xylitol, starch such as corn starch, potato starch, wheat starch, cellulose, methylcellulose, ethylcellulose, Celluloses such as sodium carboxymethylcellulose, hydroxypropylmethylcellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, magnesium stearate, mineral oil, malt, gelatin, talc, polyol, vegetable Yu etc. can be mentioned. If formulated, it may be formulated to include diluents and / or excipients, such as fillers, extenders, binders, wetting agents, disintegrants, surfactants, if necessary.
본 발명의 약제학적 조성물이 비경구용 제형으로 제조될 경우, 적합한 담체와 함께 당업계에 공지된 방법에 따라 주사제, 경피 투여제, 비강 흡입제 및 좌제의 형태로 제제화될 수 있다. 주사제로 제제화할 경우 적합한 담체로서는 멸균수, 에탄올, 글리세롤이나 프로필렌 글리콜 등의 폴리올 또는 이들의 혼합물을 사용할 수 있으며, 바람직하게는 링거 용액, 트리에탄올 아민이 함유된 PBS(phosphate buffered saline)나 주사용 멸균수, 5% 덱스트로스 같은 등장 용액 등을 사용할 수 있다. 경피 투여제로 제제화할 경우 연고제, 크림제, 로션제, 겔제, 외용액제, 파스타제, 리니멘트제, 에어롤제 등의 형태로 제제화할 수 있다. 비강 흡입제의 경우 디클로로플루오로메탄, 트리클로로플루오로메탄, 디클로로테트라플루오로에탄, 이산화탄소 등의 적합한 추진제를 사용하여 에어로졸 스프레이 형태로 제제화할 수 있으며, 좌제로 제제화할 경우 그 기제로는 위텝솔(witepsol), 트윈(tween) 61, 폴리에틸렌글리콜류, 카카오지, 라우린지, 폴리옥시에틸렌 소르비탄 지방산 에스테르류, 폴리옥시에틸렌 스테아레이트류, 소르비탄 지방산 에스테르류 등을 사용할 수 있다.When the pharmaceutical compositions of the present invention are prepared in parenteral formulations, they may be formulated in the form of injections, transdermal administrations, nasal inhalants and suppositories with suitable carriers according to methods known in the art. When formulated as an injectable, suitable carriers may be sterile water, ethanol, polyols such as glycerol or propylene glycol, or mixtures thereof, preferably PBS (phosphate buffered saline) containing Ringer's solution, triethanol amine or sterilized for injection. Water, isotonic solutions such as 5% dextrose, and the like. When formulated as a transdermal administration, it may be formulated in the form of an ointment, cream, lotion, gel, external solution, pasta, linen, aerosol, and the like. Nasal inhalants can be formulated in the form of aerosol sprays using suitable propellants, such as dichlorofluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide, etc. witepsol), tween 61, polyethylene glycols, cacao butter, laurin paper, polyoxyethylene sorbitan fatty acid esters, polyoxyethylene stearate, sorbitan fatty acid esters and the like.
약제학적 조성물의 제제화와 관련하여서는 당업계에 공지되어 있으며, 구체적으로 문헌[Remington's Pharmaceutical Sciences(19th ed., 1995)] 등을 참조할 수 있다. 상기 문헌은 본 명세서의 일부로서 간주 된다.Regarding the formulation of pharmaceutical compositions, it is known in the art and specific reference may be made to Remington's Pharmaceutical Sciences (19th ed., 1995) and the like. The document is considered part of this specification.
본 발명의 약제학적 조성물의 바람직한 투여량은 환자의 상태, 체중, 성별, 연령, 환자의 중증도, 투여 경로에 따라 1일 0.001mg/kg ~ 10g/kg 범위, 바람직하게는 0.001mg/kg ~ 1g/kg 범위일 수 있다. 투여는 1일 1회 또는 수회로 나누어 이루어질 수 있다. 이러한 투여량은 어떠한 측면으로든 본 발명의 범위를 제한하는 것으로 해석되어서는 아니 된다. Preferred dosages of the pharmaceutical compositions of the present invention range from 0.001 mg / kg to 10 g / kg per day, preferably 0.001 mg / kg to 1 g, depending on the condition, body weight, sex, age, severity of the patient and route of administration. It can range from / kg. Administration can be done once a day or divided into several times. Such dosage should not be construed as limiting the scope of the invention in any aspect.
본 발명의 조성물은 또 다른 구체적인 양태에 있어서, 모발 또는 두피에 외용적으로 적용하기 위한 모발 화장료 조성물로 파악할 수 있다.In another specific embodiment, the composition of the present invention can be understood as a hair cosmetic composition for external application to hair or scalp.
본 발명의 조성물이 모발 화장료 조성물로 파악될 경우에도 그 모발 화장료 조성물은 용액, 현탁액, 유탁액, 페이스트, 젤, 크림, 로션, 파우더, 샴푸, 비누, 계면활성제-함유 클렌징, 오일, 분말 파운데이션, 유탁액 파운데이션, 왁스 파운데이션, 스프레이 등의 제품 형태를 띨 수 있다. 구체적인 제품 형태에 있어서는 유연 화장수, 영양 화장수, 영양 크림, 마사지 크림, 에센스, 아이 크림, 클렌징 크림, 클렌징 포옴, 클렌징 워터, 팩, 스프레이 또는 파우더의 제형 등일 수 있다.Even when the composition of the present invention is regarded as a hair cosmetic composition, the hair cosmetic composition is a solution, suspension, emulsion, paste, gel, cream, lotion, powder, shampoo, soap, surfactant-containing cleansing, oil, powder foundation, Product forms include emulsion foundations, wax foundations, and sprays. In a specific product form, it may be a softening lotion, a nourishing lotion, a nourishing cream, a massage cream, an essence, an eye cream, a cleansing cream, a cleansing foam, a cleansing water, a pack, a spray or a powder formulation.
본 발명의 화장료 조성물은 그 유효성분 이외에 화장료 조성물에 통상적으로 이용되는 성분들, 예컨대, 안정화제, 용해화제, 계면활성제, 비타민, 색소 및 항료와 같은 통상적인 보조제, 및 담체를 포함할 수 있다. The cosmetic composition of the present invention may include, in addition to the active ingredient, components conventionally used in the cosmetic composition, for example, conventional adjuvants such as stabilizers, solubilizers, surfactants, vitamins, pigments and pharmaceuticals, and carriers.
본 발명의 제형이 페이스트, 크림 또는 젤인 경우에는 담체 성분으로서 동물성유, 식물성유, 왁스, 파라핀, 전분, 트라칸트, 셀룰로오스 유도체, 폴리에틸렌 글리콜, 실리콘, 벤토나이트, 실리카, 탈크 또는 산화아연 등이 이용될 수 있다.When the formulation of the present invention is a paste, cream or gel, animal oils, vegetable oils, waxes, paraffins, starches, trachants, cellulose derivatives, polyethylene glycols, silicones, bentonites, silicas, talc or zinc oxide may be used as carrier components. Can be.
본 발명의 제형이 파우더 또는 스프레이인 경우에는 담체 성분으로서 락토스, 탈크, 실리카, 알루미늄 히드록시드, 칼슘 실리케이트 또는 폴리아미드 파우더가 이용될 수 있고, 특히 스프레이인 경우에는 추가적으로 클로로플루오로히드로카본, 프로판/부탄 또는 디메틸 에테르와 같은 추진체를 포함할 수 있다.When the formulation of the present invention is a powder or a spray, lactose, talc, silica, aluminum hydroxide, calcium silicate or polyamide powder may be used, in particular in the case of a spray, additionally chlorofluorohydrocarbon, propane Propellant such as butane or dimethyl ether.
본 발명의 제형이 용액 또는 유탁액인 경우에는 담체 성분으로서 용매, 용해화제 또는 유탁화제가 이용되는데, 구체적으로 물, 에탄올, 이소프로판올, 에틸 카보네이트, 에틸 아세테이트, 벤질 알코올, 벤질 벤조에이트, 프로필렌 글리콜, 1,3-부틸글리콜 오일, 글리세롤 지방족 에스테르, 폴리에틸렌 글리콜, 소르비탄의 지방산 에스테르 등이 이용될 수 있다. 본 발명의 제형이 현탁액인 경우에는 담체 성분으로서 물, 에탄올 또는 프로필렌 글리콜과 같은 액상의 희석제, 에톡실화 이소스테아릴 알코올, 폴리옥시에틸렌 소르비톨 에스테르, 폴리옥시에틸렌 소르비탄 에스테르와 같은 현탁제, 미소결정성 셀룰로오스, 알루미늄 메타히드록시드, 벤토나이트, 아가 등이 이용될 수 있다.When the formulation of the present invention is a solution or emulsion, a solvent, solubilizing agent or emulsifying agent is used as the carrier component, specifically water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylglycol oil, glycerol aliphatic ester, polyethylene glycol, fatty acid ester of sorbitan and the like can be used. When the formulation of the present invention is a suspension, a liquid diluent such as water, ethanol or propylene glycol, a suspending agent such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester, polyoxyethylene sorbitan ester, and microcrystals are used as carrier components. Castle cellulose, aluminum metahydroxy, bentonite, agar and the like can be used.
본 발명의 제형이 계면-활성제 함유 클렌징인 경우에는 담체 성분으로서 지방족 알코올 설페이트, 지방족 알코올 에테르 설페이트, 설포숙신산 모노에스테르, 이세티오네이트, 이미다졸리늄 유도체, 메틸타우레이트, 사르코시네이트, 지방산 아미드 에테르 설페이트, 알킬아미도베타인, 지방족 알코올, 지방산 글리세리드, 지방산 디에탄올아미드, 식물성 유, 라놀린 유도체 또는 에톡실화 글리세롤 지방산 에스테르 등이 이용될 수 있다.When the formulation of the present invention is a surfactant-containing cleansing, the carrier component is aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, isethionate, imidazolinium derivative, methyltaurate, sarcosinate, fatty acid amide. Ether sulfates, alkylamidobetaines, aliphatic alcohols, fatty acid glycerides, fatty acid diethanolamides, vegetable oils, lanolin derivatives or ethoxylated glycerol fatty acid esters and the like can be used.
본 발명의 모발 화장료 조성물은 그 유효성분을 포함하는 것을 제외하고는 당업계에 통상적으로 행하여지는 화장료 조성물의 제조방법에 따라 제조할 수 있다.The hair cosmetic composition of the present invention can be prepared according to the manufacturing method of the cosmetic composition usually carried out in the art, except that the active ingredient is included.
전술한 바와 같이, 본 발명에 따르면 로즈마리 추출물을 이용한 발모 촉진 또는 탈모 억제용 조성물을 제공할 수 있다.As described above, according to the present invention can provide a composition for promoting hair growth or inhibiting hair loss using the rosemary extract.
본 발명의 조성물은 식품, 약품, 모발 화장품 등으로 제품화될 수 있다.The composition of the present invention can be commercialized into food, drugs, hair cosmetics and the like.
도 1은 로즈마리 잎 추출물 등을 사람의 모유두 세포(HHDPC)에 처리하였을 때 세포 성장 촉진 효과를 측정한 결과이다.1 is a result of measuring cell growth promoting effect when the rosemary leaf extract and the like treated to human dermal papilla cells (HHDPC).
도 2는 로즈마리 잎 추출물 등을 사람의 각질형성세포(HaCaT cell)에 처리하였을 때 세포 성장 촉진 효과를 측정한 결과이다. Figure 2 is a result of measuring the cell growth promoting effect when treated with rosemary leaf extract and the like in human keratinocytes (HaCaT cells).
도 3은 로즈마리 잎 추출물 등을 C57BL/6 마우스에 처리하였을 때 모발 생장 촉진 효과를 측정한 결과이다.Figure 3 is a result of measuring the hair growth promoting effect when the rosemary leaf extract and the like treated to C57BL / 6 mice.
도 4는 로즈마리 잎 추출물 등을 C57BL/6 마우스에 처리하였을 때 모발 밀도와 두께를 측정한 결과이다. Figure 4 is the result of measuring the hair density and thickness when the rosemary leaf extract and the like treated with C57BL / 6 mice.
도 5는 로즈마리 잎 추출물 등을 C57BL/6 마우스에 처리하였을 때 조직학적 변화를 관찰한 결과이다.Figure 5 is the result of observing histological changes when treated with rosemary leaf extract, etc. in C57BL / 6 mice.
도 6은 로즈마리 잎 추출물 등을 C57BL/6 마우스에 처리하였을 때 Bcl-2 및 Bax의 발현량을 측정한 결과이다.Figure 6 is the result of measuring the expression level of Bcl-2 and Bax when treated with rosemary leaf extract, etc. in C57BL / 6 mice.
이하 본 발명을 실시예 및 실험예를 참조하여 설명한다. 그러나 본 발명의 범위가 이러한 실시예 및 실험예에 한정되는 것은 아니다.Hereinafter, the present invention will be described with reference to Examples and Experimental Examples. However, the scope of the present invention is not limited to these examples and experimental examples.
<실시예> <Example> 시료의 준비Sample Preparation
시료는 (i) 헥산 추출법에 의한 로즈마리 잎의 정유 추출물(A), (ii) 수증기 증류법에 의한 로즈마리 잎의 정유 추출물(B), (iii) 카르노스산(C), (iv) 카르노스산과 헥산 추출법에 의한 로즈마리 잎의 정유 추출물의 1:5(카르노스산:정유, 중량비)의 혼합물(D), (v) 카르노스산과 헥산 추출법에 의한 로즈마리 잎의 정유 추출물의 1:2(카르노스산:정유, 중량비)의 혼합물(E)로 총 다섯 가지를 시료로 사용하였다.Samples were (i) essential oil extract of rosemary leaf by hexane extraction (A), (ii) essential oil extract of rosemary leaf by steam distillation (B), (iii) carnoic acid (C), (iv) carnoic acid and hexane Mixture of 1: 5 (carnoic acid: essential oil, weight ratio) of essential oil extract of rosemary leaf by extraction method (D), (v) 1: 2 (carnoic acid: essential oil) of essential oil extract of rosemary leaf by carnosic acid and hexane extraction method , A total of five were used as a sample (E).
헥산 추출법에 의한 로즈마리 잎의 정유 추출물은 로즈마리 잎 분말에 5배 중량의 헥산을 가하고 2시간 동안 70~80℃ 환류 냉각 추출한 후, 수산화나트륨으로 세척하고 핵산층을 얻은 후 그 헥산층을 무수황산나트륨으로 탈수시키고 감압농축하여 준비하였고,The essential oil extract of rosemary leaf by hexane extraction was added 5 times weight of hexane to the rosemary leaf powder, extracted by reflux at 70-80 ℃ for 2 hours, washed with sodium hydroxide, and after obtaining a nucleic acid layer, the hexane layer was dried with anhydrous sodium sulfate. Dehydrated and concentrated under reduced pressure,
수증기 증류법에 의한 로즈마리 잎의 정유 추출물은 증류장치를 이용하여 증류 장치에 로즈마리 잎을 넣고 100℃로 1시간 동안 수증기를 발생시켜 정유를 추출하고 추출된 정유를 냉각관에서 회수한 후 무수황산나트륨으로 탈수시켜 준비하였다.The essential oil extract of the rosemary leaf by steam distillation is put into the distillation apparatus using rosemary leaf, and steamed at 100 ° C for 1 hour to extract the essential oil. The extracted essential oil is recovered in a cooling tube and dehydrated with anhydrous sodium sulfate. It was prepared by.
카르노스산은 Sigma(St. Louis, MO, USA)로부터 구입하여 사용하였다.Carnosic acid was purchased from Sigma (St. Louis, MO, USA) and used.
<실험예> Experimental Example 발모 촉진 또는 탈모 억제 효과 실험Experiment to promote hair growth or inhibit hair loss
<실험예 1> Experimental Example 1 사람 모유두세포 증식 효과 평가Evaluation of human dermal papilla proliferation effect
상기 실시예의 다섯 가지 시료가 사람 모유두세포(human hair dermal papilla cell)의 세포 성장에 미치는 효과를 아래와 같이 평가하였다.The effects of the five samples of the above example on cell growth of human hair dermal papilla cells were evaluated as follows.
10% FBS(fetal bovine serum, Cambrex)가 포함된 전용 배지인 MSC(Mesenchymal Stem Cell) 배지에 사람 모유두세포(Sciencell사, cat. no. 2400)를 0.5 × 104 cells/이 되도록 하여 96 웰 플레이트에 접종하여 24 시간 동안, 37℃, 5% CO2 습윤 조건 하에서 배양하였다. 상기 배양액에 상기 실시예의 시료를 0.1, 1 및 10 ㎍/ml의 농도로 처리하여 72 시간 동안 배양한 후, 5 mg/ml의 MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide) 시약 20㎕를 상기 배양액에 넣고 37℃, 5% CO2 습윤 조건하에서 4시간 동안 배양하였다. 양성대조군으로서 미녹시딜(Minoxidil, 현대약품) 200 nM을 처리하여 상기와 동일하게 배양하였다. 배양 시, 살아있는 세포에는 보라색의 크리스탈이 형성된다. 4시간 배양 후 배지를 제거하고 DMSO 100㎕를 넣어 10분 동안 쉐이킹 인큐베이터(shaking incubator)에서 크리스탈을 녹인 후 540 nm에서 흡광도를 측정하여 세포 성장 정도를 측정하였다. 96 well plate with 0.5 × 10 4 cells / of human dermal papilla cells (Sciencell, Cat. No. 2400) in MSC (Mesenchymal Stem Cell) medium containing 10% FBS (fetal bovine serum, Cambrex) Incubated for 24 hours, incubated under 37 ° C., 5% CO 2 wet conditions. After treating the sample of Example in concentrations of 0.1, 1 and 10 μg / ml in the culture solution and incubating for 72 hours, 5 mg / ml of MTT (3- (4,5-dimethylthiazol-2-yl) -2 20 [mu] l, 5-diphenyl tetrazolium bromide) reagent was added to the culture and incubated for 4 hours at 37 ° C and 5% CO 2 wet conditions. As a positive control group was treated with 200 nM of minoxidil (Minoxidil, Hyundai drug) in the same manner as above. In culture, purple crystals form on living cells. After incubation for 4 hours, the medium was removed, 100 μl of DMSO was added, the crystals were dissolved in a shaking incubator for 10 minutes, and the absorbance was measured at 540 nm to measure cell growth.
3회 반복실험의 결과를 시료 무처리군인 음성대조군과 비교하여 백분율로 도 1에 나타내었다. 도 1을 참조하여 보면 헥산 추출법에 의한 정유 추출물(A)이 수증기 증류법에 의한 정유 추출물(B)보다 사람 모유두세포의 증식 효과가 우수하고, 카르노스산과 헥산 추출법에 의한 정유 추출물의 혼합물(D 및 E)이 카르노스산(C)이나 그 정유 추출물(A)보다 뚜렷하게 우수함을 알 수 있다. 특히 1:2의 혼합물(E)가 가장 우수하였다. The results of three replicates are shown in FIG. 1 as a percentage compared to the negative control group, which is the sample untreated group. Referring to Figure 1, the essential oil extract (A) by hexane extraction method is better than the essential oil extract (B) by steam distillation method of human dermal papilla cells proliferation effect, a mixture of essential oil extract by carnosic acid and hexane extraction method (D and It can be seen that E) is significantly superior to carnoic acid (C) or its essential oil extract (A). In particular, the mixture (E) of 1: 2 was the best.
<실험예 2> Experimental Example 2 사람 각질형성세포(HaCaT cell) 증식 효과 평가Evaluation of the proliferation effect of human HaCaT cells
상기 실시예의 다섯 가지 시료가 사람 각질형성세포의 세포 성장에 미치는 효과를, 각질형성세포(HaCaT cell)(Dr. N. E. Fusenig (Heidelberg, Germany)을 사용하여, 상기 실험예 1과 동일한 방법으로 평가하였다. 배지는 10% FBS (fetal bovine serum, Cambrex)가 포함된 RPMI 배지를 사용하였다. The effect of the five samples of the above example on the cell growth of human keratinocytes was evaluated in the same manner as in Experiment 1 using HaCaT cells (Dr. NE Fusenig (Heidelberg, Germany)). RPMI medium containing 10% FBS (fetal bovine serum, Cambrex) was used.
3회 반복실험의 결과를 시료 무처리군인 음성대조군과 비교하여 백분율로 도 2에 나타내었다. 도 2의 결과는 상기 실험예 1의 결과와 유사하게 헥산 추출법에 의한 정유 추출물(A)이 수증기 증류법에 의한 정유 추출물(B)보다 사람 모유두세포의 증식 효과가 우수하고, 카르노스산과 헥산 추출법에 의한 정유 추출물의 혼합물(D 및 E), 특히 1:2의 혼합물(E)이 가장 우수함을 보여준다.The results of three replicates are shown in FIG. 2 as a percentage compared to the negative control group, which is the sample untreated group. 2 shows that the essential oil extract (A) by the hexane extraction method is superior to the essential oil extract (B) by the steam distillation method, and the growth effect of human dermal papilla cells is superior to that of the experiment example 1, and the carnosic acid and hexane extraction method. Mixtures of essential oil extracts (D and E), especially mixtures (E) of 1: 2, show the best.
<실험예 3> Experimental Example 3 C57BL/6 마우스에서의 모발 성장 효과 평가Evaluation of Hair Growth Effect in C57BL / 6 Mice
7주령의 C57BL/6 마우스의 등 부위 털을 제거하여, 등 부위 피부가 깨끗한 마우스를 5 마리씩 4 군으로 나누었다. 마우스 등 부위의 털은 Wax-Rosin 혼합물을 이용하여 모낭을 파괴하여 제거하였다. 제 1 군은 70% 에탄올을 13일 동안 도포하였고(음성 대조군), 제 2 군은 (주)현대약품 미녹시실을 13일 동안 도포 하였으며(양성 대조군), 제 3 군은 상기 실험예 1 및 2에서 가장 활성이 우수한 것으로 나타난 카르노스산과 헥산 추출법에 의한 정유 추출물의 1:2의 혼합물(E, R3)을 DMSO에 녹인 후 70% 에탄올에 5%(v/v)의 용량으로 희석하여 13일 동안 도포하였다. The back hairs of the 7-week-old C57BL / 6 mice were removed, and five mice with clear back skin were divided into 4 groups. Hair on the back of the mouse was removed by destroying hair follicles using a Wax-Rosin mixture. The first group was applied with 70% ethanol for 13 days (negative control), the second group was applied with Hyundai drug minoxidil for 13 days (positive control), the third group was Experimental Examples 1 and 2 1: 2 mixture of carnosic acid and essential oil extract by hexane extraction (E, R3) was dissolved in DMSO and diluted to 5% (v / v) in 70% ethanol. Was applied.
제모 후 3일, 7일, 9일, 11일 및 13일에 모발의 성장과 분포 정도를 측정한 결과를 도 3에 나타내었다. 또 제모 후 3일, 11일에 모발의 근접촬영을 진행하여 모발의 밀도와 두께를 측정한 결과를 도 4에 나타내었다.3 shows the results of measuring the growth and distribution of hair at 3 days, 7 days, 9 days, 11 days and 13 days after depilation. In addition, the results of measuring the density and thickness of the hair by performing close-up photography of the hair on the 3rd and 11th day after depilation are shown in FIG. 4.
또한, 피부 내 모낭(hair follicle)의 변화, 피부 두께 및 조직의 변화를 관찰하기 위하여, 제모 후 13일째에 각 군의 마우스를 치사시킨 후, 등 쪽 피부의 발모된 부위의 조직을 채취하여 4% 포름알데히드 용액으로 고정하였다. 고정 후, 수세, 탈수과정을 거쳐 파라핀으로 포매하고, 7㎛의 절편을 제작하여 Hematoxylin & Eosin (H&E) 염색을 시행하였으며, 조직학적 변화를 관찰한 결과를 도 5에 나타내었다.In addition, in order to observe changes in hair follicle, skin thickness, and tissue in the skin, each group of mice were killed on the 13th day after hair removal, and tissues of the hairy areas of the dorsal skin were collected. Fixed with% formaldehyde solution. After fixation, washed with water, dehydrated, embedded in paraffin, 7 μm sections were prepared, and stained with Hematoxylin & Eosin (H & E). The results of the histological changes are shown in FIG. 5.
도 3, 4 및 도 5로부터, 실시예의 시료를 처리한 경우 대조군에 비하여 마우스의 등에서 피부색이 단시간에 검게 변하며 털이 빨리 자라고, 모발의 밀도와 두께도 대조군에 비해 실시예의 시료를 처리한 군에서 높게 나타났으며, 실제 피부조직에서도 모낭의 수도 실시예의 시료를 처리한 군에서 높게 나타났음을 확인할 수 있다. 3, 4 and 5, when the sample of the example was treated, the skin color of the mouse turns black in a short time and the hair grows faster than the control group, and the density and thickness of the hair are also higher in the group treated with the sample of the example than the control group. It can be seen that the number of hair follicles in the actual skin tissue was also high in the sample treated group.
<실험예 4> Experimental Example 4 C57BL/6 마우스 조직에서의 모발 성장 관련 인자의 발현량 평가Expression Evaluation of Hair Growth Related Factors in C57BL / 6 Mouse Tissues
마우스 조직에서 모발 성장 관련인자인 Bcl-2 및 Bax의 발현량을 확인하기 위하여 제모 후 13일째에 각 군의 마우스를 치사시킨 후, 등 쪽 피부의 발모된 부위의 조직을 채취하여 액체질소에 넣어 냉동시킨 후, 냉동시킨 조직으로부터 단백질을 분리하여 Western Blot 방법으로 Bcl-2 및 Bax의 발현량을 확인하였다.To confirm the expression level of hair growth-related factors Bcl-2 and Bax in mouse tissues, the mice of each group were lethal on the 13th day after hair removal, and the tissues of the hair growth areas of the dorsal skin were collected and put into liquid nitrogen. After freezing, the protein was separated from the frozen tissue, and the expression levels of Bcl-2 and Bax were confirmed by Western Blot method.
결과를 도 6에 나타내었는데, 도 6을 참조하여 보면 실시예의 시료를 처리한 경우 대조군에 비하여 마우스의 조직에서 모발 성장을 촉진하는 인자인 Bcl-2의 발현량은 증가하고, 모발 성장을 억제하는 인자인 Bax의 발현량은 줄여드는 것을 확인할 수 있다.The results are shown in FIG. 6. Referring to FIG. 6, when the sample of the example was treated, the expression level of Bcl-2, which promotes hair growth in the tissues of the mouse, was increased compared to the control group, and the hair growth was suppressed. It can be seen that the expression level of the factor Bax is reduced.

Claims (6)

  1. (i) 로즈마리 잎 추출물 또는 (ii) 카르노스산과 로즈마리 잎 정유 추출물의 혼합물을 유효성분으로 포함하는 발모 촉진 또는 탈모 억제용 조성물.A composition for promoting hair growth or inhibiting hair loss comprising (i) a rosemary leaf extract or (ii) a mixture of carnosic acid and a rosemary leaf essential oil extract as an active ingredient.
  2. 제1항에 있어서,The method of claim 1,
    상기 로즈마리 잎 정유 추출물은 로즈마리 잎을 헥산으로 환류 냉각 추출 후 탈수시켜 얻어진 추출물인 것을 특징으로 하는 발모 촉진 또는 탈모 억제용 조성물.The rosemary leaf essential oil extract is a composition for promoting hair growth or inhibiting hair loss, characterized in that the extract obtained by dehydrating the rosemary leaf after reflux extraction with hexane.
  3. 제2항에 있어서,The method of claim 2,
    상기 카르노스산과 로즈마리 잎 정유 추출물의 혼합물은 중량비로 1:5 내지 1:2(카르노스산:정유 추출물)인 것을 특징으로 하는 발모 촉진 또는 탈모 억제용 조성물.The mixture of carnosic acid and rosemary leaf essential oil extract is 1: 5 to 1: 2 (carnosic acid: essential oil extract) in weight ratio, the composition for promoting hair growth or inhibiting hair loss.
  4. 제1항 내지 제3항 중 어느 한 항에 있어서,The method according to any one of claims 1 to 3,
    상기 조성물은 식품 조성물인 것을 특징으로 하는 발모 촉진 또는 탈모 억제용 조성물.The composition for promoting hair growth or inhibiting hair loss, characterized in that the food composition.
  5. 제1항 내지 제3항 중 어느 한 항에 있어서,The method according to any one of claims 1 to 3,
    상기 조성물은 약제학적 조성물인 것을 특징으로 하는 발모 촉진 또는 탈모 억제용 조성물.The composition for promoting hair growth or inhibiting hair loss, characterized in that the pharmaceutical composition.
  6. 제1항 내지 제3항 중 어느 한 항에 있어서,The method according to any one of claims 1 to 3,
    상기 조성물은 모발 화장료 조성물인 것을 특징으로 하는 발모 촉진 또는 탈모 억제용 조성물.The composition for promoting hair growth or inhibiting hair loss, characterized in that the hair cosmetic composition.
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CN112190510A (en) * 2020-09-22 2021-01-08 广东嘉丹婷日用品有限公司 Anti-hair loss composition and preparation method thereof
CN112274463A (en) * 2020-09-22 2021-01-29 广东嘉丹婷日用品有限公司 Anti-hair loss shampoo and preparation method thereof
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KR101724510B1 (en) * 2016-06-28 2017-04-07 최한준 Composition for Hair Growth Stimulation or Hair Loss Prevention Using a Rosemary Leaf Extract
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