WO2014185610A1 - 밀기울 추출물을 유효성분으로 포함하는 탈모 방지 또는 육모 촉진 조성물 - Google Patents
밀기울 추출물을 유효성분으로 포함하는 탈모 방지 또는 육모 촉진 조성물 Download PDFInfo
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- WO2014185610A1 WO2014185610A1 PCT/KR2013/010700 KR2013010700W WO2014185610A1 WO 2014185610 A1 WO2014185610 A1 WO 2014185610A1 KR 2013010700 W KR2013010700 W KR 2013010700W WO 2014185610 A1 WO2014185610 A1 WO 2014185610A1
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- WO
- WIPO (PCT)
- Prior art keywords
- hair
- bran
- extract
- composition
- growth
- Prior art date
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- 235000010487 tragacanth Nutrition 0.000 description 1
- 239000000196 tragacanth Substances 0.000 description 1
- 229940116362 tragacanth Drugs 0.000 description 1
- WEAPVABOECTMGR-UHFFFAOYSA-N triethyl 2-acetyloxypropane-1,2,3-tricarboxylate Chemical compound CCOC(=O)CC(C(=O)OCC)(OC(C)=O)CC(=O)OCC WEAPVABOECTMGR-UHFFFAOYSA-N 0.000 description 1
- APVVRLGIFCYZHJ-UHFFFAOYSA-N trioctyl 2-hydroxypropane-1,2,3-tricarboxylate Chemical compound CCCCCCCCOC(=O)CC(O)(C(=O)OCCCCCCCC)CC(=O)OCCCCCCCC APVVRLGIFCYZHJ-UHFFFAOYSA-N 0.000 description 1
- LENZDBCJOHFCAS-UHFFFAOYSA-N tris Chemical compound OCC(N)(CO)CO LENZDBCJOHFCAS-UHFFFAOYSA-N 0.000 description 1
- COXJMKGEQAWXNP-UHFFFAOYSA-N tris(14-methylpentadecyl) 2-hydroxypropane-1,2,3-tricarboxylate Chemical compound CC(C)CCCCCCCCCCCCCOC(=O)CC(O)(C(=O)OCCCCCCCCCCCCCC(C)C)CC(=O)OCCCCCCCCCCCCCC(C)C COXJMKGEQAWXNP-UHFFFAOYSA-N 0.000 description 1
- RHNXTZDKMRCKKT-UHFFFAOYSA-N tris(6-methylheptyl) 2-hydroxypropane-1,2,3-tricarboxylate Chemical compound CC(C)CCCCCOC(=O)CC(O)(C(=O)OCCCCCC(C)C)CC(=O)OCCCCCC(C)C RHNXTZDKMRCKKT-UHFFFAOYSA-N 0.000 description 1
- 235000013799 ultramarine blue Nutrition 0.000 description 1
- 238000002137 ultrasound extraction Methods 0.000 description 1
- 239000006097 ultraviolet radiation absorber Substances 0.000 description 1
- 229960002703 undecylenic acid Drugs 0.000 description 1
- 229940099259 vaseline Drugs 0.000 description 1
- 230000024883 vasodilation Effects 0.000 description 1
- 229940124549 vasodilator Drugs 0.000 description 1
- 239000003071 vasodilator agent Substances 0.000 description 1
- 235000019871 vegetable fat Nutrition 0.000 description 1
- 235000013311 vegetables Nutrition 0.000 description 1
- 125000000391 vinyl group Chemical group [H]C([*])=C([H])[H] 0.000 description 1
- 229920002554 vinyl polymer Polymers 0.000 description 1
- 235000019155 vitamin A Nutrition 0.000 description 1
- 239000011719 vitamin A Substances 0.000 description 1
- NCYCYZXNIZJOKI-UHFFFAOYSA-N vitamin A aldehyde Natural products O=CC=C(C)C=CC=C(C)C=CC1=C(C)CCCC1(C)C NCYCYZXNIZJOKI-UHFFFAOYSA-N 0.000 description 1
- 235000010374 vitamin B1 Nutrition 0.000 description 1
- 239000011691 vitamin B1 Substances 0.000 description 1
- 235000019163 vitamin B12 Nutrition 0.000 description 1
- 239000011715 vitamin B12 Substances 0.000 description 1
- 235000019164 vitamin B2 Nutrition 0.000 description 1
- 239000011716 vitamin B2 Substances 0.000 description 1
- 235000019158 vitamin B6 Nutrition 0.000 description 1
- 239000011726 vitamin B6 Substances 0.000 description 1
- 235000011912 vitamin B7 Nutrition 0.000 description 1
- 239000011735 vitamin B7 Substances 0.000 description 1
- 235000019154 vitamin C Nutrition 0.000 description 1
- 239000011718 vitamin C Substances 0.000 description 1
- 235000001892 vitamin D2 Nutrition 0.000 description 1
- 239000011653 vitamin D2 Substances 0.000 description 1
- MECHNRXZTMCUDQ-RKHKHRCZSA-N vitamin D2 Chemical compound C1(/[C@@H]2CC[C@@H]([C@]2(CCC1)C)[C@H](C)/C=C/[C@H](C)C(C)C)=C\C=C1\C[C@@H](O)CCC1=C MECHNRXZTMCUDQ-RKHKHRCZSA-N 0.000 description 1
- 235000005282 vitamin D3 Nutrition 0.000 description 1
- 239000011647 vitamin D3 Substances 0.000 description 1
- QYSXJUFSXHHAJI-YRZJJWOYSA-N vitamin D3 Chemical compound C1(/[C@@H]2CC[C@@H]([C@]2(CCC1)C)[C@H](C)CCCC(C)C)=C\C=C1\C[C@@H](O)CCC1=C QYSXJUFSXHHAJI-YRZJJWOYSA-N 0.000 description 1
- 229940046009 vitamin E Drugs 0.000 description 1
- 235000019165 vitamin E Nutrition 0.000 description 1
- 239000011709 vitamin E Substances 0.000 description 1
- 229940045997 vitamin a Drugs 0.000 description 1
- 229940021056 vitamin d3 Drugs 0.000 description 1
- 235000020234 walnut Nutrition 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
- 239000010497 wheat germ oil Substances 0.000 description 1
- 239000000811 xylitol Substances 0.000 description 1
- 235000010447 xylitol Nutrition 0.000 description 1
- 229960002675 xylitol Drugs 0.000 description 1
- HEBKCHPVOIAQTA-SCDXWVJYSA-N xylitol Chemical compound OC[C@H](O)[C@@H](O)[C@H](O)CO HEBKCHPVOIAQTA-SCDXWVJYSA-N 0.000 description 1
- 239000011701 zinc Substances 0.000 description 1
- 229910052725 zinc Inorganic materials 0.000 description 1
- CPYIZQLXMGRKSW-UHFFFAOYSA-N zinc;iron(3+);oxygen(2-) Chemical compound [O-2].[O-2].[O-2].[O-2].[Fe+3].[Fe+3].[Zn+2] CPYIZQLXMGRKSW-UHFFFAOYSA-N 0.000 description 1
- 229910001928 zirconium oxide Inorganic materials 0.000 description 1
- 239000004711 α-olefin Substances 0.000 description 1
Images
Classifications
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- 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
- 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)
- A61K36/88—Liliopsida (monocotyledons)
- A61K36/899—Poaceae or Gramineae (Grass family), e.g. bamboo, corn or sugar cane
-
- 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]
- A61K8/9794—Liliopsida [monocotyledons]
-
- 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
- A61P17/14—Drugs for dermatological disorders for baldness or alopecia
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- 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
- A23V2250/50—Polysaccharides, gums
- A23V2250/51—Polysaccharide
- A23V2250/5106—Bran
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/30—Extraction of the material
- A61K2236/35—Extraction with lipophilic solvents, e.g. Hexane or petrol ether
Definitions
- the present invention relates to a composition for preventing hair loss or promoting hair growth comprising bran extract as an active ingredient.
- the human hair has about 100,000 to 150,000 pieces and is formed in "hair follicles". There is a papilla in the hair follicle, where the small blood vessels are distributed, providing the nutrients needed for hair growth and a seborrheic gland on the side of the teat that provides the oil to lubricate the hair.
- Each hair has a different cycle and grows and falls through the anagen, catagen and telogen phases. This cycle is repeated over three to six years, with an average of 50 to 100 hairs falling out daily.
- alopecia refers to an abnormally large number of hairs falling out due to shortening of the growth phase hair and more degenerative or resting hairs during these cycles.
- the causes of hair loss are discussed, such as hyperthermia of male hormone action, excessive sebum secretion, poor blood circulation, hypoxia caused by peroxide, bacteria, genetic factors, aging, stress.
- testosteron a type of male hormone
- DHT dihydrotestoserone
- 5 ⁇ -reductase an enzyme called 5 ⁇ -reductase
- Minoxidil was initially developed as a vasodilator for the treatment of hypertension, but it was developed as a hair restorer as hirsutism was reported as a side effect.
- the mechanism of action of minoxidil on the hair growth effect has not been clarified to date, but the effect of increased nutrient supply through vasodilation, potassium channel opening effect, and inhibition of apoptosis of dermal papilla cells The back is thought to induce hair growth.
- Finasteride developed by Merck, also inhibits the activity of 5 ⁇ -reductase, an enzyme that acts on male hormone metabolism.
- Korean Patent Publication No. 2010-0111640 mentions hair loss prevention and hair growth promoting effects of one or more compositions selected from the group consisting of brown rice, black rice, and rice. However, no prior literature mentions bran hair loss prevention and hair growth promoting effect.
- bran extract plays an important role in the growth of hair and continued the study to complete the present invention.
- It is an object of the present invention to provide a pharmaceutical composition comprising as an active ingredient bran extract that is safe for skin, prevents hair loss, and promotes hair growth.
- Another object of the present invention is to provide a cosmetic composition comprising the bran extract as an active ingredient.
- Still another object of the present invention is to provide a health functional food comprising the bran extract as an active ingredient.
- Another object of the present invention to provide a hair loss prevention and hair growth promoting method comprising the step of treating a pharmaceutical composition comprising a bran extract at the hair loss site.
- Still another object of the present invention is to provide a use of bran extract for use in the manufacture of a pharmaceutical composition for preventing hair loss and promoting hair growth.
- the present invention provides a pharmaceutical composition for preventing hair loss or promoting hair growth comprising an extract of bran as an active ingredient.
- the bran extract may be included in an amount of 0.01 to 50% by weight based on the total weight of the composition, but is not necessarily limited thereto. If outside the above range may not exhibit hair loss prevention or hair growth promoting effect.
- the pharmaceutical composition may be selected from the group consisting of tablets, capsules, injections, creams, gels, patches, sprays, ointments, warnings, lotions, linen, pasta and cataplasma.
- the present invention provides a cosmetic composition for preventing hair loss or promoting hair growth comprising an extract of bran as an active ingredient.
- the bran extract may be included in an amount of 0.01 to 50% by weight based on the total weight of the composition, but is not necessarily limited thereto. If outside the above range may not exhibit hair loss prevention or hair growth promoting effect.
- the cosmetic composition is a hair tonic, hair conditioner, hair essence, hair lotion, hair nutrition lotion, hair shampoo, hair rinse, hair treatment, hair cream, hair nutrition cream, hair moisturizing cream, hair massage cream, hair wax, hair aerosol , Hair pack, nourishment pack, hair soap, hair cleansing foam, hair oil, hair drier, hair preservative, hair dye, hair wave, hair bleach, hair gel, hair glaze, hair dresser, hair lacquer, hair moisturizer, It may be a formulation selected from the group consisting of hair mousse and hair spray.
- the present invention provides a health functional food for preventing hair loss or promoting hair growth comprising an extract of bran as an active ingredient.
- Health functional food as defined in the present invention means a food manufactured and processed using raw materials or ingredients having functional properties useful for the human body according to Act No. 6767 of the Health Functional Food Act, and "functional” means a human body It means the ingestion for the purpose of obtaining a useful effect in health use such as nutrient control or physiological action on the structure and function of.
- the bran extract may be included in an amount of 0.01 to 50% by weight based on the total weight of the composition, but is not necessarily limited thereto. If outside the above range may not exhibit hair loss prevention or hair growth promoting effect.
- the health functional food may be a powder, granule, tablet, capsule or beverage.
- the present invention provides a method for preventing hair loss and promoting hair growth, comprising treating a pharmaceutical composition including a bran extract to a hair loss site.
- the present invention provides the use of bran extract for use in the manufacture of a pharmaceutical composition for preventing hair loss and promoting hair growth.
- composition comprising the bran extract according to the present invention as an active ingredient provides an effect of inhibiting and preventing hair loss and promoting hair growth.
- the composition containing the bran extract of the present invention can be used in various fields such as pharmacy, cosmetics, food and beauty.
- Figure 1 shows the dermal papilla cell proliferation efficacy of the bran hexane fraction.
- Figure 2 is a diagram confirming the expression of cell cycle proteins and Wnt / beta-catenin signaling proteins in dermal papilla cells by the bran hexane fraction.
- Figure 3 shows the hair-fiber growth effect of the bran hexane fraction in Vibrissa follicles.
- wheat Triticum aestivum L.
- Wheat is known to have excellent anticancer and antioxidant effects.
- Brain is the remainder of the separation of flour and embryos from flour, and contains most of the seed's blood but a small amount of embryos and embryos.
- Extract as defined herein is a solvent selected from water, including purified water, lower alcohols having 1 to 4 carbon atoms or mixed solvents thereof, hexane, butylene glycol, propylene glycol, glycerin, ethyl acetate, ether, chloroform, preferably Includes extracts soluble in water or methanol mixed solvents.
- bran extract includes all of them without particular limitation, as long as it is obtained by extracting an active ingredient from bran.
- the result obtained by putting bran into water or an organic solvent and eluting an effective component through means, such as standing, stirring, pressurization, or heating, is mentioned. It also includes a freeze-dried product obtained by lyophilizing the liquid extract obtained as described above.
- any means available to those skilled in the art for extracting the active ingredient includes all the extracted extracts, including those that have been processed after extraction and freeze drying. Others include extracts obtained by conventional extraction methods such as extraction by baths or room temperature, or by conventional extraction methods described in Chinese medicine or textbooks.
- the bran extract may be used without limitation, such as being collected, commercially available, and may include, for example, using it after removing the foreign matter by washing with water to remove the foreign matter.
- the bran extract may be obtained by conventional extraction methods in the art, such as ultrasonic extraction, filtration and reflux extraction.
- the bran may be an extract extracted with water, a lower alcohol having 1 to 4 carbon atoms or a mixed solvent thereof, a solvent selected from hexane, butylene glycol, propylene glycol, glycerin, ethyl acetate, ether, chloroform.
- Air loss refers to the phenomenon of hair dropping from the scalp or the condition of the hair being thin or thin.
- Air loss prevention means preventing and suppressing hair loss as described above.
- Raising hair growth means not only promoting the production of new hair, but also ensuring that existing hair grows healthy.
- the hair cycle can be divided into three main stages, known as the growing, degenerative and resting phases.
- the growth phase hair growth occurs as hair follicles grow deep into the skin with rapid proliferation of cells.
- the next phenomenon is the degenerative phase, which is a transitional period in which the disruption of cell division is noticeable, during which hair follicles gradually degenerate and hair growth stops.
- the resting phase the degenerative hair follicles contain germs with densely packed dermal papilla cells.
- the onset of new growth phase phenomena at rest is induced by rapid cell proliferation in the embryo, swelling of the dermal papilla and the synthesis of basement membrane elements.
- hair growth occurs in the growth phase, and is promoted by induction from the resting phase to the growth phase and the delay from the growth phase to the regression phase.
- hair loss that is, to prevent hair loss or to cause hair regrowth, that is, to promote hair growth, by promoting or extending the growth phase.
- composition according to an embodiment of the present invention includes one or more of the effects of preventing existing hair from falling out, improving the thickened hair, and the like, and generating new hair.
- the composition according to the present invention can also induce hair growth by activating dermal papilla cells.
- the bran hexane fraction was tested for hair growth efficacy.
- the present invention it was investigated using rat vibrissa immortalized dermal papilla cells for the effect of the dermal papilla cells proliferation.
- the growth and proliferation of the dermal papilla cells was 115.1 ⁇ 5.2% (P ⁇ 0.01), 139.3 ⁇ 5.4% (P ⁇ 0.001), 190.5 ⁇ 6.5 % (P ⁇ 0.001) and 183.3 ⁇ 10.2% (P ⁇ 0.0001) were increased.
- the growth proliferation effect of these dermal papilla cells is higher than the growth proliferation effect of the minoxidil dermal papilla cells used as a positive control material.
- Minoxidil is known to promote cell cycle progression and induce proliferation of dermal papilla cells by increasing cyclin E and CDK2 levels and decreasing p27 kip1 levels in dermal papilla cells (Kang, JI, Kim, EJ, Kim, MK, Jeon, YJ, Kang, SM, Koh, YS, Yoo, ES and Kang, HK (2013). Mar. Drugs 11 : 1783-1799.).
- the Wnt / ⁇ -catenin signaling pathway plays an important role in hair growth and cell growth regulation (Kwack, MH, Kang, BM, Kim, MK, Kim, JC and Sung, YK (2011). J. Dermatol.
- the Wnt / ⁇ -catenin signaling pathway is regulated by a variety of factors, in particular the activation of Akt induces ⁇ -catenin phosphorylation (ser552) and GSK3 ⁇ phosphorylation (ser9), while PKA is ⁇ -catenin phosphorylation (ser552 and ser675), which in turn activates the Wnt / ⁇ -catenin signaling pathway.
- the hair growth efficacy of the bran hexane fraction treatment group was investigated.
- the bran hexane fraction treatment group showed higher growth efficiency of hair-fiber than the positive control 10 ⁇ M minoxidil.
- the bran extract can be seen that the hair loss prevention and hair growth promoting effect is excellent.
- the bran extract of the present invention is expected to act to promote the growth of hair papilla cells, which play a very important role in hair growth, to induce the growth phase of hair follicles or to maintain the growth phase.
- the pharmaceutical composition containing the bran extract of the present invention as an active ingredient is a tablet for preventing hair loss and promoting hair growth, a capsule, an injection, a cream, a gel, a patch, a spray, an ointment, a warning, a lotion, a linen, a pasta or a cata It may be prepared by using a pharmaceutical composition in the form of plasma, but is not limited thereto.
- Preferred dosages of the bran extract of the present invention vary depending on the condition and weight of the patient, the extent of the disease, the form of the drug, the route and duration of administration, and may be appropriately selected by those skilled in the art.
- the extract of the present invention is preferably administered at 0.0001 to 100 mg / kg, preferably 0.001 to 10 mg / kg per day. Administration may be administered once a day or may be divided several times. The dosage does not limit the scope of the invention in any aspect.
- the bran extract of the present invention can be used in various hair cosmetics having the effect of preventing hair loss and activating hair growth.
- products to which the composition can be added include, for example, hair tonics, hair conditioners, hair essences, hair lotions, hair nutrition lotions, hair shampoos, hair rinses, hair treatments, hair creams, hair nutrition creams, hair moisturizers.
- the cosmetic of the present invention may include a composition selected from the group consisting of water-soluble vitamins, oil-soluble vitamins, polymer peptides, polymer polysaccharides, sphingolipids and seaweed extract.
- the water-soluble vitamin may be any compound that can be incorporated into cosmetics, but preferably vitamin B1, vitamin B2, vitamin B6, pyridoxine, pyridoxine, vitamin B12, pantothenic acid, nicotinic acid, nicotinic acid amide, folic acid, vitamin C, vitamin H, and the like.
- Their salts thiamine hydrochloride, sodium ascorbate salt, etc.
- derivatives ascorbic acid-2-sodium phosphate salt, ascorbic acid-2-magnesium phosphate salt, etc.
- the water-soluble vitamins can be obtained by conventional methods such as microbial transformation, purification from microorganism culture, enzyme or chemical synthesis.
- any useful vitamin can be used as long as it can be incorporated into cosmetics, but preferably vitamin A, carotene, vitamin D2, vitamin D3, vitamin E (d1-alpha tocopherol, d-alpha tocopherol, d-alpha tocopherol) and the like.
- Derivatives thereof ascorbic palmitate, ascorbic stearate, ascorbic acid dipalmitate, dl-alpha tocopherol acetate, dl-alpha tocopherolvitamin E, DL-pantothenyl alcohol, D-pantothenyl alcohol, pantothenyl) Ethyl ether
- Oil-soluble vitamins can be obtained by conventional methods such as microbial transformation, purification of microorganism culture, enzyme or chemical synthesis.
- the polymer peptide may be any compound that can be formulated into cosmetics, but preferably collagen, hydrolyzed collagen, gelatin, elastin, hydrolyzed elastin, keratin and the like.
- Polymeric peptides can be purified and obtained by conventional methods such as purification from microbial cultures, enzymatic methods or chemical synthesis methods, or can be purified and used from natural products such as dermis and pig silk such as pigs and cattle.
- the polymer polysaccharide may be any compound as long as it can be blended into cosmetics.
- hydroxy ethyl cellulose, xanthan gum, sodium hyaluronate, chondroitin sulfate or a salt thereof (sodium salt, etc.) may be mentioned.
- chondroitin sulfate or its salt, etc. can be normally purified from a mammal or fish.
- the sphingolipid may be any compound as long as it can be blended into cosmetics.
- ceramide, phytosphingosine, sphingosaccharide lipid, etc. may be mentioned.
- Sphingo lipids can usually be purified from mammals, fish, shellfish, yeast or plants by conventional methods or obtained by chemical synthesis.
- the seaweed extract may be any compound as long as it can be blended into cosmetics.
- the seaweed extract may include brown algae extract, red algae extract, green algae extract, and the like. Colorginine, arginic acid, sodium arginate, purified from these seaweed extracts, Potassium arginate and the like are also included in the seaweed extract used in the present invention.
- Seaweed extract can be obtained by purification from seaweed by conventional methods.
- Other components that may be added include fats and oils, moisturizers, emollients, surfactants, organic and inorganic pigments, organic powders, ultraviolet absorbers, preservatives, fungicides, antioxidants, plant extracts, pH adjusters, alcohols, pigments, flavorings, Blood circulation accelerators, cooling agents, restriction agents, purified water and the like.
- Examples of the fat or oil component include ester fats, hydrocarbon fats, silicone fats, fluorine fats, animal fats, and vegetable fats and oils.
- ester fats and oils As ester fats and oils, glyceryl tri2-ethylhexanoate, cetyl 2-ethylhexanoate, isopropyl myristate, butyl mystinate, isopropyl palmitate, ethyl stearate, octyl palmitate, isocetyl isostearate, and stearic acid Butyl, ethyl linoleate, isopropyl linoleate, ethyl oleate, isocetyl acid isocetyl, isostyl acid isostearyl, isostaryl palmitate, octylate acid octyldodecyl, isostearic acid isetyl, diethyl sebacate, adipine Acid isopropyl, isoalkyl neopentane, tri (capryl, capric acid) glyceryl, tri
- hydrocarbon-based oils and fats examples include hydrocarbon oils such as squalene, liquid paraffin, alpha-olefin oligomers, isoparaffins, ceresin, paraffins, liquid isoparaffins, polybutenes, microcrystalline waxes, and vaseline.
- silicone-based oils and fats examples include polymethylsilicone, methylphenylsilicone, methylcyclopolysiloxane, octamethylpolysiloxane, decamethylpolysiloxane, dodecamethylcyclosiloxane, dimethylsiloxane, methylcetyloxysiloxane copolymer, dimethylsiloxane and methylsteoxysiloxane copolymer, and alkyl.
- Modified silicone oil amino modified silicone oil and the like.
- Perfluoro polyether etc. are mentioned as fluorine-based fats and oils.
- Animal or vegetable oils include avocado oil, almond oil, olive oil, sesame oil, rice bran oil, soybean oil, soybean oil, corn oil, rapeseed oil, almond oil, palm kernel oil, palm oil, castor oil, sunflower oil, grape seed oil. , Cottonseed oil, Palm oil, Cucumber nut oil, Wheat germ oil, Rice germ oil, Shea butter, Walnut colostrum oil, Marker demia nut oil, Meadow home oil, Egg yolk oil, Uji, Horse oil, Mink oil, Orange rape oil, Jojoba oil And animal or plant fats and oils such as candeler wax, carnava wax, liquid lanolin and hardened castor oil.
- the moisturizing agent examples include a water-soluble low molecular moisturizer, a fat-soluble molecular moisturizer, a water-soluble polymer, and a fat-soluble polymer.
- the fat-soluble low molecular humectants include cholesterol and cholesterol esters.
- the water-soluble polymer carboxyvinyl polymer, polyasparaginate, tragacanth, xanthan gum, methyl cellulose, hydroxymethyl cellulose, hydroxyethyl cellulose, hydroxypropyl cellulose, carboxymethyl cellulose, water soluble chitin, chitosan, dextrin, etc. Can be mentioned.
- fat-soluble polymers examples include polyvinylpyrrolidone-eicosene copolymers, polyvinylpyrrolidone-hexadecene copolymers, nitrocellulose, dextrin fatty acid esters, polymer silicones, and the like.
- emollient examples include long-chain acyl glutamic acid cholesteryl esters, hydroxy stearic acid cholesterol, 12-hydroxystearic acid, stearic acid, rosin acid, lanolin fatty acid cholesteryl esters, and the like.
- nonionic surfactant nonionic surfactant, anionic surfactant, cationic surfactant, amphoteric surfactant, etc. are mentioned.
- nonionic surfactant self-emulsifying glycerin monostearate, propylene glycol fatty acid ester, glycerin fatty acid ester, polyglycerol fatty acid ester, sorbitan fatty acid ester, POE (polyoxyethylene) sorbitan fatty acid ester, POE sorbitan fatty acid ester, POE Glycerin Fatty Acid Ester, POE Alkyl Ether, POE Fatty Acid Ester, POE Cured Castor Oil, POE Castor Oil, POE / POP (Polyoxyethylene / Polyoxypropylene) Copolymer, POE / POP Alkyl Ether, Polyether Modified Silicone, Laurin Acid Alkanolamide, alkylamine oxide, hydrogenated soybean phospholipid, etc. are mentioned.
- Anionic surfactants include fatty acid soaps, alpha-acyl sulfonates, alkyl sulfonates, alkyl allyl sulfonates, alkyl or phthalene sulfonates, alkyl sulfates, POE alkyl ether sulfates, alkylamide sulfates, alkyl phosphates, POE alkyl phosphorus salts, Alkylamide phosphates, alkyloylalkyltaurine salts, N-acylamino acid salts, POE alkyl ether carboxylate salts, alkyl sulfosuccinate salts, sodium alkyl sulfo acetates, acylated hydrolyzed collagen peptide salts, perfluoroalkyl phosphate esters, and the like. Can be mentioned.
- cationic surfactants include alkyltrimethylammonium chloride, stearyltrimethylammonium chloride, stearyl trimethylammonium chloride, cetostearyltrimethylammonium chloride, distearyldimethylammonium chloride, stearyldimethylbenzyl ammonium bromide, and behenyltrimethylammonium bromide.
- benzalkonium chloride diethylaminoethyl stearate, dimethylaminopropyl stearate, lanolin derivatives, quaternary ammonium salts, and the like.
- amphoteric surfactants include the carboxybetaine type, the amide betain type, the sulfobetain type, the hydroxysulfobetain type, the amide sulfobetain type, the phosphobetaine type, the aminocarboxylate type, the imidazoline derivative type, and the amideamine type.
- An amphoteric surfactant etc. are mentioned.
- Organic and inorganic pigments include silicic acid, silicic anhydride, magnesium silicate, talc, sericite, mica, kaolin, bengal, clay, bentonite, titanium film mica, bismuth oxychloride, zirconium oxide, magnesium oxide, zinc oxide, titanium oxide, aluminum oxide
- Inorganic pigments such as calcium sulfate, barium sulfate, magnesium sulfate, calcium carbonate, magnesium carbonate, iron oxide, ultramarine blue, chromium oxide, chromium hydroxide, calamine, and composites thereof; Polyamide, polyester, polypropylene, polystyrene, polyurethane, vinyl resin, urea resin, phenol resin, fluorine resin, silicon resin, acrylic resin, melamine resin, epoxy resin, polycarbonate resin, divinylbenzene, styrene copolymer, Organic pigments such as silk powder, cellulose, CI pigment yellow, CI pigment orange, and composite pigments of these inorgan
- Metal soaps such as a calcium stearate; Alkyl phosphate metal salts such as zinc sodium cetyl acid, zinc lauryl acid and calcium laurate; Acylamino acid polyvalent metal salts such as N-lauroyl-beta-alanine calcium, N-lauroyl-beta-alanine zinc, and N-lauroylglycine calcium; Amide sulfonic acid polyvalent metal salts, such as N-lauroyl-taurine calcium and N-palmitoyl-taurine calcium; N-epsilon-lauroyl-L-lysine, N-epsilon-palmitolyzine, N-alpha-paratoylol nitin, N-alpha-lauroyl arginine, N-alpha-cured fatty acid acyl arginine Acyl basic amino acids; N-acylpolypeptides, such as N-lauroyl glycy
- Examples of the ultraviolet absorber include paraaminobenzoic acid, ethyl paraaminobenzoate, amyl paraaminobenzoic acid, octyl paraaminobenzoate, ethylene glycol salicylate, phenyl salicylate, octyl salicylate, benzyl salicylate, butylphenyl salicylate, homomentyl salicylic acid and benzyl cinnamic acid.
- paraaminobenzoic acid ethyl paraaminobenzoate, amyl paraaminobenzoic acid, octyl paraaminobenzoate, ethylene glycol salicylate, phenyl salicylate, octyl salicylate, benzyl salicylate, butylphenyl salicylate, homomentyl salicylic acid and benzyl cinnamic acid.
- fungicides hinokithiol, trichloric acid, trichlorohydroxydiphenyl ether, chlorhexidine gluconate, phenoxyethanol, resorcin, isopropylmethylphenol, azulene, salicylic acid, ginxitlionone, benzalkonium chloride, photosensitive Sodium No. 301, sodium mononitrohydrol, undecylenic acid, and the like.
- antioxidants examples include butylhydroxyanisole, propyl gallic acid, and erythorbic acid.
- pH adjuster examples include citric acid, sodium citrate, malic acid, sodium malate, fmaric acid, sodium pmarate, succinic acid, sodium succinate, sodium hydroxide, sodium dihydrogen phosphate, and the like.
- Examples of the alcohol include higher alcohols such as cetyl alcohol.
- the compounding component which may be added other than this is not limited to this, Moreover, although all said components can be mix
- the cosmetic of the present invention may take the form of a solution, an emulsion, a viscous mixture, or the like.
- Ingredients included in the cosmetic composition of the present invention may include components commonly used in cosmetic compositions in addition to the extract as an active ingredient, for example, conventional stabilizers, solubilizers, vitamins, pigments and flavorings. And auxiliaries and carriers.
- the cosmetic composition of the present invention may be prepared in any formulation commonly prepared in the art, and includes, for example, milky lotion, cream, lotion, pack, foundation, lotion, essence, hair cosmetic, and the like.
- the formulation of the present invention is a paste, cream or gel, animal fibers, plant fibers, waxes, paraffins, starches, tracantes, cellulose derivatives, polyethylene glycols, silicones, bentonites, silicas, talc or zinc oxide, etc. may be used as carrier components.
- carrier components can be.
- 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.
- Solvents, solvating or emulsifying agents may be used as carrier components when the formulations of the invention are solutions or emulsions, such as water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol , Fatty acid esters of 1,3-butylglycol oil, glycerol aliphatic ester, polyethylene glycol or sorbitan, and the like.
- solvents, solvating or emulsifying agents may be used as carrier components when the formulations of the invention are solutions or emulsions, such as water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol , Fatty acid esters of 1,3-butylglycol oil, glycerol aliphatic ester, polyethylene glycol or
- liquid carrier diluents such as water, ethanol or propylene glycol
- suspension agents such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol esters and polyoxyethylene sorbitan esters, microcrystalline Cellulose, aluminum metahydroxy, bentonite, agar or tracant and the like can be used.
- the carrier component is an aliphatic alcohol sulfate, an aliphatic alcohol ether sulfate, a sulfosuccinic acid monoester, isethionate, an imidazolinium derivative, a methyltaurate, a sarcosinate, a fatty acid amide.
- Ether sulfates, alkylamidobetaines, aliphatic alcohols, fatty acid glycerides, fatty acid diethanolamides, vegetable oils, linolin derivatives or ethoxylated glycerol fatty acid esters and the like can be used.
- the present invention also provides a health functional food for preventing hair loss or promoting hair growth comprising an extract of bran as an active ingredient.
- Foods to which the bran extract of the present invention may be added include, for example, various foods, beverages, gums, teas, vitamin complexes, health supplements, and the like, and may be used in the form of powders, granules, tablets, capsules, or beverages. Can be.
- bran extract itself of the present invention Since the bran extract itself of the present invention has almost no toxicity and side effects, it is a medicament that can be used safely even when taken for long periods of time.
- the bran extract of the present invention may be added to food or beverage for the purpose of preventing and improving hair loss disease.
- the amount of the extract in the food or beverage is generally added to the health food composition of the present invention 0.01 to 50% by weight of the total food weight, the health beverage composition is 0.02 to 10 g based on 100 ml, preferably It can be added at a ratio of 0.3 to 1 g.
- the health beverage composition of the present invention has no particular limitation on the liquid component, and may contain various flavors or natural carbohydrates as additional ingredients, such as ordinary drinks.
- natural carbohydrates are conventional monosaccharides such as disaccharides such as glucose and fructose, such as maltose, sucrose and the like, and polysaccharides such as dextrin, cyclodextrin and the like.
- Sugars and sugar alcohols such as xylitol, sorbitol, and erythritol.
- natural flavoring agents tacumatin, stevia extract, rebaudioside A, glycyrrhizin, etc.
- synthetic flavoring agents sacharin, aspartame, etc.
- the proportion of such natural carbohydrates is generally about 1 to 20 g, preferably about 5 to 12 g per 100 ml of the composition of the present invention.
- the composition of the present invention includes various nutrients, vitamins, minerals (electrolytes), flavors such as synthetic flavors and natural flavors, coloring and neutralizing agents (such as cheese and chocolate), pectic acid and salts thereof, alginic acid and its Salts, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohols, carbonation agents used in carbonated beverages, and the like.
- the compositions of the present invention may also contain pulp for the production of natural fruit juices and fruit juice beverages and vegetable beverages. These components can be used independently or in combination. The proportion of such additives is not so critical but is generally selected from the range of 0 to about 20 parts by weight per 100 parts by weight of the composition of the present invention.
- the wheat bran used in the present invention is supplied from a mill of wheat bran grown in Gyeongsangbuk-do. 5 kg of total bran samples were extracted with water bath in a water bath three times for 4 hours in 90% ethanol. Then, the supernatant was collected, concentrated under reduced pressure using a rotary evaporator, and 337 g of crude extract was obtained. 300 g of the solvent was fractionated with n-hexane to obtain 133 g of an n-hexane layer, and this fraction was used as a sample for the experiment.
- Rat vibrissa immortalized dermal papilla cells isolated from rat whiskers were treated with 100 units / ml penicillin-100 ⁇ g / ml streptomycin (Gibco Inc, NY, USA) and 10% heat-inactivated fetal bovine serum.
- DMEM Hyclone Inc, USA
- FBS FBS
- Proliferation of dermal papilla cells was measured using an MTT assay. 1.0 ⁇ 10 4 cells / ml of dermal papilla cells were suspended in DMEM medium containing 1% FBS, placed in a 96 well plate and incubated for 24 hours, and then the bran hexane fractions of 0.1, 1, 10, 50 and Treatment was at a concentration of 100 ⁇ g / ml. The positive control minoxidil (minoxidil, Sigma, MO, USA) was treated at a concentration of 10 ⁇ M.
- the bran hexane fractionation treatment group at the concentration of 10, 50 and 100 ⁇ g / ml showed a proliferative effect higher than 112.3 ⁇ 2.7% (P ⁇ 0.01) of the growth effect of 10 ⁇ M minoxidil used as a positive control, the bran hexane fraction It shows that it can exhibit hair growth efficacy like minoxidil.
- bran extract hexane extract
- lysis buffer [50 mM Tris-HCl (pH 7.5), 150 mM NaCl, 2 mM EDTA, 1 mM EGTA, 1 mM NaVO3, 10 mM NaF, 1 mM dithiothreitol (DTT). ), 1 mM phenylmethylsulfonylfluoride (PMSF), 25 ⁇ g / mL aprotinin, 25 ⁇ g / mL leupeptin and 1% NP-40] were added and then cell disrupted at 4 ° C. for 30 minutes.
- PMSF phenylmethylsulfonylfluoride
- Cell lysate was centrifuged at 15,000 rpm for 15 minutes to obtain supernatant and stored at ⁇ 20 ° C. until used in the experiment. Protein concentration was quantified by the Bradford method using bovine serum albumin (BSA) as a standard. 20-30 ⁇ g of the cell lysate was denatured with 8-12% mini gel SDS-PAGE (Sodium dodecyl sulfate polyacrylamide gel electrophoresis) and at 200 mA with a polyvinylidene fluoride (PVDF) membrane (Bio-Rad, Hercules, CA, USA) Transferred for 2 hours.
- BSA bovine serum albumin
- Tween-20-TBS 50 mM Tris, pH 7.6, 150 mM NaCl, 0.1% Tween-20
- the secondary antibody (Secondary anbibody) was subjected to an antibody binding reaction at room temperature for 1 hour using an antibody diluted 1: 1: 5000 anti-rabbit IgG, HRP (Horse Radish Peroxidase) conjugated anti-rabbit IgG, anti-mouse IgG. Thereafter, the membrane was washed three times with T-TBS, and then reacted with ECL substrate (Intron, Seoul, Korea) for 1 minute and then photosensitive to X-ray film (AGFA, Mortsel, Belgium).
- ECL substrate Intron, Seoul, Korea
- bran hexane extracts proliferate the dermal papilla cells by activating Wnt / ⁇ -catenin signaling pathways through the GSK3 ⁇ , PKA and Akt pathways as well as cell cycle proteins such as cyclin D1, cyclin E, phospho-CDK2 and phospho-pRB. Suggests increased potency.
- the separated hair follicles were placed in a Petri dish containing E / P buffer and incubated at 37 ° C. in a 5% CO 2 incubator for about 1 hour.
- 2 mM L-glutamine (Gibco Inc, NY, USA) 10 ⁇ g / ml insulin (Sigma MO, USA), 50 nM hydrocortisone (Sigma MO, USA) and 100 units / Add 500 ⁇ l of William E medium (Gillco E Inc., NY, USA) containing ml penicillin-100 ⁇ g / ml streptomycin and place one hair follicle in one medium at 37 ° C., 5% CO 2 thermostat. Incubated at.
- the medium was changed every 3 days and the bran hexane fractions were treated at concentrations of 1, 10 and 50 ⁇ g / ml.
- the positive control minoxidil was treated at a concentration of 10 ⁇ M (Kang et al., 2012 Biomol Ther 20 (1), 118-124).
- hair fiber growth was 150.0 ⁇ 21.7% (P ⁇ ) when 1, 10 and 50 ⁇ g / ml were treated, respectively. 0.001), 203.6 ⁇ 7.6% (P ⁇ 0.001) and 246.4 ⁇ 19.8% (P ⁇ 0.001) (Table 2, FIG. 3).
- the hair-fiber growth increase effect of the bran hexane fraction at concentrations of 10 and 50 ⁇ g / ml was greater than that of 156.3 ⁇ 11.5% (P ⁇ 0.05) growth of minoxidil (10 ⁇ M) as a positive control. Higher.
- mice Six-week-old female C57BL / 6 mice were purchased from Orient Bio Co., Ltd. and used for experiments after adapting to the animal breeding environment for one week. Animal breeding room was maintained day and night at a temperature of 23 ⁇ 3 °C, 12 hours. Experimental animals were reared in mouse cages for isolation and were fed freely with experimental feed. The experimental group was set up in five groups, which were treated with MINOXYL TM (modern drug) and bran hexane extracts (10, 50 and 100 ⁇ g / ml), which were the control group and the positive control group. Eight mice in each experimental group were used for the experiment.
- MINOXYL TM modern drug
- bran hexane extracts 10, 50 and 100 ⁇ g / ml
- the change in skin color was observed from 22 days after hair removal in the bran hexane extract application group, and the change in skin color and hair growth was confirmed in the bran hexane extract application group at 29 and 35 days after hair removal.
- Quantitative analysis of the hair growth induction effect of the bran hexane extract application group using dotmatrix area measurement method the bran hexane extract application group at the concentration of 50 ⁇ g / ml at day 35 showed significantly higher growth induction effect than the control group It was confirmed that it is shown (Fig. 4).
- the bran hexane extract induces hair follicle growth phase or growth phase through the effect of promoting cell growth proliferation by activation of cell cycle proteins, Wnt / ⁇ -catenin signaling pathway in the dermal papilla cells that play an important role in hair growth. It is maintained.
- the above ingredients were added to the indicated contents, mixed uniformly, stirred and granulated. After drying, using a tableting machine, a desired tablet containing 33.3 wt% of the bran extract, which is an active ingredient, was prepared.
- the ingredients are added to a given content, mixed uniformly, and then filled into gelatin capsules of an appropriate size so that 33 wt% of the bran extract, an active ingredient, is contained per capsule. Prepared.
- composition ratio of the above-mentioned vitamin and mineral mixtures is mixed with a component suitable for a health food in a preferred embodiment, the compounding ratio may be arbitrarily modified, and the above ingredients are mixed according to a conventional health food manufacturing method.
- the granules may be prepared and used for preparing a health food according to a conventional method.
- composition ratio is a composition that is relatively suitable for the preferred beverage in a preferred embodiment
- compounding ratio may be arbitrarily modified according to regional and ethnic preferences such as demand hierarchy, demand country, and usage.
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Abstract
본 발명은 밀기울 추출물을 유효성분으로 포함하는 탈모 방지 또는 육모 촉진용 조성물에 관한 것이다. 본 발명에 따른 밀기울 추출물을 유효성분으로 포함하는 약학 조성물은 탈모를 억제 및 예방하며 육모를 촉진하는 효능이 인정되고, 약학, 화장품, 식품 및 미용 등의 분야에서 다양하게 활용 가능하다.
Description
본 발명은 밀기울 추출물을 유효성분으로 포함하는 탈모 방지 또는 육모 촉진을 위한 조성물에 관한 것이다.
인체의 모발은 약 10만∼15만 개 정도이며 “모낭”에서 형성된다. 모낭에는 유두가 있는데, 이 부위에는 작은 혈관이 분포되어 모발의 성장에 필요한 영양분을 공급하고 유두 위의 옆으로는 모발에 윤기를 주는 기름을 공급해 주는 지루샘이 있다.
각각의 모발은 서로 다른 주기를 가지며 성장기(anagen), 퇴행기(catagen), 휴지기(telogen)를 거쳐서 성장하고 탈락한다. 이러한 주기는 3∼6년에 걸쳐 반복되는데, 일일 평균 50∼100개의 모발이 정상적으로 탈락하게 된다.
일반적으로 탈모증이라 함은 이러한 주기 중에서 성장기 모발의 비율이 짧아지고 퇴행기 또는 휴지기 모발이 많아져 탈락하는 모발의 숫자가 비정상적으로 많아지는 것을 일컫는다.
탈모의 원인으로는 남성호르몬 작용 과잉설, 피지분비 과잉설, 혈액순환 불량설, 과산화물, 세균 등에 의한 두피기능 저하설, 유전적 요인, 노화, 스트레스 등이 논의되고 있다.
구체적으로 남성호르몬의 일종인 테스토스테론(testosteron)은 5α-리덕타아제(5α-reductase)라는 효소에 의해 디하이드로테스토스테론(dihydrotestoserone; DHT)으로 활성화된다. 이 DHT가 특정 수용체와 결합하고 탈모를 일으키는 단백질을 유도하여 탈모가 일어난다. 또한 이와 같은 기작에 의하여 피지를 과잉 생성하기도 하며 여드름이나 지루성 피부염 등을 일으켜서 두피에 염증을 동반한 탈모가 나타나기도 한다.
지금까지 탈모를 방지하여 주고 발모촉진 및 모발 생장 효과를 가지고 있다고 알려져 있는 약물에는 미녹시딜(minoxidil)이나 트리코사카라이드(trichosaccharide), 피나스테라이드(Finasteride) 등이 있다. 미녹시딜은 처음에 고혈압 치료를 위한 혈관확장제로 개발되었으나, 부작용으로 다모증이 보고되면서 발모제로 개발되었다. 미녹시딜의 발모효과에 대한 작용기전은 현재까지 명확히 밝혀지지 않았지만, 혈관확장을 통한 영양공급 증가, 포타슘 채널 오프닝(potassium channel opening) 효과, 및 모유두 세포(dermal papilla cells)의 세포자살(apoptosis) 억제 효능 등이 모발성장을 유도하는 것으로 생각되고 있다. 또한 머크(Merck)에서 개발한 피나스테라이드는 남성호르몬 대사에 작용하는 효소인 5α-리덕타아제의 활성을 억제시키는 물질이다. 처음에는 전립선 비대증 치료제로 개발되었으나 후에 모발의 성장을 촉진시킴이 알려지면서 발모제로 개발되었다. 그러나 이러한 제제들의 경우, 뚜렷한 효능의 부재 및 인체 안정성, 피부자극 유발 등의 부작용 문제가 대두되고 있어 안전성 및 효능이 확보된 조성물 개발이 시급한 실정이다.
한국 공개특허 제 2010-0111640호에서 현미, 흑미, 쌀로 이루어진 군에서 선택되는 하나 이상의 조성물의 탈모 예방 및 발모 촉진 효과를 언급하고 있다. 그러나 그 어떤 선행문헌에서도 밀기울의 탈모 방지 및 육모 촉진 효과와 관련하여서 언급하고 있지 않다.
이에 본 발명자들은 밀기울 추출물이 모발의 성장에 중요한 역할을 하는 것을 발견하고 연구를 계속하여 본 발명을 완성하였다.
본 발명의 목적은 피부에 안전하고 탈모를 방지하며 육모를 촉진하는 밀기울 추출물을 유효성분으로 포함하는 약학 조성물을 제공하는 것이다.
본 발명의 다른 목적은 밀기울 추출물을 유효성분으로 포함하는 화장료 조성물을 제공하는 것이다.
본 발명의 또 다른 목적은 밀기울 추출물을 유효성분으로 포함하는 건강기능성 식품을 제공하는 것이다.
본 발명의 또 다른 목적은 탈모 부위에 밀기울 추출물을 포함한 약학 조성물을 처리하는 단계를 포함하는 탈모 방지 및 육모 촉진 방법을 제공하는 것이다.
본 발명의 또 다른 목적은 탈모 방지 및 육모 촉진용 약학 조성물의 제조에 사용하기 위한, 밀기울 추출물의 용도를 제공하는 것이다.
본 발명은 밀기울의 추출물을 유효성분으로 포함하는 탈모 방지 또는 육모 촉진용 약학 조성물을 제공한다.
상기 밀기울 추출물은 조성물 전체 중량에 대해 0.01 내지 50% 중량으로 포함될 수 있으나, 반드시 이로 제한되는 것은 아니다. 상기 범위를 벗어나는 경우 탈모 방지 또는 육모 촉진 효과가 나타나지 않을 수 있다.
상기 약학 조성물은 정제, 캅셀제, 주사제, 크림, 젤, 패취, 분무제, 연고제, 경고제, 로션제, 리니멘트제, 파스타제 및 카타플라스마제로 이루어진 군에서 선택될 수 있다.
다른 측면에서 본 발명은 밀기울의 추출물을 유효성분으로 포함하는 탈모 방지 또는 육모 촉진용 화장료 조성물을 제공한다.
상기 밀기울 추출물은 조성물 전체 중량에 대해 0.01 내지 50% 중량으로 포함될 수 있으나, 반드시 이로 제한되는 것은 아니다. 상기 범위를 벗어나는 경우 탈모 방지 또는 육모 촉진 효과가 나타나지 않을 수 있다.
상기 화장료 조성물은 헤어토닉, 헤어컨디셔너, 헤어에센스, 헤어로션, 헤어영양로션, 헤어샴푸, 헤어린스, 헤어트리트먼트, 헤어크림, 헤어영양크림, 헤어모이스처크림, 헤어맛사지크림, 헤어왁스, 헤어에어로졸, 헤어팩, 헤어영양팩, 헤어비누, 헤어클렌징폼, 머릿기름, 모발건조제, 모발보존처리제, 모발염색제, 모발용 웨이브제, 모발탈색제, 헤어겔, 헤어글레이즈, 헤어드레싱어, 헤어래커, 헤어모이스처라이저, 헤어무스 및 헤어스프레이로 이루어진 군에서 선택된 제형일 수 있다.
다른 측면에서 본 발명은 밀기울의 추출물을 유효성분으로 포함하는 탈모 방지 또는 육모 촉진용 건강기능성 식품을 제공한다.
본 발명에서 정의되는 “건강기능성 식품”은 건강기능식품에 관한 법률 제6727호에 따른 인체에 유용한 기능성을 가진 원료나 성분을 사용하여 제조 및 가공한 식품을 의미하며, "기능성"이라 함은 인체의 구조 및 기능에 대하여 영양소를 조절하거나 생리학적 작용 등과 같은 보건 용도에 유용한 효과를 얻을 목적으로 섭취하는 것을 의미한다.
상기 밀기울 추출물은 조성물 전체 중량에 대해 0.01 내지 50% 중량으로 포함될 수 있으나, 반드시 이로 제한되는 것은 아니다. 상기 범위를 벗어나는 경우 탈모 방지 또는 육모 촉진 효과가 나타나지 않을 수 있다.
상기 건강기능성 식품은 분말, 과립, 정제, 캡슐 또는 음료일 수 있다.
다른 측면에서 본 발명은 탈모 부위에 밀기울 추출물을 포함한 약학 조성물을 처리하는 단계를 포함하는 탈모 방지 및 육모 촉진 방법을 제공한다.
또 다른 측면에서 본 발명은 탈모 방지 및 육모 촉진용 약학 조성물의 제조에 사용하기 위한, 밀기울 추출물의 용도를 제공한다.
본 발명에 따른 밀기울 추출물을 유효성분으로 포함하는 조성물은 탈모를 억제 및 예방하며 육모를 촉진하는 효과를 제공한다. 본 발명의 밀기울 추출물을 포함하는 조성물은 약학, 화장품, 식품 및 미용 등의 분야에서 다양하게 활용 가능하다.
도 1은 밀기울 헥산 분획물의 모유두 세포(dermal papilla cell) 증식 효능을 나타낸 것이다.
도 2는 밀기울 헥산 분획물에 의한 모유두 세포에서의 세포주기 단백질 및 Wnt/베타-카테닌 신호전달 단백질들의 발현을 확인한 도이다.
도 3은 비 모낭(Vibrissa follicles)에서 밀기울 헥산 분획물의 헤어-섬유(hair-fiber) 성장 효과를 나타낸 것이다.
도 4는 밀기울 헥산 분획물에 의한 육모 효과를 in vivo로 확인한 도이다:
A: 쥐에서의 대조군, 밀기울 헥산 분획물 처리군 및 미녹시딜 처리군(MINOXYLTM)에서의 시간에 따른 육모 효과; 및
B: 전체 피부에 대한 털이 난 피부 부위 (검은 부분) %.
이하 본 발명을 상세히 설명한다.
본 발명에서 “밀(Triticum aestivum L.)”은 외떡잎식물 벼목 화본과에 속하는 한해살이 풀이다. 밀은 항암 및 항산화 효과가 우수한 것으로 알려져 있다.
“밀기울”은 제분밀로부터 밀가루와 배아를 분리한 나머지의 것으로, 종자의 피가 대부분이지만 소량의 배유부, 배아부를 함유하고 있다.
본 발명에서 정의되는 “추출물”은 정제수를 포함한 물, 탄소수 1 내지 4의 저급 알코올 또는 이들의 혼합 용매, 헥산, 부틸렌글리콜, 프로필렌글리콜, 글리세린, 초산에틸, 에테르, 클로로포름으로부터 선택된 용매, 바람직하게는 물 또는 메탄올 혼합용매에 가용한 추출물을 포함한다.
“밀기울 추출물”이라 함은, 밀기울로부터 유효성분을 추출하여 얻어진 것이라면 특별히 한정하지 않고 모두 포함한다. 예컨대, 밀기울을 물이나 유기용매에 넣고 정치, 교반, 가압 또는 가열 등의 수단을 통해 유효성분을 용출함으로써 얻어진 결과물을 들 수 있다. 또한, 상기와 같이 얻어진 액상 추출물을 동결건조함으로써 얻어진 동결건조물을 포함한다.
또한, 그러한 동결건조물을 분쇄한 분말을 포함한다. 그 외에도 유효성분을 추출하기 위해 이 기술분야에서 통상의 기술자가 이용 가능한 수단이라면 무엇이든지 가리지 않고 추출된 모든 추출물을 포함하며, 추출된 후 동결 건조 등의 가공을 거친 것까지도 모두 포함된다. 기타, 중탕이나 상온에 의한 추출법과 같이 예로부터 전해 내려오거나, 한의서 또는 교과서에 기재되어 있는 통상적인 추출법에 의한 추출물을 포함한다.
또한, 특정 활성 화합물들을 분리하기 위한 특별한 추출방법인 Stass Otto 추출법이나 각종 칼럼 크로마토그래피 방법 등을 통하여 얻어진 분획 추출물을 포함한다.
상기 밀기울 추출물은 채취한 것, 시판되는 것 등 제한 없이 사용할 수 있으며, 일례로 채취한 후 물로 세척하여 이물질을 제거한 후 건조하여 사용하는 것을 포함할 수 있다.
상기 밀기울 추출물은 초음파 추출법, 여과법 및 환류추출법 등 이 기술분야의 통상적인 추출방법으로 얻을 수 있다.
바람직하게는 밀기울을 물, 탄소수 1 내지 4의 저급 알코올 또는 이들의 혼합 용매, 헥산, 부틸렌글리콜, 프로필렌글리콜, 글리세린, 초산에틸, 에테르, 클로로포름으로부터 선택된 용매로 추출한 추출물일 수 있다.
“탈모”는 두피로부터 모발이 탈락하는 현상 또는 모발이 성기거나 가늘어지는 상태를 의미한다. “탈모 방지”라 함은 상술한 바와 같은 탈모 현상을 예방하고 억제하는 것을 의미한다. “육모 촉진”이라 함은 새로운 모발 생성의 촉진뿐만 아니라 기존 모발이 건강하게 자라도록 하는 것을 의미한다.
모발주기는 성장기, 퇴행기, 휴지기로 알려져 있는 3개의 주요 단계들로 나눌 수 있다. 성장기에는 세포의 빠른 증식과 함께 피부 안으로 깊이 모낭이 성장하면서 모발형성이 이루어진다. 다음 현상은 퇴행기인데 이것은 세포 분열의 중단이 두드러지는 과도기이며, 이 과정에서 모낭은 점차 퇴행하며 육모가 중단된다. 다음 현상인 휴지기에서는 퇴행 모낭은 조밀하게 찬 모유두(dermal papilla) 세포를 갖는 배(germ)를 포함한다. 휴지기에서 새로운 성장기 현상의 개시는 상기 배에서 빠른 세포 증식, 모유두의 팽창 및 기저막 요소의 합성에 의해 유도된다.
일반적으로“육모”는 성장기에서 이루어지며, 휴지기에서 성장기로의 유도 및 성장기에서 퇴행기로의 지연에 의하여 촉진된다.
따라서 성장기의 촉진 또는 연장에 의해 모발의 손실 방지, 즉 탈모를 방지하거나 모발의 재성장을 유발, 즉 육모를 촉진하는 것이 필요하다.
본 발명의 일실시예에 따른 조성물은 이미 존재하고 있는 모발이 빠지지 않게 하는 효과, 이미 존재하는 모발을 굵게 하는 등 개선시키는 효과, 및 새로운 모발을 생성하는 효과 중 하나 이상을 포함한다. 또한 본 발명에 따른 조성물은 모유두 세포를 활성화하여 모발 성장을 유도할 수 있다.
본 발명의 구체적 실시예에서는 모낭에서 유래된 모유두 세포와 쥐의 비 모낭(rat vibrissa follicles)의 배양을 이용하여 밀기울 헥산(hexane) 분획이 육모 효능이 있는지 실험하였다.
본 발명의 일 실시예에서 모유두 세포 증식 효과가 있는지에 대해 불멸화된 모유두 세포(rat vibrissa immortalized dermal papilla cell)을 사용하여 조사하였다. 1, 10, 50 및 100 μg/ml의 농도로 밀기울 헥산 분획을 처리하였을 때, 모유두 세포의 성장 증식이 115.1±5.2%(P<0.01), 139.3±5.4%(P<0.001), 190.5±6.5%(P<0.001)및 183.3±10.2%(P<0.0001) 증가되었다. 이런 모유두 세포의 성장 증식 효과는 양성대조 물질로 사용한 미녹시딜(minoxidil)의 모유두 세포의 성장 증식 효과보다 높은 것이다.
본 발명의 일 실시예에서 모유두 세포에서 모발 성장, 세포 증식 조절에서 중요한 역할을 하는 신호 전달 경로에 관여하는 단백질들의 발현을 확인하였다. 포유류 세포는 세포증식의 주요 과정인 세포 주기 진행을 위해 cyclin E/CDK2 complex의 활성화, cyclin D1 증가 및 pRB의 인산화 증가 등을 필요로 함이 알려져 있다 (Sherr, C. J. (1996). Science274:1672-1677., Sherr, C. J. and Roberts, J. M. (1999). GenesDev.13:1501-1512. Prall, O. W., Sarcevic, B., Musgrove, E. A., Watts, C. K. and Sutherland, R. L. (1997). J.Biol.Chem.272:10882-10894.). Minoxidil은 모유두 세포에서 cyclin E 및 CDK2의 level 증가, p27kip1level감소를 통해 cell cycle 진행을 촉진하고 모유두 세포의 증식을 유도하는 것으로 알려져 있다 (Kang, J. I., Kim, E. J., Kim, M. K., Jeon, Y. J., Kang, S. M., Koh, Y. S., Yoo, E. S. and Kang, H.K. (2013). Mar.Drugs
11:1783-1799.). 또한, Wnt/β-catenin 신호전달 경로는 모발성장, 세포증식 조절에서 중요한 역할을 한다 (Kwack, M. H., Kang, B. M., Kim, M. K., Kim, J. C. and Sung, Y. K. (2011). J.Dermatol.Sci.62: 154-159., Wangefjord, S., Braedstedt, J., Ericson Lindquist, K., Nodin, B., Jirstroem, K. and Eberhard, J. (2013). Diagn.Pathol.
8:10.). Wnt/β-catenin 신호전달 경로는 다양한 인자에 의해 조절되며 특히, Akt의 활성화는 β-catenin의 인산화(ser552) 및 GSK3β의 인산화(ser9)를 유도하며, PKA는 β-catenin의 인산화(ser552 및 ser675)를 유도하여 결국 Wnt/β-catenin 신호전달 경로를 활성화 하게 된다. 그 이후에 β-catenin의 안정화 및 세포핵으로의 이동 촉진 등의 과정이 일어나 target gene의 발현을 조절하게 된다. Lee 등은 minoxidil이 human DPCs 및 C3H mouse에서 β-catenin level에 영향을 미치지 않음을 보고한 반면 Kwack 등은 minoxidil이 human DPCs에서PKA, Akt 및 GSK3β의 활성화를 경유한 β-catenin 신호전달 경로의 조절을 통해 육모 효능을 나타냄을 보고하고 있다.
또한, 본 발명의 일 실시예에서 3주 동안 성장기 쥐의 비 모낭(rat vibrissa follicles)을 배양하면서 밀기울 헥산 분획 처리군의 육모효능을 조사하였다. 특히, 10 및 50 μg/ml의 농도에서 밀기울 헥산 분획 처리군은 양성대조군인 10 μM 미녹시딜보다 더 높은 헤어-섬유(hair-fiber)의 성장 효능을 나타내었다.
상기 실시예를 통하여 밀기울 추출물은 탈모 방지 및 육모 촉진 효능이 우수함을 알 수 있다.
결론적으로 본 발명의 밀기울 추출물은 모발성장에 매우 중요한 역할을 하는 모유두 세포의 성장 증식을 촉진하여 모낭의 성장기를 유도하거나 성장기를 유지시키는 데 작용할 것으로 생각된다.
본 발명의 밀기울 추출물을 유효성분으로 함유하는 약학 조성물은 탈모 방지 및 육모 촉진용 정제, 캅셀제, 주사제, 크림, 젤, 패취, 분무제, 연고제, 경고제, 로션제, 리니멘트제, 파스타제 또는 카타플라스마제 형태의 약학 조성물로 제조하여 사용할 수 있으나, 이에 한정하는 것은 아니다.
본 발명의 밀기울 추출물의 바람직한 투여량은 환자의 상태 및 체중, 질병의 정도, 약물 형태, 투여 경로 및 기간에 따라 다르지만, 통상의 기술자에 의해 적절하게 선택될 수 있다. 그러나 바람직한 효과를 위해서, 본 발명의 추출물은 1일 0.0001 내지 100 ㎎/㎏으로, 바람직하게는 0.001 내지 10 ㎎/㎏으로 투여하는 것이 좋다. 투여는 하루에 한번 투여할 수도 있고, 수회 나누어 투여할 수도 있다. 상기 투여량은 어떠한 면으로든 본 발명의 범위를 한정하는 것은 아니다.
또한, 본 발명의 밀기울 추출물은 탈모 방지 및 발모를 활성시키는 효과를 갖는 모발 화장품에 다양하게 이용될 수 있다. 본 조성물을 첨가할 수 있는 제품으로는, 예를 들어, 헤어토닉, 헤어컨디셔너, 헤어에센스, 헤어로션, 헤어영양로션, 헤어샴푸, 헤어린스, 헤어트리트먼트, 헤어크림, 헤어영양크림, 헤어모이스처크림, 헤어맛사지크림, 헤어왁스, 헤어 에어로졸, 헤어팩, 헤어영양팩, 헤어비누, 헤어클렌징폼, 머릿기름, 모발건조제, 모발보존처리제, 모발염색제, 모발용 웨이브제, 모발탈색제, 헤어겔, 헤어글레이즈, 헤어드레싱어, 헤어래커, 헤어모이스처라이저, 헤어무스 또는 헤어스프레이 등과 같은 화장품류 등이 있다.
본 발명의 화장료는 수용성 비타민, 유용성 비타민, 고분자 펩티드, 고분자 다당, 스핑고지질 및 해초 엑기스로 이루어진 군에서 선택된 조성물을 포함할 수 있다.
수용성 비타민으로서는 화장품에 배합 가능한 것이라면 어떠한 것이라도 되지만, 바람직하게는 비타민 B1, 비타민 B2, 비타민 B6, 피리독신, 염산피리독신, 비타민 B12, 판토텐산, 니코틴산, 니코틴산아미드, 엽산, 비타민 C, 비타민 H 등을 들 수 있으며, 그들의 염(티아민염산염, 아스코르빈산나트륨염 등)이나 유도체(아스 코르빈산-2-인산나트륨염, 아스코르빈산-2-인산마그네슘염 등)도 본 발명에서 사용할 수 있는 수용성 비타민에 포함된다. 수용성 비타민은 미생물 변환법, 미생물의 배양물로부터의 정제법, 효소법 또는 화학 합성법 등의 통상의 방법에 의해 수득할 수 있다.
*유용성 비타민으로서는 화장품에 배합 가능한 것이라면 어떠한 것이라도 되지만, 바람직하게는 비타민 A, 카로틴, 비타민 D2, 비타민 D3, 비타민 E(d1-알파 토코페롤, d-알파 토코페롤, d-알파 토코페롤) 등을 들 수 있으며, 그들의 유도체(팔미틴산아스코르빈, 스테아르산아스코르빈, 디팔미틴산아스코르빈, 아세트산 dl-알파 토코페롤, 니코틴산 dl-알파 토코페롤비타민 E, DL-판토테닐알코올, D-판토테닐알코올, 판토테닐에틸에테르 등) 등도 본 발명에서 사용되는 유용성 비타민에 포함된다. 유용성 비타민은 미생물 변환법, 미생물의 배양물로부터의 정제법, 효소 또는 화학 합성법 등의 통상의 방법에 의해 취득할 수 있다.
*고분자 펩티드로서는 화장품에 배합 가능한 것이라면 어떠한 것이라도 되지만, 바람직하게는 콜라겐, 가수 분해 콜라겐, 젤라틴, 엘라스틴, 가수 분해 엘라스틴, 케라틴 등을 들 수 있다. 고분자 펩티드는 미생물의 배양액으로부터의 정제법, 효소법 또는 화학 합성법 등의 통상의 방법에 의해 정제 취득할 수 있으며, 또는 통상 돼지나 소 등의 진피, 누에의 견섬유 등의 천연물로부터 정제하여 사용할 수 있다. 고분자 다당으로서는 화장품에 배합 가능한 것이라면 어떠한 것이라도 되지만, 바람직하게는 히드록시 에틸셀룰로오스, 크산탄검, 히알루론산나트륨, 콘드로이틴 황산 또는 그 염(나트륨염 등) 등을 들 수 있다. 예를 들어, 콘드로이틴 황산 또는 그 염 등은 통상 포유동물이나 어류로부터 정제하여 사용할 수 있다.
스핑고 지질로서는 화장품에 배합 가능한 것이라면 어떠한 것이라도 되지만, 바람직하게는 세라미드, 피토스핑고신, 스핑고당지질 등을 들 수 있다. 스핑고 지질은 통상 포유류, 어류, 패류, 효모 또는 식물 등으로 부터 통상의 방법에 의해 정제하거나 화학 합성법에 의해 취득할 수 있다.
해초 엑기스로는 화장품에 배합 가능한 것이라면 어떠한 것이라도 되지만, 바람직하게는 갈조 엑기스, 홍조 엑기스, 녹조 엑기스 등을 들 수 있으며, 또, 이들의 해초 엑기스로부터 정제된 칼라기난, 아르긴산, 아르 긴산나트륨, 아르긴산칼륨 등도 본 발명에서 사용되는 해초 엑기스에 포함된다. 해초 엑기스는 해초로부터 통상의 방법에 의해 정제하여 취득할 수 있다.
본 발명의 화장료에는 상기 필수 성분과 더불어 필요에 따라 통상 화장료에 배합되는 다른 성분을 배합해도 된다.
이외에 첨가해도 되는 배합 성분으로서는 유지 성분, 보습제, 에몰리엔트제, 계면 활성제, 유기 및 무기 안료, 유기 분체, 자외선 흡수제, 방부제, 살균제, 산화 방지제, 식물 추출물, pH 조정제, 알콜, 색소, 향료, 혈행 촉진제, 냉감제, 제한(制汗)제, 정제수 등을 들 수 있다.
유지 성분으로서는 에스테르계 유지, 탄화수소계 유지, 실리콘계 유지, 불소계 유지, 동물 유지, 식물 유지 등을 들 수 있다.
에스테르계 유지로서는 트리2-에틸헥산산글리세릴, 2-에틸헥산산세틸, 미리스틴산이소프로필, 미리스틴산부틸, 팔미틴산이소프로필, 스테아르산에틸, 팔미틴산옥틸, 이소스테아르산이소세틸, 스테아르산부틸, 리놀레산에틸, 리놀레산이소프로필, 올레인산에틸, 미리스틴산이소세틸, 미리스틴산이소스테아릴, 팔미틴산이소스테아릴, 미리스틴산옥틸도데실, 이소스테아르산이소세틸, 세바신산디에틸, 아디핀산디이소프로필, 네오펜탄산이소알킬, 트리(카프릴, 카프린산)글리세릴, 트리2-에틸헥산산트리메틸롤프로판, 트리이소스테아르산트리메틸롤프로판, 테트라2-에틸헥산산펜타엘리슬리톨, 카프릴산세틸, 라우린산데실, 라우린산헥실, 미리스틴산데실, 미리스틴산미리스틸, 미리스틴산세틸, 스테아르산스테아릴, 올레인산데실, 리시노올레 인산세틸, 라우린산이소스테아릴, 미리스틴산이소트리데실, 팔미틴산이소세틸, 스테아르산옥틸, 스테아르산이소세틸, 올레인산이소데실, 올레인산옥틸도데실, 리놀레산옥틸도데실, 이소스테아르산이소프로필, 2-에틸헥산산세 토스테아릴, 2-에틸헥산산스테아릴, 이소스테아르산헥실, 디옥탄산에틸렌글리콜, 디올레인산에틸렌글리콜, 디카프린산프로필렌글리콜, 디(카프릴,카프린산)프로필렌글리콜, 디카프릴산프로필렌글리콜, 디카프린산네오펜틸글 리콜, 디옥탄산네오펜틸글리콜, 트리카프릴산글리세릴, 트리운데실산글리세릴, 트리이소팔미틴산글리세릴, 트리 이소스테아르산글리세릴, 네오펜탄산옥틸도데실, 옥탄산이소스테아릴, 이소노난산옥틸, 네오데칸산헥실데실, 네오데칸산옥틸도데실, 이소스테아르산이소세틸, 이소스테아르산이소스테아릴, 이소스테아르산옥틸데실, 폴리글리세린올레인산에스테르, 폴리글리세린이소스테아르산에스테르, 시트르산트리이소세틸, 시트르산트리이소알킬, 시트르산트리이소옥틸, 락트산라우릴, 락트산미리스틸, 락트산세틸, 락트산옥틸데실, 시트르산트리에틸, 시트르산아세틸트리에틸, 시트르산아세틸트리부틸, 시트르산트리옥틸, 말산디이소스테아릴, 히드록시스테아르산 2-에틸헥실, 숙신산디2-에틸헥실, 아디핀산디이소부틸, 세바신산디이소프로필, 세바신산디옥틸, 스테아르산콜레스테릴, 이소스테아르산콜레스테릴, 히드록시스테아르산콜레스테릴, 올레인산콜레스테릴, 올레인산디히드로콜레스테릴, 이소스테아르산피트스테릴, 올레인산피트스테릴, 12-스테알로일히드록시스테아르산이소 세틸, 12-스테알로일히드록시스테아르산스테아릴, 12-스테알로일히드록시스테아르산이소스테아릴 등의 에스테르계 등을 들 수 있다.
탄화수소계 유지로서는 스쿠알렌, 유동 파라핀, 알파-올레핀올리고머, 이소파라핀, 세레신, 파라핀, 유동 이소파라핀, 폴리부덴, 마이크로크리스탈린왁스, 와셀린 등의 탄화 수소계 유지 등을 들 수 있다.
실리콘계 유지로서는 폴리메틸실리콘, 메틸페닐실리콘, 메틸시클로폴리실록산, 옥타메틸폴리실록산, 데카메틸폴리실록산, 도데카메틸시클로실록산, 디메틸실록산ㆍ메틸세틸옥시실록산 공중합체, 디메틸실록산ㆍ메틸스테알록시실록산 공중합체, 알킬 변성 실리콘유, 아미노 변성 실리콘유 등을 들 수 있다.
불소계 유지로서는 퍼플루오로폴리에테르 등을 들 수 있다.
동물 또는 식물 유지로서는 아보카도유, 아르몬드유, 올리브유, 참깨유, 쌀겨유, 새플라워유, 대두유, 옥수수유, 유채유, 행인(杏仁)유, 팜핵유, 팜유, 피마자유, 해바라기유, 포도종자유, 면실유, 야자유, 쿠쿠이너트유, 소맥배아유, 쌀 배아유, 시아버터, 월견초유, 마커데이미아너트유, 메도홈유, 난황유, 우지(牛脂), 마유, 밍크유, 오렌지라피유, 호호바유, 캔데리러왁스, 카르나바왁스, 액상 라놀린, 경화피마자유 등의 동물 또는 식물 유지를 들 수 있다.
보습제로서는 수용성 저분자 보습제, 지용성 분자 보습제, 수용성 고분자, 지용성 고분자 등을 들 수 있다. 수용성 저분자 보습제로서는 세린, 글루타민, 솔비톨, 만니톨, 피롤리돈-카르복실산나트륨, 글리세린, 프로필렌글리콜, 1,3-부틸렌글리콜, 에틸렌글리콜, 폴리에틸렌글리콜B(중합도 n = 2 이상), 폴리프로필렌글리콜(중합도 n = 2 이상), 폴리글리세린B(중합도 n = 2 이상), 락트산, 락트산염 등을 들 수 있다.
지용성 저분자 보습제로서는 콜레스테롤, 콜레스테롤에스테르 등을 들 수 있다. 수용성 고분자로서는 카르복시비닐폴리머, 폴리아스파라긴산염, 트라가칸트, 크산탄검, 메틸셀룰로오스, 히드록시메틸셀룰로오스, 히드록시에틸셀룰로오스, 히드록시프로필셀룰로오스, 카르복시메틸셀 룰로오스, 수용성 키틴, 키토산, 덱스트린 등을 들 수 있다.
지용성 고분자로서는 폴리비닐피롤리돈ㆍ에이코센 공중합체, 폴리비닐피롤리돈ㆍ헥사데센 공중합체, 니트로셀룰로오스, 덱스트린지방산에스테르, 고분자 실리콘 등을 들 수 있다.
에몰리엔트제로서는 장쇄아실글루타민산콜레스테릴에스테르, 히드록시스테아르산콜레스테릴, 12-히드록시스테아르산, 스테아르산, 로진산, 라놀린지방산콜레스테릴에스테르 등을 들 수 있다.
계면 활성제로서는 비이온성 계면 활성제, 음이온성 계면 활성제, 양이온성 계면 활성제, 양성 계면 활성제 등을 들 수 있다.
비이온성 계면 활성제로서는 자기 유화형 모노스테아르산글리세린, 프로필렌글리콜지방산에스테르, 글리세린지방산에스테르, 폴리글리세린지방산에스테르, 솔비탄지방산에스테르, POE(폴리옥시에틸렌) 솔비탄지방산에스테르, POE 솔비트지방산에스테르, POE 글리세린지방산에스테르, POE 알킬에테르, POE 지방산에스테르, POE 경화피마자유, POE 피마자유, POEㆍPOP(폴리옥시에틸렌ㆍ폴리옥시프로필렌) 공중합체, POEㆍPOP 알킬에테르, 폴리에테르변성실리콘, 라우린산알카놀아미드, 알킬아민옥시드, 수소첨가대두인지질 등을 들 수 있다.
음이온성 계면 활성제로서는 지방산비누, 알파-아실술폰산염, 알킬술폰산염, 알킬알릴술폰산염, 알킬나 프탈렌술폰산염, 알킬황산염, POE 알킬에테르황산염, 알킬아미드황산염, 알킬인산염, POE 알킬인삼염, 알킬아미드인산염, 알킬로일알킬타우린염, N-아실아미노산염, POE 알킬에테르카르복실산염, 알킬술포숙신산염, 알킬술포아세트산나트륨, 아실화 가수분해 콜라겐펩티드염, 퍼플루오로알킬인산에스테르 등을 들 수 있다.
양이온성 계면 활성제로서는 염화알킬트리메틸암모늄, 염화스테아릴트리메틸암모늄, 브롬화스테아릴트 리메틸암모늄, 염화세토스테아릴트리메틸암모늄, 염화디스테아릴디메틸암모늄, 염화스테아릴디메틸벤질암모늄, 브롬화베헤닐트리메틸암모늄, 염화벤잘코늄, 스테아르산디에틸아미노에틸아미드, 스테아르산디메틸아미노프로필 아미드, 라놀린 유도체 제 4급 암모늄염 등을 들 수 있다.
양성 계면 활성제로서는 카르복시베타인형, 아미드베타인형, 술포베타인형, 히드록시술포베타인형, 아미드술포베타인형, 포스포베타인형, 아미노카르복실산염형, 이미다졸린 유도체형, 아미드아민형 등의 양성 계면 활성제 등을 들 수 있다.
유기 및 무기 안료로서는 규산, 무수규산, 규산마그네슘, 탤크, 세리사이트, 마이카, 카올린, 벵갈라, 클레이, 벤토나이트, 티탄피막운모, 옥시염화비스무트, 산화지르코늄, 산화마그네슘, 산화아연, 산화티탄, 산화 알루미늄, 황산칼슘, 황산바륨, 황산마그네슘, 탄산칼슘, 탄산마그네슘, 산화철, 군청, 산화크롬, 수산화크롬, 칼라민 및 이들의 복합체등의 무기 안료; 폴리아미드, 폴리에스테르, 폴리프로필렌, 폴리스티렌, 폴리우레탄, 비닐수지, 요소수지, 페놀수지, 불소수지, 규소수지, 아크릴수지, 멜라민수지, 에폭시수지, 폴리카보네이트수지, 디비닐벤젠ㆍ스티렌 공중합체, 실크파우더, 셀룰로오스, CI 피그먼트옐로우, CI 피그먼트 오렌지 등의 유기 안료 및 이들의 무기 안료와 유기 안료의 복합 안료 등을 들 수 있다.
유기 분체로서는 스테아르산칼슘 등의 금속비누; 세틸린산아연나트륨, 라우릴린산아연, 라우릴린산칼 슘 등의 알킬인산금속염; N-라우로일-베타-알라닌칼슘, N-라우로일-베타-알라닌아연, N-라우로일글리신칼슘 등의 아실아미노산 다가금속염; N-라우로일-타우린칼슘, N-팔미토일-타우린칼슘 등의 아미드술폰산 다가금속염; N-엡실론-라우로일-L-리진, N-엡실론-팔미토일리진, N-알파-파리토일올니틴, N-알파-라우로일아르기닌, N-알파-경화우지지방산아실아르기닌 등의 N-아실염기성아미노산; N-라우로일글리실글리신 등의 N-아실폴리펩티드; 알파-아미노카프릴산, 알파-아미노라우린산 등의 알파-아미노지방산 ; 폴리에틸렌, 폴리프로필렌, 나일론, 폴리메 틸메타크릴레이트, 폴리스티렌, 디비닐벤젠ㆍ스티렌 공중합체, 사불화에틸렌 등을 들 수 있다.
자외선 흡수제로서는 파라아미노벤조산, 파라아미노벤조산에틸, 파라아미노벤조산아밀, 파라아미노벤조산옥틸, 살리실산에틸렌글리콜, 살리신산페닐, 살리신산옥틸, 살리신산벤질, 살리신산부틸페닐, 살리신산호모멘틸, 계피산벤질, 파라메톡시계피산-2-에톡시에틸, 파라메톡시계피산옥틸, 디파라메톡시계피산모노-2-에틸헥산글리세릴, 파라메톡시계피산이소프로필, 디이소프로필ㆍ디이소프로필계피산에스테르 혼합물, 우로카닌산, 우로카닌산 에틸, 히드록시메톡시벤조페논, 히드록시메톡시벤조페논술폰산 및 그 염, 디히드록시메톡시벤조페논, 디히드록 시메톡시벤조페논디술폰산나트륨, 디히드록시벤조페논, 테트라히드록시벤조페논, 4-tert-부틸-4'-메톡시디벤조일메탄, 2,4,6-트리아닐리노-p-(카르보-2'-에틸헥실-1'-옥시)-1,3,5-트리아진, 2-(2-히드록시-5-메틸페닐)벤조 트리아졸 등을 들 수 있다.
살균제로서는 히노키티올, 트리클로산, 트리클로로히드록시디페닐에테르, 크로르헥시딘글루콘산염, 페녹시에탄올, 레조르신, 이소프로필메틸페놀, 아줄렌, 살리칠산, 진크필리티온, 염화벤잘코늄, 감광소 301호, 모노니트로과이어콜나트륨, 운데시렌산 등을 들 수 있다.
산화방지제로서는 부틸히드록시아니솔, 갈릭산프로필, 엘리소르빈산 등을 들 수 있다.
pH 조정제로서는 시트르산, 시트르산나트륨, 말산, 말산나트륨, 프말산, 프말산나트륨, 숙신산, 숙신산 나트륨, 수산화나트륨, 인산일수소나트륨 등을 들 수 있다.
알코올로서는 세틸알코올(Cetyl Alcohol) 등의 고급 알코올을 들 수 있다.
또한, 이외에 첨가해도 되는 배합 성분은 이에 한정되는 것은 아니며, 또, 상기 어느 성분도 본 발명의 목적 및 효과를 손상시키지 않는 범위 내에서 배합 가능하지만, 총 중량에 대하여 바람직하게는 0.01 - 50 % 중량 백분율로 배합될 수 있다.
본 발명의 화장료는 용액, 유화물, 점성형 혼합물 등의 형상을 취할 수 있다.
본 발명의 화장료 조성물에 포함되는 성분은 유효성분으로서 상기 추출물 이외에 화장료 조성물에 통상 적으로 이용되는 성분들을 포함할 수 있으며, 예를 들면, 안정화제, 용해화제, 비타민, 안료 및 향료와 같은 통 상적인 보조제 및 담체를 포함할 수 있다.
본 발명의 화장료 조성물은 당업계에서 통상적으로 제조되는 어떠한 제형으로도 제조될 수 있으며, 예를 들어 유액, 크림, 화장수, 팩, 파운데이션, 로션, 미용액, 모발화장료 등을 들 수 있다.
본 발명의 제형이 페이스트, 크림 또는 겔인 경우에는 담체 성분으로서 동물섬유, 식물섬유, 왁스, 파라핀, 전분, 트라칸트, 셀룰로오스 유도체, 폴리에틸렌글리콜, 실리콘, 벤토나이트, 실리카, 탈크 또는 산화아연 등이 이용될 수 있다.
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 디메틸에테르와 같은 추진체를 포함할 수 있다.
본 발명의 제형이 용액 또는 유탁액의 경우에는 담체 성분으로서 용매, 용매화제 또는 유탁화제가 이용될 수 있고, 예컨대 물, 에탄올, 이소프로판올, 에틸 카보네이트, 에틸 아세테이트, 벤질 알코올, 벤질 벤조에이트, 프로필렌글리콜, 1,3-부틸글리콜 오일, 글리세롤 지방족 에스테르, 폴리에틸렌 글리콜 또는 소르비탄의 지방산 에스테르 등일 수 있다.
본 발명의 제형이 현탁액인 경우에는 담체 성분으로서 물, 에탄올 또는 프로필렌 글리콜과 같은 액상 희석제, 에톡실화 이소스테아릴 알코올, 폴리옥시에틸렌 소르비톨 에스테르 및 폴리옥시에틸렌 소르비탄 에스테르와 같은 현탁제, 미소결정성 셀룰로오스, 알루미늄 메타히드록시드, 벤토나이트, 아가 또는 트라칸트 등이 이용될 수 있다.
본 발명의 제형이 계면-활성제 함유 클린징인 경우에는 담체 성분으로서 지방족 알코올 설페이트, 지방족 알코올 에테르설페이트, 설포숙신산 모노에스테르, 이세티오네이트, 이미다졸리늄 유도체, 메틸타우레이트, 사르코시네이트, 지방산 아미드 에테르 설페이트, 알킬아미도베타인, 지방족 알코올, 지방산 글리세리드, 지방산 디에탄올아미드, 식물성유, 리놀린 유도체 또는 에톡실화 글리세롤 지방산 에스테르 등이 이용될 수 있다.
또한, 본 발명은 또한 본 발명은 밀기울의 추출물을 유효성분으로 포함하는 탈모 방지 또는 육모 촉진용 건강기능성 식품을 제공한다.
본 발명의 밀기울 추출물을 첨가할 수 있는 식품으로는, 예를 들어, 각종 식품류, 음료, 껌, 차, 비타민 복합제, 건강보조식품류 등이 있고, 분말, 과립, 정제, 캡슐 또는 음료인 형태로 사용할 수 있다.
본 발명의 밀기울 추출물 자체는 독성 및 부작용은 거의 없으므로 예방 목적으로 장기간 복용시에도 안심하고 사용할 수 있는 약제이다.
본 발명의 밀기울 추출물은 탈모 질환의 예방 및 개선을 목적으로 식품 또는 음료에 첨가될 수 있다. 이때, 식품 또는 음료 중의 상기 추출물의 양은 일반적으로 본 발명의 건강식품조성물은 전체 식품 중량의 0.01 내지 50 중량%로 가할 수 있으며, 건강 음료 조성물은 100 ㎖를 기준으로 0.02 내지 10 g, 바람직하게는 0.3 내지 1 g의 비율로 가할 수 있다.
본 발명의 건강 음료 조성물은 지시된 비율로 필수 성분으로서 상기 추출물을 함유하는 것 외에 액체성분에는 특별한 제한점은 없으며 통상의 음료와 같이 여러 가지 향미제 또는 천연 탄수화물 등을 추가 성분으로서 함유할 수 있다. 상술한 천연 탄수화물의 예는 모노사카라이드, 예를 들어, 포도당, 과당 등의 디사카라이드, 예를 들어 말토스, 슈크로스 등의 및 폴리사카라이드, 예를 들어 덱스트린, 시클로덱스트린 등과 같은 통상적인 당 및 자일리톨, 소르비톨, 에리트리톨 등의 당알콜이다. 상술한 것 이외의 향미제로서 천연 향미제(타우마틴, 스테비아 추출물, 레바우디오시드 A, 글리시르히진등) 및 합성 향미제(사카린, 아스파르탐 등)를 유리하게 사용할 수 있다. 상기 천연 탄수화물의 비율은 본 발명의 조성물 100 ㎖당 일반적으로 약 1 내지 20g, 바람직하게는 약 5 내지 12g이다.
상기 외에 본 발명의 조성물은 여러 가지 영양제, 비타민, 광물(전해질), 합성 풍미제 및 천연 풍미제 등의 풍미제, 착색제 및 중진제(치즈, 초콜릿 등), 펙트산 및 그의 염, 알긴산 및 그의 염, 유기산, 보호성 콜로이드 증점제, pH 조절제, 안정화제, 방부제, 글리세린, 알콜, 탄산 음료에 사용되는 탄산화제 등을 함유할 수 있다. 그밖에 본 발명의 조성물들은 천연 과일 쥬스 및 과일 쥬스 음료 및 야채 음료의 제조를 위한 과육을 함유할 수 있다. 이러한 성분은 독립적으로 또는 조합하여 사용할 수 있다. 이러한 첨가제의 비율은 그렇게 중요하진 않지만 본 발명의 조성물 100 중량부 당 0 내지 약 20 중량부의 범위에서 선택되는 것이 일반적이다.
이하, 본 발명의 이해를 돕기 위하여 실시예를 들어 상세하게 설명하기로 한다. 다만 하기의 실시예는 본 발명의 내용을 예시하는 것일 뿐 본 발명의 범위가 하기 실시예에 한정되는 것은 아니다. 본 발명의 실시예는 이 기술 분야에서 평균적인 지식을 가진 자에게 본 발명을 보다 완전하게 설명하기 위해 제공되는 것이다.
<실시예 1> 밀기울의 획득 및 추출
본 발명에서 사용한 밀기울은 경상북도에서 재배된 우리밀의 밀기울을 정미소에서 공급받은 것이다. 총 밀기울 시료 5 kg을 90 % 에탄올에 4 시간씩 3회 워터 배스(Water Bath)에서 중탕으로 추출하였다. 그 다음 상층액을 회수하고 회전감압농축기(rotary evaporator)를 사용하여 감압 농축한 후 조추출물을 337 g수득하였다. 이중 300 g은 n-헥산(n-hexane)을 이용하여 용매 분획하여 n-헥산층 133 g을 얻었고 이 분획물을 실험의 시료로 사용하였다.
<실시예 2> 모유두 세포의 증식 효능
<실시예 2-1> 모유두 세포의 증식 확인
흰쥐 수염에서 분리된 불멸화된 모유두 세포(Rat vibrissa immortalized dermal papilla cell)를 100 units/ml 페니실린-100 μg/ml 스트렙토마이신(Gibco Inc, NY, USA)과 10% 열-불활성화 우태혈청( heat-inactivated fetal bovine serum, FBS; Gibco Inc, NY, USA)이 함유된 DMEM(Hyclone Inc, USA) 배지를 사용하여 37℃, 5% CO2 항온기에서 배양하였으며, 3 일에 한 번씩 계대배양 하였다.
모유두 세포(dermal papilla cell)의 증식은 MTT 에세이를 이용하여 측정하였다. 모유두 세포(dermal papilla cell) 1.0×104 cells/ml를 1% FBS를 포함한 DMEM 배지에 혼탁하여 96 웰 플레이트(well plate)에 넣고 24시간 배양 후 밀기울 헥산 분획을 0.1, 1, 10, 50 및 100 μg/ml의 농도로 처리하였다. 양성 대조군인 미녹시딜(minoxidil, Sigma, MO, USA)는 10 μM의 농도로 처리하였다.
4 일 동안 배양한 후 50 ㎕의 MTT(Sigma, MO, USA)을 첨가하고 4 시간 동안 반응시켰다. 상층액은 제거하고 DMSO 200 ㎕을 가하여 침전물을 용해시킨 후 VERSAmax 마이크로플레이트 리더(VERSAmax microplate reader, Molecular Devices, CA, USA)를 사용하여 540 nm에서 흡광도를 측정하였다. 각 시료 첨가군에 대한 평균 흡광도 값을 구하였으며, 대조군의 흡광도 값과 비교하여 증식 정도를 조사하였다.
모든 측정 결과는 평균 ± 표준편차로 나타내었으며, 통계학적 유의성 검정은 스튜던트 t-검정(Student's t-test)으로 검정하였으며, p-값이 0.05 이하일 경우 유의성을 인정하였다. 통계 처리는 윈도우용 SPSS 12.0K(Release 12.0.1. SPSS Inc. USA)를 사용하였다.
그 결과 밀기울 헥산 분획을 0.1, 1, 10, 50 및 100 μg/ml의 농도로 처리하였을 때, 대조군에 비하여 각각 106.9±11.5%, 115.1±5.2%(P<0.01), 139.3±5.4%(P<0.001), 190.5±6.5%(P<0.001)및 183.3±10.2%(P<0.001)로 모유두 세포(dermal papilla cell)의 증식이 증가하였다(표 1, 도 1).
표 1
시 료 | 농 도(/ml) | 모유두 세포 증식률(%) |
미녹시딜 | 10 M | 112.3±2.7(P<0.01) |
밀기울 헥산 분획 | 0.1 | 106.9±11.5(P<0.01) |
1 | 115.1±5.2(P<0.01) | |
10 | 139.3±5.4(P<0.001) | |
50 | 190.5±6.5(P<0.001) | |
100 | 183.3±10.2(P<0.001) |
특히, 10, 50 및 100 μg/ml의 농도에서 밀기울 헥산 분획 처리 실험군은 양성 대조군으로 사용한 10 μM 미녹시딜의 증식 효과인 112.3±2.7%(P<0.01) 보다 높은 증식 효능을 나타내었으며, 밀기울 헥산 분획이 미녹시딜 같은 육모 효능을 나타낼 수 있음을 보여준다.
<실시예 2-2> 모유두 세포에서의 세포 주기 단백질 발현 확인
밀기울헥산(hexane) 추출물이 모유두 세포의 성장증식을 증가시키는 분자적 기전을 밝히기 위해 세포 주기(cell cycle) 단백질의 발현을 관찰하였다. 구체적으로, 모유두 세포(Dermal papilla cell) (1.0×105 cell/ml in 100 mm 디쉬)를 1% FBS를 포함하는 DMEM 배지에서 24 시간 동안 배양한 후에 밀기울 추출물(1, 10 및 50 μg/ml) 및 10 μM minoxidil을 각각 처리한 다음 24시간 동안 배양하였다. 세포는 PBS로 2회 세척한 후 200 ㎕의 lysis buffer[50 mM Tris-HCl(pH 7.5), 150 mM NaCl, 2 mM EDTA, 1 mM EGTA, 1 mM NaVO3, 10 mM NaF, 1 mM dithiothreitol(DTT), 1 mM phenylmethylsulfonylfluoride(PMSF), 25 μg/mL aprotinin, 25 μg/mL leupeptin 및 1% NP-40]를 첨가한 다음, 4℃에서 30 분 동안 세포파쇄하였다. 세포 파쇄액(lysate)을 15,000 rpm에서 15분 동안 원심분리하여 상층액을 얻었고 실험에 사용하기 전까지 -20℃에서 보관하였다. 단백질 농도는 BSA(bovine serum albumin)를 표준물질로 사용하여 브래드포드(Bradford) 방법으로 정량하였다. 20-30μg의 세포파쇄액을 8-12% mini gel SDS-PAGE(Sodium dodecyl sulfate polyacrylamide gel electrophoresis)로 변성분리하여 PVDF(polyvinylidene fluoride) 멤브레인(Bio-Rad, Hercules, CA, USA)으로 200 mA에서 2시간 동안 트랜스퍼(transfer)하였다. 또한, 멤브레인의 blocking은 5% nonfat dried milk가 함유된 Tween-20-TBS(T-TBS)(50 mM Tris, pH 7.6, 150 mM NaCl, 0.1% Tween-20) 용액에서 2시간 동안 실시하였다. 그 후, 상기 멤브레인에 Cyclin D1(1:1000), phospho-CDK2(1:1000), Cyclin E(1:2000), phospho(ser780)-pRB(1:1000), phospho-ERK1/2(1:1000), ERK1/2(1:1000), phospho(ser552)-β-catenin(1:1000), phospho(ser675)-β-catenin(1:1000), β-catenin(1:2000), phospho(ser9)-GSK3β(1:1000), GSK3β(1:1000) 및 β-actin(1:5000)에 대한 일차 항체(primary antibody)를 첨가한 뒤 4℃에서 오버나이트하였다. 이차 항체(Secondary anbibody)는 HRP(Horse Radish Peroxidase)가 결합된 항-rabbit IgG, 항-mouse IgG를 1:5000으로 희석한 항체를 이용하여 1시간 동안 상온에서 항체 결합 반응을 진행하였다. 그 뒤, T-TBS로 멤브레인을 3회 세척한 후, ECL 기질(Intron, Seoul, Korea)로 1분 동안 반응시킨 다음 X-ray film(AGFA, Mortsel, Belgium)에 감광하였다.
그 결과, 밀기울 헥산 추출물을 모유두 세포에 24시간 동안 1, 10 및 50 μg/ml 농도로 처리하였을 때, cyclin D1, cyclin E, phospho-CDK2 및 phospho-pRB의 발현이 증가함을 확인할 수 있었고, minoxidil의 처리 역시 밀기울 헥산 추출물과 비슷한 결과를 보임을 확인하였다(도 2A). 또한, 밀기울 헥산 추출물 처리에서는 Phospho(ser552)-β-catenin, Phospho(ser675)-β-catenin, Phospho(ser9)-GSK3β의 활성화가 증가함을 확인하였고, minoxidil의 처리는 Wnt/β-catenin 경로를 다소 활성화시킴을 관찰하였다(도 2B). 이러한 결과들은 밀기울 hexane 추출물이 cyclin D1, cyclin E, phospho-CDK2, phospho-pRB같은 cell cycle 단백질뿐만 아니라 GSK3β, PKA및 Akt 경로를 통한 Wnt/β-catenin 신호전달 경로의 활성화에 의해 모유두 세포의 증식증가 효능을 나타냄을 시사한다.
<실시예 3> 육모 효능 측정: 헤어-섬유(Hair-fiber) 길이 성장 측정
<실시예 3-1> 쥐의 비 모낭(Rat vibrissa follicles)의 분리 및 배양
생후 3주령인 위스터 랫(Wistar rat) 수컷(Japan SLC, Hamamatsu, Janpan)을 ㈜ 중앙실험동물로부터 구입하여 에틸 에테르(ethyl ether)로 마취 후 경추도살하였다. 랫(Rat) 왼쪽과 오른쪽 미스타셜 패드(mystacial pads)를 분리하여 100 units/ml 페니실린-100 μg/ml 스트렙토마이신(streptomycin, Gibco Inc, NY, USA)이 함유된 E/P buffer [Earle's balanced salts solution(EBSS, Sigma MO, USA) + phosphate-buffered saline(PBS, Sigma MO, USA)]에 넣었다. 해부 현미경으로 관찰하며 비 모낭(vibrissa follicles)을 조심스럽게 분리하였다. 모낭이 모두 분리될 때까지 E/P buffer를 넣은 페트리 디쉬에 분리된 모낭을 넣고 37℃, 5% CO2 항온기에서 1시간 정도 배양하였다. 24 well plate의 각 well에 2 mM L-글루타민(Gibco Inc, NY, USA), 10 μg/ml 인슐린(Sigma MO, USA), 50 nM 하이드로콜티손(hydrocortisone, Sigma MO, USA)과 100 units/ml 페니실린-100 μg/ml 스트렙토마이신을 포함하는 500 ㎕의 윌리엄 E 미디엄(William E medium, Gibco Inc, NY, USA)을 넣고, 하나의 배지에 하나씩의 모낭을 넣어서 37℃, 5% CO2 항온기에서 배양하였다. 배양 중에 배지는 3 일마다 교환하였고 밀기울 헥산 분획은 1, 10 및 50 μg/ml의 농도로 처리하였다. 양성 대조물질인 미녹시딜을 10 μM의 농도로 처리하였다(Kang et al., 2012 Biomol Ther 20(1), 118-124).
<실시예 3-2> 비 모낭(Vibrissa follicles) 성장의 측정
배양중인 비 모낭(vibrissa follicle) 형태는 현미경(Olympus, Japan)을 사용하여 촬영하였다. 모낭 길이는 이미지 분석 프로그램(image analyzer, DP controller; Olympus, Japan)를 사용하여 측정하였다. 모낭 길이 변화의 평균값을 구하고 대조군의 평균길이와 비교하여 성장 정도를 측정하였다.
모든 측정결과는 평균 ± 표준오차로 나타내었으며, 통계학적 유의성 검정은 스튜던트 t-검정(Student's t-test)으로 검정하였으며, p-값이 0.05 이하일 경우 유의성을 인정하였다. 통계 처리는 윈도우용 SPSS 12.0K(Release 12.0.1. SPSS Inc. USA)를 사용하였다.
21일째 밀기울 헥산 분획 처리군과 대조군과의 모낭의 길이 차이(%)를 비교한 결과, 1, 10 및 50 μg/ml을 각각 처리하였을 때 대조군보다 hair fiber의 성장이 150.0±21.7%(P<0.001), 203.6±7.6%(P<0.001) 및 246.4±19.8%(P<0.001)로 증가하였다(표 2, 도 3).
표 2
시 료 | 농 도(/ml) | hair fiber 성장 증가율(%) |
미녹시딜 | 10 M | 156.3±11.5(P<0.05) |
밀기울 헥산 분획 | 1 | 150.0±21.7(P<0.001) |
10 | 203.6±7.6(P<0.001) | |
50 | 246.4±19.8(P<0.001) |
특히 10 및 50 μg/ml의 농도에서 밀기울 헥산 분획의 헤어-섬유(hair-fiber) 성장 증가 효과는 양성대조 물질로 사용한 미녹시딜(10 μM)의 156.3±11.5%(P<0.05) 성장 증가 효과보다 더 높았다.
<실시예 4> 육모 효능 측정: in vivo 실험
<실시예 4-1> 실험동물
생후 6주된 암컷 C57BL/6 마우스를 ㈜ 오리엔트바이오로부터 구입하여 1주간 동물 사육실 환경에 적응 시킨 후 실험에 사용하였다. 동물 사육실은 온도 23±3℃, 12 시간씩 밤낮을 유지하였다. 실험동물은 격리용 마우스 케이지에서 사육하였고, 실험용 사료를 자유로이 급여하였다. 실험군은 5개의 군으로 설정하여 대조군, 양성 대조군인 MINOXYLTM (현대약품) 과 밀기울 헥산 추출물(10, 50 및 100 μg/ml) 처리군으로 하였다. 한 실험군에 각 8마리의 마우스를 실험에 이용하였다.
<실시예 4-1> 육모 효능 측정
육모 효과를 살펴보기 위해, 등 쪽 피부의 색이 분홍색인 휴지기 체모의 7주령을 사용하였다. mouse 등 부위를 소동물용 clipper로 털을 완전히 제거하였다. 하루에 한 번씩 시료를 200 ㎕씩 제모부위에 떨어뜨린 다음에 두드리면서 도포하고 대조군에는 50% 에탄올을 도포하였다. 실험 시작 후 1일, 15일 22일, 29일 및 35일에 털이 자라는 상태를 확인하기 위하여 사진촬영을 실시하였고, 정량적 평가를 위해 dotmatrix planimetry를 수행하였다.
그 결과, 밀기울헥산 추출물 도포군에서 제모 후 22일부터 피부색의 변화가 관찰되었고, 제모 후 29 일 및 35일째에 밀기울 헥산 추출물 도포군은 피부색의 변화 및 모발의 성장을 확인할 수 있었다. 밀기울 헥산 추출물 도포군의 모발의 성장기 유도 효능을 dotmatrix 면적측정법을 이용하여 정량적으로 분석한 결과, 35일째에 50 μg/ml의 농도에서 밀기울 헥산 추출물 도포군은 대조군에 비해 유의하게 높은 성장기 유도 효능을 나타냄을 확인하였다 (도 4). 또한, 양성 대조군인 MINOXYLTM(5% minoxidil 함유)도포군은 제모 후 15일 이후부터 피부색상의 변화 및 뚜렷한 모발 성장 효과를 확인 할 수 있었다 (도 4). 이러한 결과는 밀기울 hexane 추출물이 in vivo에서 모발의 성장기를 유도할 수 있음을 시사한다.
<실시예 5> 통계 분석
모든 측정결과는 평균 ± 표준오차로 나타내었으며, 통계학적 유의성 검정은 student's t-test으로 검정하였으며, p-value가 0.05이하일 경우 유의성을 인정하였다. 통계처리는 SPSS 12.0K for Windows(Release 12.0.1. SPSS Inc. USA)를 사용하였다.
본 발명에서는 밀기울 hexane 추출물이 모발의 성장에서 중요한 역할을 하는 모유두 세포에서 cell cycle proteins, Wnt/β-catenin 신호전달 경로의 활성화에 의한 세포의 성장증식을 촉진 효과를 통해 모낭의 성장기를 유도하거나 성장기를 유지시킴을 밝혔다. 이와 같은 연구결과는 밀기울 헥산 추출물이 탈모치료 및 예방제로 이용될 수 있는 가능성을 가지고 있다는 근거를 제시하는 것이다.
하기에 본 발명의 일 실시예에 따른 조성물의 제제예를 설명하나, 본 발명은 이를 한정하고자 함이 아니라 단지 구체적으로 설명하고자 함이다.
제제예 1. 헤어토닉의 제조
제제예 2. 헤어컨디셔너의 제조
제제예 3. 헤어로션의 제조
제제예 4. 헤어비누의 제조
제제예 5. 친수성 연고제
제제예 6. 정제의 제조
통상적인 정제의 제조 방법에 따라, 상기 성분들을 제시된 함량으로 첨가하여 균일하게 혼합하고, 교반한 후, 과립화하였다. 건조 후 타정기를 사용하여 1 정당 유효 성분인 밀기울 추출물이 33.3 중량%씩 포함되어 있는 목적하는 정제를 제조하였다.
제제예 7. 캅셀제의 제조
통상적인 캅셀제의 제조 방법에 따라, 상기 성분들을 제시된 함량으로 첨가하여 균일하게 혼합한 후, 1 캅셀 당 유효 성분인 밀기울 추출물이 33 중량%씩 포함되도록 적절한 크기의 젤라틴 캅셀에 충진하여 목적하는 캅셀제를 제조하였다.
제제예 8. 액제의 제조
통상의 액제의 제조방법에 따라 정제수에 각각의 성분을 가하여 용해시키고 레몬향을 적량 가한 다음 상기의 성분을 혼합한 다음 정제수를 가하여 전체를 정제수를 가하여 전체 100㎖로 조절한 후 갈색병에 충진하여 멸균시켜 액제를 제조한다.
제제예 9. 건강 식품의 제조
상기의 비타민 및 미네랄 혼합물의 조성비는 비교적 건강식품에 적합한 성분을 바람직한 실시예로 혼합 조성하였지만, 그 배합비를 임의로 변형 실시하여도 무방하며, 통상의 건강식품 제조방법에 따라 상기의 성분을 혼합한 다음, 과립을 제조하고, 통상의 방법에 따라 건강식품 제조에 사용할 수 있다.
제제예 10. 건강 음료의 제조
통상의 건강음료 제조방법에 따라 상기의 성분을 혼합한 다음, 약 1시간 동안 85℃에서 교반 가열한 후, 만들어진 용액을 여과하여 멸균된 2ℓ 용기에 취득하여 밀봉 멸균한 뒤 냉장 보관한 다음 본 발명의 건강음료 조성물제조에 사용한다.
상기 조성비는 비교적 기호음료에 적합한 성분을 바람직한 실시예로 혼합 조성하였지만, 수요계층, 수요국가, 사용용도 등 지역적, 민족적 기호도에 따라서 그 배합비를 임의로 변형 실시하여도 무방하다.
Claims (11)
- 밀기울의 추출물을 유효성분으로 포함하는 탈모 방지 또는 육모 촉진용 약학 조성물.
- 제 1항에 있어서,상기 밀기울 추출물은 조성물 전체 중량에 대해 0.01 내지 50 중량%로 포함되는 것인 약학 조성물.
- 제 1항 또는 제 2항에 있어서,상기 조성물은 정제, 캅셀제, 주사제, 크림, 젤, 패취, 분무제, 연고제, 경고제, 로션제, 리니멘트제, 파스타제 및 카타플라스마제로 이루어진 군에서 선택된 제형인 것을 특징으로 하는 탈모 방지 또는 육모 촉진용 약학 조성물.
- 밀기울의 추출물을 유효성분으로 포함하는 탈모 방지 또는 육모 촉진용 화장료 조성물.
- 제 4항에 있어서,상기 밀기울 추출물은 조성물 전체 중량에 대해 0.01 내지 50 중량%로 포함되는 것인 화장료 조성물.
- 제 4항 또는 제 5항에 있어서,상기 조성물은 헤어토닉, 헤어컨디셔너, 헤어에센스, 헤어로션, 헤어영양로션, 헤어샴푸, 헤어린스, 헤어트리트먼트, 헤어크림, 헤어영양크림, 헤어모이스처크림, 헤어맛사지크림, 헤어왁스, 헤어에어로졸, 헤어팩, 헤어영양팩, 헤어비누, 헤어클렌징폼, 머릿기름, 모발건조제, 모발보존처리제, 모발염색제, 모발용 웨이브제, 모발탈색제, 헤어겔, 헤어글레이즈, 헤어드레싱어, 헤어래커, 헤어모이스처라이저, 헤어무스 및 헤어스프레이로 이루어진 군에서 선택된 제형인 것을 특징으로 하는 탈모 방지 또는 육모 촉진용 화장료 조성물.
- 밀기울의 추출물을 유효성분으로 포함하는 탈모 방지 및 육모 촉진용 건강기능성 식품.
- 제 7항에 있어서,상기 밀기울 추출물은 조성물 전체 중량에 대해 0.01 내지 50 중량%로 포함되는 것인 건강기능성 식품.
- 제 7항 또는 제 8항에 있어서,상기 건강기능성 식품은 분말, 과립, 정제, 캡슐 또는 음료인 탈모 방지 및 육모 촉진용 건강기능성 식품.
- 탈모 부위에 밀기울 추출물을 포함한 약학 조성물을 처리하는 단계를 포함하는 탈모 방지 및 육모 촉진 방법.
- 탈모 방지 및 육모 촉진용 약학 조성물의 제조에 사용하기 위한, 밀기울 추출물의 용도.
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- 2013-05-16 KR KR20130056013A patent/KR101510939B1/ko active IP Right Grant
- 2013-11-22 WO PCT/KR2013/010700 patent/WO2014185610A1/ko active Application Filing
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KR20140135552A (ko) | 2014-11-26 |
CN105392492B (zh) | 2020-04-21 |
CN105392492A (zh) | 2016-03-09 |
KR101510939B1 (ko) | 2015-04-10 |
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