WO2024034866A1 - Composition for preventing hair loss and promoting hair growth including complex fermentation product containing natural vitamins - Google Patents
Composition for preventing hair loss and promoting hair growth including complex fermentation product containing natural vitamins Download PDFInfo
- Publication number
- WO2024034866A1 WO2024034866A1 PCT/KR2023/009580 KR2023009580W WO2024034866A1 WO 2024034866 A1 WO2024034866 A1 WO 2024034866A1 KR 2023009580 W KR2023009580 W KR 2023009580W WO 2024034866 A1 WO2024034866 A1 WO 2024034866A1
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- WO
- WIPO (PCT)
- Prior art keywords
- fermentation product
- fermentation
- rye
- vitamin
- hair
- Prior art date
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- 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]
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- 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
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q5/00—Preparations for care of the hair
-
- 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
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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
- A61K2236/10—Preparation or pretreatment of starting material
- A61K2236/19—Preparation or pretreatment of starting material involving fermentation using yeast, bacteria or both; enzymatic treatment
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2300/00—Mixtures or combinations of active ingredients, wherein at least one active ingredient is fully defined in groups A61K31/00 - A61K41/00
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
- A61K2800/40—Chemical, physico-chemical or functional or structural properties of particular ingredients
- A61K2800/59—Mixtures
- A61K2800/591—Mixtures of compounds not provided for by any of the codes A61K2800/592 - A61K2800/596
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
- A61K2800/80—Process related aspects concerning the preparation of the cosmetic composition or the storage or application thereof
- A61K2800/85—Products or compounds obtained by fermentation, e.g. yoghurt, beer, wine
Definitions
- the present invention relates to cosmetic compositions.
- hair grows from hair follicles on the scalp, and throughout life, the cycle of growth (Anagen), catagen (catagen), and telogen (Telogen) is repeated continuously and goes through creation, growth, and shedding.
- the catagen phase of hair follicles the growth of the hair follicle stops and apoptosis and reorganization of the extracellular matrix occur.
- hair falls out and does not grow again, resulting in hair loss.
- Hair is divided into a visible hair shaft and an invisible hair root.
- the root from which hair grows is called the hair bulb, and the hollow part at the bottom is called the dermal papilla.
- the dermal papilla is the most important part in the creation and growth of hair. It contains blood vessels and nerves, which promote hair growth by delivering nutrients and growth factors to the hair. In other words, since the death and growth reduction of dermal papilla cells leads to a decrease in the number of hairs and hair loss, proliferation of dermal papilla cells is necessary to maintain and increase normal hair.
- vascular endothelial growth factor is a glycoprotein that promotes angiogenesis by inducing the growth of vascular endothelial cells.
- VEGF attaches to VEGF receptor-2 (VEGFR-2) present in the dermal papilla and acts as a VEGF- It induces proliferation of dermal papilla cells by activating the 2/ERK pathway.
- fibroblast growth factor 7 FGF 7 is a proliferation factor that regulates the hair production cycle by participating in the growth of the outer part of the hair follicle and the part of the hair that appears on the scalp surface. Therefore, increasing gene expression of VEGF and FGF7 in dermal papilla cells can be of great help in inducing hair growth.
- the present inventors applied bioconversion technology to soy milk, rye, and soybeans to ferment them and then completed a complex fermentation product by mixing them at a certain ratio, which consists of 7 types of vitamin B group and separated from natural products. Since it contains various active ingredients and has the effect of increasing the expression of VEGF and FGF7 in human dermal papilla cells (HFDPC), it is expected to be used as a cosmetic composition for preventing hair loss and promoting hair growth.
- HFDPC human dermal papilla cells
- One aspect provides a cosmetic composition for preventing hair loss and/or promoting hair growth, which includes as an active ingredient a complex fermentation product obtained by bioconverting and mixing soymilk, rye, and soybeans.
- compositions for external skin application for preventing hair loss and/or promoting hair growth comprising the complex fermentation product as an active ingredient.
- Another aspect provides a pharmaceutical composition for preventing or treating hair loss, comprising the complex fermentation product as an active ingredient.
- Another aspect provides a method for preventing hair loss and promoting hair growth, comprising administering to an individual a composition containing the complex fermentation product as an active ingredient.
- One aspect is to provide a cosmetic composition for preventing hair loss and/or promoting hair growth, which includes as an active ingredient a complex fermentation product obtained by bioconverting soymilk, rye, and soybeans and then mixing them.
- soymilk in this specification refers to a colloidal (milk) beverage made by grinding soybeans. It can be made by grinding soybeans soaked in water, pouring water, boiling, and then filtering, or processed commercially available products. You can use soy milk.
- rye Greek cereale
- seed in this specification refers to the annual herb of the legume family and its fruit, and is widely cultivated as a food crop.
- bioconversion in this specification refers to a method of converting an initial material into a desired material using microorganisms or enzymes.
- natural products such as soy milk, rye and soybeans are mixed with Lactobacillus .
- the term “complex fermentation product” may refer to a mixture of fermentation products obtained by fermenting two or more natural products.
- “fermented product” refers to the result of inoculating natural products such as soy milk, rye, and soybeans with microorganisms and fermenting them under predetermined conditions.
- the fermented product is a fermented extract extracted with a specific solvent and filtered by physical/chemical methods. Includes all fermentation filtrate, etc.
- the complex fermentation product may refer to a mixture of fermentation product, fermentation extract, or fermentation filtrate obtained by fermenting soy milk, rye, and soybeans with lactic acid bacteria or Bacillus subtilis, respectively.
- the complex fermentation product may be a useful substance itself produced by extracting the fermentation product, a fermentation product obtained by centrifuging and removing fermentation microorganisms from a culture medium in which the fermentation microorganisms and the fermentation product coexist, and fermentation from the culture medium.
- a fermented product obtained by filtering microorganisms, a fermented product obtained by sterilizing and filtering fermented microorganisms from the culture medium, a diluted liquid obtained by diluting the fermented product, a material produced by extracting the dried product obtained by drying the fermented product, or any combination thereof. Can include all.
- strains of the Lactobacillus genus or Bacillus genus may be used.
- the Lactobacillus genus strains include, for example, Lactobacillus casei , Lactobacillus aci. Lactobacillus acidophilus , Lactobacillus plantarum, Lactobacillus fermentum , Lactobacillus curvatus , Lactobacillus confusus , Lactobacillus gasseri ( It may be Lactobacillus gasseri ), Lactobacillus rhamnosus , or any combination thereof, and more specifically, it may be Lactobacillus plantarum, but is not particularly limited thereto.
- Bacillus genus strains include, for example, Bacillus subtilis , Bacillus cereus , Bacillus licheniformis , Bacillus toyoi , and Bacillus coagulans . ), Bacillus polyfermenticus, Bacillus amyloliquefaciens , or any combination thereof, and more specifically, Bacillus subtilis, but is not particularly limited thereto.
- riboflavin (Vitamin B2), pyridoxine (Vitamin B6), and cyanocobalamin (Vitamin B12) are contained in the lactic acid bacteria/soymilk fermentation filtrate fermented by inoculating soymilk with Lactobacillus plantarum. ), Lactic acid bacteria fermented by inoculating rye powder with Lactobacillus plantarum/thiamine (Vitamin B1), nicotinic acid (Vitamin B3), and pantothenic acid (Vitamin B5) in the fermented rye powder, and Bacillus subtilis fermented on soybeans. It was confirmed that folic acid (Vitamin B9) was contained in the Bacillus subtilis/soybean fermentation extract, and a complex fermentation product was prepared by mixing the fermented products at different ratios.
- the complex fermentation product includes cyanocobalamin, pyridoxine, riboflavin, thiamine, nicotinic acid, pantothenic acid, and folic acid.
- the complex fermentation product may include soymilk fermentation filtrate, rye fermentation, and soybean fermentation extract.
- the complex fermentation product may include 1 to 5 parts by weight of soy milk fermentation filtrate, 1 to 5 parts by weight of rye fermentation product, and 1 to 5 parts by weight of soybean fermentation extract, based on 100 parts by weight of the total complex fermentation product, More specifically, each fermented product may be included in 1:1:1 parts by weight. According to one example, it was confirmed that the effect of increasing VEGF and FGF7 gene expression in human hair papilla cells (HFDPC) was the best when soy milk fermentation filtrate, rye fermentation, and soybean fermentation extract were included in equal parts by weight.
- HFDPC human hair papilla cells
- the term "natural vitamin” refers to vitamins manufactured through the process of extracting and refining vitamins contained in natural products such as grains, vegetables, and fish, without going through chemical processes, and containing artificial flavors, synthetic colorings, or synthetic preservatives. It is characterized by not doing so.
- the complex fermented product contains natural vitamins thiamine (Vitamin B1), riboflavin (Vitamin B2), nicotinic acid (Vitamin B3), pantothenic acid (Vitamin B5), pyridoxine (Vitamin B6), and folic acid ( Vitamin B9) and cyanocobalamin (Vitamin B12).
- the term 'natural vitamin' may be used interchangeably with 'fermented vitamin'.
- the term “Alopecia” refers to the phenomenon of hair falling out from the scalp or the condition in which hair becomes sparse or thin
- the term “Hair Loss Prevention” refers to suppressing or preventing hair loss caused by a decrease in the proportion of hair in the growth phase. Or it means any act of treatment.
- the hair loss includes Alopecia Areata, Androgenetic Alopecia, Telogen Effluvium, Traumatic Alopecia, Trichotillomania, Pressure Alopecia, Anagen Alopecia, Alopecia Pityriasis, and Alopecia Pityriasis. It may be one or more selected from the group consisting of Alopecia Syphlltiac, Alopecia Seborrhecia, symptomatic alopecia, cicatricial alopecia and congenital alopecia.
- hair growth promotion refers to any act that promotes the production and growth of hair and ultimately increases the proportion of growing hair in the total hair, and includes all acts of preventing or treating hair loss.
- the cosmetic composition for preventing hair loss and promoting hair growth is effective in preventing hair loss and promoting hair growth caused by exogenous or endogenous factors.
- the extrinsic factors include excessive pressure, pulling, hair treatment, ultraviolet rays, etc.
- the endogenous factors include genetic factors. Factors include hormonal changes, cell death of dermal papilla cells caused by telomeres, and oxidative stress due to cell function decline.
- the cosmetic composition may further include functional additives and ingredients included in general cosmetic compositions, and may include commonly used antioxidants, stabilizers, solubilizers, vitamins, pigments, fragrances, etc. It may further contain auxiliary ingredients and carriers.
- the cosmetic composition may further include auxiliary ingredients such as glycerin, butylene glycol, polyoxyethylene hydrogenated castor oil, tocopheryl acetate, citric acid, squalane, sodium citrate, and allantoin, and the solvent may include Hexanediol, purified water, etc. may be included.
- the functional additive may include ingredients selected from the group consisting of water-soluble vitamins, oil-soluble vitamins, polymer peptides, polymer polysaccharides, sphingolipids, and seaweed extract.
- Other ingredients included include oils and fats, moisturizers, emollients, surfactants, organic and inorganic pigments, organic powders, ultraviolet absorbers, preservatives, disinfectants, antioxidants, plant extracts, pH adjusters, alcohol, pigments, fragrances, and blood circulation agents.
- the cosmetic composition according to one aspect includes, for example, hair tonic, hair conditioner, hair essence, hair lotion, hair nutrition lotion, hair shampoo, hair rinse, hair treatment, hair cream, hair nutrition cream, hair moisture cream, Hair massage cream, hair wax, hair aerosol, hair pack, hair nutrition pack, hair soap, hair cleansing foam, hair oil, hair dryer, hair preservation treatment, hair dye, hair waving agent, hair bleach, hair gel, hair glaze, hair dressing.
- hair lacquer, hair moisturizer, hair mousse, or hair spray and specifically, shampoo, cleanser, conditioner, treatment, hair pack, rinse, scalp pack, tonic, essence, lotion, cream, oil, It may be a mist or spray formulation, but is not particularly limited thereto.
- the formulation of the cosmetic composition according to one aspect is a paste, cream or gel, animal fiber, plant fiber, wax, paraffin, starch, tracant, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc or oxidized carrier ingredient. Zinc, etc. may be used.
- the formulation of the cosmetic composition according to one aspect is a powder or spray
- lactose, talc, silica, aluminum hydroxide, calcium silicate, or polyamide powder may be used as the carrier ingredient, and in the case of a spray, chlorofluoro may be additionally used.
- chlorofluoro may be additionally used.
- propellants such as hydrocarbons, propane/butane or dimethyl ether.
- a solvent, solvating agent, or emulsifying agent is used as a carrier component, such as water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, and benzyl benzoate.
- a carrier component such as water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, and benzyl benzoate.
- propylene glycol, 1,3-butyl glycol oil, glycerol aliphatic ester, polyethylene glycol, or sorbitan fatty acid ester, etc. can be used.
- the carrier component may include water, a liquid diluent such as ethanol or propylene glycol, a suspending agent such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester, and polyoxyethylene sorbitan ester, Microcrystalline cellulose, aluminum metahydroxide, bentonite, agar, or tracant may be used.
- a liquid diluent such as ethanol or propylene glycol
- a suspending agent such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester, and polyoxyethylene sorbitan ester
- Microcrystalline cellulose aluminum metahydroxide, bentonite, agar, or tracant may be used.
- Another aspect is to provide a composition for external skin application for preventing hair loss and/or promoting hair growth, comprising the complex fermentation product as an active ingredient.
- the same parts as described above also apply to the external skin composition.
- the external skin composition may include anything applied from the outside of the skin, for example, a group consisting of creams, gels, patches, sprays, ointments, warning agents, lotions, linear agents, pasta agents, and cataplasma agents. It may be any one selected from, but is not particularly limited thereto.
- the external skin preparations include, for example, aqueous ingredients, oil-based ingredients, powder ingredients, alcohols, moisturizers, thickeners, ultraviolet absorbers, whitening agents, preservatives, antioxidants, surfactants, fragrances, colorants, skin nutrients or these. Combinations may be included, and these may be appropriately combined by a person skilled in the art as needed.
- the skin external preparations include metal sequestrants such as disodium edetate, trisodium edetate, sodium citrate, sodium polyphosphate, sodium metaphosphate, and gluconic acid, caffeine, tannin, belafamil, licorice extract, glablidin, and calin.
- hot water extract of fruits various herbal medicines, medicines such as tocopherol acetate, glythylitic acid, tranexamic acid and its derivatives or salts, vitamin C, magnesium ascorbate phosphate, ascorbate glucoside, arbutin, kojic acid, glucose, frug Sugars such as toss and trehalose may be mixed, but are not particularly limited thereto.
- medicines such as tocopherol acetate, glythylitic acid, tranexamic acid and its derivatives or salts, vitamin C, magnesium ascorbate phosphate, ascorbate glucoside, arbutin, kojic acid, glucose, frug Sugars such as toss and trehalose may be mixed, but are not particularly limited thereto.
- Another aspect is to provide a pharmaceutical composition for preventing or treating hair loss, comprising the complex fermentation product as an active ingredient.
- the same parts as described above also apply to the pharmaceutical composition.
- the pharmaceutical composition may further contain pharmaceutical auxiliaries such as preservatives, stabilizers, wetting agents or emulsification accelerators, salts and/or buffers for osmotic pressure adjustment, and other therapeutically useful substances.
- pharmaceutical auxiliaries such as preservatives, stabilizers, wetting agents or emulsification accelerators, salts and/or buffers for osmotic pressure adjustment, and other therapeutically useful substances.
- the oral administration agents include, for example, tablets, pills, hard and soft capsules, solutions, suspensions, emulsifiers, syrups, powders, powders, fine granules, granules, pellets, etc., and these dosage forms contain, in addition to the active ingredient, the surface Activators, diluents (e.g. lactose, dextrose, sucrose, mannitol, sorbitol, cellulose and glycine), lubricants (e.g. silica, talc, stearic acid and its magnesium or calcium salts and polyethylene glycol) may include.
- diluents e.g. lactose, dextrose, sucrose, mannitol, sorbitol, cellulose and glycine
- lubricants e.g. silica, talc, stearic acid and its magnesium or calcium salts and polyethylene glycol
- the tablet may contain a binder such as magnesium aluminum silicate, starch paste, gelatin, tragacanth, methylcellulose, sodium carboxymethylcellulose and polyvinylpyrrolidine, and may additionally contain a binder such as starch, agar, alginic acid or its sodium salt. It may contain pharmaceutical additives such as disintegrants, absorbents, colorants, flavoring agents, and sweeteners. Additionally, the tablets can be manufactured by conventional mixing, granulating or coating methods.
- a binder such as magnesium aluminum silicate, starch paste, gelatin, tragacanth, methylcellulose, sodium carboxymethylcellulose and polyvinylpyrrolidine
- a binder such as starch, agar, alginic acid or its sodium salt. It may contain pharmaceutical additives such as disintegrants, absorbents, colorants, flavoring agents, and sweeteners.
- the tablets can be manufactured by conventional mixing, granulating or coating methods.
- the parenteral administration may be in a transdermal dosage form, for example, injections, drops, ointments, lotions, gels, creams, sprays, suspensions, emulsions, suppositories, patches, etc. However, it is not particularly limited thereto.
- the pharmaceutical composition may be administered parenterally, rectally, topically, transdermally, subcutaneously, etc., and more specifically, may be administered topically to the scalp.
- the daily dosage can be appropriately selected by a person skilled in the art in consideration of various factors such as the degree of progression, onset time, age, health condition, and complications of the subject to be administered. there is.
- Another aspect provides a method for preventing hair loss and promoting hair growth, comprising administering to an individual a composition containing as an active ingredient a complex fermentation product obtained by bioconverting soymilk, rye, and soybeans.
- a composition containing as an active ingredient a complex fermentation product obtained by bioconverting soymilk, rye, and soybeans.
- the composition containing the complex fermentation product of the present invention may be administered to an individual as a cosmetic composition, external skin preparation composition, pharmaceutical composition, or quasi-drug composition.
- the administration method is not particularly limited and may be appropriately selected depending on the formulation of the composition and those skilled in the art.
- the composition can be administered orally, parenterally, rectally, topically, transdermally, intravenously, intramuscularly, intraperitoneally, subcutaneously, etc., and more specifically, it can be applied to the scalp or administered topically.
- the cosmetic composition according to one aspect increases the expression of VEGF and FGF7 in human follicular dermal papilla cells (HFDPC), promotes nutrient delivery to dermal papilla cells, induces growth of hair root cells, and induces proliferation of dermal papilla cells. This can be expected to promote hair growth and prevent hair loss.
- HFDPC human follicular dermal papilla cells
- Figure 1 shows the results confirming the effect of increasing VEGF gene expression according to the concentration of the complex fermentation product of the present invention compared to the positive control group AS2P.
- Figure 2 shows the results confirming the effect of increasing FGF7 gene expression according to the concentration of the complex fermentation product of the present invention compared to the positive control group AS2P.
- Figure 3 shows the results of confirming the effect of increasing VEGF gene expression of the complex fermentation products of Examples 1 to 4 compared to the lactic acid bacteria or Bacillus subtilis culture and natural raw materials (soy milk, rye powder, soybean).
- Figure 4 shows the results of confirming the effect of increasing FGF7 gene expression of the complex fermentation products of Examples 1 to 4 compared to lactic acid bacteria or Bacillus subtilis culture and natural raw materials (soy milk, rye powder, soybean).
- Figure 5 shows the results confirming the effect of increasing VEGF gene expression of the complex fermented product of Example 1 compared to the vitamin B group (Vitamin B1, B2, B3, B5, B6, B9, B12) and their mixed group.
- Figure 6 shows the results confirming the effect of increasing FGF7 gene expression of the complex fermentation of Example 1 compared to the vitamin B group (Vitamin B1, B2, B3, B5, B6, B9, B12) and their mixed group.
- Fermented vitamins were manufactured by introducing bio-conversion technology, which combines microbial fermentation with natural products.
- Soy milk was used by adding 10 times the weight of water to soybeans, pulverizing them in a blender, and filtering them through a 150 mesh filter (104 micron).
- the fermentation medium was prepared with the following ingredients: 2.0% glucose, 1.0% Na 2 HPO 4 , and 0.5% NaH 2 PO 4 , and then sterilized by adding 90% soy milk. Lactobacillus plantarum was inoculated into the fermentation medium, and stationary fermentation was performed at 30°C for 24 hours. After completion of fermentation, insoluble substances such as remaining bacterial cells were removed using a continuous centrifuge. The recovered fermented product was neutralized with 2N KOH (pH 6.0), preservative, and filtered for sterilization. Afterwards, it was confirmed that the fermentation filtrate contained riboflavin (Vitamin B2), pyridoxine (Vitamin B6), and cyanocobalamin (Vitamin B12).
- Rye powder was wet-sterilized after adding 5 times the weight of water. Afterwards, in order to decompose the starch in the rye powder, 0.1% of the weight was added using amylase, and the starch was decomposed at 50-60°C for 24 hours and then used for fermentation. Fermentation medium was prepared with the following ingredients: 2.0% glucose, 1.0% Na 2 HPO 4 , and 0.5% NaH 2 PO 4 , and then 90% rye powder was added and sterilized. Lactobacillus plantarum was inoculated into the fermentation medium, and stationary fermentation was performed at 30°C for 48 hours. After completion of fermentation, insoluble substances such as remaining bacterial cells were removed using a continuous centrifuge.
- the recovered fermented product was neutralized with 2N KOH (pH 6.0), preservative, and filtered for sterilization. Afterwards, it was confirmed that the fermented product contained thiamine (Vitamin B1), nicotinic acid (Vitamin B3), and pantothenic acid (Vitamin B5).
- soybeans were added to purified water and heated at 100°C for 1 hour. Afterwards, it was cooled to remove the solid content and only the extract was recovered. Fermentation medium was prepared with ingredients of 2.0% glucose, 1.0% Na 2 HPO 4 and 0.5% NaH 2 PO 4 and sterilized by adding 90% soybean extract and 0.5% folic acid. Bacillus subtilis was inoculated into the fermentation medium, and stationary fermentation was performed at 45°C for 72 hours. After completion of fermentation, insoluble substances such as remaining bacterial cells were removed using a continuous centrifuge, and the pH was adjusted to 4 using 2M HCl. Afterwards, 2 times the amount of ethanol was added to recover the solid content of the Bacillus subtilis/soybean fermentation extract. The recovered solid content was neutralized with 2N KOH (pH 6.0), preservative, and filtered for sterilization. Afterwards, it was confirmed that the fermented extract contained high molecular form folic acid (Vitamin B9).
- HFDPC human hair papilla cells
- C1000 Thermal Cycler Bio- cDNA was synthesized at Rad, USA.
- VEGF F One 5'-GTGCCCACTGAGGAGTTCAAC-3' After activating the polymerase at 94°C for 5 minutes, polymerization reaction was performed in 40 cycles at 95°C for 30 seconds, 60°C for 30 seconds, and 72°C for 30 seconds.
- R 2 5'-CCCTATGTGCTGGCCTTGAT-3' FGF7
- F 3 5'-TCCTGCCAACTTTGCTCTACA-3'
- R 4 5'-CAGGGCTGGAACAGTTCACAT-3'
- R 6 5'-GAGACCTTCAACACCCCAGC-3'
- the complex fermented products of Examples 1 to 4 which were mixed with the fermented products of Experimental Example 1 at different ratios, had a significantly better effect of increasing VEGF and FGF7 gene expression compared to the lactic acid bacteria or Bacillus subtilis culture and natural raw materials (soy milk, rye powder, soybeans). It was confirmed that, except for Example 2, there was a superior effect of increasing VEGF and FGF7 gene expression compared to AS2P, which was a positive control (FIGS. 3 and 4).
- each vitamin B group (Vitamin B1, B2, B3, B5, B6, B9, B12) and their mixture group (1 each: It was confirmed that there was a significantly better effect of increasing VEGF and FGF7 gene expression compared to (included in 1:1:1:1:1:1 parts by weight) (FIGS. 5 and 6).
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Abstract
The present invention pertains to a cosmetic composition. A cosmetic composition according to an aspect increases the expression of VEGF and FGF7 in human follicular dermal papilla cells (HFDPCs), and thus can promote the delivery of nutrients to follicular dermal papilla cells and induce the growth of hair root cells and the proliferation of follicular dermal papilla cells. Accordingly, the cosmetic composition is expected to promote hair growth and prevent hair loss.
Description
본 발명은 화장료 조성물에 관한 것이다.The present invention relates to cosmetic compositions.
인체에 있어서 모발은 두피에 있는 모낭에서 자라나며, 일생 동안 성장기(Anagen), 퇴행기(Catagen), 휴지기(Telogen) 등 주기가 지속적으로 반복되며 생성, 성장 및 탈락을 거치게 된다. 모낭의 퇴행기에는 모낭의 성장이 중단되고 세포사멸 및 세포외 기질의 재구성이 발생하기 때문에, 퇴행기 및 휴지기 상태의 모낭 세포가 많아질수록 머리카락이 빠지고 다시 자라나지 않는 탈모 현상이 나타내게 된다.In the human body, hair grows from hair follicles on the scalp, and throughout life, the cycle of growth (Anagen), catagen (catagen), and telogen (Telogen) is repeated continuously and goes through creation, growth, and shedding. In the catagen phase of hair follicles, the growth of the hair follicle stops and apoptosis and reorganization of the extracellular matrix occur. As the number of hair follicle cells in the catagen and resting phases increases, hair falls out and does not grow again, resulting in hair loss.
모발은 눈에 보이는 모간부와 눈에 보이지 않는 모근부로 나누어지는데, 모근부 중 모발이 자라는 뿌리를 모구(Hair bulb)라고 부르며, 그 아랫부분에 움푹 들어간 부분을 모유두라고 한다. 모유두는 모발의 생성 및 성장에 있어 가장 중요한 부분으로, 혈관 및 신경이 있어 모발에 영양분과 성장인자 등을 전달하여 모발의 성장을 촉진한다. 즉, 모유두세포의 사멸 및 생장 감소는 모발 수의 감소 및 탈모로 이어지기 때문에, 정상적인 모발의 유지 및 증가를 위해서는 모유두세포의 증식이 필요하다.Hair is divided into a visible hair shaft and an invisible hair root. Among the hair roots, the root from which hair grows is called the hair bulb, and the hollow part at the bottom is called the dermal papilla. The dermal papilla is the most important part in the creation and growth of hair. It contains blood vessels and nerves, which promote hair growth by delivering nutrients and growth factors to the hair. In other words, since the death and growth reduction of dermal papilla cells leads to a decrease in the number of hairs and hair loss, proliferation of dermal papilla cells is necessary to maintain and increase normal hair.
한편, 혈관 내피 성장인자(Vascular endothelial growth factor: VEGF)는 혈관 내피 세포의 성장을 유도하여 혈관 신생을 촉진시키는 당단백으로, VEGF는 모유두에 존재하는 VEGF 수용체-2(VEGFR-2)에 붙어 VEGF-2/ERK 경로를 활성화함으로써 모유두세포의 증식을 유도한다. 또한, 섬유아세포 성장인자(Fibroblast growth factor 7: FGF 7)는 모낭 바깥 부분과 두피 표면으로 드러나는 모발 부분의 생장에 관여함으로써 모발 생성 주기를 조절하는 증식 인자이다. 따라서 모유두세포에서 VEGF 및 FGF7의 유전자 발현의 증가는 모발 생장을 유도하는 데 큰 도움을 줄 수 있다.Meanwhile, vascular endothelial growth factor (VEGF) is a glycoprotein that promotes angiogenesis by inducing the growth of vascular endothelial cells. VEGF attaches to VEGF receptor-2 (VEGFR-2) present in the dermal papilla and acts as a VEGF- It induces proliferation of dermal papilla cells by activating the 2/ERK pathway. In addition, fibroblast growth factor 7 (FGF 7) is a proliferation factor that regulates the hair production cycle by participating in the growth of the outer part of the hair follicle and the part of the hair that appears on the scalp surface. Therefore, increasing gene expression of VEGF and FGF7 in dermal papilla cells can be of great help in inducing hair growth.
이러한 배경 하에, 본 발명자들은 두유, 호밀 및 대두에 대해 바이오컨버전(bioconversion) 기술을 적용하여 발효시킨 후 일정 비율로 혼합한 복합발효물을 완성하였으며, 이는 7종의 비타민 B군 및 천연물로부터 분리된 다양한 유효성분을 포함하여 인간 모유두세포(HFDPC)에서 VEGF 및 FGF7의 발현을 증가시키는 효과가 있는 바, 탈모 방지 및 발모 촉진용 화장료 조성물 등으로 활용할 수 있을 것으로 기대한다.Under this background, the present inventors applied bioconversion technology to soy milk, rye, and soybeans to ferment them and then completed a complex fermentation product by mixing them at a certain ratio, which consists of 7 types of vitamin B group and separated from natural products. Since it contains various active ingredients and has the effect of increasing the expression of VEGF and FGF7 in human dermal papilla cells (HFDPC), it is expected to be used as a cosmetic composition for preventing hair loss and promoting hair growth.
일 양상은 두유, 호밀 및 대두를 바이오컨버전(bioconversion)시킨 후 혼합한 복합발효물을 유효성분으로 포함하는, 탈모 방지 및/또는 발모 촉진용 화장료 조성물을 제공한다.One aspect provides a cosmetic composition for preventing hair loss and/or promoting hair growth, which includes as an active ingredient a complex fermentation product obtained by bioconverting and mixing soymilk, rye, and soybeans.
다른 양상은 상기 복합발효물을 유효성분으로 포함하는, 탈모 방지 및/또는 발모 촉진용 피부외용제 조성물을 제공한다.Another aspect provides a composition for external skin application for preventing hair loss and/or promoting hair growth, comprising the complex fermentation product as an active ingredient.
또 다른 양상은 상기 복합발효물을 유효성분으로 포함하는, 탈모 예방 또는 치료용 약학적 조성물을 제공한다.Another aspect provides a pharmaceutical composition for preventing or treating hair loss, comprising the complex fermentation product as an active ingredient.
또 다른 양상은 상기 복합발효물을 유효성분으로 포함하는 조성물을 개체에 투여하는 단계를 포함하는, 탈모 방지 및 발모 촉진 방법을 제공한다.Another aspect provides a method for preventing hair loss and promoting hair growth, comprising administering to an individual a composition containing the complex fermentation product as an active ingredient.
일 양상은 두유, 호밀 및 대두를 바이오컨버전(bioconversion)시킨 후 혼합한 복합발효물을 유효성분으로 포함하는, 탈모 방지 및/또는 발모 촉진용 화장료 조성물을 제공하는 것이다.One aspect is to provide a cosmetic composition for preventing hair loss and/or promoting hair growth, which includes as an active ingredient a complex fermentation product obtained by bioconverting soymilk, rye, and soybeans and then mixing them.
본 명세서에서의 용어 "두유"는 콩을 갈아서 만든 콜로이드(밀크) 상태의 음료를 의미하는 것으로, 물에 불린 콩을 간 다음 물을 붓고 끓인 후 여과하여 제조된 것을 사용하거나, 시중에 판매하는 가공두유를 사용할 수 있다. The term "soymilk" in this specification refers to a colloidal (milk) beverage made by grinding soybeans. It can be made by grinding soybeans soaked in water, pouring water, boiling, and then filtering, or processed commercially available products. You can use soy milk.
본 명세서에서의 용어 "호밀(Secale cereale)"은 Triticeae과의 일년생 동작물로서, 보다 구체적으로는 호밀 열매의 가루(호밀분말)를 의미할 수 있다. The term "rye (Secale cereale)" in this specification refers to an annual crop of the Triticeae family, and more specifically, may refer to the powder of rye fruit (rye powder).
본 명세서에서의 용어 "대두"는 콩과의 일년생 초본 및 그 열매이며 식용작물로 널리 재배된다. The term “soybean” in this specification refers to the annual herb of the legume family and its fruit, and is widely cultivated as a food crop.
본 명세서에서의 용어 "바이오컨버전(bioconversion"은 미생물 또는 효소 등을 이용하여 초기 물질을 목적하는 물질로 전환시키는 방법을 의미하는 것으로, 본 발명에서는 두유, 호밀 및 대두와 같은 천연물에 락토바실러스(Lactobacillus) 속 또는 바실러스(Bacillus) 속 균주 등에 의한 발효를 통해 발효물을 얻는 과정을 의미할 수 있다.The term "bioconversion" in this specification refers to a method of converting an initial material into a desired material using microorganisms or enzymes. In the present invention, natural products such as soy milk, rye and soybeans are mixed with Lactobacillus . ) may refer to the process of obtaining fermented products through fermentation by strains of the genus or Bacillus .
본 명세서에서의 용어 "복합발효물"은 2 이상의 천연물을 발효시킨 발효물의 혼합물을 의미할 수 있다. 여기서 "발효물"은 두유, 호밀 및 대두와 같은 천연물에 미생물을 접종하여 소정의 조건에서 발효시킨 결과물을 의미하는 것으로, 상기 발효물을 특정 용매로 추출한 발효 추출물 및 물리적/화학적인 방법으로 여과한 발효 여과물 등을 모두 포함한다. 일 실시예에 따르면, 상기 복합발효물은 두유, 호밀 및 대두를 각각 유산균 또는 고초균으로 발효시킨 발효물, 발효 추출물 또는 발효 여과물의 혼합물을 의미하는 것일 수 있다.As used herein, the term “complex fermentation product” may refer to a mixture of fermentation products obtained by fermenting two or more natural products. Here, “fermented product” refers to the result of inoculating natural products such as soy milk, rye, and soybeans with microorganisms and fermenting them under predetermined conditions. The fermented product is a fermented extract extracted with a specific solvent and filtered by physical/chemical methods. Includes all fermentation filtrate, etc. According to one embodiment, the complex fermentation product may refer to a mixture of fermentation product, fermentation extract, or fermentation filtrate obtained by fermenting soy milk, rye, and soybeans with lactic acid bacteria or Bacillus subtilis, respectively.
보다 구체적으로, 상기 복합발효물은 상기 발효물을 추출하여 생성된 유용한 물질 자체일 수 있으며, 발효 미생물 및 발효물이 공존하는 배양 배지로부터 발효 미생물을 원심분리하여 제거한 발효물, 상기 배양 배지로부터 발효 미생물을 여과시킨 발효물, 상기 배양 배지로부터 발효 미생물을 멸균 처리하고 이를 여과시킨 발효물, 상기 발효물을 희석시킨 희석액, 상기 발효물을 건조시킨 건조물을 추출하여 생성된 물질 또는 이들의 임의의 조합을 모두 포함할 수 있다.More specifically, the complex fermentation product may be a useful substance itself produced by extracting the fermentation product, a fermentation product obtained by centrifuging and removing fermentation microorganisms from a culture medium in which the fermentation microorganisms and the fermentation product coexist, and fermentation from the culture medium. A fermented product obtained by filtering microorganisms, a fermented product obtained by sterilizing and filtering fermented microorganisms from the culture medium, a diluted liquid obtained by diluting the fermented product, a material produced by extracting the dried product obtained by drying the fermented product, or any combination thereof. Can include all.
상기 발효물 또는 복합발효물을 제조하기 위한 균주로 락토바실러스 속 또는 바실러스 속 균주 등이 사용될 수 있으며, 구체적으로 상기 락토바실러스 속 균주는 예를 들어, 락토바실루스 카제이(Lactobacillus casei), 락토바실루스 아시도필루스(Lactobacillus acidophilus), 락토바실루스 플란타룸(Lactobacillus plantarum), 락토바실루스 퍼멘텀(Lactobacillus fermentum), 락토바실루스 커바투스(Lactobacillus curvatus), 락토바실루스 콘푸수스(Lactobacillus confusus), 락토바실루스 가세리(Lactobacillus gasseri), 락토바실루스 람노서스(Lactobacillus rhamnosus) 또는 이들의 임의의 조합일 수 있으며, 보다 구체적으로는 락토바실루스 플란타룸일 수 있으나, 이에 특별히 제한되는 것은 아니다.As a strain for producing the fermented product or complex fermented product, strains of the Lactobacillus genus or Bacillus genus may be used. Specifically, the Lactobacillus genus strains include, for example, Lactobacillus casei , Lactobacillus aci. Lactobacillus acidophilus , Lactobacillus plantarum, Lactobacillus fermentum , Lactobacillus curvatus , Lactobacillus confusus , Lactobacillus gasseri ( It may be Lactobacillus gasseri ), Lactobacillus rhamnosus , or any combination thereof, and more specifically, it may be Lactobacillus plantarum, but is not particularly limited thereto.
상기 바실러스 속 균주는 예를 들어, 바실러스 서브틸리스(Bacillus subtilis), 바실러스 세레우스(Bacillus cereus), 바실러스 리케니포미스(Bacillus licheniformis), 바실러스 토요이(Bacillus toyoi), 바실러스 코어귤런스(Bacillus coagulans), 바실러스 폴리퍼멘티쿠스(Bacillus polyfermenticus), 바실러스 아밀로리쿼페시언스(Bacillus amyloliquefaciens) 또는 이들의 임의의 조합일 수 있으며, 보다 구체적으로는 바실러스 서브틸리스일 수 있으나, 이에 특별히 제한되는 것은 아니다. The Bacillus genus strains include, for example, Bacillus subtilis , Bacillus cereus , Bacillus licheniformis , Bacillus toyoi , and Bacillus coagulans . ), Bacillus polyfermenticus, Bacillus amyloliquefaciens , or any combination thereof, and more specifically, Bacillus subtilis, but is not particularly limited thereto.
일 실시예에 따르면, 본 발명에서는 두유에 락토바실러스 플란타룸(Lactobacillus plantarum)을 접종하여 발효시킨 유산균/두유 발효 여과물 내에 리보플라빈(Vitamin B2), 피리독신(Vitamin B6) 및 시아노코발라민(Vitamin B12), 호밀분말에 락토바실러스 플란타룸을 접종하여 발효시킨 유산균/호밀분말 발효물 내에 티아민(Vitamin B1), 니코틴산(Vitamin B3) 및 판토텐산(Vitamin B5), 대두에 고초균(Bacillus subtilis)을 발효시킨 고초균/대두 발효 추출물 내에 엽산(Vitamin B9)을 포함하고 있는 것을 확인하였으며, 상기 발효물을 서로 다른 비율로 혼합하여 복합발효물을 제조하였다. According to one embodiment, in the present invention, riboflavin (Vitamin B2), pyridoxine (Vitamin B6), and cyanocobalamin (Vitamin B12) are contained in the lactic acid bacteria/soymilk fermentation filtrate fermented by inoculating soymilk with Lactobacillus plantarum. ), Lactic acid bacteria fermented by inoculating rye powder with Lactobacillus plantarum/thiamine (Vitamin B1), nicotinic acid (Vitamin B3), and pantothenic acid (Vitamin B5) in the fermented rye powder, and Bacillus subtilis fermented on soybeans. It was confirmed that folic acid (Vitamin B9) was contained in the Bacillus subtilis/soybean fermentation extract, and a complex fermentation product was prepared by mixing the fermented products at different ratios.
상기 복합발효물에는 시아노코발아민, 피리독신, 리보플라빈, 티아민, 니코틴산, 판토텐산 및 엽산이 포함되어 있고, 일 예로 상기 복합발효물은 두유 발효 여과물, 호밀 발효물 및 대두 발효 추출물을 포함할 수 있다. 보다 구체적인 일 예로 상기 복합발효물은 복합발효물 전체 100 중량부를 기준으로 두유 발효 여과물 1 내지 5 중량부, 호밀 발효물 1 내지 5 중량부 및 대두 발효 추출물 1 내지 5 중량부로 포함할 수 있으며, 보다 더 구체적으로는 각각의 발효물을 1:1:1 중량부로 포함하는 것일 수 있다. 일 실시예에 따르면, 두유 발효 여과물, 호밀 발효물 및 대두 발효 추출물을 동일한 중량부로 포함하는 경우 인간모유두세포(HFDPC)에서 VEGF 및 FGF7 유전자 발현 증가 효과가 가장 우수함을 확인하였다.The complex fermentation product includes cyanocobalamin, pyridoxine, riboflavin, thiamine, nicotinic acid, pantothenic acid, and folic acid. For example, the complex fermentation product may include soymilk fermentation filtrate, rye fermentation, and soybean fermentation extract. . As a more specific example, the complex fermentation product may include 1 to 5 parts by weight of soy milk fermentation filtrate, 1 to 5 parts by weight of rye fermentation product, and 1 to 5 parts by weight of soybean fermentation extract, based on 100 parts by weight of the total complex fermentation product, More specifically, each fermented product may be included in 1:1:1 parts by weight. According to one example, it was confirmed that the effect of increasing VEGF and FGF7 gene expression in human hair papilla cells (HFDPC) was the best when soy milk fermentation filtrate, rye fermentation, and soybean fermentation extract were included in equal parts by weight.
본 명세서에서 용어 "천연 비타민"은 곡물, 채소, 생선 등 천연물에 함유된 비타민을 추출하고 정제하는 과정을 통해 제조된 비타민으로, 화학적 공정을 거치지 않으며, 인공향, 합성착색료 또는 합성보존료 등을 포함하지 않는 것을 특징으로 한다. 상기 복합발효물은 두유, 호밀 또는 대두의 발효를 통해 제조된 천연 비타민 티아민(Vitamin B1), 리보플라빈(Vitamin B2), 니코틴산(Vitamin B3), 판토텐산(Vitamin B5), 피리독신(Vitamin B6), 엽산(Vitamin B9) 및 시아노코발라민(Vitamin B12)을 포함할 수 있고, 본 명세서에서 용어 '천연 비타민'은 '발효 비타민'과 혼용될 수 있다.In this specification, the term "natural vitamin" refers to vitamins manufactured through the process of extracting and refining vitamins contained in natural products such as grains, vegetables, and fish, without going through chemical processes, and containing artificial flavors, synthetic colorings, or synthetic preservatives. It is characterized by not doing so. The complex fermented product contains natural vitamins thiamine (Vitamin B1), riboflavin (Vitamin B2), nicotinic acid (Vitamin B3), pantothenic acid (Vitamin B5), pyridoxine (Vitamin B6), and folic acid ( Vitamin B9) and cyanocobalamin (Vitamin B12). In this specification, the term 'natural vitamin' may be used interchangeably with 'fermented vitamin'.
본 명세서에서 용어 "탈모(Alopecia)"는 두피로부터 모발이 탈락하는 현상 또는 모발이 성기거나 가늘어지는 상태를 의미하며, "탈모 방지"는 성장기 모발의 비중이 감소함에 따라 유발되는 탈모를 억제하거나 예방 또는 치료하는 모든 행위를 의미한다. 상기 탈모는 원형 탈모증(Alopecia Areata), 유전성 안드로겐 탈모증(Androgenetic Alopecia), 휴지기 탈모증(Telogen Effluvium), 외상성 탈모증, 발모벽(Trichotillomania), 압박성 탈모증(Pressure Alopecia), 생장기 탈모증, 비강성 탈모증, 매독성 탈모증(Alopecia Syphlltiac), 지루 탈모증(Alopecia Seborrhecia), 증후성 탈모증, 반흔성 탈모증 및 선천성 탈모증으로 이루어진 군으로부터 선택되는 하나 이상일 수 있다.As used herein, the term "Alopecia" refers to the phenomenon of hair falling out from the scalp or the condition in which hair becomes sparse or thin, and the term "Hair Loss Prevention" refers to suppressing or preventing hair loss caused by a decrease in the proportion of hair in the growth phase. Or it means any act of treatment. The hair loss includes Alopecia Areata, Androgenetic Alopecia, Telogen Effluvium, Traumatic Alopecia, Trichotillomania, Pressure Alopecia, Anagen Alopecia, Alopecia Pityriasis, and Alopecia Pityriasis. It may be one or more selected from the group consisting of Alopecia Syphlltiac, Alopecia Seborrhecia, symptomatic alopecia, cicatricial alopecia and congenital alopecia.
본 명세서에서의 용어 "발모 촉진"은 모발의 생성 및 성장을 촉진하여 궁극적으로는 전체 모발에서 성장기 모발의 비중을 증가시키는 모든 행위로서, 탈모의 예방 또는 치료하는 행위를 모두 포함한다. 상기 탈모 방지 및 발모 촉진용 화장료 조성물은 외인성 또는 내인성 요인에 의해 유발되는 탈모 방지 및 발모 촉진에 효과적이며, 상기 외인성 요인은 과도한 압력, 당김과 헤어시술, 자외선 등을 포함하고, 내인성 요인은 유전적 요인, 호르몬 변화, 텔로미어에 의한 모유두세포의 세포사 및 세포 기능 저하에 따른 산화 스트레스 등을 포함한다.The term “hair growth promotion” as used herein refers to any act that promotes the production and growth of hair and ultimately increases the proportion of growing hair in the total hair, and includes all acts of preventing or treating hair loss. The cosmetic composition for preventing hair loss and promoting hair growth is effective in preventing hair loss and promoting hair growth caused by exogenous or endogenous factors. The extrinsic factors include excessive pressure, pulling, hair treatment, ultraviolet rays, etc., and the endogenous factors include genetic factors. Factors include hormonal changes, cell death of dermal papilla cells caused by telomeres, and oxidative stress due to cell function decline.
상기 화장료 조성물에는 본 명세서에 개시된 유효성분 이외에 기능성 첨가물 및 일반적인 화장료 조성물에 포함되는 성분이 추가로 포함될 수 있으며, 통상적으로 사용되는 항산화제, 안정화제, 용해화제, 비타민, 안료, 향료 등과 같은 통상적인 보조성분 및 담체를 더 포함할 수 있다. 예를 들어, 상기 화장료 조성물에는 글리세린, 부틸렌글리콜, 폴리옥시에칠렌 경화피마자유, 토코페릴 아세테이트, 시트릭산, 스쿠알란, 소듐 시트레이트, 알란토인 등의 보조성분이 추가로 더 포함될 수 있고, 용제로는 헥산디올, 정제수 등이 포함될 수 있다. 또한, 상기 기능성 첨가물로는 수용성 비타민, 유용성 비타민, 고분자 펩티드, 고분자 다당, 스핑고 지질 및 해초 엑기스로 이루어진 군에서 선택된 성분을 포함할 수 있다. 이외에 포함되는 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 등이 있을 수 있다.In addition to the active ingredients disclosed herein, the cosmetic composition may further include functional additives and ingredients included in general cosmetic compositions, and may include commonly used antioxidants, stabilizers, solubilizers, vitamins, pigments, fragrances, etc. It may further contain auxiliary ingredients and carriers. For example, the cosmetic composition may further include auxiliary ingredients such as glycerin, butylene glycol, polyoxyethylene hydrogenated castor oil, tocopheryl acetate, citric acid, squalane, sodium citrate, and allantoin, and the solvent may include Hexanediol, purified water, etc. may be included. Additionally, the functional additive may include ingredients selected from the group consisting of water-soluble vitamins, oil-soluble vitamins, polymer peptides, polymer polysaccharides, sphingolipids, and seaweed extract. Other ingredients included include oils and fats, moisturizers, emollients, surfactants, organic and inorganic pigments, organic powders, ultraviolet absorbers, preservatives, disinfectants, antioxidants, plant extracts, pH adjusters, alcohol, pigments, fragrances, and blood circulation agents. There may be accelerators, cooling agents, restrictors, purified water, etc.
일 양상에 따른 상기 화장료 조성물은, 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 헤어스프레이의 제형을 포함할 수 있으며, 구체적으로 샴푸, 클렌져, 컨디셔너, 트리트먼트, 헤어팩, 린스, 두피팩, 토닉, 에센스, 로션, 크림, 오일, 미스트 또는 스프레이 제형일 수 있으나, 이에 특별히 제한되는 것은 아니다.The cosmetic composition according to one aspect includes, for example, hair tonic, hair conditioner, hair essence, hair lotion, hair nutrition lotion, hair shampoo, hair rinse, hair treatment, hair cream, hair nutrition cream, hair moisture cream, Hair massage cream, hair wax, hair aerosol, hair pack, hair nutrition pack, hair soap, hair cleansing foam, hair oil, hair dryer, hair preservation treatment, hair dye, hair waving agent, hair bleach, hair gel, hair glaze, hair dressing. It may include formulations of hair lacquer, hair moisturizer, hair mousse, or hair spray, and specifically, shampoo, cleanser, conditioner, treatment, hair pack, rinse, scalp pack, tonic, essence, lotion, cream, oil, It may be a mist or spray formulation, but is not particularly limited thereto.
일 양상에 따른 상기 화장료 조성물의 제형이 페이스트, 크림 또는 겔인 경우에는 담체 성분으로서 동물섬유, 식물섬유, 왁스, 파라핀, 전분, 트라칸트, 셀룰로오스 유도체, 폴리에틸렌 글리콜, 실리콘, 벤토나이트, 실리카, 탈크 또는 산화아연 등이 이용될 수 있다.When the formulation of the cosmetic composition according to one aspect is a paste, cream or gel, animal fiber, plant fiber, wax, paraffin, starch, tracant, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc or oxidized carrier ingredient. Zinc, etc. may be used.
일 양상에 따른 상기 화장료 조성물의 제형이 파우더 또는 스프레이인 경우에는 담체 성분으로서 락토스, 탈크, 실리카, 알루미늄히드록시드, 칼슘 실리케이트 또는 폴리아미드 파우더가 이용될 수 있고, 스프레이인 경우에는 추가적으로 클로로플루오로히드로카본, 프로판/부탄 또는 디메틸 에테르와 같은 추진체를 포함할 수 있다.If the formulation of the cosmetic composition according to one aspect is a powder or spray, lactose, talc, silica, aluminum hydroxide, calcium silicate, or polyamide powder may be used as the carrier ingredient, and in the case of a spray, chlorofluoro may be additionally used. May contain propellants such as hydrocarbons, propane/butane or dimethyl ether.
일 양상에 따른 상기 화장료 조성물의 제형이 용액 또는 유탁액의 경우에는 담체 성분으로서 용매, 용매화제 또는 유탁화제가 이용되고, 예컨대 물, 에탄올, 이소프로판올, 에틸 카보네이트, 에틸 아세테이트, 벤질 알코올, 벤질 벤조에이트, 프로필렌 글리콜, 1,3-부틸글리콜 오일, 글리세롤 지방족 에스테르, 폴리에틸렌 글리콜 또는 소르비탄의 지방산 에스테르 등이 이용될 수 있다.When the formulation of the cosmetic composition according to one aspect is a solution or emulsion, a solvent, solvating agent, or emulsifying agent is used as a carrier component, such as water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, and benzyl benzoate. , propylene glycol, 1,3-butyl glycol oil, glycerol aliphatic ester, polyethylene glycol, or sorbitan fatty acid ester, etc. can be used.
일 양상에 따른 조성물의 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.When the formulation of the composition according to one aspect is a suspension, the carrier component may include water, a liquid diluent such as ethanol or propylene glycol, a suspending agent such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester, and polyoxyethylene sorbitan ester, Microcrystalline cellulose, aluminum metahydroxide, bentonite, agar, or tracant may be used.
다른 양상은 상기 복합발효물을 유효성분으로 포함하는, 탈모 방지 및/또는 발모 촉진용 피부외용제 조성물을 제공하는 것이다. 상기에서 설명한 내용과 동일한 부분은 상기 피부외용제 조성물에도 공히 적용된다.Another aspect is to provide a composition for external skin application for preventing hair loss and/or promoting hair growth, comprising the complex fermentation product as an active ingredient. The same parts as described above also apply to the external skin composition.
상기 피부외용제 조성물은 피부 외부에서 도포되는 어떠한 것이라도 포함할 수 있으며, 예를 들어, 크림, 젤, 패치, 분무제, 연고제, 경고제, 로션제, 리니먼트제, 파스타제 및 카타플라스마제로 이루어진 군으로부터 선택되는 어느 하나일 수 있으나, 이에 특별히 제한되는 것은 아니다.The external skin composition may include anything applied from the outside of the skin, for example, a group consisting of creams, gels, patches, sprays, ointments, warning agents, lotions, linear agents, pasta agents, and cataplasma agents. It may be any one selected from, but is not particularly limited thereto.
상기 피부외용제는 유효성분 이외에, 예를 들어, 수성성분, 유성성분, 분말성분, 알코올류, 보습제, 증점제, 자외선흡수제, 미백제, 방부제, 산화방지제, 계면활성제, 향료, 색제, 피부 영양제 또는 이들의 조합을 포함할 수 있으며, 이들은 필요에 따라서 당업자에 의해 적절하게 배합될 수 있다. 또한, 상기 피부외용제에는 에데트산이나트륨, 에데트산삼나트륨, 시트르산나트륨, 폴리인산나트륨, 메타인산나트륨, 글루콘산 등의 금속봉쇄제, 카페인, 탄닌, 벨라파밀, 감초추출물, 글라블리딘, 칼린의 과실의 열수추출물, 각종생약, 아세트산토코페롤, 글리틸리틴산, 트라넥삼산 및 그 유도체 또는 그 염 등의 약제, 비타민 C, 아스코르브산인산마그네슘, 아스코르브산글루코시드, 알부틴, 코지산, 글루코스, 프룩토스, 트레할로스 등의 당류가 배합될 수 있으나, 이에 특별히 제한되는 것은 아니다.In addition to the active ingredients, the external skin preparations include, for example, aqueous ingredients, oil-based ingredients, powder ingredients, alcohols, moisturizers, thickeners, ultraviolet absorbers, whitening agents, preservatives, antioxidants, surfactants, fragrances, colorants, skin nutrients or these. Combinations may be included, and these may be appropriately combined by a person skilled in the art as needed. In addition, the skin external preparations include metal sequestrants such as disodium edetate, trisodium edetate, sodium citrate, sodium polyphosphate, sodium metaphosphate, and gluconic acid, caffeine, tannin, belafamil, licorice extract, glablidin, and calin. hot water extract of fruits, various herbal medicines, medicines such as tocopherol acetate, glythylitic acid, tranexamic acid and its derivatives or salts, vitamin C, magnesium ascorbate phosphate, ascorbate glucoside, arbutin, kojic acid, glucose, frug Sugars such as toss and trehalose may be mixed, but are not particularly limited thereto.
또 다른 양상은 상기 복합발효물을 유효성분으로 포함하는, 탈모 예방 또는 치료용 약학적 조성물을 제공하는 것이다. 상기에서 설명한 내용과 동일한 부분은 상기 약학적 조성물에도 공히 적용된다.Another aspect is to provide a pharmaceutical composition for preventing or treating hair loss, comprising the complex fermentation product as an active ingredient. The same parts as described above also apply to the pharmaceutical composition.
상기 약학적 조성물은 본 명세서에 개시된 유효성분 이외에 방부제, 안정화제, 수화제 또는 유화 촉진제, 삼투압 조절을 위한 염 및/또는 완충제 등의 약제학적 보조제 및 기타 치료적으로 유용한 물질을 추가로 함유할 수 있으며, 통상적인 방법에 따라 다양한 경구 투여제 또는 비경구 투여제 형태로 제형화될 수 있다.In addition to the active ingredients disclosed herein, the pharmaceutical composition may further contain pharmaceutical auxiliaries such as preservatives, stabilizers, wetting agents or emulsification accelerators, salts and/or buffers for osmotic pressure adjustment, and other therapeutically useful substances. , can be formulated into various oral or parenteral dosage forms according to conventional methods.
상기 경구 투여제는, 예를 들어, 정제, 환제, 경질 및 연질 캅셀제, 액제, 현탁제, 유화제, 시럽제, 분제, 산제, 세립제, 과립제, 펠렛제 등이 있으며, 이들 제형은 유효성분 이외에 계면 활성제, 희석제(예를 들어, 락토즈, 덱스트로즈, 수크로즈, 만니톨, 솔비톨, 셀룰로오스 및 글리신), 활택제(예를 들어, 실리카, 탈크, 스테아르산 및 그의 마그네슘 또는 칼슘염 및 폴리에틸렌 글리콜)를 포함할 수 있다. 상기 정제는 마그네슘 알루미늄 실리케이트, 전분페이스트, 젤라틴, 트라가칸스, 메틸셀룰로오스, 나트륨 카복시메틸셀룰로오스 및 폴리비닐피롤리딘과 같은 결합제를 포함할 수 있으며, 추가적으로 전분, 한천, 알긴산 또는 그의 나트륨 염과 같은 붕해제, 흡수제, 착색제, 향미제, 및 감미제 등의 약제학적 첨가제를 포함할 수 있다. 또한, 상기 정제는 통상적인 혼합, 과립화 또는 코팅 방법에 의해 제조될 수 있다.The oral administration agents include, for example, tablets, pills, hard and soft capsules, solutions, suspensions, emulsifiers, syrups, powders, powders, fine granules, granules, pellets, etc., and these dosage forms contain, in addition to the active ingredient, the surface Activators, diluents (e.g. lactose, dextrose, sucrose, mannitol, sorbitol, cellulose and glycine), lubricants (e.g. silica, talc, stearic acid and its magnesium or calcium salts and polyethylene glycol) may include. The tablet may contain a binder such as magnesium aluminum silicate, starch paste, gelatin, tragacanth, methylcellulose, sodium carboxymethylcellulose and polyvinylpyrrolidine, and may additionally contain a binder such as starch, agar, alginic acid or its sodium salt. It may contain pharmaceutical additives such as disintegrants, absorbents, colorants, flavoring agents, and sweeteners. Additionally, the tablets can be manufactured by conventional mixing, granulating or coating methods.
상기 비경구 투여제는 경피 투여형 제형일 수 있으며, 예를 들어, 주사제, 점적제, 연고, 로션, 겔, 크림, 스프레이, 현탁제, 유제, 좌제(坐劑), 패치 등의 제형일 수 있으나, 이에 특별히 제한되는 것은 아니다.The parenteral administration may be in a transdermal dosage form, for example, injections, drops, ointments, lotions, gels, creams, sprays, suspensions, emulsions, suppositories, patches, etc. However, it is not particularly limited thereto.
상기 약학적 조성물은 비경구, 직장, 국소, 경피, 피하 등으로 투여될 수 있으며, 보다 구체적으로는 두피에 국소 투여될 수 있다.The pharmaceutical composition may be administered parenterally, rectally, topically, transdermally, subcutaneously, etc., and more specifically, may be administered topically to the scalp.
상기 약학적 조성물의 투여량 결정은 당업자의 기술 수준 내에 있으며, 1일 투여 용량은 투여하고자 하는 대상의 진행 정도, 발병 시기, 연령, 건강상태, 합병증 등의 다양한 요인을 고려하여 당업자가 적절히 선택할 수 있다.Determination of the dosage of the pharmaceutical composition is within the skill level of a person skilled in the art, and the daily dosage can be appropriately selected by a person skilled in the art in consideration of various factors such as the degree of progression, onset time, age, health condition, and complications of the subject to be administered. there is.
또 다른 양상은 두유, 호밀 및 대두를 바이오컨버전(bioconversion)시킨 후 혼합한 복합발효물을 유효성분으로 포함하는 조성물을 개체에 투여하는 단계를 포함하는, 탈모 방지 및 발모 촉진 방법을 제공한다. 상기에서 설명한 내용과 동일한 부분은 상기 방법에도 공히 적용된다.Another aspect provides a method for preventing hair loss and promoting hair growth, comprising administering to an individual a composition containing as an active ingredient a complex fermentation product obtained by bioconverting soymilk, rye, and soybeans. The same parts as described above also apply to the above method.
구체적으로, 본 발명의 복합발효물을 포함하는 조성물은 화장료 조성물, 피부외용제 조성물, 약학적 조성물 또는 의약외품 조성물 등으로 개체에 투여될 수 있다. 상기 투여하는 단계에 있어서 투여 방법은 특별히 제한되지 않으며, 상기 조성물의 제형 및 당업계 통상의 기술자에 따라 적절히 선택될 수 있다. 구체적으로, 상기 조성물은 경구, 비경구, 직장, 국소, 경피, 정맥 내, 근육 내, 복강 내, 피하 등으로 투여될 수 있으며, 보다 구체적으로는 두피에 도포하거나 국소 투여될 수 있다.Specifically, the composition containing the complex fermentation product of the present invention may be administered to an individual as a cosmetic composition, external skin preparation composition, pharmaceutical composition, or quasi-drug composition. In the administration step, the administration method is not particularly limited and may be appropriately selected depending on the formulation of the composition and those skilled in the art. Specifically, the composition can be administered orally, parenterally, rectally, topically, transdermally, intravenously, intramuscularly, intraperitoneally, subcutaneously, etc., and more specifically, it can be applied to the scalp or administered topically.
일 양상에 따른 화장료 조성물은 인간 모유두세포(Human Follicular Dermal Papilla Cells: HFDPC)에서 VEGF 및 FGF7의 발현을 증가시켜 모유두세포로의 영양분 전달 촉진, 모근부 세포의 생장을 유도하고 모유두세포의 증식을 유도할 수 있으며, 이를 통해 발모의 촉진 및 탈모의 예방 효과를 기대할 수 있다.The cosmetic composition according to one aspect increases the expression of VEGF and FGF7 in human follicular dermal papilla cells (HFDPC), promotes nutrient delivery to dermal papilla cells, induces growth of hair root cells, and induces proliferation of dermal papilla cells. This can be expected to promote hair growth and prevent hair loss.
도 1은 양성대조군 AS2P 대비 본 발명 복합발효물의 농도에 따른 VEGF 유전자 발현 증가 효과를 확인한 결과를 나타낸 것이다.Figure 1 shows the results confirming the effect of increasing VEGF gene expression according to the concentration of the complex fermentation product of the present invention compared to the positive control group AS2P.
도 2는 양성대조군 AS2P 대비 본 발명 복합발효물의 농도에 따른 FGF7 유전자 발현 증가 효과를 확인한 결과를 나타낸 것이다.Figure 2 shows the results confirming the effect of increasing FGF7 gene expression according to the concentration of the complex fermentation product of the present invention compared to the positive control group AS2P.
도 3은 유산균 또는 고초균 배양액, 천연물 원물(두유, 호밀분말, 대두) 대비 실시예 1 내지 4 복합발효물의 VEGF 유전자 발현 증가 효과를 확인한 결과를 나타낸 것이다.Figure 3 shows the results of confirming the effect of increasing VEGF gene expression of the complex fermentation products of Examples 1 to 4 compared to the lactic acid bacteria or Bacillus subtilis culture and natural raw materials (soy milk, rye powder, soybean).
도 4는 유산균 또는 고초균 배양액, 천연물 원물(두유, 호밀분말, 대두) 대비 실시예 1 내지 4 복합발효물의 FGF7 유전자 발현 증가 효과를 확인한 결과를 나타낸 것이다.Figure 4 shows the results of confirming the effect of increasing FGF7 gene expression of the complex fermentation products of Examples 1 to 4 compared to lactic acid bacteria or Bacillus subtilis culture and natural raw materials (soy milk, rye powder, soybean).
도 5은 비타민 B군(Vitamin B1, B2, B3, B5, B6, B9, B12) 및 이들의 혼합군 대비 실시예 1 복합발효물의 VEGF 유전자 발현 증가 효과를 확인한 결과를 나타낸 것이다.Figure 5 shows the results confirming the effect of increasing VEGF gene expression of the complex fermented product of Example 1 compared to the vitamin B group (Vitamin B1, B2, B3, B5, B6, B9, B12) and their mixed group.
도 6는 비타민 B군(Vitamin B1, B2, B3, B5, B6, B9, B12) 및 이들의 혼합군 대비 실시예 1 복합발효물의 FGF7 유전자 발현 증가 효과를 확인한 결과를 나타낸 것이다.Figure 6 shows the results confirming the effect of increasing FGF7 gene expression of the complex fermentation of Example 1 compared to the vitamin B group (Vitamin B1, B2, B3, B5, B6, B9, B12) and their mixed group.
이하 실험예를 통하여 보다 상세하게 설명한다. 그러나, 이들 실험예는 예시적으로 설명하기 위한 것으로 본 발명의 범위가 이들 실험예에 한정되는 것은 아니다.Hereinafter, it will be described in more detail through an experimental example. However, these experimental examples are for illustrative purposes only and the scope of the present invention is not limited to these experimental examples.
실험예 1: 발효 추출물로부터 비타민 제조Experimental Example 1: Preparation of vitamins from fermented extract
천연물에 미생물 발효를 접목한 바이오컨버전(Bio-conversion) 기술을 도입하여 발효 비타민을 제조하였다. Fermented vitamins were manufactured by introducing bio-conversion technology, which combines microbial fermentation with natural products.
실험예 1-1: 유산균/두유 발효 여과물Experimental Example 1-1: Lactic acid bacteria/soymilk fermentation filtrate
두유는 대두(콩)를 중량의 10배의 물을 첨가한 후 믹서기로 분쇄하여 150 메쉬 여과망(104 micron)으로 여과하여 사용하였다. 발효 배지는 2.0 % glucose, 1.0 % Na2HPO4, 0.5% NaH2PO4의 성분으로 제조하였고, 이후 90% 두유를 첨가하여 멸균하였다. 락토바실러스 플란타룸(Lactobacillus plantarum)을 발효배지에 접종하고, 30 ℃에서 24시간 동안 정치발효를 진행하였다. 발효 종료 후 연속 원심분리기를 이용하여 남아있는 균체 등 불용성 물질을 제거하였다. 회수한 발효물은 2N KOH로 중화(pH 6.0)하고, 방부 후 제균 여과하였다. 이후 해당 발효 여과물이 리보플라빈(Vitamin B2), 피리독신(Vitamin B6) 및 시아노코발라민(Vitamin B12)을 함유하고 있는 것을 확인하였다.Soy milk was used by adding 10 times the weight of water to soybeans, pulverizing them in a blender, and filtering them through a 150 mesh filter (104 micron). The fermentation medium was prepared with the following ingredients: 2.0% glucose, 1.0% Na 2 HPO 4 , and 0.5% NaH 2 PO 4 , and then sterilized by adding 90% soy milk. Lactobacillus plantarum was inoculated into the fermentation medium, and stationary fermentation was performed at 30°C for 24 hours. After completion of fermentation, insoluble substances such as remaining bacterial cells were removed using a continuous centrifuge. The recovered fermented product was neutralized with 2N KOH (pH 6.0), preservative, and filtered for sterilization. Afterwards, it was confirmed that the fermentation filtrate contained riboflavin (Vitamin B2), pyridoxine (Vitamin B6), and cyanocobalamin (Vitamin B12).
실험예 1-2: 유산균/호밀분말 발효물Experimental Example 1-2: Lactic acid bacteria/rye powder fermentation product
호밀분말을 중량의 5배의 물을 첨가한 후 습윤멸균시켰다. 이후 호밀분말의 전분을 분해하기 위하여 전분분해효소(amylase)를 이용하여 중량 대비 0.1%를 투입하고, 50~60 ℃에서 24시간 동안 전분분해 후 발효에 사용하였다. 발효배지를 2.0 % glucose, 1.0 % Na2HPO4, 0.5% NaH2PO4의 성분으로 제조하고, 이후 90% 호밀분말을 첨가하여 멸균하였다. 락토바실러스 플란타룸(Lactobacillus plantarum)을 발효배지에 접종하고, 30 ℃에서 48시간 동안 정치발효를 진행하였다. 발효 종료 후 연속 원심분리기를 이용하여 남아있는 균체 등 불용성 물질을 제거하였다. 회수한 발효물은 2N KOH(pH 6.0)로 중화하고, 방부 후 제균 여과하였다. 이후 해당 발효물이 티아민(Vitamin B1), 니코틴산(Vitamin B3) 및 판토텐산(Vitamin B5)을 함유하고 있는 것을 확인하였다.Rye powder was wet-sterilized after adding 5 times the weight of water. Afterwards, in order to decompose the starch in the rye powder, 0.1% of the weight was added using amylase, and the starch was decomposed at 50-60°C for 24 hours and then used for fermentation. Fermentation medium was prepared with the following ingredients: 2.0% glucose, 1.0% Na 2 HPO 4 , and 0.5% NaH 2 PO 4 , and then 90% rye powder was added and sterilized. Lactobacillus plantarum was inoculated into the fermentation medium, and stationary fermentation was performed at 30°C for 48 hours. After completion of fermentation, insoluble substances such as remaining bacterial cells were removed using a continuous centrifuge. The recovered fermented product was neutralized with 2N KOH (pH 6.0), preservative, and filtered for sterilization. Afterwards, it was confirmed that the fermented product contained thiamine (Vitamin B1), nicotinic acid (Vitamin B3), and pantothenic acid (Vitamin B5).
실험예 1-3: 고초균/대두 발효 추출물Experimental Example 1-3: Bacillus subtilis/soybean fermentation extract
정제수에 대두 5.0 %를 넣고 100 ℃에서 1시간 동안 가열하였다. 이후 냉각하여 고형분을 제거하여 추출물만 회수하였다. 발효배지를 2.0 % glucose, 1.0 % Na2HPO4, 0.5% NaH2PO4의 성분으로 제조하고, 90% 대두 추출물, 엽산 0.5%를 첨가하여 멸균하였다. 고초균(Bacillus subtilis)을 발효배지에 접종하고, 45 ℃에서 72시간 동안 정치발효를 진행하였다. 발효 종료 후 연속 원심분리기를 이용하여 남아있는 균체 등 불용성 물질을 제거하고, 2M HCl을 사용하여 pH 4까지 조정하였다. 이후 2 배수의 에탄올을 투입하여 고초균/대두 발효추출물 고형분을 회수하였다. 회수한 고형분을 2N KOH로 중화(pH 6.0)하고, 방부 후 제균 여과하였다. 이후 해당 발효 추출물 내 고분자 형태의 엽산(Vitamin B9)을 함유하고 있는 것을 확인하였다.5.0% soybeans were added to purified water and heated at 100°C for 1 hour. Afterwards, it was cooled to remove the solid content and only the extract was recovered. Fermentation medium was prepared with ingredients of 2.0% glucose, 1.0% Na 2 HPO 4 and 0.5% NaH 2 PO 4 and sterilized by adding 90% soybean extract and 0.5% folic acid. Bacillus subtilis was inoculated into the fermentation medium, and stationary fermentation was performed at 45°C for 72 hours. After completion of fermentation, insoluble substances such as remaining bacterial cells were removed using a continuous centrifuge, and the pH was adjusted to 4 using 2M HCl. Afterwards, 2 times the amount of ethanol was added to recover the solid content of the Bacillus subtilis/soybean fermentation extract. The recovered solid content was neutralized with 2N KOH (pH 6.0), preservative, and filtered for sterilization. Afterwards, it was confirmed that the fermented extract contained high molecular form folic acid (Vitamin B9).
각각의 발효물을 서로 다른 비율로 혼합한 복합발효물에 대해 발모 촉진 효과를 확인하고자 하였으며, 이에 비타민 B군을 포함하는 각 복합발효물(실시예 1 내지 4)의 조성비를 아래 표 1에 나타내었다.We attempted to confirm the effect of promoting hair growth in complex fermentations in which each fermentation product was mixed at different ratios. Accordingly, the composition ratios of each complex fermentation product (Examples 1 to 4) containing vitamin B group are shown in Table 1 below. It was.
유산균/두유 발효물Lactic acid bacteria/soy milk fermentation product | 유산균/호밀분말 발효물Lactic acid bacteria/fermented rye powder | 고초균/대두 발효물Bacillus subtilis/soybean fermentation | |
실시예 1Example 1 | 1One | 1One | 1One |
실시예 2Example 2 | 22 | 1One | 1One |
실시예 3Example 3 | 1One | 22 | 1One |
실시예 4Example 4 | 1One |
1 |
22 |
실험예 2: 복합발효물의 모발 성장인자(VEGF 및 FGF7) 발현 증가 효과 확인Experimental Example 2: Confirmation of the effect of increasing the expression of hair growth factors (VEGF and FGF7) of complex fermentation
실험예 2-1: HFDPC에서 Real-time PCR(실시간 중합효소 연쇄반응)을 이용한 VEGF 및 FGF7 발현 증가 확인Experimental Example 2-1: Confirmation of increased expression of VEGF and FGF7 using Real-time PCR (real-time polymerase chain reaction) in HFDPC
실험예 1에서 제조한 각각의 복합발효물을 처리함에 따라 인간모유두세포(HFDPC)에서 모발 성장을 유도하는 인자인 VEGF 및 FGF7의 발현이 유도될 수 있는지 여부를 평가하기 위해, 실시간 중합효소연쇄반응(Quantitative Real-Time Polymerase Chain Reaction: qRT-PCR)을 수행하였다. 먼저 HFDPC를 6웰 플레이트에 3×105 세포/웰씩 분주한 후, 세포 배양 조건에서 24시간 배양하였다. 배양 24시간 후 배지를 버리고 PBS로 세척한 다음, 배지 보충물을 포함하지 않은 배지를 사용하여 세포를 기아 상태로 만들어 준 후, 다음날 아무것도 포함하지 않은 배지와 실시예 1 내지 4의 복합발효물을 포함하는 배지로 교체해 준 후, 24시간 동안 추가 배양하였다. 이후 RNeasy Mini Kit(Qiagen)을 이용하여 세포에서 RNA를 분리한 뒤, 나노드롭(nanodrop)을 이용하여 260nm에서 RNA를 정량한 후, 각각 2㎍의 RNA를 사용하여 증폭기(C1000 Thermal Cycler, Bio-Rad, 미국)에서 cDNA를 합성하였다. 합성된 cDNA에 타겟 단백질인 VEGF, FGF7 프라이머와 시아닌 염료인 사이버그린(SYBR Green supermis, Applied Biosystems, 미국)을 첨가한 혼합물을 이용하여 real-time PCR 기계에서 실시간 중합효소 연쇄반응을 실시함으로써, 최종적으로 VEGF, FGF7 유전자의 발현 정도를 평가하였다. 사용된 프라이머의 서열 및 반응 조건은 아래 표 2에 나타냈으며, 각 유전자의 발현량은 β-actin 유전자에 대한 보정을 통해 최종적으로 분석하였다. 양성 대조군으로는 AS2P(Ascorbic-acid 2-phosphate, 2.5mM)을 사용하였다.To evaluate whether the expression of VEGF and FGF7, factors that induce hair growth in human hair papilla cells (HFDPC), can be induced by processing each complex fermentation product prepared in Experimental Example 1, real-time polymerase chain reaction was used. (Quantitative Real-Time Polymerase Chain Reaction: qRT-PCR) was performed. First, HFDPC was dispensed into a 6-well plate at 3×10 5 cells/well and then cultured for 24 hours under cell culture conditions. After 24 hours of incubation, the medium was discarded and washed with PBS, and the cells were starved using medium containing no medium supplements. The next day, medium containing nothing and the complex fermentation products of Examples 1 to 4 were used. After replacing with the medium containing the culture, additional culture was performed for 24 hours. Afterwards, RNA was isolated from cells using the RNeasy Mini Kit (Qiagen), RNA was quantified at 260 nm using a nanodrop, and 2 ㎍ of RNA was used each to be cycled using an amplifier (C1000 Thermal Cycler, Bio- cDNA was synthesized at Rad, USA). By performing a real-time polymerase chain reaction on a real-time PCR machine using a mixture of synthesized cDNA with target protein VEGF and FGF7 primers and cyanine dye Cybergreen (SYBR Green supermis, Applied Biosystems, USA), the final The expression levels of VEGF and FGF7 genes were evaluated. The sequences of the primers used and reaction conditions are shown in Table 2 below, and the expression level of each gene was finally analyzed through correction for the β-actin gene. AS2P (Ascorbic-acid 2-phosphate, 2.5mM) was used as a positive control.
프라이머 서열primer sequence | 서열번호sequence number | 서열order | 반응 조건reaction conditions | |
VEGFVEGF | FF | 1One | 5'-GTGCCCACTGAGGAGTTCAAC-3'5'-GTGCCCACTGAGGAGTTCAAC-3' | 94℃5분 동안 중합 효소 활성화 후 95℃30초, 60℃30초, 72℃30초 조건에서 40 사이클로 중합반응After activating the polymerase at 94°C for 5 minutes, polymerization reaction was performed in 40 cycles at 95°C for 30 seconds, 60°C for 30 seconds, and 72°C for 30 seconds. |
R |
22 | 5'-CCCTATGTGCTGGCCTTGAT-3'5'-CCCTATGTGCTGGCCTTGAT-3' | ||
FGF7 | FF | 33 | 5'-TCCTGCCAACTTTGCTCTACA-3'5'-TCCTGCCAACTTTGCTCTACA-3' | |
R |
44 | 5'-CAGGGCTGGAACAGTTCACAT-3'5'-CAGGGCTGGAACAGTTCACAT-3' | ||
β-actinβ-actin | FF | 55 | 5'-GGCCATCTCTTGCTCGAAGT-3'5'-GGCCATCTCTTGCTCGAAGT-3' | |
R |
66 | 5'-GAGACCTTCAACACCCCAGC-3'5'-GAGACCTTCAACACCCCAGC-3' |
실험예 2-2: 복합발효물의 농도에 따른 VEGF 및 FGF7 발현 증가 확인Experimental Example 2-2: Confirmation of increase in VEGF and FGF7 expression according to concentration of complex fermentation product
도 1 및 도 2에서 확인되는 바와 같이, 본 발명의 복합발효물(실시예 1)을 처리한 군에서 VEGF 및 FGF7 유전자 발현 증가가 확인되었고, 특히 복합발효물을 1% 농도로 처리하였을 때 양성 대조군 AS2P와 비교하여 현저한 수준의 VEGF 및 FGF7 유전자 발현 증가 효과가 나타나는 것을 확인하였다.As confirmed in Figures 1 and 2, an increase in VEGF and FGF7 gene expression was confirmed in the group treated with the complex fermentation product of the present invention (Example 1), and in particular, when the complex fermentation product was treated at a concentration of 1%, it was positive. It was confirmed that there was a significant increase in VEGF and FGF7 gene expression compared to the control AS2P.
실험예 2-3: 두유 발효 여과물, 호밀 발효물 및 대두 발효 추출물의 함량 비율에 따른 VEGF 및 FGF7 발현 증가의 상승 효과 확인Experimental Example 2-3: Confirmation of the synergistic effect of increasing VEGF and FGF7 expression according to the content ratio of soymilk fermentation filtrate, rye fermentation, and soybean fermentation extract
실험예 1의 발효물을 서로 다른 비율로 혼합한 실시예 1 내지 4의 복합발효물은 유산균 또는 고초균 배양액, 천연물 원물(두유, 호밀분말, 대두) 대비 현저히 우수한 VEGF 및 FGF7 유전자 발현 증가 효과가 있는 것으로 나타났으며, 실시예 2를 제외하고는 모두 양성 대조군인 AS2P와 비교하여서도 더 우수한 VEGF 및 FGF7 유전자 발현 증가 효과가 있음을 확인하였다(도 3 및 도 4). 또한, 그 중 가장 효과가 우수한 것으로 나타는 실시예 1의 복합발효물의 경우, 각각의 비타민 B군(Vitamin B1, B2, B3, B5, B6, B9, B12) 및 이들의 혼합군(각각 1:1:1:1:1:1:1 중량부로 포함) 대비 현저히 우수한 VEGF 및 FGF7 유전자 발현 증가 효과가 있음을 확인하였다(도 5 및 도 6).The complex fermented products of Examples 1 to 4, which were mixed with the fermented products of Experimental Example 1 at different ratios, had a significantly better effect of increasing VEGF and FGF7 gene expression compared to the lactic acid bacteria or Bacillus subtilis culture and natural raw materials (soy milk, rye powder, soybeans). It was confirmed that, except for Example 2, there was a superior effect of increasing VEGF and FGF7 gene expression compared to AS2P, which was a positive control (FIGS. 3 and 4). In addition, in the case of the complex fermentation product of Example 1, which was shown to be the most effective among them, each vitamin B group (Vitamin B1, B2, B3, B5, B6, B9, B12) and their mixture group (1 each: It was confirmed that there was a significantly better effect of increasing VEGF and FGF7 gene expression compared to (included in 1:1:1:1:1:1 parts by weight) (FIGS. 5 and 6).
전술한 본 발명의 설명은 예시를 위한 것이며, 본 발명이 속하는 기술분야의 통상의 지식을 가진 자는 본 발명의 기술적 사상이나 필수적인 특징을 변경하지 않고서 다른 구체적인 형태로 쉽게 변형이 가능하다는 것을 이해할 수 있을 것이다. 그러므로 이상에서 기술한 실시예들은 모든 면에서 예시적인 것이며 한정적이 아닌 것으로 이해해야만 한다.The description of the present invention described above is for illustrative purposes, and those skilled in the art will understand that the present invention can be easily modified into other specific forms without changing the technical idea or essential features of the present invention. will be. Therefore, the embodiments described above should be understood in all respects as illustrative and not restrictive.
Claims (12)
- 두유, 호밀 및 대두를 바이오컨버전(bioconversion)시킨 복합발효물을 유효성분으로 포함하는, 탈모 방지 및 발모 촉진용 화장료 조성물.A cosmetic composition for preventing hair loss and promoting hair growth, comprising as an active ingredient a complex fermentation product obtained by bioconversion of soy milk, rye, and soybeans.
- 제1항에 있어서, 상기 복합발효물은 두유 발효 여과물, 호밀 발효물 및 대두 발효 추출물을 포함하는 것인, 화장료 조성물.The cosmetic composition of claim 1, wherein the complex fermentation product includes soymilk fermentation filtrate, rye fermentation product, and soybean fermentation extract.
- 제2항에 있어서, 상기 복합발효물은 상기 두유 발효 여과물 1 내지 5 중량부, 상기 호밀 발효물 1 내지 5 중량부 및 상기 대두 발효 추출물 1 내지 5 중량부를 포함하는 것인, 화장료 조성물.The cosmetic composition of claim 2, wherein the complex fermentation product includes 1 to 5 parts by weight of the soymilk fermentation filtrate, 1 to 5 parts by weight of the rye fermentation product, and 1 to 5 parts by weight of the soybean fermentation extract.
- 제2항에 있어서, 상기 복합발효물은 상기 두유 발효 여과물, 호밀 발효물 및 대두 발효 추출물을 1:1:1의 중량부로 포함하는 것인, 화장료 조성물.The cosmetic composition according to claim 2, wherein the complex fermentation product includes the soymilk fermentation filtrate, rye fermentation product, and soybean fermentation extract in a weight ratio of 1:1:1.
- 제2항에 있어서, 상기 두유 발효 여과물은 상기 두유에 락토바실러스 속(Lactobacillus sp.) 균주를 접종하여 발효시킨 것인, 화장료 조성물. The cosmetic composition of claim 2, wherein the soymilk fermentation filtrate is obtained by inoculating the soymilk with Lactobacillus sp. and fermenting it.
- 제2항에 있어서, 상기 호밀 발효물은 호밀분말에 락토바실러스 속(Lactobacillus sp.) 균주를 접종하여 발효시킨 것인, 화장료 조성물.The cosmetic composition according to claim 2, wherein the fermented rye product is fermented by inoculating rye powder with a Lactobacillus sp. strain.
- 제2항에 있어서, 상기 대두 발효 추출물은 상기 대두에 바실러스 속(Bacillus sp.) 균주를 접종하여 발효시킨 것인, 화장료 조성물. The cosmetic composition of claim 2, wherein the fermented soybean extract is fermented by inoculating the soybean with a Bacillus sp. strain.
- 제1항에 있어서, 상기 복합발효물은 천연 비타민 티아민(Vitamin B1), 리보플라빈(Vitamin B2), 니코틴산(Vitamin B3), 판토텐산(Vitamin B5), 피리독신(Vitamin B6), 엽산(Vitamin B9) 및 시아노코발라민(Vitamin B12)을 포함하는 것인, 화장료 조성물.The method of claim 1, wherein the complex fermentation product contains natural vitamins thiamine (Vitamin B1), riboflavin (Vitamin B2), nicotinic acid (Vitamin B3), pantothenic acid (Vitamin B5), pyridoxine (Vitamin B6), folic acid (Vitamin B9), and cyanoacid. A cosmetic composition containing nocobalamin (Vitamin B12).
- 제1항에 있어서, 상기 복합발효물은 인간 모유두세포(Human Follicular Dermal Papilla Cells: HFDPC)에서 VEGF 및 FGF7의 발현을 증가시키는 것인, 화장료 조성물.The cosmetic composition of claim 1, wherein the complex fermentation product increases the expression of VEGF and FGF7 in human follicular dermal papilla cells (HFDPC).
- 두유, 호밀 및 대두를 바이오컨버전시킨 복합발효물을 유효성분으로 포함하는, 탈모 방지 및 발모 촉진용 피부외용제 조성물.A composition for external application for skin for preventing hair loss and promoting hair growth, comprising as an active ingredient a complex fermentation product obtained by bioconverting soymilk, rye, and soybeans.
- 두유, 호밀 및 대두를 바이오컨버전시킨 복합발효물을 유효성분으로 포함하는, 탈모 예방 또는 치료용 약학적 조성물. A pharmaceutical composition for preventing or treating hair loss, comprising as an active ingredient a complex fermentation product obtained by bioconverting soymilk, rye, and soybeans.
- 두유, 호밀 및 대두를 바이오컨버전(bioconversion)시킨 후 혼합한 복합발효물을 유효성분으로 포함하는 조성물을 개체에 투여하는 단계를 포함하는, 탈모 방지 및 발모 촉진 방법.A method for preventing hair loss and promoting hair growth, comprising administering to an individual a composition containing as an active ingredient a complex fermentation product obtained by bioconverting soymilk, rye, and soybeans.
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PCT/KR2023/009580 WO2024034866A1 (en) | 2022-08-11 | 2023-07-06 | Composition for preventing hair loss and promoting hair growth including complex fermentation product containing natural vitamins |
Country Status (2)
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KR (1) | KR20240023456A (en) |
WO (1) | WO2024034866A1 (en) |
Citations (7)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20050089499A1 (en) * | 2002-01-15 | 2005-04-28 | Philippe Moussou | Active substances for use in cosmetic and/or pharmaceutical products, obtainable from the fermentation of plant components and/or plant extracts |
JP2009234942A (en) * | 2008-03-26 | 2009-10-15 | Toyobo Co Ltd | Cosmetic composition containing vegetable extract |
JP2012176931A (en) * | 2011-01-31 | 2012-09-13 | Angfa Co Ltd | Hair tonic |
WO2013143989A1 (en) * | 2012-03-26 | 2013-10-03 | Otc Gmbh | Hair conditioning composition for permanent and semi-permanent hair coloration applications |
JP2013538187A (en) * | 2010-07-12 | 2013-10-10 | ジュリアーニ ソシエタ ペル アチオニ | Fermented soy-based mixture comprising isoflavone-aglycone, equol and lunacil, its preparation method and use in the food, pharmaceutical and cosmetic fields |
KR101779480B1 (en) * | 2013-12-18 | 2017-09-27 | (주)제니트리 | Composition for prevention or treatment of depilation or improvement of hair growth having fermented peptone, and formulation having the same |
KR102069712B1 (en) * | 2019-06-21 | 2020-01-23 | 주식회사 티에스트릴리온 | Cosmetic Composition for Preventing Alopecia and Promoting Hair Growth Containing Natural Extract |
-
2022
- 2022-08-11 KR KR1020220100589A patent/KR20240023456A/en unknown
-
2023
- 2023-07-06 WO PCT/KR2023/009580 patent/WO2024034866A1/en unknown
Patent Citations (7)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20050089499A1 (en) * | 2002-01-15 | 2005-04-28 | Philippe Moussou | Active substances for use in cosmetic and/or pharmaceutical products, obtainable from the fermentation of plant components and/or plant extracts |
JP2009234942A (en) * | 2008-03-26 | 2009-10-15 | Toyobo Co Ltd | Cosmetic composition containing vegetable extract |
JP2013538187A (en) * | 2010-07-12 | 2013-10-10 | ジュリアーニ ソシエタ ペル アチオニ | Fermented soy-based mixture comprising isoflavone-aglycone, equol and lunacil, its preparation method and use in the food, pharmaceutical and cosmetic fields |
JP2012176931A (en) * | 2011-01-31 | 2012-09-13 | Angfa Co Ltd | Hair tonic |
WO2013143989A1 (en) * | 2012-03-26 | 2013-10-03 | Otc Gmbh | Hair conditioning composition for permanent and semi-permanent hair coloration applications |
KR101779480B1 (en) * | 2013-12-18 | 2017-09-27 | (주)제니트리 | Composition for prevention or treatment of depilation or improvement of hair growth having fermented peptone, and formulation having the same |
KR102069712B1 (en) * | 2019-06-21 | 2020-01-23 | 주식회사 티에스트릴리온 | Cosmetic Composition for Preventing Alopecia and Promoting Hair Growth Containing Natural Extract |
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KR20240023456A (en) | 2024-02-22 |
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