KR20200029323A - Cosmetic Composition For Preventing Hair Loss Containing Arctium Lappa Seed Extract - Google Patents

Abstract

The present invention relates to a cosmetic composition for preventing hair loss, which comprises arctin, arctigenin, lappaol, and lappaol C as Arctium lappa extracts in a predetermined ratio. The cosmetic composition for preventing hair loss is effective in suppressing dandruff-causing bacteria, Malassezia furfur, inhibiting 5a-reductase activity, propagating dermal papilla cells, and regulating hair-growth signalling of cells (β-catenin level and TGF-β level). The cosmetic composition for preventing hair loss is extracted with an extraction solvent selected from the group consisting of water, anhydrous or hydrous lower alcohol having 1-4 carbon atoms, acetone, ethyl acetate, and butyl acetate, and has a formulation selected from the group consisting of a solution, a suspension, an emulsion, a paste, a gel, a cream, a lotion, powder, a soap, a surfactant-containing cleanser, an oil, a powder foundation, an emulsion foundation, a wax foundation, and a spray.

Description

Cosmetic composition for preventing hair loss, which includes the extract of Ubangja as an active ingredient {Cosmetic Composition For Preventing Hair Loss Containing Arctium Lappa Seed Extract}

The present invention is an anti-hair loss cosmetic composition containing as an active ingredient an extract of Arctium lappa , actin, arctigenin, and lappaol or lappaol C, which is an extract of the acquaintance. It is about.

In general, hair loss refers to a condition in which no hair is present in a region where hair should normally be present, and means that the hair of the scalp falls out. In normal people, about 50 to 70 hairs per day drop out, and when more than 100 hairs fall out, symptoms of hair loss can be suspected.

Clinically, hair loss is classified into alopecia with scar formation and alopecia with no scar formation. It is caused by various causes, but genetic cause and testosterone, the male hormone, are considered to be important factors.

Testosterone is known to be involved in skeletal muscle growth, genital genital organs, scrotum growth, sperm formation, etc. Testosterone is known to be involved in hair loss, acne, and prostatic hypertrophy by being reduced to dihydrotestosterone by reductase (Diane et al; JID 775 -778, 1995. Bruchovsky, Net al; JBC 243,2112-2121, 1968).

In more detail, it is known that dihydrotestosterone causes hair loss by reducing the proliferation of hair follicle cells by lowering protein synthesis around the hair follicle.

-catenin 신호 전달 조절, TGF-

신호전달 조절의 효능을 가지고 있다.Existing treatments used to alleviate hair loss and promote hair growth are finasteride or minoxidil, and two types of drugs are widely used. The main mechanisms of these therapies are 5a-reductase selective inhibition, promoting papillary cell proliferation, ATP-sensitive K + -channel opening, Wnt /

-catenin signaling regulation, TGF-

It has the effect of regulating signal transmission.

Korean Patent Registration No. 10-131097 discloses a herbal extract for preventing hair loss and a hair product composition comprising the same. Republic of Korea Patent Registration No. 10-121834 discloses a composition containing a plant extract or fraction, such as Ubangja, its use, and a method for preparing the extract.

However, the known therapeutic agent has a temporary effect, and the effect may be different depending on the individual, so the need for new materials is increasing worldwide in relation to alleviation and hair loss.

The object of the present invention is to search for various skin physiological activities of natural plants and extracts thereof to alleviate hair loss and promote hair growth, and confirm the efficacy from extracts of natural plant extracts (Arctium lappa), its active ingredient actin, The present invention was completed by identifying actigenin, rapao and rapaol C.

Provided is a cosmetic composition for preventing hair loss, which uses actin, arctigenin, and rapaol derived from an extract of Arctium lappa as an active ingredient.

Preferably, it is a cosmetic composition for preventing hair loss, characterized in that it contains actin, actigenin, and rapaol in a certain proportion.

Preferably, the cosmetic composition for preventing hair loss is a cosmetic composition for preventing hair loss, which is characterized by inhibiting malesage puffer, a dandruff bacterium.

Preferably, the cosmetic composition for preventing hair loss is a cosmetic composition for preventing hair loss, characterized by inhibiting the activity of 5a-reductase.

-catenin level, TGF-

level)을 조절하는 탈모 방지용 화장료 조성물이다.Preferably, the cosmetic composition for preventing hair loss has the effect of proliferating the dermal papilla cells and signaling the hair proliferation of the cells (

-catenin level, TGF-

level) is a cosmetic composition for preventing hair loss.

Preferably, the yeonbangja extract is a cosmetic composition for preventing hair loss, containing 0.01 to 30.0% by weight of the total cosmetic composition weight.

Preferably, the cosmetic composition for preventing hair loss is extracted from an acquaintance with an extraction solvent selected from the group consisting of water, anhydrous or hydrous lower alcohol having 1 to 4 carbon atoms, acetone, ethyl acetate and butyl acetate. It is a composition.

Preferably, the type of the rapaol is a cosmetic composition for preventing hair loss, characterized in that Rapaol C.

Preferably, the cosmetic composition for preventing hair loss, characterized in that it is Rapaol C, is characterized in that it inhibits dandruff bacterium maleesia puffer.

Preferably, it contains 0.01 to 0.1% by weight of rapaol C with respect to the total weight of the cosmetic composition for preventing hair loss, characterized in that it is Rapaol C.

Preferably, the cosmetic composition for preventing hair loss, characterized in that it is Rapaol C, is characterized by inhibiting the activity of 5a-reductase.

-catenin level, TGF-

level)을 조절한다.Preferably, the cosmetic composition for preventing hair loss, characterized in that it is Rapaol C, has the effect of proliferating the dermal papilla cells and signaling the hair proliferation of the cells (

-catenin level, TGF-

level).

-catenin level, TGF-

level)을 조절하는 효과를 가진다.The present invention is characterized by preventing hair loss by a cosmetic composition comprising actin extract, actin, actigenin or rapol and rapol C. The cosmetic composition for preventing hair loss suppresses dandruff bacterium malesace puffer, inhibits the activity of 5a-reductase, proliferates the effect of dermal papilla cells, and signals the hair proliferation of cells (

-catenin level, TGF-

level).

All technical terms used in the present invention, unless defined otherwise, have the following definitions and conform to the meaning as commonly understood by those skilled in the art in the relevant field of the present invention. In addition, although a preferred method or sample is described herein, similar or equivalent ones are included in the scope of the present invention.

The term “about” means 30, 25, 20, 15, 10, 9, 8, 7, 6, 5, for reference amount, level, value, number, frequency, percent, dimension, size, amount, weight or length It means a quantity, level, value, number, frequency, percentage, dimension, size, quantity, weight or length that varies by 4, 3, 2 or 1%.

Throughout this specification, the terms "comprises" and "comprising", unless otherwise required by context, include the steps or components presented, or groups of steps or components, but any other steps or components, or It should be understood that it implies that the group of steps or components is not excluded.

The present invention relates to a cosmetic composition for preventing hair loss comprising actin, actigenin, rapaol, and rapaol C as active ingredients obtained from an extract of a visitor.

Ubangja used in the present invention is the fruit of the burdock, a perennial plant of the Chrysanthemum family, and is also known as a nickname such as Daedoja, Uchaeja, and Black satire. In September, the fruit tree hangs and ripens to grayish brown, with brown hairs attached and black seeds coming out from inside. The main efficacy of the friend is effective in alleviating symptoms such as sea water, sputum, and sore throat caused by wind fever.

Ubangja extract of the present invention can be prepared according to conventional methods known in the art, that is, under the conditions of conventional temperature and pressure, using a conventional solvent, but preferably (a) water, carbon number 1- 4, Anhydrous or hydrous lower alcohols (eg methanol, ethanol, propanol and butanol), propylene glycol, 1,3-butylene glycol, glycerin, acetone, diethyl ether, ethyl acetate, butyl acetate, dichloromethane, chloroform , Solvent extraction method using a solvent selected from the group consisting of hexane and mixtures thereof (b) extraction using a supercritical extraction method under reduced pressure and high temperature with carbon dioxide or (c) an ultrasonic extraction method. In the present invention, a solvent extraction method using an extraction solvent is preferably used. In the present invention, the extract of Ubangja is various extraction solvents, for example, water, anhydrous or hydrous lower alcohols having 1 to 4 carbon atoms (for example, methanol, ethanol, propanol and butanol), propylene glycol, 1,3-butylene glycol. , Glycerin, acetone, diethyl ether, ethyl acetate, butyl acetate, dichloromethane, chloroform, hexane and mixtures thereof, and can be obtained using at least one extraction solvent selected from the group consisting of ethanol, preferably 70% (v / v) ethanol or water, most preferably 70% (v / v) ethanol.

On the other hand, it is apparent to those skilled in the art that the extract of the ally of the present invention can obtain an extract having substantially the same effect using other extraction solvents as well as the above-described extraction solvents. In addition, the extract of thebangbang of the present invention includes not only extracts by the above-described extraction solvent, but also extracts that have undergone conventional purification and fermentation processes. For example, reduced pressure by carbon dioxide, extraction by supercritical extraction method by high temperature, extraction by ultrasonic extraction method, separation using an ultrafiltration membrane having a constant molecular weight cut-off value, various chromatography (size, charge, hydrophobicity or affinity) Active fractions obtained through various purification and extraction methods, such as separation by sex) or extraction by fermentation products using natural or various microorganisms, are also included in the extract of the present invention.

Ubangja extract according to the present invention also includes an extract that has been subjected to a fermentation process, and the fermentation extract of the Ubangja can be prepared as follows. The lysate is crushed finely to about 100 to 500 mesh, and then 1-50 g / L of a conventional microbial culture solution is added, and microorganisms such as yeast strains or lactic acid bacteria are added in an amount of 10,000 to 100,000 cfu / L. The culture temperature is cultured under normal microorganism culture conditions of 30 ~ 37 ℃. The pH is 5 to 7 and cultured for about 5 to 10 days under aerobic or normal anaerobic conditions. It can then be obtained through aging and filtration.

According to the present invention, the yeonbangja extract is contained 0.001 to 30.0% by weight relative to the total weight of the cosmetic composition, more preferably, the baebangja fermentation extract is characterized in that containing 0.01 to 10% by weight relative to the total weight of the cosmetic composition Is done. When the content of the extract is less than 0.001% by weight, the effect of promoting hair loss and promoting hair growth does not appear, and when it exceeds 30.0% by weight, there is a problem in the safety and stability of the formulation and it is not economical.

The components included in the cosmetic composition of the present invention as active ingredients may include ingredients commonly used in cosmetic compositions in addition to the active ingredients, for example, antioxidants, stabilizers, solubilizers, vitamins, pigments and fragrances Adjuvants, and carriers.

The cosmetic composition of the present invention can be prepared in any formulation conventionally prepared in the art, for example, solution, suspension, emulsion, paste, gel, cream, lotion, powder, soap, surfactant-containing cleansing , Oil, pack, massage cream and spray, but may be formulated, but is not limited thereto.

When the formulation of the present invention is a paste, cream or gel, animal oil, vegetable oil, wax, paraffin, starch, trakant, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc or zinc oxide, etc. may be used as a carrier component. You can.

When the formulation of the present invention is a solution or emulsion, a solvent, solubilizer or emulsifier is used as a carrier component, such as water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1 , 3-butyl glycol oil, glycerol aliphatic ester, polyethylene glycol or sorbitan fatty acid ester.

When the formulation of the present invention is a suspension, liquid diluents such as water, ethanol or propylene glycol as carrier components, ethoxylated isostearyl alcohol, suspensions such as polyoxyethylene sorbitol esters and polyoxyethylene sorbitan esters, microcrystals Sex cellulose, aluminum metahydroxide, bentonite, agar or trakant, etc. can be used.

When the formulation of the present invention is a powder or a spray, lactose, talc, silica, aluminum hydroxide, calcium silicate or polyamide powder can be used as a carrier component, and in the case of a spray, additionally chlorofluorohydrocarbon, propane / Propellant such as butane or dimethyl ether.

When the formulation of the present invention is a surfactant-containing cleansing, aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, isethionate, imidazolinium derivatives, methyltaurate, sarcosinate, fatty acid amide ether as carrier components Sulfates, alkylamidobetaines, aliphatic alcohols, fatty acid glycerides, fatty acid diethanolamides, vegetable oils, lanolin derivatives or ethoxylated glycerol fatty acid esters and the like can be used.

When the cosmetic composition of the present invention is a soap, a surfactant-containing cleansing formulation, or a surfactant-free cleansing formulation, it may be wiped, peeled off, or washed with water after application to the skin. As a specific example, the soap is liquid soap, powdered soap, solid soap and oil soap, and the surfactant-containing cleansing formulation is a cleansing foam, cleansing water, a cleansing towel and a cleansing pack, and the surfactant-free cleansing formulation is a cleansing cream , Cleansing lotions, cleansing water and cleansing gels, but are not limited thereto.

Hereinafter, the present invention will be described in more detail through examples. These examples are only intended to illustrate the present invention in more detail, and according to the gist of the present invention, the scope of the present invention is not limited by these examples.

In the present invention, actin, actigenin, and rapaol, which are active ingredients isolated from the extract of the stool extract, are shown in the following formula.

[Formula 1] Actin

[Formula 2] Actigenin

[Formula 3] Rapaol

[Formula 4] Rapaol C

Preferably, in the mixture of actin, actigenin, lapaol, and lapaol C, the content of actin relative to the total weight of the cosmetic composition is 0.01 to 0.1% by weight, the content of actigenin is 0.01 to 0.1% by weight, and the content of lapaol is 0.01 to 0.1% by weight, the content of Rapaol C is 0.01 to 0.1% by weight.

In the present invention, the "fermented extract" is an extract obtained through fermentation of the extract of the ally, and all formulations that can be formed using an extract, such as a diluent or a concentrate of the extract, a dried product obtained by drying the extract, a crude product or a purified product of the extract. Means an extract.

Preparation Example 1: Preparation of Ubangja extract

After washing and drying 20kg of cleansers with clean water, crushed to a certain size (100mesh), refluxed 3 times for 5 hours with 70% (V / V) ethanol aqueous solution, quenched, filtered through Whatman # 4 filter paper Using a concentrator, 1.2 kg of the extract of Ubangja was prepared.

Preparation Example 2: Preparation of fermented ferment extract

The fruit extract obtained from Preparation Example 1 was added to the lactic acid bacteria culture solution to be 20g / 1L, and dipotassium phosphate was added as a glucose and pH regulator as a carbon nutrient. Lactobacillus acidophilus (KTCT 3140) and Bifidobacterium lactis (5854) were added to the culture medium at the same ratio of 50,000 cfu / L, and then cultured at 37 ° C for 7 days at a pH of 6. After incubation, the culture medium was filtered using a 0.25 μM filter, and the filtrate was concentrated to prepare 1.0 kg of fermentation extract.

Preparation Example 3: Preparation of actin and actinin

After obtaining and dispersing 10 L of 10% ethanol in 1.0 kg of fermented ferment extract obtained through Preparation Example 2, non-polar components were removed using 1 L of n-hexane (repeated 3 times), and then methylene in the water layer. The layer fractionated by adding 10 L of chloride was concentrated under reduced pressure to obtain a fraction of 50 g. This was separated into active ingredients using a silica gel (60 to 120 mesh) column, and separated and purified using a mixture of methanol and methylene chloride (1:20-> 1: 5) as a mobile phase to recover all 9 fractions. From two fractions with cell proliferation effect 2.14g and 1.84g were recovered and separated and identified using NMR. NMR results were compared with literature data to confirm that the components identified and separated were actin and actigenin (Lu, H., Sun, Z., Shan, H., & Song, J. (2015) .Microwave-assisted extraction and Purification of arctiin and arctigenin from fructus arctii by high-speed countercurrent chromatography.Journal of chromatographic science, 54 (3), 472-478.) Actin and actigenin were isolated from Preparation Example 3, and the specific chemical formula is 1 It is shown in Section 2.

Production Example 4: Rapaol production

After obtaining and dispersing 10 L of 10% ethanol in 1.0 kg of fermented ferment extract obtained through Preparation Example 2, 1 L of non-polar component of n-hexane (hexane) was removed (repeat 3 times), and then methylene chloride was added to the water layer. , Solvent fractionation was performed in the order of ethyl acetate. The ethyl acetate fraction layer was filtered and concentrated to obtain 340 g of ethyl acetate fraction, and separation of the active ingredients was performed using a silica gel (200-400 mesh) column to separate the active ingredients. As a mobile phase, a total of 15 fractions were recovered using a mixture of chloroform acetone, and 10.99 g was recovered by concentrating the fractions having proliferative effects on the dermal papilla cells. The recovered fractions were separated and purified by column chromatography (mobile phase methanol) using sephadex-LH20 to obtain 1.02 g, and structural identification was performed using NMR. The NMR results were compared with the literature data, and it was confirmed that the component identified and separated was rapaol. (Ichihara, A., Numata, Y., Kanai, S., & Sakamura, S. (1977) .New sesquilignans from Arctium lappa L. The structure of lappaol C, D and E. Agricultural and Biological Chemistry, 41 (9 ), 1813-1814.)

Preparation Example 5: Preparation of Rapaol C

After obtaining and dispersing 10 L of 10% ethanol in 1.5 kg of fermented ferment extract obtained through Preparation Example 2, 1 L of non-polar component of n-hexane (hexane) was removed (repeat 3 times), and then methylene chloride was added to the water layer. , Solvent fractionation was performed in the order of ethyl acetate. The ethyl acetate fraction layer was filtered and concentrated to obtain 340 g of ethyl acetate fraction, and separation of the active ingredients was performed using a silica gel (200-400 mesh) column to separate the active ingredients. As a mobile phase, a total of 5 fractions were recovered using a mixture of chloroformmethanol, and the fractions having proliferative effects on dermal papilla cells were concentrated to recover 67.24 g. The recovered fractions were recovered by performing column chromatography using sephadex-LH20 (mobile phase: chloroform acetone mixture) to recover four fractions, and fractions having proliferative effects on dermal papilla cells were obtained, 10.02 g, and structural identification using NMR. Was conducted. NMR results were compared with literature data to confirm that the component identified and identified was Rapaol C (Umehara K., Sugawa A., Kuroyanagi M., Ueno A., Taki T. , Chem. Pharm. Bull ., 41, 1774 -1779 (1993)., Ichihara A., Numata Y., Kanai S., Sakamura S., Agric. Biol. Chem ., 41, 1813-1814 (1977).

Preparation Example 6: Preparation of actin, actigenin, and rapaol mixtures

A mixture was prepared by mixing Preparation Example 3 and Preparation Example 4 in a ratio of 1: 1.

Experimental Example 1: 5a-reductase activity inhibition effect test

The 24 hour fasted mature Sprogue-Dawley rats (7-8 weeks old) were killed with diethylcer, and the liver was ice-cold with Krebs-Ringer phosphate buffer (pH 7.2). To this, 5 times the amount of ice-cold Tris HCL buffer (Tri-HCL buffer, pH 7.2) was added, homogenized, and centrifuged at 900Xg for 10 minutes. The upper layer was centrifuged at 5000Xg for 10 minutes, and the upper layer obtained when centrifuging at 9000Xg for 10 minutes was used as an enzyme solution. The enzyme solution was cryopreserved at 80 ° C. Tris HCL buffer (Tris-Hcl buffer, pH 7.2) 1.0 ml of testosterone (500 ug / ml), 0.3 ml of the sample prepared through the preparation example was mixed with 0.2 ml of enzyme and 1.0 ml of enzyme solution, and NADPH (0.77 mg / ml) ) 0.5 ml was added to start the reaction, and reacted at 37 ° C for 30 minutes. The reaction was stopped by adding 5 ml of dichloromethane, and 0.5 ml of an internal standard (p-hydroxybenzoic acid n-hexyl ester, 0.1 mg / ml) was added, shaken for 10 minutes, and centrifuged at 3000 rpm for 10 minutes. After removing the upper layer, a dichloromethane layer was left there, and 5 ml of methanol was added thereto to prepare a sample for high-speed liquid chromatography (HPLC). The 5a-reductase inhibition rate was calculated according to the calculation formula of Equation 1 below after measuring the amount of testosterone of each experimental group using an HPLC method, and the results are shown in Table 1 when used alone, and in Table 2, Preparation Example 4 The results for the synergistic effect with and Preparation Example 3 were shown.

Sample name Concentration (ppm) 5a-reductase inhibition rate (%) Ubangja Extract 1000 27.44 ± 1.35 100 15.12% ± 2.04 10 8.08% ± 1.84 Fermenter extract 1000 45.49% ± 2.11 100 29.72% ± 1.47 10 14.84% ± 1.91 Actin 100 23.24% ± 2.31 10 12.91% ± 1.74 One 9.11% ± 2.01 Actinine 100 36.14% ± 1.69 10 20.67% ± 1.53 One 8.58% ± 2.28 Rapaol 100 39.27% ± 1.78 50 30.39% ± 1.87 10 16.70% ± 1.09 One 9.84% ± 2.37 Actin, Actigenin, Raphaol 100 57.70% ± 2.93 10 41.84% ± 2.71 One 14.75% ± 1.85 Lapaol C 100 45.21% ± 1.22 50 32.19% ± 1.07 10 18.74% ± 1.59 One 12.44% ± 2.31

Sample name Concentration (ppm) Sample name Concentration (ppm) 5a-reductase inhibition rate (%) Rapaol 10 Actin 10 27.36% ± 1.15 One 22.74% ± 1.27 One Actin 10 12.08% ± 1.08 One 9.91% ± 0.94 10 Actinine 10 35.44% ± 1.73 One 20.49% ± 1.51 One Actinine 10 19.47% ± 1.37 One 10.02% ± 2.13

Experimental Example 2: Sterilizing power test for active dandruff maleesia puffer

The bactericidal force test for the active dandruff malesezia furfur (Malassezia furfur. KCTC 7743) was performed as follows. After inoculating the dandruff bacteria in a petri dish (bacterial culture dish), the cells were cultured in an aerobic culture tank for 3 days, and the number of colonies of the dandruff bacteria was measured. The extracts and active ingredients obtained through the preparation example are dissolved in an aqueous solution of a certain concentration, dropped into a petri dish, incubated in an aerobic culture tank for at least 3 days, the colonies are measured again, and sterilization against dandruff bacteria according to Equation 2 below. Rate (%) was calculated.

Sample name (100 ppm each) Early dandruff bacteria 3 days later
Dandruff bacteria Sterilization power against dandruff bacteria (%) Ubangja Extract 1.59 * 10 ⁶ 1.31 * 10 ⁶ 17.61% Fermenter extract 1.24 * 10 ⁶ 22.01% Actin 1.18 * 10 ⁶ 25.78% Actinine 1.08 * 10 ⁶ 32.07% Rapaol 1.11 * 10 ⁶ 30.18% Actin, Actigenin, Raphaol 9.81 * 10 ^ 5 38.30% Lapaol C 9.83 * 10 ^ 5 39.10%

Experimental Example 3: Cell proliferation effect of dermal papilla cells

Human dermal papilla cells are cultured using DMEM medium containing 10% FBS and 1% antibiotics (100 units / ml penicillin and 100 ug / ml streptomycin). Using the 3-4th generation of papillary cells in the experiment, the growth proliferation efficacy of the papillary cells was confirmed by MTT assay. In a DMEM medium containing 1% FBS in a 96-well microplate, papillary cells (1 × 04 cells / ml) were added, pre-incubated for 24 hours, and the samples obtained from the preparation were treated at various concentrations, and then cultured for 4 days. 3- [4,5-dimethylthiazol-2-yl] -2,5-diphenyltetrazolium bromide (MTT) ) Was added to 50 μl and incubated for 4 hours, and then centrifuged to remove the supernatant. Here, dimethyl sulfoxide was added to dissolve the precipitate, and then the absorbance was measured at 540 nm to confirm the proliferation effect of human-derived dermal papilla cells to the extent of formation of formazan precipitate following reduction of MTT. Table 4 shows the results when used alone, and Table 5 shows the results of the synergistic effect between Production Example 4 and Production Example 3.

Sample name Concentration (ppm) Cell proliferation effect (%) (compared to control) Ubangja Extract 10 101.82% ± 3.51 Fermenter extract 100 115.40% ± 2.43 10 102.18% ± 2.11 Actin 100 129.85% ± 2.41 10 121.11% ± 3.37 One 109.47% ± 1.41 Actinine 100 120.04% ± 2.09 10 113.76% ± 2.68 One 105.57% ± 2.70 Rapaol 100 139.52% ± 2.96 10 116.12% ± 2.83 One 107.09% ± 1.25 Actin, actigenin, and rapaol (production example 6) 100 147.25% ± 2.44 10 132.49% ± 1.94 One 113.27% ± 1.07 Lapaol C 100 147.29% ± 3.07 10 126.57% ± 3.17 One 118.47% ± 1.55

Sample name Concentration (ppm) Sample name Concentration (ppm) Cell proliferation effect (%) (compared to control) Rapaol 10 Actin 10 132.54% ± 2.45 One 109.27% ± 1.07 One Actin 10 120.41% ± 2.10 One 104.70% ± 1.03 10 Actinine 10 126.29% ± 1.91 One 114.79% ± 1.67 One Actinine 10 120.03% ± 1.43 One 111.49% ± 0.96

Experimental Example 4: Hair cycle regulation signal transmission of human-derived dermal papilla cells

-카테닌 수준(

-catenin level), TGF-

수준(TGF-

level)을 확인하였다.Samples were treated with human-derived dermal papilla cells alone or pre-treated with DHT and then cultured for a period of time to prepare. The cells were harvested and washed three times with PBS, 500 μl of lysis buffer was added, followed by lysis for 1 hour, followed by centrifugation at 12,000 rpm for 15 minutes to obtain a supernatant, and the protein concentration was BSA (bovine). Serum albumin) was used as a standard and quantified using Protein Assay Kit (BIO-RAD, HC, USA). 20-30 μg of lysate is denatured and separated by 12% mini gel SDS-PAGE (Sodium dodecyl sulfate-polyacrylamide gel electrophoresis) to 200 mA with a polyvinylidene difluoride membrane (BIO-RAD, HC, USA) Transfer was carried out for 2 hours at. Blocking of the membrane was performed overnight in a TTBS (TBS + 0.1% Tween 20) solution containing 5% skim milk. The primary antibody was used diluted in TTBS solution, and the reaction was performed at room temperature for 2 hours. As a secondary antibody, HPR (Horse Radish Peroxidase) -coupled anti-rabbit IgG (Amersham Prarmacia Biotech, NY, USA) and anti-mouse IgG (Amersham Prarmacia Biotech, NY, USA) were diluted (1: 5000). Thus, the reaction proceeded at room temperature for 30 minutes. Then, the membrane was washed 3 times with TTBS, reacted with the ECL substrate for 1 to 3 minutes, and then photosensitive to the X-ray film.

-Catenin levels (

-catenin level), TGF-

Level (TGF-

level).

Table 6 shows the results when used alone, and Table 7 shows the results of the synergy between Production Example 3 and Production Example 4.

Sample name Concentration (ppm)

-catenin level
(control 대비)

-catenin level
(compared to control) TGF-

level
(compared to control) Flk-1 level
(compared to control) Ubangja Extract 10 5.71% 2.17% 4.02% Fermenter extract 100 13.45% 3.91% 2.47% 10 2.09% - - One - - - Actin 100 15.11% 16.07% 7.18% 10 9.54% 12.13% - One 6.47% 5.70% - Actinine 100 20.09% 15.71% 6.09% 10 13.31% 9.94% - One 4.97% 6.10% - Rapaol 100 31.07% 24.29% 10.84% 10 13.07% 8.97% 5.15% One 5.73% 2.47% - Actin, Actigenin, Raphaol 100 37.03% 35.13% 9.45% 10 32.76% 30.99% 4.66% One 8.44% 7.91% - Lapaol C 100 45.13% 38.83% 27.44% 10 33.46% 29.44% 17.18% One 12.84% 14.41% 10.75%

Sample name Concentration (ppm) Sample name Concentration (ppm)

-catenin level
(control 대비)

-catenin level
(compared to control) TGF-

level
(compared to control) Rapaol 10 Actin 10 26.05% 27.46% One 15.97% 16.09% One Actin 10 14.75% 13.18% One 5.10% 6.73% 10 Actinine 10 20.44% 14.49% One 12.88% 10.72% One Actinine 10 10.11% 11.14% One 6.05% 5.46%

Prescription Example 1: Scalp Essence

Prescription examples of the essence in the cosmetic composition containing the manufacturing example containing the fermented extract of Ubangja made through the above-described preparation example and the active ingredient thereof are shown in Table 5 below, and are not limited to this prescription example. More specifically, it can be prepared in the form of flexible lotion, nutrition lotion, nutrition cream, massage cream, essence, eye cream, shampoo, treatment, scalp ampoule, cleansing cream, cleansing foam, cleansing water, pack, spray or powder. have.

ingredient weight% Formulation Example 1 Formulation Example 2 Formulation Example 3 Formulation Example 4 Formulation Example 5 Formulation Example 6 Preparation Example 1
glycerin
Betaine
Fiji 1500
Allantoin
DL-panthenol
EDTA-2Na
Benzophenone-9
Hydroxyethyl cellulose
Sodium hyaluronate
Carboxyvinyl polymer
Triethanolamine
Octyldodecanol
Octyldodeseth-16
ethanol
Preservative, fragrance, coloring
Distilled water Preparation Example 2
glycerin
Betaine
Fiji 1500
Allantoin
DL-panthenol
EDTA-2Na
Benzophenone-9
Hydroxyethyl cellulose
Sodium hyaluronate
Carboxyvinyl polymer
Triethanolamine
Octyldodecanol
Octyldodeseth -16
ethanol
Preservative, fragrance, coloring
Distilled water Preparation Example 3
glycerin
Betaine
Fiji 1500
Allantoin
DL-panthenol
EDTA-2Na
Benzophenone-9
Hydroxyethyl cellulose
Sodium hyaluronate
Carboxyvinyl polymer
Triethanolamine
Octyldodecanol
Octyldodeseth -16
ethanol
Preservative, fragrance, coloring
Distilled water Preparation Example 4
glycerin
Betaine
Fiji 1500
Allantoin
DL-panthenol
EDTA-2Na
Benzophenone-9
Hydroxyethyl cellulose
Sodium hyaluronate
Carboxyvinyl polymer
Triethanolamine
Octyldodecanol
Octyldodeseth -16
ethanol
Preservative, fragrance, coloring
Distilled water Preparation Example 5
glycerin
Betaine
Fiji 1500
Allantoin
DL-panthenol
EDTA-2Na
Benzophenone-9
Hydroxyethyl cellulose
Sodium hyaluronate
Carboxyvinyl polymer
Triethanolamine
Octyldodecanol
Octyldodeseth -16
ethanol
Preservative, fragrance, coloring
Distilled water Preparation Example 6
glycerin
Betaine
Fiji 1500
Allantoin
DL-panthenol
EDTA-2Na
Benzophenone-9
Hydroxyethyl cellulose
Sodium hyaluronate
Carboxyvinyl polymer
Triethanolamine
Octyldodecanol
Octyldodeseth -16
ethanol
Preservative, fragrance, coloring
Distilled water 2%
3.0
10.0
5.0
2.0
0.1
0.3
0.02
0.04
0.1
8.0
0.2
0.18
0.3
0.4
6.0
a very small amount
Balance Sum 100

Example 1: Anti-hair loss effect of cosmetics

Example 1 was tested using the scalp essence Prescription Example 1 containing a certain concentration of the manufacturing example containing the extract of the stool obtained through the manufacturing example and its separation compound, and recruited 20 subjects in their 20s to 30s for each group A The group does not contain extract, formulation B contains formulation example 1 (extractor extract), group C contains formulation example 2 (fertilizer fermentation extract), group D contains formulation example 3 (actin, actigenin), group E contains formulation examples Formulation Example 5 (Rapaol C) was used for 4 (rapaol), F group, and Formulation Example 6 (actin, actigenin, rapaol) was used for G group. It was used twice a day for 8 weeks after washing the scalp in the morning and evening. After use, the sensory evaluation of the subjects was surveyed.

Group A Group B Group C Group D Group E Group F Group G Hair loss improvement 2 4 7 7 8 9 12 Little improvement 5 7 5 5 8 8 6 no effect 13 9 8 8 4 3 2

Example 2: Effect of preventing cosmetic hair loss 2

Example 1 was tested using a scalp shampoo containing a certain concentration of the manufacturing example containing the fermented extract of the stool obtained through the manufacturing example and its separation compound, and recruited 20 subjects in their 20s and 30s for each group. A small dot of 1 mm diameter was tattooed on the scalp to be evaluated by performing gram. Formulation without extract in group A, Formulation Example 1 in group B (extractor extract), Formulation Example 2 in group C (fermenter extract from fertilizer), Formulation Example 3 in group D (actin, actigenin), Formulation in group E Example 4 (Rapaol), Formulation Example 5 (Lapaol C) for the F group, and Formulation Example 6 (Actin, Actigenin, Rapaol) were used for the G group. Twice a day, it was used for 8 weeks for scalp cleaning in the morning and evening. Before use, after 4 weeks of use, and after 8 weeks of use, the change in the number of hairs and the diameter of the hairs in the corresponding phototrigram was measured using the IS-3000C (Besutopia, Korea).

Changes in the number of hairs in the phototrigram 0 weeks 4 weeks 8 weeks Group A 2.954 2.988 2.976 Group B 2.771 2.841 2.774 Group C 3.011 3.269 3.454 Group D 3.198 3.751 4.912 Group E 3.374 4.108 5.097 Group F 3.207 3.788 4.107 Group G 3.207 4.387 5.178

Change of hair diameter in phototrigram 0 weeks 4 weeks 8 weeks Group A 0.0772 0.0798 0.0803 Group B 0.0811 0.0818 0.0822 Group C 0.0864 0.0891 0.0941 Group D 0.0862 0.0890 0.929 Group E 0.0772 0.0829 0.0894 Group F 0.0742 0.0809 0.0884 Group G 0.0827 0.0910 0.0932

The manufacturing example of the present invention confirmed the effectiveness in the cell unit in relation to the symptoms of hair loss through the experimental example, and through the example of the formulation containing the manufacturing example, it is helpful in increasing the number of hairs in the phototricogram and improving the thickness of the hair. It was confirmed that it was helpful in alleviating hair loss symptoms.

Claims (9)

Cosmetic composition for preventing hair loss, which uses actin, arctigenin and rapaol derived from the extract of Arctium lappa as an active ingredient. The cosmetic composition for preventing hair loss according to claim 1, wherein the composition contains actin, actigenin, and rapaol in a mixing ratio of 1 to 10: 1 to 10: 1 to 10. A cosmetic composition for preventing hair loss using rapaol C derived from the extract of Ubangja as an active ingredient. The cosmetic composition for preventing hair loss according to any one of claims 1 to 3, wherein the cosmetic composition for preventing hair loss suppresses malesacea puff, which is a dandruff bacterium. The cosmetic composition for preventing hair loss according to claim 1, wherein the cosmetic composition for preventing hair loss suppresses the activity of 5a-reductase. According to claim 1 to claim 3, wherein the cosmetic composition for preventing hair loss is the effect of proliferating the dermal papilla cells and signaling of hair proliferation of cells (

-catenin level, TGF-

level) is a cosmetic composition for preventing hair loss. The cosmetic composition for preventing hair loss according to claim 1, wherein the actin extract-derived actin, actigenin, and rapaol contain 0.01 to 30.0% by weight of the total cosmetic composition. According to claim 1 to claim 3, wherein the cosmetic composition for preventing hair loss is extracted from a friend with an extraction solvent selected from the group consisting of water, anhydrous or hydrous lower alcohol having 1 to 4 carbon atoms, acetone, ethyl acetate and butyl acetate. A cosmetic composition for preventing hair loss. The cosmetic composition for preventing hair loss according to claim 3, wherein the cosmetic composition contains 0.01 to 0.1% by weight of Rapaol C with respect to the total weight.