WO2014003232A1 - Composition comprising dendropanax morbifera léveille extract as active ingredient for promoting hair growth - Google Patents

Description

Hair growth promoting composition comprising the Hwangchil-tree extract as an active ingredient

The present invention relates to a composition for promoting hair growth comprising hwangchil wood extract as an active ingredient.

Dendropanax morbifera Lev. Belonging to the family Elmaceae is an evergreen broad-leaved arboreous tree that grows in the southern coastal areas of Jeju Island and Jeju Island. .

Hwangchil has been used as a rare paint that emits the golden color of emperor's armor, helmets, and other metal ornaments since the Three Kingdoms.It is a Korean envoy, written in the Goryeo Dynasty, China's Guilin History, Guilin, and Haedong History. It is written on the back, and it remains in the Tangbu History Book Book District, which is a special product of Baekje. In addition, it is reported that Hwangchil wood is effective in eliminating heat, detoxification, treating eye and jaundice, and leprosy and harmless to the human body (Lee Si-jin, Herbal Medicine, Chinese Moonlight Book, 1590).

Related arts related to the conventional Hwangchil wood describe Hwangchil wood extract having anticancer activity in Korean Patent Publication No. 2000-0004499, and Hwangchil extract, Hwangchil fraction and these having a hepatoprotective effect in Korea Patent Publication No. 2003-0079205 A pharmaceutical composition containing the same is described, and Korean Patent Laid-Open Publication No. 2004-01077853 describes a hwangchil extract that inhibits ethanol-induced liver damage, and Korean Patent Laid-Open Publication No. 2004-0107852 discloses a hwangchil extract and a hwangchil fraction having a skin whitening effect. It is described in the Korean Patent Laid-Open Publication No. 2005-0036093 discloses a sunscreen cosmetic composition containing the sap of sakchi wood as an active ingredient.

However, the molecular mechanisms involved in the regulation of 15-PGDH (15-hydroxyprostaglandin dehydrogenase) and PGE ₂ activity of Hwangchil-tree extract have not been elucidated. Has not been reported at all.

Thus, the present inventors completed the present invention by confirming that the hexane extract of the yellow lacquer tree leaf has an activity of promoting 15-PGDH enzymatic activity and promoting hair growth by increasing PGE2.

Therefore, it is an object of the present invention to provide a cosmetic composition for promoting hair growth comprising the extract of Hwangchil wood as an active ingredient.

It is another object of the present invention to provide a dietary supplement or pharmaceutical composition for hair loss prevention, hair growth promotion and scalp improvement.

In order to solve the above problems, the present invention provides a cosmetic composition for promoting hair growth comprising hwangchil wood extract as an active ingredient.

In one embodiment of the present invention, the hwangchil wood extract may be a crude extract of the hwangchil wood, polar solvent soluble extract or non-polar solvent soluble extract.

In one embodiment of the present invention, the crude extract may be an extract available from a solvent selected from water, methanol, ethanol, butanol or a mixed solvent thereof, including purified water.

In one embodiment of the present invention, the non-polar solvent soluble extract may be an extract soluble in hexane, chloroform, dichloromethane or ethyl acetate.

In one embodiment of the present invention, the hwangchil wood extract, (i) 15-PGDH enzyme activity inhibition; (ii) increased PGE ₂ intracellularly and extracellularly; (iii) increased expression of cyclooxygenase-1 (COX-1) and multidrug resistance-associated protein4 (MRP4) genes; (iv) inhibition of prostaglandin transporter (PGT) expression; (v) Inhibition of 5α reductase (5α-R) expression; And (vi) androgen receptor (AR) expression inhibitory activity.

The present invention provides a pharmaceutical composition for preventing hair loss, promoting hair growth, and improving the scalp, comprising the extract of Hilchi chinensis as an active ingredient.

In one embodiment of the present invention, the pharmaceutical composition may be selected from the group consisting of creams, gels, patches, sprays, ointments, warnings, lotions, linens, pasta and cataplasma as external skin preparations. have.

In another aspect, the present invention provides a health functional food for hair loss prevention, hair growth promotion and scalp improvement comprising an extract of Hwangchil as an active ingredient.

In one embodiment of the invention, the food is selected from the group consisting of beverages, meat, chocolate, foods, confectionery, pizza, ramen, other noodles, gums, candy, ice cream, alcoholic beverages, vitamin complexes and health supplements Can be.

The present invention relates to a cosmetic composition for promoting hair growth comprising the extract of Hwangchil-tree as an active ingredient, a pharmaceutical composition for preventing hair loss, promoting hair growth and improving the scalp, and a functional food, and the extract of Hwangchil-tree according to the present invention has 15-PGDH activity. By inhibiting the activity of increasing PGE ₂ is excellent, it is confirmed that the effect of preventing hair loss and promoting hair growth is excellent, which can be usefully used in the manufacture of cosmetics, medicines and functional foods for promoting hair growth.

1 is a schematic diagram showing the biosynthesis process of various PG (prostaglandins).

2 is a stereostructure of the complex in which 15-PGDH and PGE ₂ are bound.

3 is a picture of a hwangchil wood.

Figure 4 is a schematic diagram showing one process for producing a hwangchil wood extract of the present invention.

5 is a schematic diagram of a PGEX-2T expression vector containing recombinant 15-PGDH.

6 is a standard curve graph of NADH.

7 is a schematic of an experimental process for measuring PGE ₂ using an ELISA kit.

8 is a graph of 15-PGDH inhibitory activity (ED ₅₀ ) of the Hwangchil-tree extract.

Figure 9 is a graph of the cytotoxicity evaluation of Hwangchil-tree extract in HaCaT cells by MTT assay.

10 is a graph showing the evaluation of cytotoxicity of Hilchi chinensis extract in LNCaP.FGC cells by MTT assay.

11 is a graph showing the effect of increasing the concentration-dependent PGE ₂ by the extract of Hilchi chinensis in A549 cells.

FIG. 12 is a graph showing the change of time-dependent extracellular PGE ₂ concentration affected by Hwangchil extract DMHE in HaCaT cells.

FIG. 13 is a graph showing changes in intra-cellular and extra-cellular PGE ₂ concentrations by H. chinensis extract DMHE in HaCaT cells.

Figure 14 is a graph showing the effect of DMHE on COX-1 / 2, MRP4, 15-PGDH and PGT mRNA expression in HaCaT cells.

15 is a photograph confirming the hair growth promoting effect of DMHE using C57BL / 6 mice.

Figure 16 is a graph showing the effect on the expression of prostate specific antigen (PSA) mRNA of the Hwangchil-tree extract in LNCaP.FGC cells.

Figure 17 is a graph showing the effect on the expression of 5α-R 1/2 mRNA of Hwangchil-tree extract in LNCaP.FGC cells.

FIG. 18 is a graph showing the effects on AR, 15-PGDH and PSA mRNA expression of DMHE and flutamide in LNCaP.FGC cells. FIG.

FIG. 19 is a graph showing DMHE concentration-dependent effects on DHT-induced overexpression of AR and their target genes (15-PGDH and PSA) in LNCaP.FGC cells.

20 is a graph showing the effect of PSA mRNA inhibition, which is a target gene of AR by DMHE, is restored by high concentration of DHT, and it is a graph showing the competitive inhibition with DHT for AR.

21 is a graph showing that the DMHE of the present invention has an activity of inhibiting the concentration of 5αR-1 gene in primary hair follicle dermal papilla cells (HFDPC).

FIG. 22 is a graph showing that DMHE of the present invention has an activity of concentration-dependently inhibiting 5αR-2 gene expression in primary hair follicle dermal papilla cells (HFDPC).

FIG. 23 is a graph showing that DMHE of the present invention has activity of inhibiting expression of androgen receptor (AR) in HFDPC (primary hair follicle dermal papilla cells).

24 is a graph showing that the DMHE of the present invention has an activity of inhibiting the expression of 15-PGDH, a target gene of androgen receptor (AR), in a concentration-dependent manner in primary hair follicle dermal papilla cells (HFDPC).

Definitions of terms used in the present invention are as follows.

"Extract" is characterized in that crude extract, polar solvent soluble extract or non-polar solvent soluble extract of yellow lacquer tree.

"Crude extract" is a solvent selected from water containing purified water, lower alcohols having 1 to 4 carbon atoms such as methanol, ethanol, butanol, or a mixed solvent thereof, preferably a mixed solvent of water and methanol, more preferably 50 to 100% Extracts soluble in methanol.

"Polar solvent soluble extract" includes extracts soluble in water, methanol, butanol or a mixed solvent thereof, preferably water or methanol, more preferably methanol.

"Non-polar solvent soluble extract" includes extracts soluble in hexane, chloroform, dichloromethane, or ethyl acetate, preferably hexane, dichloromethane or ethyl acetate, more preferably in hexane or ethyl acetate solvent.

By "pharmaceutical composition" is meant a mixture of the yellow lacquer extract of the present invention with other chemical components such as diluents or carriers.

A "carrier" is defined as a compound that facilitates the addition of a compound into a cell or tissue. For example, dimethyl sulfoxide (DMSO) is a commonly used carrier that facilitates the incorporation of many organic compounds into cells or tissues of an organism.

A "diluent" is defined as a compound that not only stabilizes the biologically active form of a compound of interest, but also is diluted in water to dissolve the compound. Salts dissolved in buffer solutions are used as diluents in the art. A commonly used buffer solution is phosphate buffered saline, because it mimics the salt state of human solutions. Because buffer salts can control the pH of a solution at low concentrations, buffer diluents rarely modify the biological activity of a compound.

An "effective amount" is an appropriate amount that affects a beneficial or desirable clinical or biochemical result. An effective amount can be administered once or more. For the purposes of the present invention, an effective amount is an amount suitable to temporarily alleviate, ameliorate, stabilize, reverse, slow or slow the progression of a disease state. If the recipient animal can withstand the administration of the composition or if the administration of the composition to that animal is suitable,

Hereinafter, the present invention will be described in detail.

All technical terms used in the present invention, unless defined otherwise, are used in the meaning as commonly understood by those skilled in the art in the related field of the present invention. Also described herein are preferred methods or samples, but similar or equivalent ones are within the scope of the present invention. The contents of all publications described herein by reference are incorporated into the present invention.

The present invention relates to the use of the Hwangchil-tree extract, and relates to the exercise of the specific physiological activity and function contained in the Hwangchil-tree extract.

Hwangchil- tree ( Dendropanox morbifera LEV.) Is an evergreen broad-leaved arborescent tree belonging to the elm tree, and is a native species of Korea that grows on the south and west coasts such as Jeju Island, Wando, Bogildo, and Haenam (Fig. 3). Cloves contained in Hwangchil Tree contain small amount of Terpene and Sesquiterpene, which are different depending on the time and place of collection, but germacrene-d, β-selinene, α-amorphene , α-selinene, δ-cadinene, γ-cadinene, T-muurolol, β-elemene, bicyclo [4,4,0] dec-1 -en-2 -isopropyl-5-methyl-9-methylene, β-cadinene , germacrene-B, α-copaene, α-humulene, α-cadinene and small amounts of linalool L, α-terpinene, α-cubebene, α-ylangene, (+)-calarene, 3,7-guaiadine, (-)- Contains isoledene, β-cubebene, limonene, aromadendrene, cadina-1,4-diene.

The part of the hwangchil wood that can be used in the present invention is not limited to leaves, stems, bark, etc., but preferably leaves can be used.

Hwangchil tree extract can be prepared and used by methods known in the art, modified methods thereof or the method according to the invention. As one embodiment, it can manufacture by the following method.

Hwangchil-tree extract or crude extract of the present invention is water, methanol, ethanol, butanol and the like containing about 1 to 30-fold volume, preferably 5 to 15-volume (w / v%) of purified water, A solvent selected from a lower alcohol having 1 to 4 carbon atoms or a mixed solvent thereof, preferably a water and ethanol mixed solvent, more preferably 50 to 100% ethanol, is added at a temperature of about 0 to 100 ° C, preferably 10 to room temperature. 60 hours, preferably 30 to 50 hours extraction method such as cold extraction, hot water extraction, ultrasonic extraction, reflux cooling extraction, or heat extraction method, preferably extracted by hot water extraction method, filtered and concentrated under reduced pressure to the yellow lacquer of the present invention Tree crude extract can be obtained.

In addition, the polar solvent or non-polar solvent soluble extract of the present invention is about 1 to 150 times the weight of the crude extract, preferably 50 to 100% ethanol crude extract, preferably 5 to 100 times the volume (w / v% ), Hexane, ethyl acetate and butanol are sequentially added to a volume of about 1 to 10 times, preferably 1 to 5 times, and fractionated 1 to 5 times, preferably 2 to 4 times. The polar solvent and nonpolar solvent soluble extract of the invention can be obtained. Preferably hexane extract can be obtained and used.

The concentrate of the extracts can be obtained in powder form through -80 ℃ freeze drying or 50 ℃ vacuum decompression.

The present invention includes a method for producing the hwangchil wood extract. The preparation method is merely an exemplary method thereof, and may be suitably modified by various methods based on the art. For example, the non-exemplified extraction method according to the present invention may be used. Successful modifications can be made by those skilled in the art.

If those skilled in the art to which the present invention pertains, specific reaction conditions for the production of the Hwangchil-tree extract according to the present invention can be confirmed through examples to be described later, and a detailed description thereof will be omitted.

The present invention is based on the discovery of the molecular mechanisms of 15-PGDH activity and PGE ₂ related mechanisms of the extract of Hilchi chinensis.

(1) Hwangchil-tree extract of the present invention inhibits the activity of NAD ⁺ dependent 15-PGDH to increase the intracellular production of PGE ₂ .

Prostaglandin (PG) is a mediator known as eicosanoid, produced from arachidonic acid liberated in cell membranes by physical, chemical, and specific cytokine growth factors and other stimuli. Biosynthetic processes of various PGs are shown in FIG. 1. In particular, PGE ₂ is known as an important mediator of wound healing, peptic ulcer, eyebrow and hair formation, and bone formation. However, PGE ₂ is rapidly metabolized by NAD ⁺ dependent 15-PGDH, so the half-life is very short in the body.

Hwangchil extract of the present invention has an excellent activity of increasing PGE ₂ by inhibiting the 15-PGDH.

In one embodiment of the present invention, 15-PGDH enzyme activity inhibitory effect (ED ₅₀ ) of the yellow leaves extract (methanol, n-hexane, n-butanol and water) was 1.74 μg / mL to 661.8 μg / mL In particular, n-hexane and ethyl acetate extracts showed ED ₅₀ values of 1.74 μg / mL and 10.6 μg / mL, respectively. Further, hwangchil leaf hexane extract sikyeotgo the PGE ₂ concentration-dependent increase in extracellular page A549 cancer cells, HaCaT cell line was increased in the cells and PGE ₂ concentration outside the cell.

(2) In addition, the hwangchil-tree extract of the present invention also affects the expression mechanism of other specific genes related to the mechanism.

That is, the Hwangchil-tree extract of the present invention increased the expression of COX-1 and MRP4 mRNA, and decreased the mRNA expression of PGT and 15-PGDH.

(3) In addition, the Hwangchil-tree extract has a hair growth promoting effect by inhibiting 15-PGDH activity.

Hair loss may be caused by disease, hair loss caused by hormonal causes, hair loss caused by nutritional imbalance, or hair loss caused by stress. Hair loss caused by the disease may be inherited, autoimmune diseases, burns, chemotherapy, psoriasis, Fungal infections, radiation exposure, or tumors may be the cause, and hair loss caused by the hormonal cause may be due to drugs, pre and postnatal pregnancy, menopause, diabetes, pituitary dysfunction, parathyroid or thyroid dysfunction.

There are also hair loss is related to the degradation of PGE _2, PGE ₂ promotes cell division and the head of the people in the body and to smooth the hair nutrition by expanding the blood in my veins accelerate the growth of hair. Essential fatty acids are degraded to arachidonic acid by the PLA 2 enzyme and back to prostaglandins by the COX-2 enzyme, where 15-PGDH degrades prostaglandins in a NAD ⁺ dependent manner. Therefore, by inhibiting the action of 15-PGDH it is possible to inhibit the degradation of prostaglandins to promote hair growth or to prevent hair loss. In addition, PGE ₂ is also involved in the inflammatory response of the scalp to remove dandruff (Michelet JF et al., 2008, Exp. Dermatol, 17 (10), 821-8).

In one embodiment of the present invention was confirmed the excellent hair growth effect of the Hwangchil leaves hexane extract in the hair loss model depilation C57BL / 6 black mice entered the resting period.

(4) as well as the hwangchil wood extract of the present invention has anti-androgen properties that inhibit the protein synthesis of AR.

In other words, the Hwangchil-tree extract of the present invention is involved in the suppression of AR signaling known as the main mechanism of androgenetic alopecia. 5α-reductase is an important enzyme that accelerates signaling to AR by converting testosterone to 5α dihydroxytestosterone (DHT). Finasteride, a 5α-reductase inhibitor, is used for the treatment of male hair loss (Propecia) and for the treatment of benign prostatic hyperplasia (BPH) (Proscar). Hwangchil-tree extract of the present invention inhibits 5α-reductase 1 and 2 and AR signaling by AR gene expression reduction and AR receptor blockade, resulting in a quantitative decrease of their target enzyme 15-PGDH. In addition, DHT was shown to act as a competitive inhibition (AR) pattern for AR. That is, by interfering with the protein synthesis and activity of AR, the expression of 15-PGDH mRNA was reduced along with the target gene PSA. In the case of male hair loss, increased activity of AR and 5α-reductase is reported as a major factor, and the hwangchil-tree extract of the present invention has a great effect on male hair loss.

It can be seen that this mechanism can have therapeutic and prophylactic effects even in diseases related to AR activity and 15-PGDH activity. For example, over-synthesized DHT binds to AR in prostate cells, leading to prostatic hypertrophy and even prostate cancer. 5α-R is also present in hair follicles of the scalp as well as converting testosterone to DHT for hair loss. Induced, it can be useful in the treatment of such diseases and hair-related cosmetic composition.

As such, the hwangchil wood extract of the present invention inhibits 15-PGDH enzyme activity; Increasing intracellular and extracellular PGE ₂ concentrations; Increased expression of cyclooxygenase-1 (COX-1) and multidrug resistance-associated protein4 (MRP4) genes; Inhibition of prostaglandin transporter (PGT) expression; And 15-PGDH activity based on this molecular mechanism because it inhibits 5α-reductase gene expression, inhibits protein synthesis of androgen receptor (AR), also inhibits 15-PGDH expression, and is not toxic in laboratory animals. And the prevention and treatment of PGE ₂ related diseases.

Therefore, the present invention provides a composition for hair growth, preferably a cosmetic composition for promoting hair growth, containing the extract of Hilchi chinensis as an active ingredient.

Alopecia is a disease associated with 15-PGDH activity, which refers to a symptom of hair loss, shortening the growth period in the hair cycle, and decreasing the proportion of growth hair in the total hair. In the case of androgenetic alopecia, the mechanism of its development is not known, but it depends on male hormones, and the proportion of resting hair follicles increases and the hair follicles become dwarfed.

The cosmetic composition according to the present invention may include components commonly used in cosmetic compositions, as well as extracts from the hwangchil wood, such as conventional auxiliaries such as antioxidants, stabilizers, solubilizers, vitamins, pigments and flavors, And a carrier.

In addition, the composition of the present invention can be used in addition to the above-mentioned Hwangchil-tree extract, a wool or hair-grown agent that has been conventionally used insofar as it does not impair the effect of hair loss (hair loss) and hair improvement by the Hwangchil-tree extract.

In addition, the cosmetic composition of the present invention may be prepared in any formulation commonly prepared in the art, for example, hair tonic, hair conditioner, hair essence, hair lotion, hair nutrition lotion, hair shampoo, hair rinse, Hair Treatment, Hair Cream, Nutritional 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 Preservative, Hair Dye, Hair wave agent, hair bleach, hair gel, hair glaze, hair dresser, hair lacquer, hair moisturizer, hair mousse and hair spray can be prepared in various forms, but is not limited thereto.

According to a preferred embodiment of the present invention, the content of the hwangchil wood extract of the present invention is 0.00001-30% by weight, preferably 0.5-20%, more preferably 1.0-10% by weight based on the total weight of the composition. When the content of the Hwangchil-tree extract is less than 0.00001% by weight, the hair loss prevention and hair growth improvement effect by the Hwangchil-tree extract is greatly reduced, if it exceeds 30% by weight may cause skin irritation, and the problem of formulation have.

In addition, the composition of the present invention may be a pharmaceutical composition comprising a hwangchil wood extract as an active ingredient.

The pharmaceutical composition of the present invention may be prepared using a pharmaceutically suitable and physiologically acceptable adjuvant in addition to the active ingredient, and the adjuvant may include excipients, disintegrants, sweeteners, binders, coating agents, swelling agents, lubricants, Lubricants, flavors and the like can be used.

The pharmaceutical composition may be preferably formulated into a pharmaceutical composition including one or more pharmaceutically acceptable carriers in addition to the above-described active ingredient for administration.

Formulation forms of the pharmaceutical composition may be granules, powders, tablets, coated tablets, capsules, suppositories, solutions, syrups, juices, suspensions, emulsions, drops or injectable solutions. For example, for formulation in the form of tablets or capsules, the active ingredient may be combined with an oral, nontoxic, pharmaceutically acceptable inert carrier such as ethanol, glycerol, water and the like. In addition, if desired or necessary, suitable binders, lubricants, disintegrants and coloring agents may also be included in the mixture. Suitable binders include but are not limited to natural and synthetic gums such as starch, gelatin, glucose or beta-lactose, corn sweeteners, acacia, trackercance or sodium oleate, sodium stearate, magnesium stearate, sodium Benzoate, sodium acetate, sodium chloride and the like. Disintegrants include, but are not limited to, starch, methyl cellulose, agar, bentonite, xanthan gum, and the like. Acceptable pharmaceutical carriers in compositions formulated in liquid solutions are sterile and physiologically compatible, including saline, sterile water, Ringer's solution, buffered saline, albumin injectable solutions, dextrose solution, maltodextrin solution, glycerol, ethanol and One or more of these components may be mixed and used, and other conventional additives such as antioxidants, buffers and bacteriostatic agents may be added as necessary. Diluents, dispersants, surfactants, binders and lubricants may also be added in addition to formulate into injectable formulations, pills, capsules, granules or tablets such as aqueous solutions, suspensions, emulsions and the like. Furthermore, the method disclosed in Remington's Pharmaceutical Science, Mack Publishing Company, Easton PA can be formulated according to each disease or component, as appropriate in the art.

In one embodiment of the present invention, the hwangchil wood extract of the present invention may be included in 0.00001 to 30% by weight relative to the total weight of the composition.

The external skin pharmaceutical composition of the present invention is a skin external preparation having the effect of preventing hair loss, promoting hair growth and improving the scalp, and includes cream, gel, patch, spray, ointment, warning, lotion, linen, pasta or cataplasma. It may be prepared and used as a pharmaceutical composition in the form of external preparations for skin, but is not limited thereto.

The composition of the present invention may also be a food composition, which may contain various flavors or natural carbohydrates as additional ingredients, as well as ordinary food compositions, in addition to the extract of Hilchi chinensis as an active ingredient.

Examples of the above-mentioned natural carbohydrates include monosaccharides such as glucose, fructose and the like; Disaccharides such as maltose, sucrose and the like; And conventional sugars such as polysaccharides such as dextrin, cyclodextrin, and sugar alcohols such as xylitol, sorbitol, and erythritol. The aforementioned flavoring agents can advantageously be used natural flavoring agents (tautin), stevia extracts (for example rebaudioside A, glycyrrhizin, etc.) and synthetic flavoring agents (saccharin, aspartame, etc.).

The food composition of the present invention may be formulated in the same manner as the pharmaceutical composition, used as a functional food, or added to various foods. Foods to which the composition of the present invention may be added include, for example, beverages, meat, chocolate, foods, confectionery, pizza, ramen, other noodles, gums, candy, ice creams, alcoholic beverages, vitamin complexes, and health supplements. There is this.

In addition, the food composition is a variety of nutrients, vitamins, minerals (electrolytes), synthetic flavors and natural flavoring agents, colorants and neutralizing agents (cheese, chocolate, etc.), pectic acid and its Salts, alginic acid and salts thereof, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohols, carbonation agents used in carbonated drinks and the like. In addition, the food composition of the present invention may contain a fruit flesh for producing natural fruit juice and fruit juice beverage and vegetable beverage.

Hwangchil wood extract as an active ingredient of the present invention as a natural substance has little toxicity and side effects, so can be used with confidence even for long-term use for the purpose of preventing hair loss, promoting hair growth and improving the scalp.

The present invention also provides a health functional food for hair loss prevention, hair growth promotion and scalp improvement comprising the extract of Hilchi chinensis as an active ingredient.

The health functional food of the present invention can be produced and processed in the form of tablets, capsules, powders, granules, liquids, pills and the like for the purpose of preventing hair loss, promoting hair growth and improving the scalp.

In the present invention, "health functional food" refers to a food manufactured and processed using raw materials or ingredients having functional properties useful for the human body according to the Health Functional Food Act No. 6767, and nutrients for the structure and function of the human body. It is meant to be consumed for the purpose of regulating or obtaining a useful effect for health use such as physiological action.

The health functional food of the present invention may include a conventional food additive, and the suitability as a food additive, unless otherwise specified, in accordance with the General Regulations of the Food Additives and General Test Methods approved by the Food and Drug Administration, etc. Judging by the standards and standards.

Examples of the items listed in the "Food Additive Reduction" include chemical compounds such as ketones, glycine, calcium citrate, nicotinic acid, and cinnamic acid; Natural additives such as dark blue pigment, licorice extract, crystalline cellulose, high color pigment and guar gum; And mixed preparations such as sodium L-glutamate, algae additives, preservatives and tar dyes.

For example, the health functional food in the form of a tablet is granulated in a conventional manner by mixing the mixture of excipients, hwangchil wood extract, an active ingredient of the present invention with excipients, binders, disintegrants and other additives, and then compressed with a lubricant and the like Or the mixture can be directly compression molded. In addition, the health functional food in the form of tablets may contain a mating agent or the like as necessary.

Hard capsules among the health functional foods in the form of capsules may be prepared by filling a mixture of a mixture of additives such as excipients with the sulfur extract of the present invention in a conventional hard capsules, soft capsules are excipients, etc. The mixture mixed with the additive of may be prepared by filling in a capsule base such as gelatin. The soft capsule agent may contain a plasticizer such as glycerin or sorbitol, a colorant, a preservative, and the like, as necessary.

The health functional food in the form of a cyclic form may be prepared by molding a mixture of Hwangchil-tree extract, an active ingredient of the present invention, an excipient, a binder, a disintegrant, and the like by a conventionally known method. It may be avoided, or the surface may be coated with materials such as starch, talc.

The health functional food in the form of granules can be prepared by granulating a mixture of Hwangchil-tree extract, an active ingredient of the present invention, an excipient, a binder, a disintegrant, and the like by a conventionally known method. And the like.

The health functional food may be beverages, meat, chocolate, foods, confectionery, pizza, ramen, other noodles, gum, candy, ice cream, alcoholic beverages, vitamin complexes and health supplements.

In another aspect, the present invention provides a use of the composition comprising a hwangchil wood extract for the production of cosmetics, medicine or food for preventing hair loss, promoting hair growth and scalp improvement. The composition of the present invention containing the above-mentioned Hwangchil wood extract as an active ingredient can be used for the manufacture of cosmetics, medicine or food for preventing hair loss, promoting hair growth and improving the scalp.

In another aspect, the present invention provides a method for preventing or treating hair loss, comprising administering the extract of Hilchi chinensis to a mammal.

As used herein, the term "mammal" refers to a mammal that is the subject of treatment, observation or experimentation, preferably human.

As used herein, the term “therapeutically effective amount” means an amount of an active ingredient or pharmaceutical composition that induces a biological or medical response in a tissue system, animal or human, as thought by a researcher, veterinarian, doctor or other clinician, which Amounts that induce alleviation of the symptoms of the disease or disorder being treated. It will be apparent to those skilled in the art that the therapeutically effective dosages and frequency of administrations for the active ingredients of the present invention will vary depending on the desired effect. Therefore, the optimal dosage to be administered can be easily determined by one skilled in the art and includes the type of disease, the severity of the disease, the amount of active ingredients and other ingredients contained in the composition, the type of formulation, and the age, weight, general health of the patient. , Sex and diet, time of administration, route of administration and rate of composition, duration of treatment, and drugs used concurrently. In the treatment method of the present invention, in the case of an adult, when the hwangchil-tree extract of the present invention is administered once to several times a day, it is preferable to administer at a dose of 1 mg / kg to 250 mg / kg.

In the method of treatment of the present invention, the composition comprising the Hwangchil-tree extract of the present invention as an active ingredient may be administered through oral, rectal, intravenous, intraarterial, intraperitoneal, intramuscular, intrasternal, transdermal, topical, intraocular or intradermal routes. It may be administered in a conventional manner.

Example

Hereinafter, the present invention will be described in more detail with reference to Examples. These examples are only for illustrating the present invention, it will be apparent to those skilled in the art that the scope of the present invention is not to be construed as being limited by these examples. In addition, each experiment was performed three or more times, and each data was expressed as mean ± SE. Statistical significance was determined by paired Student's t test. P <0.05 was considered statistically significant.

Materials and utensils

The leaves of the Hwangchil tree (Dendropanax morbifera) were collected from a grain farm in Muan, Korea.

PGE ₂ , NAD +, NADH, glutathione-Sepharose 4B, dithiothreitol (DTT), sodium dodecylsulfate (EDS), EDTA, reduced glutathione, SC560 (COX-1 inhibitor), celecoxib (celecoxib, COX-2 inhibitor), naproxen (non-selective COX inhibitor), 5α-dihydrotestosterone (DHT), mitomycin, sucralfate, carboxy methyl cellulose (CMC), and minoxidil to Sigma (St. Louis, MO, USA) and TGF-β1 was obtained from Biovision Pharmacia Co. (New Jersey, USA).

The cDNA of human 15-PGDH was cloned from the human placental cDNA library (Cho and Tai, 2002). UV spectra were obtained using a Shimadzu RF-5301 IPC Fluorescence Spectrophotometer (Shimadzu, Japan).

PGE ₂ enzyme immunoassay kit was purchased from Thermo Scientific (Rockford, IL, USA) and real-time PCR was performed with a Light Cycler 2.0 Instrument (Roche, Mannheim, Germany).

In vitro wound scratches were photographed with a reversed-phase transmission electron microscope (Hitachi, Tokyo, Japan).

Analytical HPLC was performed using a Waters SunFire ™ (4.6 × 150 mm, 5 μm) column on an Agilent HP1100 series consisting of a degasser, binary mixing pump, column oven and PDA detector. Semi-preparative HPLC was performed using a Waters SunFire ™ Prep C18 (10 × 250 mm and 19 × 150 mm, 5 μm) column with a Waters multisolvent delivary system combined with a DECASSIT ™ 6342 degaser. All solvents used for plant extraction were HPLC grade

Example 1 Preparation of Hwangchil Tree Leaf Extract

(1) extract manufacturing

Collected D. morbifera leaves were dried in the shade at room temperature.

The dried leaves were extracted three times with methanol at room temperature. The methanol extract was filtered using Whatman No. 1 filter paper and the combined methanol extract was concentrated in vacuo with a rotary evaporator. The methanol extract was suspended in water and then fractionated respectively by polarity with hexane, diethyl ether, ethyl acetate, n-butanol and water. Each extract was concentrated with a rotary vacuum evaporator and this process is shown in FIG. 4.

(2) fractionation and separation process

Of hexane extract with Isolera Flash Purification system (BIOTAGE, Sweden) using gradient solvent system [CHCl3 (0.1% HCOOH) 100% through CHCl3 (0.1% HCOOH) -MeOH (0.1% HCOOH) / 55: 45 for 120 min] Activity-guided fractionation was carried out under 254 and 280 nm to obtain 10 fractions (DEE01-DEE10), each fraction using HPLC in a gradient MeCN-H ₂ O (0.1% HCOOH) solvent system. Monitored and detected each compound.

Spots were also detected by thin layer chromatography under UV light (254 nm) or sprayed with vanillin-H ₂ SO ₄ and then heated to 110 ° C.

Gradient solvent system of MeCN / H ₂ O with 0.1% formic acid (30:70 → 100% MeCN), repeated semi-precision using SunFire ™ Prep C18 (10 x 250 mm and 19 x 150 mm, 5 μm) columns A -preparative HPLC experiment was performed for 40 minutes, and finally compounds were separated from this fraction as major components.

(3) results

The yield of the Hilchi chinensis extract according to the extraction solvent is shown in Table 1 below.

Table 1 menstruum Final product (g) yield(%) Methanol 230.1 13.6 n- Hexane 34.6 2.1 Ethyl acetate 31.0 1.9 n -Butanol 46.6 2.8 Water 98.4 5.8

As a result of extraction, the yield of methanol extract was the highest, while the yield using ethyl acetate was somewhat lower.

Example 2 Confirmation of Expression of 15-PGDH

(1) Expression and Purification of 15-PGDH

Escherichia coli BL-21 DE3 cells were transformed with a vector as shown in FIG. 5, ie, a PGEX-2T expression vector containing recombinant 15-PGDH between the BamH I and EcoR I positions. Cells were grown in 500 mL medium containing 50 μg / mL ampicillin under conditions of 37 ° C. and 220 rpm to incubate with an OD of 0.6 at 600 nm. Then IPTG (isoprophyl β-D-thiogalactoside, 1M stock solution) was added and the cells were incubated at 25 ° C. for 12 hours. Cells are then obtained by centrifugation at 4000 xg at 4 ° C for 30 minutes and the cell pellets are resuspended in 20 mL of cold lysis buffer (1 x PBS buffer pH 7.4 containing 1 mM EDTA and 0.1 mM DTT) and sonicated. Decompose (4 x 10 s at 4 ° C). Crushed cells were centrifuged at 4000 xg for 20 minutes at 4 ° C.

The supernatant was then slowly applied to a Glutathione-Sepharose 4B column and equilibrated with lysis buffer at 4 ° C. The column was washed with lysis buffer from O.D 280 up to 0.005 O.D. Thereafter, 15-PGDH was eluted from the glutathione-Sepharose 4B column using an elution buffer (50 mM Tris-HCl pH 8.0 containing 10 mM reduced glutathione, 1 mM EDTA and 0.1 mM DTT) for 5 minutes at room temperature. The concentration of 15-PGDH was measured by Bradford method (Schleicher and Wieland, 1978), and the purity of 15-PGDH (molecular weight 29KD) was confirmed by SDS-PAGE (sodium dodecyl sulfate polyacryl amide gel electrophoresis) and coomasine blue staining.

(2) Coomassie blue dyeing

SDS-PAGE gels were stained with staining solution (0.1% Coomassie brilliant blue R 250, 50% methanol and 10% glacial acetic acid in water) for 20 minutes. The gel was washed several times with destaining solution (10% methanol, 7% glacial acetic acid in water) until the background of the gel was completely decolorized. Finally, the gel was stored in stock solution (5% glacial acetic acid in water) and scanned for detection.

[Correction under Rule 91 06.08.0013]

(3) 15-PGDH Activity Assay

Activity analysis of 15-PGDH inhibitors was performed by measuring the formation of NADH at 468 nm and 340 nm using a fluorescence spectrophotometer.

Tris-HCl buffer (50 mM, pH 7.5) containing 0.1 mM DTT, 0.25 mM NAD +, purified enzyme (10 μg), 21 μM PGE2 and various concentrations of 15-PGDH inhibitor was added to each reaction mixture. Each concentration was analyzed in triplicate. The absorbance of the reaction mixture was corrected for 15-PGDH inhibitor activity from the standard curve of NADH in FIG. 6.

The results are shown in Table 2 and FIG. 8.

ED ₅₀ values for 15-PGDH ranged from 1.7 μg / mL to 661.8 μg / mL depending on the extraction solvent.

[Correction under Rule 91 06.08.0013]
TABLE 2

Example 3: Cell Viability Assay

(1) cell culture

HaCaT cells, human keratinocyte cell lines, were incubated in Dulbecco's modified Eagle's media (DMEM) supplemented with 10% heat inactivated FBS, 100 μg / mL antibiotics, in a 5% CO ₂ incubator at 37 ° C. A549 cells derived from adecarcarnoma-like human alveolar type II were cultured in a 5% CO ₂ incubator at 37 ° C. with RPMI medium. And, LNCaP.FGC (androgen-dependent prostate cancer) cells were cultured in a 5% CO ₂ incubator at 37 ° C. with RPMI medium. All cultured media were supplemented with 10% heat inactivated FBS and 100 μg / m L penicillin.

 (2) cell viability evaluation

 Cell viability was measured by MTT assay (Mosmann, 1983).

HaCaT cells (1 × 10 4 / mL), LNCaP.FGC cells (4 × 10 4 / mL) per 90 μL of DMEM medium were seeded in 96 well plates. After overnight incubation, the drugs were treated for 72 hours and incubated for 4 hours with 10 μL of MTT (5 mg / mL stock solution). Thereafter, the medium was removed and formazan was dissolved by adding 150 μL of DMSO. Absorbance was measured at 540 nm using an ELISA microplate reader (PerkinElmer, Calif ornia, USA).

The results are shown in Table 3 below, and FIGS. 9 and 10.

TABLE 3 menstruum HaCaT IC ₅₀ (μg / mL) LNCaP / FGC IC ₅₀ (μg / mL) n -Hexane 39.86 169 Ethyl acetate 246.27 88.3 n -Butanol 471.40 562 Water > 1000.00 > 800

As can be seen from the table above, IC ₅₀ of hexane, ethyl acetate, methanol and water extracts of yellow lacquer tree ranged from 39.86 μg / mL to> 1000 μg / mL in HaCaT cells (FIG. 9), 88 μg / in LNCaP.FGC cells. It was found to be from mL to> 800 μg / mL (Figure 10). In other words, the hydrophobic extract showed relatively high toxicity, but it was not fatal to cell viability.

Example 4 Protein Quantitation and PGE ₂ Release Assay

COX and microsomal PGE synthase-1 (mPGES-1) -derived PGs-like PGE ₂ are considered to be important regulators of pulmonary function. In particular, PGE ₂ production in infected sites modulates immune and inflammatory responses and is reported to be free by lung epithelial cells (N'Guessan et al., 2007).

Protein concentrations were determined by Bio-Rad protein assay based on Bradford (Schleicher and Wieland, 1978). Standard curves were prepared using serial dilution of bovine serum albumin (BSA), and Bio-Ra d protein assay staining reagents were diluted 1: 4 in water. 4 μL of standard and samples were added to 1 mL of diluted staining reagent and the absorbance was measured at 595 nm. Sample protein concentrations were determined from standard curves prepared with BSA.

Meanwhile, to determine PGE ₂ release, HaCaT cells and A549 cells were seeded in 6-well culture plates in each medium of DMEM and RPMI containing FBS and antibiotics at 37 ° C. and in a wet 5% CO ₂ incubator (5 x 10 ⁵ cells / well), after treatment with different concentrations of 15-PGDH inhibitor, the supernatants were collected at specific times after media treatment. Intracellular and extracellular PGE ₂ levels were measured using a PGE ₂ enzyme immunoassay kit (Thermo Scientific, Rockford, IL, USA).

As a result, as shown in FIG. 11, DMEA (1-, 2-, 3-fold ED ₅₀ ), DMW (1-, 10-, 100-fold ED ₅₀ ), DMHE (1-, 10-, 100-fold ED ₅₀ ) and DMME (0.5-, 1- 2-fold ED ₅₀ ) increased PGE ₂ in a concentration dependent manner. D. morbifera hexane extract (DMHE) inhibited 15-PGDH more efficiently.

And time-dependent change of the extracellular PGE ₂ was confirmed. PGE ₂ levels rapidly increased to 1200% of the control at 24 hours, and then rapidly decreased after that time (Table 4, Figure 12). These results confirm that the appropriate time for PGE ₂ detection is 12 hours, DMHE (17.4 μg / mL) was treated for 12 hours, and intracellular and extracellular PGE ₂ levels were also measured and 165% compared with the control group, respectively. And up to 315% (Table 5, Figure 13).

Table 4 time Sample PGE ₂ (pg / mL) 0 hours Control 143.6 DMHE 143.6 12 hours Control 420.4 DMHE 3967.1 24 hours Control 442.9 DMHE 5373.23 48 hours Control 323.28 DMHE 687.28

Table 5 Sample Intracellular (ng / mg) (mean ± SD) Extracellular (pg / mL) (mean ± SD) Control 1.96 ± 0.18 393.67 ± 12.46 DMHE (17.4 μg / mL) 3.24 ± 0.53 * 1238.04 ± 91.76 *

* p <0.05

Example 5: Quantitative RT-PCR Analysis

Intra and extra PGE ₂ levels are functionally related to gene expression levels of C OX-1 / 2, MRP4, 15-PGDH and PGT. To determine the effect of D. morbifera on the genes, mRNA expression of COX-1 / 2, MRP4 and PGT was measured after DMHE (17.4 μg / mL) treatment in HaCaT cells. The DMHE (17.4 μg / mL, 10 × ED50) concentration used in the assay used a concentration that showed sufficient 15-PGDH inhibitory activity without cytotoxicity from previous experiments. To this end, total cell RNA was isolated from the cells using TRI reagents (RNAiso Plus, Takara). 20 μL of cDNA was synthesized from each RNA sample (Invitrogen, USA). PCR reactions included 4 μL of 1: 5 diluted cDNA, 4 mM MgCl ₂ , 10 p mole of each primer and 4 μL of Fast Starter Mix buffer (dNTPs, SYBR Green dye and Tag polymerase). Primers and conditions utilized for RT-PCR are as described in the table below. 5R 1 and 2 primers not listed in the table were purchased from Qiagen Korea Ltd. (Seoul, Korea) and used. Each amplicon size was 185 bp, 119 bp and the annealing temperature was 5 seconds at 60 ° C., extension. Was carried out at 72 ° C. for 8 seconds.

[Correction under Rule 91 06.08.0013]
Table 6

[Correction under Rule 91 06.08.0013]
TABLE 7

As a result, as shown in Figure 14, Real-time PCR analysis showed that DMH increased the expression of COX-1 and MRP4 mRNA, and decreased the expression of MRP4, PGT and 15-PGDH.

Example 6 hair growth promoting effect

 Eight weeks-old C57BL / 6 mice that were kept in the resting phase (synchronized-telogen stage) were used to investigate the effect of promoting hair growth, and the animal experiments performed in the examples of the present invention were approved by the Chonnam National University Animal Research Ethics Committee (IRB). (CNU IACUC-YB-R-2012-8) was performed and the assay pigment was considered evidence of the transition of hair follicles from resting phase to growth phase (Peters et al., 2002).

(1) hair growth effect

In order to confirm the hair growth promoting effect of the yellow lacquer tree, healthy C57BL / 6 mice (7 weeks old, female or male) were prepared by Damul Science Co. Purchased from (Daejeon, Korea)

A week later, animal hair clippers were used to push the hair on all the backs of animals so that all hair follicles were in the resting phase. Animals were randomly divided into three groups: negative control (vehicle), positive control (minosidyl) and experimental group (DMHE). Three days after shaving, DMHE (8.7 μg / 25 μL), 0.5% minoxidil (25 μL) or vehicle was applied daily to the dorsal skin of C57BL / 6 mice. In addition to drug treatment, Vaseline (petroleum jelly cream) was applied to help the drug absorb. Visible hair growth was recorded one and two weeks after drug treatment.

As a result, as shown in FIG. 15, after one week, dark pigment was detected in the DMHE- and minoxidil-treated groups, but not in the vehicle control group. Two weeks later, in the DMHE- and minoxidil-treated groups, the shaved back was completely covered with hair and returned to its original state.

(2) anti-androgen activity

Since it is well known that androgen is involved in hair loss, we further verified whether DM HE has anti-androgen activity. To determine the effect on androgen receptor signaling, LNCaP.FGC cells, androgen-dependent prostate cancer cell lines were used. In prostate cancer, prostate s pecific antigen (PSA) is one of the AR target genes.

As a result, as shown in Figure 16, the hwangchil extract of the present invention inhibited DHT-induced expression of PSA mRNA: n-Hex>EA>n-But> H ₂ O extract in order.

 In addition, as in FIGS. 17-19, non-flutamide DMHE also reduced the target genes, including PSA and 15-PDGH expression, by decreasing 5αRI / 2 enzyme in DHT synthesis.

This was inhibited by the addition of large amounts of DHT, which suggests that the anti-androgen effect of DMHE stems from a competitive mechanism with DHT on AR (FIG. 20). In addition, DMHE downregulated gene expression of AR was also observed in flutamide-treated cells (FIG. 18).

Example 7 AR Signaling Inhibitory Effect of Hair Follicle Dermal Papilla Cells (HFDPC)

To examine the anti-androgen effect of the Hwangchil extract identified through the androgen-dependent LNCaP / FGC prostate cancer cell line, and to determine whether it is effective for male-type alopecia known as the progression of AR-dependent alopecia, HFDPC derived from human scalp Primary hair follicle dermal papilla cells) were used to verify the effect of the following experiment. At this time, HFDPC was used to purchase Primary Human Follicle Dermal Papilla Cells (HFDPC) (Cat.No. C-12071, PromoCell) from PromoCell (Heidelberg, Germany). Stain positively on alkaline phosphatase. In addition, HFDPC has an androgen receptor (AR) and can be used for AR signaling studies. The primary HFDPCs used in this experiment were all 3 4 passages, and the Follicle Dermal Papilla Cell growth medium (C-26501). , PromoCell) was cultured in a 37 ° C incubator with 5% CO _2, water. The jongbun to the culture ^{HFDPC (5 x 10 5/6} well) DMHE then the cells are attached was administered 24 hours. Subsequently, the effects on the mRNA expression of 5αR-1 and 5αR-2 and AR involved in amplifying sensitivity to AR were examined by converting intracellular AR target genes, 15-PGDH and testosterone, into DHT.

As a result, as shown in Figure 21 and 22, the hwangchil extract of the present invention was shown to inhibit the expression of 5αR-1 and 5α-R2 concentration-dependently, in particular showed a stronger inhibitory effect on 5αR-2. In the case of minoxidil or bimatoprost used as a positive control also showed an inhibitory effect on 5αR-1 or 5αR-2, but did not show a concentration-dependent inhibitory effect like the hwangchil extract of the present invention (results not shown).

In addition, as shown in Figure 23, the expression of AR also appeared to have the effect of inhibiting the expression in the case of minoxidil and yellow lacquer extract, Hwangchil extract concentration-dependent strong inhibitory effect on the expression of 15-PGDH target gene of AR (See FIG. 24).

Therefore, the present inventors have found that the hwangchil extract of the present invention has a sufficient role as an inhibitor of 15-PGDH activity and at the same time inhibits 15-PGDH expression by targeting AR signaling. In other words, hair loss progression due to hyperactivation of AR signaling, a major factor of male hair loss, simultaneously brings about 15-PGDH activity, a target gene, leading to a decrease in PGE2 around the scalp. Therefore, it can be seen that the hwangchil extract can be used to treat male hair loss as a substance having an excellent effect of blocking this mechanism and suppressing male hair loss mechanism.

Preparation Example 1 Preparation of Pharmaceutical Formulation

1. Preparation of Tablets

Tablets containing Hwangchil-tree extract of the present invention obtained in the above Example as an active ingredient were prepared by the following method.

Lactose, starch and pregelatinized corn starch were mixed with the extract of Hilchi chile, and then a suitable volume of purified water was added and granulated into powder. The granules were dried and then mixed with magnesium stearate and compressed to obtain tablets.

The components of the tablet are as follows.

Hwangchil Tree Extract 5.0 mg

Lactose BP 150.0 mg

Starch BP 30.0 mg

Pregelatinized Corn Starch BP 15.0 mg

1.0 mg magnesium stearate

2. Preparation of Capsule

Capsules containing the hwangchil wood extract of the present invention as an active ingredient were prepared by the following method.

The hwangchil wood extract of the present invention was mixed with a certain amount of excipient and magnesium stearate. The resulting mixture was filled into gelatin capsules to obtain capsules.

The components of the capsule are as follows.

Hwangchil Tree Extract 5.0 mg

Starch 1500 100.0 mg

Magnesium Stearate BP 1.0 mg

3. Preparation of Injection

Injection solution containing the hwangchil wood extract of the present invention as an active ingredient was prepared by the following method.

The plant extract was dissolved in an appropriate volume of sodium chloride BP for injection, and the pH of the resulting solution was adjusted to pH 3.5 with dilute hydrochloric acid BP, then the volume was adjusted with sodium chloride BP for injection and thoroughly mixed. The solution was filled into a 5 ml Type I ampoule made of transparent glass, and the glass was dissolved and enclosed under an upper grid of air, followed by autoclaving at 120 ° C. for at least 15 minutes to obtain an injection solution.

The components of the injection solution are as follows.

100 µg / ml Hwangchil Tree Extract

Dilute hydrochloric acid BP until pH 3.5

Injectable sodium chloride BP up to 1 ml

<Preparation Example 2> Preparation of the external preparation composition

1. Preparation of Cream

Cetostearyl alcohol 2.8 parts by weight

2.6 parts by weight of beeswax

Stearic acid 1.4 parts by weight

2 parts by weight of glycerol phosphate monostearate

Fiji-100 Stearate 1 part by weight

Sesquioleate sorbitan 1.4 parts by weight

4 parts by weight of jojoba oil

Squalane 3.8 parts by weight

Polysorbate 60 1.1 parts by weight

2 parts by weight of macadamia oil

Tocopherol acetate 0.2 parts by weight

0.4 parts by weight of methylpolysiloxane

0.1 parts by weight of ethyl paraben

0.1 parts by weight of propylparaben

Euxyl K-400 0.1 part by weight

1,3-butylene glycol 7 parts by weight

Methyl paraben 0.05 parts by weight

Glycerin 6 parts by weight

0.2 parts by weight of d-pandenol

4.6 parts by weight of Hwangchil tree extract

0.2 parts by weight of triethanolamine

pt 41891 0.2 parts by weight

pH ₂ O 46.05 parts by weight

2. Preparation of Lotion

Cetostearyl alcohol 1.6 parts by weight

Stearic acid 1.4 parts by weight

Lipophilic monostearic acid 1.8 parts by weight

Fiji-100 Stearate 2.6 parts by weight

0.6 parts by weight of sesquioleate sorbate

4.8 parts by weight of squalene

2 parts by weight of macadamia oil

2 parts by weight of jojoba oil

Tocopherol Acetate 0.4 parts by weight

0.2 parts by weight of methylpolysiloxane

0.1 parts by weight of ethyl paraben

0.1 parts by weight of propylparaben

1,3-butylene glycol 4 parts by weight

0.1 parts by weight of methyl paraben

Xanthan Gum 0.1 parts by weight

Glycerin 4 parts by weight

0.15 parts by weight of d-pandenol

Allantoin 0.1 part by weight

Hwangchil Tree Extract 3.5 parts by weight

4 parts by weight of carbone (2% aq.Sol)

0.15 parts by weight of triethanolamine

3 parts by weight of ethanol

pt 41891 0.1 parts by weight

pH ₂ 0 48.3 parts by weight

Preparation Example 3 Preparation of Patch

1 ~ 25 parts by weight of hwangchil

1 ~ 35 parts by weight of drug release regulator (fatty acid ester)

50 to 95 parts by weight of polymer adhesive (acrylate)

Moisturizer (polyethylene glycol 200) 0.5-25 parts by weight

Preparation Example 4 Preparation of Hair Products

1. Manufacture of Shampoo

The shampoo composition of the present invention was prepared using the following formulation.

Sodium Lauret-7-Sulfate 16 parts by weight

Lauroyl diethanolamide 2.0 parts by weight

0.5 part by weight of Hwangchil-tree extract

DI water 81.5 parts by weight

2. Preparation of Rinse

The rinse composition of the present invention was prepared using the following formulation.

Bariku Art 638 2.0 parts by weight

Lauroyl diethanolamide 2.0 parts by weight

0.5 parts by weight of silicone oil (molecular weight of 10,000)

0.5 part by weight of Hwangchil wood extract

DI water 95.0 parts by weight

So far I looked at the center of the preferred embodiment for the present invention. Those skilled in the art will appreciate that the present invention can be implemented in a modified form without departing from the essential features of the present invention. Therefore, the disclosed embodiments should be considered in descriptive sense only and not for purposes of limitation. The scope of the present invention is shown in the claims rather than the foregoing description, and all differences within the scope will be construed as being included in the present invention.

Claims (9)

Hair growth promoting cosmetic composition comprising an extract of Hwangchil as an active ingredient. The method of claim 1, The hwangchil wood extract is a crude extract of the hwangchil, polar solvent soluble extract or non-polar solvent soluble extract characterized in that the cosmetic composition for promoting hair growth. The method of claim 2, The crude extract is a cosmetic composition for promoting hair growth, characterized in that the extract is available from a solvent selected from water, methanol, ethanol, butanol or a mixed solvent thereof, including purified water. The method of claim 2, The non-polar solvent soluble extract is a cosmetic composition for promoting hair growth, characterized in that the extract soluble in hexane, chloroform, dichloromethane or ethyl acetate.  The method of claim 1, The hwangchil wood extract, (i) inhibition of 15-PGDH enzyme activity; (i) increased PGE ₂ intracellularly and extracellularly; (iii) increased expression of cyclooxygenase-1 (COX-1) and multidrug resistance-associated protein4 (MRP4) genes; (iv) inhibiting the expression of prostaglandin transporter (PGT); (v) inhibition of 5α-reductase expression; And (vi) Cosmetic composition for promoting hair growth characterized by having androgen receptor (AR) expression inhibiting activity. A pharmaceutical composition for preventing hair loss, promoting hair growth, and improving scalp, comprising the extract of Hilchi chinensis as an active ingredient. The method of claim 6, The pharmaceutical composition is a composition for external skin preparations selected from the group consisting of creams, gels, patches, sprays, ointments, warnings, lotions, linings, pastas and cataplasmas as external skin preparations. Pharmaceutical composition. Health functional foods for hair loss prevention, hair growth promotion and scalp improvement comprising the extract of Hwangchil-tree. The method of claim 8, The food is a health functional food, characterized in that selected from the group consisting of beverages, meat, chocolate, food, confectionery, pizza, ramen, other noodles, gum, candy, ice cream, alcoholic beverages, vitamin complexes and health supplements.

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