KR102612128B1 - A cosmetic composition for hair containing an extract of Spiraea prunifolia - Google Patents

Abstract

The present invention relates to a composition for preventing hair loss and promoting hair growth containing the crude extract of the Spiraea tree as an active ingredient, which contains 0.0001 to 10% of the total weight of the cosmetic composition for preventing hair loss as the active ingredient. It is characterized by According to the present invention, it not only helps in the proliferation of hair cells, but also has the effect of inhibiting the expression of human SRD5A1 and SRD5A2, which are genes that promote activation of DHT (Dehydrotestosterone) by 5a-reductase, and VEGF-A, an angiogenesis promoting factor. It is possible to provide a composition for preventing hair loss that can alleviate hair loss by increasing blood vessels in the hair. Therefore, this composition can be effectively used as an anti-hair loss agent to prevent baldness, alopecia areata, and male pattern baldness.

Description

Composition for preventing hair loss and hair growth containing extract of Spiraea prunifolia as an active ingredient {A cosmetic composition for hair containing an extract of Spiraea prunifolia}

The present invention relates to a composition for preventing hair loss or promoting hair growth containing extract of the spirea tree as an active ingredient.

A human hair consists of approximately 150,000 hairs, and each hair goes through a different hair cycle, repeating the process of hair growth and loss. The hair growth cycle is largely divided into three stages: anagen, when the hair grows (anagen), maintenance phase (catagen), when the hair is maintained after growth, and telogen, when the dermal papilla shrinks and hair follicles gradually shrink and hair loss occurs. Repeat step 3 periodically (Herbal Extracts Induce Dermal Papilla Cell Proliferation of Human Hair Follicles, Ann Dermatol, 2015:667-675). The hair growth cycle varies depending on internal factors such as family history, genetics, constitution, and hormonal regulation, and external factors such as nutritional status, aging, surrounding environment, and stress. As hair loss progresses, the growth cycle is abnormally shortened and hair length decreases. You can observe hair follicles becoming shorter, thinner, and shriveled.

The causes of hair loss include not only genetic factors but also environmental factors. Recently, hair loss due to environmental factors has been increasing, and these environmental factors are closely related to free radicals. Active oxygen is oxygen with strong oxidizing power generated during various metabolic processes. It is generated excessively due to environmental factors such as fatigue, poor diet, ultraviolet rays, chemicals, and stress. It accelerates aging and reduces hair vitality through oxidation. . In addition, active oxygen combines with cholesterol to turn into lipid peroxide and attaches to the blood vessel wall, narrowing the scalp blood vessels and blocking pores, reducing blood flow, making it difficult to supply nutrients to the hair roots, and attacking the cell membrane of the cells that make up the hair follicles, causing cell death. It causes scalp inflammation and hair loss. Therefore, in developing a hair loss treatment, the antioxidant ability to suppress the generation of free radicals in the scalp and remove or reduce the activity of excessively generated free radicals can be an important indicator.

In addition, one of the biggest causes of hair loss is male hormones. When testosterone, a male hormone, undergoes the action of 5α-reductase in hair follicles, it is converted into DHT (dehyrotestosterone), and the converted DHT is an androgen in the dermal papilla cell membrane in the frontal and parietal areas. When combined with the receptor, a cell destruction signal is transmitted to the DNA of hair mother cells, and cell necrosis factors BMP, DKK-1, and TGF-β are generated. The cell necrosis factor produced in this way attacks and destroys hair cells, shortening the hair growth period and causing the hair to go into catagen at an early age. There are two types of 5α-reductase: type 1, which is mainly secreted in the sebaceous glands, and type 2, which is distributed in the outer hair root sheath and dermal papilla of hair follicles. Type 2 enzyme is the main cause of hair loss. Therefore, if 5α-reductase can be inhibited or removed, androgenetic alopecia such as male pattern baldness can be prevented or treated.

In addition, external stresses such as childbirth, disease, diet, blood circulation disorders, and peroxidation have been reported to affect hair loss. Recently, interest in hair loss is increasing as hair loss is occurring in younger age groups and women. Hair loss is viewed as a disease and various treatments for this are being released. Minoxidil is an FDA-approved hair loss treatment. ) and finasteride. Minoxidil, a representative product, acts on the blood vessels to increase blood flow and facilitate the supply of nutrients to promote hair growth. It acts directly on the hair follicles to induce hair growth, but it is inconvenient to continuously apply it twice a day. Since it takes about 3 hours to take effect, there are side effects such as scalp dryness, scalp itching, and scalp irritation. Although it helps grow down hair, it is difficult to expect the effect of growing thick hair long. Meanwhile, finasteride delays the progression of hair loss and induces hair growth by suppressing the male hormone DHT, but it has the problem of returning to its original state and hair loss occurring again when you stop taking it, and causes erectile dysfunction in men due to decreased libido. In women, its use is restricted due to the rare side effect of causing birth defects. As such, the effectiveness of existing hair loss treatments, including minoxidil and finasteride, is impaired when use is discontinued, so hair loss treatment that requires long-term treatment requires minimizing side effects. Accordingly, there is a need to develop hair loss treatments derived from new natural substances that minimize side effects rather than chemical treatments.

Meanwhile, spirea is a dicotyledonous plant of the Rosaceae family that grows wild throughout Korea. Its young leaves, fruits, and roots have been reported to be used as a diuretic, detoxifier, inflammation treatment, and painkiller in China since ancient times, and in Korea, it has been reported to be used as a treatment for fever, cough, fever, etc. It has been used as a therapeutic agent (Inhibitory activity of plant extracts on nitric oxide synthesis in LPS-activated macrophages, Phytother. Res., 17(5), 2003:485-489, Pharmacognostic studies on the folk medicine, Kor. J. Pharmacogn. , 2012:193-197).

Accordingly, the present inventors have made efforts to alleviate the above side effects and provide a hair loss treatment with high effectiveness. As a result, the extract of the spirea tree promotes the proliferation of dermal papilla cells and inhibits the expression of enzyme genes that induce the activation of DHT, a male hormone, preventing hair loss and The present invention was completed by confirming that it was effective in promoting hair growth.

KR 10-0998992 B1

The present invention was devised to solve the above problems, and the purpose of the present invention is to provide a composition for preventing hair loss or promoting hair growth by promoting the proliferation of human hair papilla cells and regulating gene expression, which contains an extract isolated from the spirea tree as an active ingredient. Do it as

In order to achieve the above object, the present invention provides a pharmaceutical composition for preventing hair loss and promoting hair growth, or a cosmetic composition for preventing hair loss and improving hair growth, containing crude extract of Spiraea tree as an active ingredient. It is preferable to use leaves, stems, or a mixture of the two for the spirea, and the crude extract is obtained by hot water extracting the ethanol extract of the spirea, and then fractionating the 30% ethanol fraction into 60% ethanol. More preferably, the 60% ethanol fraction is used as a 60% ethanol fraction separated after fractionation with 80% ethanol. The crude extract reduces 5α-reductase gene expression and promotes human hair papilla cell proliferation.

In addition, the present invention provides a composition in which the crude extract of the spirea tree is added in an amount of 0.0001 to 10% by weight based on 100% by weight of the total composition.

As another example, the present invention includes the first step of drying the leaves, stems, or a mixture of the two of the spirea tree, extracting them with ethanol, filtering them, and then concentrating them under reduced pressure; Step 2 of suspending the concentrate in water and then sequentially extracting it twice using purified water, 30% ethanol, 60% ethanol, and 80% ethanol to prepare a Fraction of Spiraea tree corresponding to each solvent; And a third step of concentrating under reduced pressure the 60% ethanol fraction of Spiraea tree prepared in step 2 above. It provides a method for producing a crude extract of Spiraea tree for preventing hair loss and promoting hair growth, including a step. More specifically, in step 2, extraction was repeated twice with purified water, then extracted twice with 30% ethanol, extracted twice with 60% ethanol, extracted twice with 80% ethanol, and then extracted with 60% ethanol. Recover and proceed to step 3.

The composition of the present invention contains meadowsweet extract as an active ingredient and is effective in promoting hair growth and preventing hair loss by influencing the hair growth cycle by promoting the proliferation of human hair papilla cells and controlling the amount of gene expression, and is effective in preventing baldness disease and alopecia areata. It can be usefully used as a pharmaceutical or cosmetic composition for the prevention, alleviation, and treatment of.

Figure 1 shows (a) the process of producing a spirea concentrate and (b) the process of producing a spirea fraction.
Figure 2 shows the chromatographic pattern of separation of the active ingredient from the 60% ethanol fraction of Spearmint tree.
Figure 3 shows the promotion of hair papilla cell proliferation and gene activity.
Figure 4 shows the results of a test on the expression level of genes involved in the hair cycle.

Hereinafter, the present invention will be described in detail.

The present invention provides a pharmaceutical composition for preventing hair loss and promoting hair growth, or a cosmetic composition for alleviating hair loss and improving hair growth, comprising a crude extract of the spirea tree as an active ingredient.

The crude extract is characterized by reducing 5α-reductase gene expression and increasing proliferation of human hair papilla cells.

The Spiraea prunifolia is a plant that grows wild throughout most of Korea and has antioxidant effects (Korean Patent 10-1453609 B1, Korean Patent 10-1176526 B1) and anti-inflammatory activity (Korean J.Plant REs. 30(4) ), 2017:335-342) was reported to exist. This spirea tree has the effect of inhibiting the expression of human SRD5A1 and SRD5A2, genes that promote activation of DHT by 5α-reductase, and increases VEGF-A, an angiogenesis-promoting factor, to create blood vessels in the hair and prevent hair loss. It was confirmed that it could be alleviated.

More specifically, the present invention involves the first step of preparing a sapphire extract by drying the leaves, stems, or a mixture of the two, extracting them with ethanol, filtering, and then concentrating under reduced pressure; Step 2: Suspend the sapphire extract prepared in step 1 in water, sequentially add purified water, 30% ethanol, 60% ethanol, and 80% ethanol, and extract each repeatedly twice to prepare a fraction corresponding to each solvent. step; and a third step of concentrating the 60% ethanol fraction of the Fraction of the Spiraea tree prepared in step 2 under reduced pressure.

In the first step, it is preferable that the spirea extract is extracted at room temperature with a 70% ethanol aqueous solution containing purified water.

In the present invention, the composition is a cosmetic, and the cosmetic formulation includes hair tonic, hair ampoule, hair gel, hair cream, hair lotion, hair skin, hair conditioner, hair shampoo, and hair rinse.

In addition, the composition of the present invention can be manufactured into conventional formulations such as cream, lotion, tonic, solution, suspension, emulsion, skin, and ampoule by additionally containing a cosmetically acceptable carrier. The composition of the present invention usually contains a base, and other ingredients generally used as cosmetic raw materials may be appropriately mixed as needed. The base of the composition of the present invention includes purified water, ethanol, glycerin, butylene glycol, biotin, and sodium hyaluronate, and examples of other ingredients blended in the composition of the present invention include surfactants, emulsifiers, solubilizers, It includes, but is not limited to, thickeners, preservatives, antioxidants, fragrances, etc.

In addition to the above ingredients, the composition of the present invention may contain ingredients that can serve to supply nutrients to hair follicles or commonly used hair growth promotion auxiliary ingredients, and may include growth factors, vitamins, peptides, extracts, etc. Examples may be given, but are not limited to these. Their amount is preferably 0.0001 to 5% by weight of the total composition, especially 0.001 to 1% by weight.

The composition of the present invention can be used as a skin cosmetic formulation by applying it directly to the hair or scalp. The hair to which the composition of the present invention is applied refers to the hair roots and hair follicles of the head, hair roots and hair follicles throughout the body, such as hair and eyebrows. It means area.

The content of the composition according to the present invention can be appropriately determined considering gender, age, hair loss symptoms, hair condition, etc. The 60% ethanol fraction according to the present invention, which has the function of promoting human papilla cell proliferation and regulating gene expression, can be added as a raw material for cosmetics for external skin use. At this time, the 60% ethanol fraction is preferably added in an amount of 0.0001 to 5% by weight, more preferably 0.001 to 1% by weight, based on the dry weight of the external skin composition.

Below, preferred embodiments are presented to aid understanding of the present invention. However, the following examples are provided to make the present invention easier to understand, and the content of the present invention is not limited by the examples.

[Preparation Example 1] Preparation of spirea extract

10 kg of the mixture of leaves and stems of the spirea tree was dried at room temperature for 168 hours, and then 70% ethanol was added to 6.8 kg of the mixture of dried leaves and stems of the spirea tree and left at room temperature for 168 hours for extraction. Then, it was filtered and concentrated under reduced pressure to obtain 250 g of a 70% ethanol extract of Pine Tree. (See Figure 1(a))

[Preparation Example 2] Preparation of spirea crude extract

After sufficiently suspending the 70% ethanol extract of Spearmint tree prepared in Preparation Example 1 in water, 10 times the volume of purified water, 7 times the volume of 30% ethanol, 5 times the volume of 60% ethanol, and 4 times the volume of 80% ethanol were sequentially added. and repeated extraction twice to separate the water layer, 30% ethanol layer, 60% ethanol layer, and 80% ethanol layer, and obtain spirea fractions corresponding to each solvent. (See Figure 1(b))

[Test Example 1] Evaluation of hair papilla cell growth promotion and gene expression regulation

Dermal palilla cells are key cells responsible for hair growth. The dermal papilla is connected to the capillaries and receives oxygen and nutrients, thereby growing hair. Hair cells that receive resupply of nutrients from the dermal papilla divide and proliferate to produce keratin protein. Hair loss is caused by androgens or active oxygen stimulating dermal papilla cells, which then secrete substances that destroy hair follicles or block the secretion of growth factors that grow hair.

Accordingly, in order to confirm the effect of the extract of the present invention on promoting the growth of dermal papilla cells and regulating gene expression, human dermal papilla cells (HDPC, human hair papilla cells) purchased from Cefobio were cultured in a 96 well plate at 1×10 ⁴ cells/well. After seeding, the extracts and fractions of Sporitifolia obtained in Preparation Examples 1 and 2 were treated using serum-free medium at the concentrations according to Table 2 below. After culturing in a 5% CO ₂ incubator at 37°C for 24 hours, absorbance was measured at 450 nm using WST-1 reagent.

Treatment concentration (㎍/㎖) Incubation time (hr) Spearmint tree extract (in DMSO)
2, 10, 50, 100, 200
24 Spearmint 30% ethanol fraction (in DMSO) Spearmint 60% ethanol fraction (in DMSO)

As a result, looking at Figure 3, almost no cytotoxicity was observed even when all three samples were treated and cultured at each concentration, and when the 60% ethanol fraction was treated at 100 μg/ml, the relatively highest survival rate was observed. The results of promoting hair papilla cell proliferation were confirmed. When the 60% ethanol fraction was treated at 200 ㎍/㎖, the survival rate was somewhat low. This cytotoxic result was due to the osmotic pressure or acidity due to the large volume of the 60% ethanol fraction in the culture medium rather than the toxicity of the 60% ethanol fraction itself. It is expected to have an effect on cells. Through the above results, it was confirmed that the sapphire fraction fraction promotes the proliferation of human hair papilla cells and also has a positive effect on the regulation of gene expression.

Meanwhile, the expression levels of genes involved in the hair cycle were confirmed in human hair papilla cells to determine the effect of each fraction of the spirea extract on the hair cycle.

[Test Example 2]

Human hair papilla cells were seeded at 1.5x10^5cells/well, cultured in a CO2 incubator for 24 hours, and the medium was replaced with serum free DMEM. Each sample was treated at a concentration of 50ug/ml and cultured in a CO2 incubator for 24 hours. Afterwards, the cells were disrupted to obtain RNA. RT (Reverse transcription) was performed to obtain cDNA using this RNA as a template. Primers for each gene were prepared using the cDNA created in this way as a template, and PCR (polymerase chain reaction) was performed. After PCR, electrophoresis was performed on 1.5% agarose gel and detection was performed using a UV lamp.

As a result of the experiment, in cells treated with the 60% ethanol fraction, the genes of hSRD5A1 and hSRD5A2 decreased, and the genes of hBAX and hVEGF-A increased, which affects the hair cycle, which provides indirect evidence that it ultimately alleviates hair loss. ( (see Figure 4)

The pharmaceutical composition according to the present invention may contain 0.1 to 99% by weight of the crude extract based on the total weight of the composition.

The pharmaceutical composition containing the crude extract of the present invention may further include appropriate carriers, excipients, and diluents commonly used in the preparation of the composition.

The pharmaceutical dosage form of the crude extract of the present invention may be used in the form of its pharmaceutically acceptable salt, and may also be used alone or in combination with other pharmaceutically active compounds, as well as in an appropriate collection.

The pharmaceutical composition containing the crude extract according to the present invention can be prepared in oral dosage forms such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, external preparations, suppositories and sterile injection solutions according to conventional methods. It can be formulated and used in a form.

Carriers, excipients, and diluents that may be included in the composition containing the crude extract include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, gum acacia, alginate, gelatin, calcium phosphate, Examples include calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, and mineral oil.

When formulated, it is prepared using diluents or excipients such as commonly used fillers, extenders, binders, wetting agents, disintegrants, and surfactants.

Solid preparations for oral administration include tablets, pills, powders, granules, capsules, etc. These solid preparations include the crude extract with at least one excipient, such as starch, calcium carbonate, and sucrose. ) or prepared by mixing lactose, gelatin, etc. Additionally, in addition to simple excipients, lubricants such as magnesium stearate and talc are also used.

Liquid preparations for oral administration include suspensions, oral solutions, emulsions, and syrups. In addition to the commonly used simple diluents such as water and liquid paraffin, various excipients such as wetting agents, sweeteners, fragrances, and preservatives may be included. there is.

Preparations for parenteral administration include sterile aqueous solutions, non-aqueous solutions, suspensions, emulsions, lyophilized preparations, and suppositories. Non-aqueous agents and suspensions may include propylene glycol, polyethylene glycol, vegetable oil such as olive oil, and injectable ester such as ethyl oleate. As a base for suppositories, witepsol, macrogol, tween, cacao, laurel, glycerogelatin, etc. can be used.

The preferred dosage of the crude extract of the present invention varies depending on the patient's condition and weight, degree of disease, drug form, administration route and period, but can be appropriately selected by a person skilled in the art. However, for desirable effects, the crude extract of the present invention is preferably administered at 0.0001 to 100 mg/kg per day, preferably at 0.001 to 100 mg/kg. Administration may be administered once a day, or may be administered several times. The above dosage does not limit the scope of the present invention in any way.

Pharmaceutical Example 1: mountain offering manufacturing

Pine tree crude extract 20 mg

Lactose 100 mg

Talc 10 mg

The above ingredients were mixed and filled into an airtight bubble to prepare a powder.

Pharmaceutical example 2: Preparation of tablets

Pine tree crude extract 10 mg

Corn starch 100 mg

Lactose 100 mg

Magnesium stearate 2 mg

After mixing the above ingredients, tablets were manufactured by tableting according to a conventional tablet manufacturing method.

cosmetic Formulation example 3. Hair tonic manufacturing

Hair tonic was prepared according to a conventional method with the content shown in Table 3 below.

content (%) Purified water 27.20 panthenol 1.00 Spearmint tree extract 1.00 Niacinamide 2.00 sphinganin 0.10 Capryloyl Salicylic Acid 0.20 ethanol 65.00 PEG-60 Hydrogenated Castor Oil 0.50 Butylene glycol 3.00

cosmetic Formulation example 4. hair ampoule manufacturing

Hair ampoule was prepared according to a conventional method with the content shown in Table 4 below.

content (%) Purified water 96.4228 Cooper Tripeptide-1 0.0070 Sodium Hyaluronate 0.4000 biotin 0.1000 arginine 0.0700 Oligopeptide-1 0.0001 Oligopeptide-3 0.0001 Spearmint tree extract 3.0000

Above, the present invention has been described by way of example, and those skilled in the art will be able to make various modifications without departing from the essential characteristics of the present invention. Accordingly, the embodiments disclosed in this specification are for illustrative purposes rather than limiting the present invention, and the spirit and scope of the present invention are not limited by these embodiments. The scope of protection of the present invention should be interpreted in accordance with the claims below, and all technologies within the equivalent scope should be interpreted as being included in the scope of rights of the present invention.

Claims (8)

Pharmaceutical composition for preventing hair loss and promoting hair growth containing crude extract of the spirea tree as an active ingredient
The pharmaceutical composition according to claim 1, wherein the spirea is a leaf, a stem, or a mixture of the two.
The pharmaceutical composition according to claim 1, wherein the crude extract is obtained by hot water extracting the ethanol extract of the spirea tree and then fractionating the 30% ethanol fraction into 60% ethanol.
The pharmaceutical composition according to claim 3, wherein the crude extract is a 60% ethanol fraction separated after fractionating the 60% ethanol fraction with 80% ethanol.
According to paragraph 1,
The crude extract is a pharmaceutical composition characterized in that it reduces 5α-reductase gene expression.
The pharmaceutical composition according to claim 1, wherein the crude extract of the spirea tree is added in an amount of 0.0001 to 10% by weight based on 100% by weight of the total composition.
Cosmetic composition for alleviating hair loss and improving hair growth containing crude extract of the spirea tree as an active ingredient


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