KR101530415B1 - Composition for preventing hair loss or promoting hair growth comprising extract of Platycarya strobilacea - Google Patents

Abstract

The present invention relates to a composition for preventing hair loss or hair growth comprising extract of Aspergillus oryzae as an active ingredient and more particularly to a cosmetic composition and a pharmaceutical composition containing an extract of Aspergillus oryzae indicative of hair loss prevention or hair growth promoting effect as an active ingredient to provide. Platycarya strobilacea extract is confirmed that the antioxidant activity over the polyphenols, flavonoids, DPPH scavenging activity measurement, and in vivo (in vivo test was conducted to show an excellent effect on hair loss or hair growth.

Description

[0001] The present invention relates to a composition for preventing hair loss or promoting hair growth, which comprises extract of Aspergillus oryzae as an active ingredient,

The present invention relates to a composition for preventing hair loss or for promoting hair growth comprising extract of Aspergillus oryzae as an active ingredient.

Recent rapid economic growth and improvement of living standards have improved the quality of life of the people, but due to pollution and environmental pollution, modern people are exposed to external stimuli and diseases. In particular, it has been found that alopecia is caused by local infection, genetic predisposition, and endocrine disorders related to hair, but recent studies have shown that various factors related to hair loss such as stress, adverse effects due to environmental pollution, . In the mechanism of hair loss, immune cells and neutrophils are activated by the environment such as stress, causing an inflammatory reaction, destroying hair follicle tissue, causing hair cell death, and causing hair loss.

Hair loss is not limited to men but is increasing in women as well, and teenagers are increasingly worried about hair loss. The domestic hair loss population has increased from about 5 million in 2006 to 8 million in 2007 and reached about 9 million in 2008 and increased by 200% in 2006 to over 10 million by the end of 2011. From 2001 to 2008, the proportion of female hair loss patients increased by 73%, male hair loss patients increased by 49%, and the rate of hair loss disease among young people in their 20s and 30s increased significantly (National Health Insurance Corporation, February ; 2012).

Currently, there are two types of minoxidil and finasteride approved by the FDA as a hair growth promoting agent. Minoxidil has been used as an oral antihypertensive drug, but it has been observed that hirsutism appears in patients taking this drug, and is currently used for alopecia areata for scalp application. Minoxidil is a pyrimidine derivative that is widely used as a treatment for hair loss because it enlarges the blood vessels of the scalp, locally increases blood flow, activates the hair cells, slows the progress of hair loss, and has the effect of growing fluff. However, according to several clinical studies, minoxidil has been reported to be effective only in less than 50% of patients, and it is effective in alopecia, but when it is discontinued, hair loss again occurs. Finasteride is the first oral therapeutic for the treatment of male androgenetic alopecia that prevents hair loss by inhibiting type Ⅱ 5α-reductase and thereby promotes hair growth. Approximately 2.6 million people around the world are currently using this drug after approval by the FDA in 1997 .

However, minoxidil has been associated with weight gain, edema, increased heart rate, angina pectoris, dermatitis, itching, and side effects of such a hair growth promoting agent. Finastelide is reported to be a serious example of adverse effects such as male sexual dysfunction and birth defects. There is a problem that is limited or the patients themselves show resistance to drugs. As a result, consumers are interested in hair loss preventive substances derived from natural materials, and various studies on plant extracts are being conducted.

Platycarya Strobilacea S. et Z. ) is a deciduous broad-leaved arboreous tree belonging to the spiny tree family and distributed throughout Korea. Flowers bloom in May-June, fruits ripen in September, and fruits and roots have been used medicinally (Korean botanical book, Angora, 2003. p.181). In the study on the sorghum wood, 5-hydroxy-2-methoxy-1,4-naphthoquinone (5-hydroxy-2-methoxy-1,4-naphthoquinone) And showed strong activity against solid cancer cells in the compound isolated from the leaves of the leaves.

Korean Patent No. 10-0881754 discloses a pharmaceutical composition for treating and preventing inflammatory bowel disease, which contains an extract of Aspergillus oryzae as an active ingredient. The extract of Aspergillus oryzae has low adverse effects as a natural substance, And suppresses the expression of inflammatory cytokines such as MCP-1 and IL-8, thereby being useful as a therapeutic agent for inflammatory bowel disease. However, there is no mention of prevention of hair loss or stimulation of hair growth.

The object of the present invention is to provide Platycarya strobilacea extract of the present invention as an active ingredient.

Another object of the present invention is to provide a cosmetic composition and a pharmaceutical composition containing an extract of Aspergillus oryzae as an effective ingredient for preventing hair loss or promoting hair growth.

In order to achieve the above object, the inventors of the present invention conducted studies on polyphenol, flavonoid, DPPH scavenging ability and frap to examine the effect of the extract of Aspergillus oryzae on the human hair growth, Antioxidant activity was measured by In vivo (in vivo test was conducted to evaluate the use of the extract as a therapeutic agent for hair growth and hair loss, and the present invention was completed.

The present invention relates to Platycarya strobilacea extract as an active ingredient. Specifically, the extract of Aspergillus oryzae is extracted with a solvent selected from the group consisting of water, C1 to C4 lower alcohols, dimethyl sulfoxide (DMSO), and a mixed solvent thereof. In addition, the composition of the present invention is characterized by comprising 0.1 to 50 parts by weight of the extract of Aspergillus oryzae on 100 parts by weight of the composition.

In addition, the composition of the present invention may be provided in various forms selected from a cosmetic composition or a pharmaceutical composition.

When the composition of the present invention is a cosmetic composition, the composition of the cosmetic composition may be prepared by any of the formulations conventionally produced in the art. For example, hair tonic, hair conditioner, hair essence, hair lotion, Hair shampoo, hair rinse, hair treatment, hair cream, hair nutrition cream, hair moisturizing cream, hair massage cream, hair wax, hair aerosol, hair pack, hair nutrition pack, hair soap, hair cleansing foam, But are not limited to, hair preserving agents, hair dyes, hair bleaching agents, hair bleaching agents, hair gels, hair glazes, hair dressing agents, hair lacquers, hair moisturizers, hair mousses and hair sprays.

When the composition of the present invention is a pharmaceutical composition, the pharmaceutical composition may be formulated into a cream, a gel, a patch, a spray, an ointment, a warning agent, a lotion, a liniment, a pasta agent and a cataplasma agent. In addition, the pharmaceutical composition may contain a pharmaceutically acceptable carrier other than the extract of Aspergillus oryzae. Such pharmaceutically acceptable carriers are those conventionally used in pharmaceutical preparations, and include lactose, dextrose, sucrose, But are not limited to, sorbitol, mannitol, starch, acacia gum, calcium phosphate, alginate, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, methylcellulose, methylhydroxybenzoate, Tartrate, magnesium stearate, mineral oil, and the like, but is not limited thereto. In addition, the pharmaceutical composition may further include a lubricant, a wetting agent, a sweetening agent, a flavoring agent, an emulsifying agent, a suspending agent, a preservative, etc. as an additive.

The pharmaceutical composition may be administered according to the severity of depilation, and local administration is usually preferred. The dosage of the active ingredient in the pharmaceutical composition may vary depending on the route of administration, the severity of the disease, the age, sex, and weight of the patient, and may be administered once to several times per day.

On the other hand, the extract of Aspergillus oryzae, which is used in the composition of the present invention, is a natural product with little toxicity and side effects, and can be safely used for prolonged administration for preventive purposes.

The invention there is related to a hair loss prevention or hair growth promoting composition containing Platycarya strobilacea extract as an active ingredient, Platycarya strobilacea extract the antioxidant activity over the polyphenols, flavonoids, DPPH scavenging activity measurement, in vivo (in vivo test was conducted to find that there was an excellent effect on hair loss or hair growth. Therefore, it is considered that the extract of Aspergillus oryzae can be used as a treatment for hair growth and hair loss.

In order to compare hair growth effects, FIG. 1 shows the effect of a solvent (DMSO) on the hair-removed C57BL / 6 mouse, a Platycarya extract of 20mg / 20ml / DMSO concentration strobilacea extracts, PSE) and 5% minoxidil (MXD).
FIG. 2 is a graph showing the effect of a solvent (DMSO) on hair-removed C57BL / 6 mice, a Platycarya extract of 20mg / 20ml / DMSO concentration strobilacea extracts, PSE) and 5% minoxidil (MXD).
FIG. 3 is a graph showing the effect of the solvent (DMSO) on the hair-removed C57BL / 6 mouse, the extract of Platycarya (20mg / 20ml / DMSO) strobilacea extracts, PSE) and 5% minoxidil (MXD).
FIG. 4 is a graph showing the effect of the solvent (DMSO) on the hair-removed C57BL / 6 mice, Platycarya extract of 20mg / 20ml / DMSO concentration strobilacea extracts, PSE) and 5% minoxidil (MXD).
Figure 5 shows the results of treatment with DMSO, Platycarya strobilacea extracts (PSE) and 5% minoxidil (MXD) at a concentration of 20 mg / 20 ml / DMSO and after three weeks, H & E staining in C57BL / 6 mice It is the result of observing the shape and depth of hair follicles.
Figure 6 shows mast cell counts in C57BL / 6 mice after 3 weeks of treatment with DMSO, 20 mg / 20 ml / DMSO, Platycarya strobilacea extracts (PSE) and 5% minoxidil This is a result.
Figure 7 shows the effect of stem cell factor (DMSO) on C57BL / 6 mice after 3 weeks of treatment with DMSO, 20 mg / 20 ml / DMSO, Platycarya strobilacea extracts (PSE) and 5% minoxidil Immunohistochemistry of stem cell factor (SCF) antigen is shown.

Hereinafter, the present invention will be described in more detail with reference to the following examples. However, the present invention is not limited by these examples.

< Example  1> Bark tree  Extract preparation

Distilled water (DW) was extracted from the Korean plant extract bank. 10 ml of 50% methanol (MeOH; MeOH) was added to 20 mg of the sample, and the mixture was extracted at 37 ° C for 24 hours, followed by filtration through a 0.2 μm syringe filter (PALL, Life Sciences). The filtered samples were diluted 2,000 and 4,000 times, respectively.

< Example  2> Bark tree  Extract Antioxidant ability  Measure

1. Total polyphenol content measurement

The total polyphenol content was determined by slightly modifying the Folin-Denis method. 0.5 mL of a 2,000-fold dilution of 50% MeOH extract was added to each well and 0.5 mL of a Folin-Denis reagent (Fluka, Switzerland) was added thereto. The mixture was mixed and left at room temperature for 3 minutes. After 3 minutes, the mixture was added to a 10% sodium carbonate solution (Samchun, Korea) and allowed to stand for 1 hour. Then, the supernatant was taken and analyzed by UV-VIS spectrophotometer (7315, JENWAY, Bibby Scientific Ltd, UK Absorbance at 760 nm. The polyphenol content was determined from a standard curve prepared using tannic acid (Yakuri Pure Chemicals Co., Ltd., Japan). The tannic acid standard curve was prepared by dissolving 0.01 g of tannic acid in 10 mL of 70% methanol and adjusting the final concentrations to be 0.1%, 0.05%, 0.025%, 0.0125% and 0.00625% nm and absorbance was measured.

2. Total flavonoid content measurement

To determine the total flavonoid content, 0.5 mL of a 2000-fold diluted sample was extracted with 50% MeOH, and 5 mL of diethylene glycol (Junsei Chemicals, Japan) was added thereto. After that, 0.5 mL of 1N NaOH (Duksan Pure Chemicals, Korea) was added and mixed well. The mixture was left in a 37 ° C water bath for 1 hour. Absorbance was measured at 420 nm with a UV-VIS spectrophotometer (7315, JENWAY, Bibby Scientific Ltd, UK) exactly one hour later. A standard calibration curve was prepared using naringin (Tokyo Kasei Kogyo Co., Ltd., Japan). After 0.01% of naringin, 10 mL of 99.8% EtOH was added to make 0.1%, and the absorbance was measured at 420 nm by diluting the solution to 0.05%, 0.025% and 0.0125%, and the absorbance of the sample was measured % &Lt; / RTI &gt; naringin equivalent.

3. Measurement of DPPH radical scavenging ability (electron donating ability measurement)

For the measurement of electron donating ability, the sample was diluted with 50% methanol to a concentration of 25 mg / 100 ml. The DPPH reagent was prepared by dissolving in ethanol at a concentration of 0.1 mM while blocking the light. The absorbance at 517 nm was measured by spectrophotometer (7315, JENWAY, Bibby Scientific Ltd, UK) using 0.5 ml of sample and 3 ml of DPPH reagent. (EDA,%) values were calculated.

Electron donating ability (EDA,%) = [(Blank absorbance - sample absorbance) / blank absorbance] × 100

4. Results

The results of total polyphenol, flavonoid, and electron donating ability of the extract of Aspergillus species are shown in Table 1. The total polyphenol content of P. japonica extract was 366.44 ± 6.60 mg / g. The results of other studies showed that the polyphenol content of P. vivax was 176.34 mg / g and that of the leaves was 124.4 mg / g (2008) 18 (2): 213-219), and the total polyphenol content of the extract of Aspergillus oryzae is higher than that of both plants Compared with the extracts. The content of total flavonoids in polyphenol was 152.65 ± 0.36 mg / g. The results of the electron donating ability test showed 73.73 ± 0.11%, which is higher than that of the previous studies (18.4%) and the leaves (70%). (Journal of Life Science. The DPPH radical scavenging activity of P. thunbergii was higher than that of the two previous plant extracts (Fig. 1).

Total Polyphenol Content of Extract Product Name Total polyphenol
(mg / weight of extract, g) Total flavonoid
(mg / weight of extract, g) Electron donating ability (%)
(25 mg / 100 ml) Bark tree 366.44 + - 6.60 152.65 + - 0.36 73.73 ± 0.11

< Example  3> In vivo ( in vivo ) exam

1. Experimental animals and treatment

Four weeks old male C57BL / 6 mice were purchased from the Central laboratory animal, and after the adaptation period for 2 weeks in the breeding room, general symptoms were observed and the experiment was carried out at the 6th week of rest. The experimental group consisted of 4 groups [experimental group: Platycarya (DMC ) application group, strobilacea extracts (PSE) application group, positive control group (PC): minoxidil (MXD) application group and solvent control group (VC): solvent (DMSO) application group). The experimental animals were housed 2 cages per cage. The environmental conditions of the cage were adjusted to room temperature of 22 ± 3 ℃, relative humidity of 55 ± 10%, illumination time of 12 hours (7:00 am to 7:00 pm) Was freely ingested. The experimental animals were treated with CO ₂ anesthesia on the third day of the experiment, and the skin was immediately frozen in liquid nitrogen and stored frozen in a deep freezer.

2. Sample preparation and application

DMSO (1 mL) was added to 20 mg of distilled water (D.W.) extracted from the Korean plant extract bank, and the mixture was extracted at 37 ° C for 24 hours and filtered through a 0.2 μm syringe filter (PALL, Life Sciences). The filtered sample was diluted 1000 times, blocked with light and stored in a refrigerator.

To observe the effects of hair growth, the animal's back hair was removed from the mice one day before the start of the experiment using an electric animal epilator. The samples were pipetted (200 μl per day, once per day, 6 times per week) ) For 3 weeks. 5% minoxidil (Hyundai Pharm Co., Ltd, Korea) was used as a positive control and DMSO was used as a normal group.

3. Gross observation

After the start of the experiment, the hair was anesthetized with ether for 4 times (0, 7, 14, 21 days) during the experiment to observe the state of hair growth. The hair growth effect of each group was visually determined for each animal.

4. Optical microscopic observation after hematoxylin and eosin (H & E) staining

On the last day after the start of the experiment, the part to be coated with the test substance was extracted and fixed with formalin. After dehydration with alcohol and xylene, Respectively. A 3 μm tissue slice was prepared using a microtome (Leica RM 2145, Germany) and stained with hematoxylin-eosin (Sigma Chem. Co., St. Louis, Mo., USA) Leica light microscope, Leica DM 500, Germany). The thickness of the dermis and the depth of the hair follicle were measured using the scale bar of the microscope program.

5. Optical microscopic observation after staining with toluidine blue

Tissue sections were prepared in the same manner as the H & E staining method, and after dehydration with alcohol and xylene according to the usual treatment steps, toluidine blue (Sigma Chem. Co., St. Louis, MO , USA) for 2 minutes, and the number of viable dermis and subcutaneous mast cells was counted by a 200 magnification optical microscope.

6. Stem cell factor (SCF)

To observe the cytokine associated with hair growth, a primary antibody diluted 1:50 in stem cell factor (SCF) was added to the tissue section and reacted at room temperature for 12 hours. At this time, dilution of the primary antibody was performed using 0.1% PBTS and 1% normal goat serum (Vector Laboratories Ins., USA). After that, tissue sections were washed with 0.1M PBTS three times for 5 minutes at room temperature, and 3% H ₂ O _{2 was added} to each tissue for 5 minutes and left for 10 minutes in PBTS. The secondary antibody was prepared in a ratio of 1: 50 to 0.1 M PBTS in a polyclonal goat anti-mouse immunoglobulin HRP (D-2600, DAKO, Denmark) And then washed three times for 5 minutes with PBTS. 3-3 'diaminobenzidine (Roche Diagnostics GmbH, Germany) was dissolved in 0.1 M PBS, and each tissue was reacted with hematoxylin (Sigma Chem. Co., St. Louis, MO, USA) for 20 sec. After dewatering and transparency according to the usual method, they were sealed and observed with an optical microscope for the degree of staining in hair follicles, mast cells and dermis.

7. Results

There were no specific abnormalities in the gross observations of each group, but 5% minoxidil (MXD) application group and Platycarya strobilacea extracts, PSE). This phenomenon is due to the contact between individuals, and it seems that the first week of the experiment did not affect the results of the wound healing. From 7 days after application of epilation after each episode, the skin began to turn blue and the fluff was observed to grow (Fig. 2). (DMSO) application group and 5% minoxidil (MXD) application group and the extract of Platycarya strobilacea extracts, PSE), but the shoots of the PSE - coated group showed partial hairy growth. This tendency was observed until the 10th day of application, and at 3 weeks after the experiment, hair growth was observed in all groups (Fig. 4).

On the other hand, the morphology and depth of the hair follicles were observed by H & E staining of mouse tissues after 3 weeks of application, and PSE group was superior to the other groups (Fig. 5). The depth of the hair follicle was measured using a microscope program of a 100 magnification optical microscope (Leica DM 500, Germany). The depth of the hair follicle was 472.67 ± 333.55 ㎛ in the DMSO group and the MXD (5% Minoxidil) group showed 423 ± 199.52 ㎛ similar results in both groups. On the other hand, PSE ( Platycarya strobilacea extracts) group was 843.33 ± 146.06 ㎛, which was significantly higher than DMSO group and MXD group (Table 2). The depth of hair follicles in the experimental group was higher than that of the solvent control group and the positive control group, and the dermal and hair follicles of the experimental group were found to be more robust than those of the other two groups (Fig. 5).

Measurement of hair follicle depth in C57BL / 6 mice after 3 weeks of application parking DMSO MXD GSE 3 472.67 占 333.55 占 퐉 423 占 199.52 占 퐉 843.33 占 146.06 占 퐉

In addition, the results of observation of mast cells in mouse tissues of each group in the third week of administration through toluidine blue staining are shown in FIG. When mast cells were observed at 200 magnification using an optical microscope (Leica DM 500, Germany), blue-stained mast cells appeared around the hair follicles in the DMSO and MXD groups, whereas in the PSE group, But mast cells showed a significantly lower distribution.

Finally, expression of stem cell factor (SCF) by immunohistochemical staining was observed in hair follicles, mast cells, and dermal layer (Fig. 7). Immunoreactivity to SCF was moderate in the dermis and hair follicles in the DMSO group and the MXD group, and the immunoreactivity to the SCF in the DMSO group and the MXD group was moderate in the dermal layer and the hair follicle. Mild response in mast cells. On the other hand, PSE group showed strong immune responses in hair follicles, mast cells, sebaceous glands and mast cells.

Claims (7)

A composition for preventing hair loss or promoting hair growth comprising extracts of Platycarya strobilacea with water and then extracting the extract with dimethyl sulfoxide (DMSO) as an active ingredient. delete The composition for promoting hair loss prevention or hair growth according to claim 1, wherein the composition comprises 0.1 to 50 parts by weight of the extract of Aspergillus oryzae to 100 parts by weight of the composition. The hair cosmetic composition according to claim 1, wherein the composition is a cosmetic composition. The cosmetic composition may be at least one selected from the group consisting of hair tonic, hair conditioner, hair essence, hair lotion, hair nutrition lotion, hair shampoo, hair conditioner, hair treatment, Hair Moisture Cream, Hair Massage Cream, Hair Wax, Hair Aerosol, Hair Pack, Hair Nutrition Pack, Hair Soap, Hair Cleansing Foam, Hair Oil, Hair Drying Agent, Hair Preservative, Hair Dye, Hair Dye, Hair Decolorant, Hair Gel, Hair Wherein the composition has a formulation selected from the group consisting of glaze, hair dresser, hair lacquer, hair moisturizer, hair mousse, and hair spray. delete The composition according to claim 1, wherein the composition is a pharmaceutical composition, wherein the pharmaceutical composition is at least one selected from the group consisting of creams, gels, patches, sprays, ointments, alerts, lotions, liniments, pastes and cataplasms Wherein the composition for preventing hair loss or for promoting hair growth is characterized in that the composition for promoting hair loss is hair growth promoting composition. delete

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