KR102482119B1 - The cosmetic composition containing Chrysanthemum boreale extract and its fermentation product - Google Patents

Abstract

The present invention relates to a cosmetic composition containing chrysanthemum chrysanthemum extract and fermented chrysanthemum chrysanthemum as active ingredients. The composition according to the present invention uses natural substances as active ingredients, resulting in fewer side effects, as well as skin regeneration ability and collagen synthesis according to optimal fermentation. It has excellent anti-aging effect due to its excellent ability, and it has excellent efficacy in hair growth by increasing the dermal papilla cell proliferation ability and the expression level of IGF and VEGF.

Description

The cosmetic composition containing Chrysanthemum boreale extract and its fermentation product}

The present invention relates to a cosmetic composition containing chrysanthemum extract and fermented product as active ingredients, and more particularly, to a composition using the above ingredients to have excellent anti-aging effect and hair growth effect.

The composition according to the present invention uses natural substances as an active ingredient and has few side effects, as well as excellent skin regeneration ability and collagen synthesis ability according to optimal fermentation, so it has excellent anti-aging effect, dermal papilla cell proliferation ability and expression of IGF and VEGF It has an excellent effect on hair growth by increasing the amount.

Chrysanthemum boreale is a type of wild chrysanthemum and is also called Gaehwa. grows in the mountains It is about 1m high. The rhizome is long, and the stems are gathered and stand upright. It has white hairs and many branches are divided. The leaves attached to the root die when the flower blooms. The leaves attached to the stem are alternate, long oval, egg-shaped, 5-7cm long and 4-7cm wide. It is deeply divided into a pinnate shape and has sharp sawtooth on the edge. The petiole is 1-2 cm long. The flowers bloom in yellow in September-October, and the head flower is about 1.5cm in diameter and hangs similarly to the umbel at the end of the branch and stem. The gun is about 4mm long, and the gun pieces are lined up in 3-4 lines, and the outer pieces are line-shaped or narrow and long oval. The corolla is tubular and the end is split into 5 branches. The fruit is an achene, ripens in October to November, and is about 1mm long. The flowers have sedative, detoxifying, and soothe effects, so they are used for headaches and dizziness. It is planted for ornamental purposes, and young shoots are eaten as vegetables. Distributed in Korea, northern China, and Japan.

Skin aging can be divided into two types. The first is intrinsic aging, which refers to the aging phenomenon that cannot be avoided as time passes. The second is photoaging, which refers to the aging phenomenon observed on the skin of the face, back of the hand, and back of the neck exposed to sunlight for a long time.

Sunlight plays a role in budding life and is indispensable for maintaining life for humans, supplying energy to plants and animals, inducing vitamin-D synthesis in our skin, and being used in the treatment of psoriasis or vitiligo. However, repeated exposure to strong light causes aging of the skin. Therefore, the best way to prevent photoaging is to avoid exposure to UV rays. On the other hand, endogenous aging is mainly observed in skin that is not exposed to sunlight, and since it is caused by aging as we age, continuous care is required to prevent aging. Since there are various causes such as blood vessels and hormones, exercise, stress care, and dietary It can be prevented in the same way as the method to reduce general aging phenomena such as environmental improvement.

The clinical characteristics of endogenous aging are more fine wrinkles than photoaging, and compared to photoaging, hyperpigmentation of skin pigment or abnormal pigmentation caused by concentration of pigment in one place rarely occurs, but rather, the skin is often pale due to less pigment. .

Compared to endogenous aging, the clinical features of photoaging are more severe and observed earlier. Compared to endogenous aging, thick and deep wrinkles occur, and many fine wrinkles occur. Irregular pigmentation occurs on the skin exposed to sunlight, and pigment diseases such as solar lentigo increase. The skin becomes very rough, dry, and its elasticity is reduced, resulting in sagging skin in severe cases.

Several studies have reported that skin aging occurs through various molecular mechanisms, including oxidative stress, DNA damage and mutation, telomere shortening, inflammation, and accumulation of advanced glycation end products (AGEs). Furthermore, reactive oxygen species (ROS) stimulate the expression of Matrix Metallo-Proteinases (MMPs), which cause skin aging by decomposing various components of Extracellular Matrix (ECM) Proteins such as Collagen, Fibronectin, Elastin, and Proteoglycans. causes aging

In recent years, as the aging population has increased significantly, interest in anti-aging products has increased rapidly. As a result, interest in anti-aging skin care products has increased, and in particular, the cosmetics market is gradually thriving. Therefore, research leading to the development of functional anti-aging cosmetics by developing natural ingredients that are neither toxic nor irritating to the skin is being actively pursued.

On the other hand, hair loss refers to an increase in the number of hairs that fall out abnormally due to a higher ratio of hairs growing from hair follicles in the catagen or telogen phase than hairs growing in the follicles in the anagen phase during the growth cycle of the hair follicles. There are two main types of alopecia, the first of which is called male pattern baldness, alopecia seniles or androgenic alopecia, which is common in middle-aged men and starts in the frontal and parietal regions. do. Second, it is called alopecia areata, and there is alopecia that can occur in adolescence with round or oval hair loss with relatively clear boundaries. Alopecia is not a disease only in men, but also occurs in women, and most of the alopecia is androgenetic alopecia, which occurs regardless of gender. Alopecia areata also occurs in both men and women, but occurs less frequently in women than in men.

The causes of such hair loss include the deterioration or inhibition of proliferation of dermal papilla cells and geminal matrix cells related to hair cycle control, excessive action of male hormones, decrease in blood flow to hair follicles, excessive sebum secretion, and peroxides. It is known that various causes work alone or in combination.

Recently, the occurrence of alopecia is increasing in modern people due to environmental factors such as air pollution and mental stress, and the age of occurrence is gradually decreasing. Such alopecia causes severe stress to the parties concerned and can cause interpersonal problems, so interest in hair loss treatment and prevention is increasing.

In the treatment and prevention of alopecia, various attempts have been made so far, and external methods include scalp plastic surgery by scalp flapping, hair loss area reduction and tissue expansion, and recently developed autologous hair transplantation. Although these surgical methods are excellent in terms of effectiveness, they have a disadvantage in that they are difficult to be widely used because the economic and psychological burden on the patient for the surgery is very great.

For this reason, various external preparations are used. Currently, the most widely used is Upjohn's minoxidil-containing preparation approved by the US FDA, and recently, Merck's Propecia (finasteride), whose main ingredient is finasteride, which has an inhibitory effect on Type 2 5 alpha-reductase, is used. Propecia) has been released and is being used. However, in the case of minoxidil, side effects such as itching and irritation occur, and in the case of finasteride, when use is stopped, the problem returns to its original state within 2 months, and side effects of sexual function decline such as loss of libido and erectile dysfunction occur, and women give birth to deformed children. There is a downside to not being able to take it because there is a risk.

In addition, studies are being conducted on natural extracts such as cnidium, Phellinus Phellinus, chamomile extracts, sagebrush, calamus, pomegranate, and ginkgo leaf extracts as natural substances with fewer side effects, but when these hair restorers are used in clinical practice, they act on the scalp. effect is not very good. Accordingly, research on hair loss and hair growth using natural products with less stimulation is continuously required.

An object of the present invention is to provide active ingredients exhibiting excellent effects on skin regeneration and hair growth. In particular, the present inventors paid attention to fermented chrysanthemum chrysanthemum extract or fermented chrysanthemum chrysanthemum, which provides effective and safe skin regeneration and hair growth effects.

To achieve the above purpose,

The present invention provides a cosmetic composition containing chrysanthemum extract or a fermented product thereof as an active ingredient.

The chrysanthemum extract is characterized in that it is obtained through hot water extraction at 100 to 150 ° C.

In addition, the fermented product of chrysanthemum extract is characterized in that it is obtained through complex fermentation of two strains of Bacillus methylotrophicus and Bacillus subtilis.

The present invention also provides a cosmetic composition for anti-aging and for preventing hair loss and promoting hair growth, comprising an extract of chrysanthemum chrysanthemum or fermented chrysanthemum chrysanthemum as an active ingredient.

The composition according to the present invention not only has fewer side effects because it uses natural substances as active ingredients, but also has excellent effects on cell regeneration ability, skin regeneration ability, and hair growth promotion through the increase in efficacy according to optimal extraction and fermentation.

Hereinafter, the present invention will be described with examples.

1. Manufacture of chrysanthemum extract

To prepare the extract, 200 g of fermented chrysanthemum was added with 2 Kg of purified water (DI-Water), extracted under the conditions shown in Table 1 below, and filtered through 300 mesh, followed by yield measurement.

The highest yield of 72% was shown under the condition of 120 ℃, 8 hr, but there was no significant difference in yield, so 120 ℃, 6 hr was used as the standard for extraction to save energy. At this time, the Brix of the extract was measured in the range of 5 to 8.

2. Manufacture of fermented product

Fermentation of chrysanthemum extract was carried out using two microorganisms isolated from fermented foods. Two microorganisms, Bacillus methylotrophicus (Apep microrganism S001) and Bacillus subtilis (Apep microrganism K007), which were screened and selected as optimal fermentation strains by the present inventors, were used. The pre-culture was cultured for 24 hours in a shaking incubator at 37° C. for each of the two fermentation strains using Nutrient broth (Difco, Detroit, MI, USA).

The composition used during fermentation of sanguk extract was sanguk extract, Glucose 0.6%, Yeast Extract 0.3%, and Soytone 0.1%. The chrysanthemum extract was quantified based on the solid content and used during fermentation. The medium for fermentation was sterilized at 121 ° C for 20 minutes by putting the medium ingredients except for the chrysanthemum extract in a fermentor. Components of the extract were not destroyed by heat. Here, two types of fermenting microorganisms were inoculated. The concentration of fermenting microorganisms was measured with a spectrophotometer at a concentration of about 1.0 X 109 cfu/ml and used. After inoculation was completed, fermentation was performed by shaking culture at 37°C for 3 days.

After completion of fermentation, the fermented product was centrifuged to filter out microorganisms, and then filtered once more with a 0.2 μm filter to completely remove microorganisms.

3. Comparison of material changes before and after fermentation

The extracted extract was fractionated according to the permittivity of the organic solvent, and after confirming the analysis of the solvent having physiological activity, it was concentrated again, and the separation solvent for TLC was selected according to the characteristics of the compound. TLC is a method in which the adsorbent is made into a uniform thin layer. A thin layer of Silica Gel is often used. For highly polar compounds, a thin layer of C18, Polyamide, and Cellulose is used. C is a developing solvent with good resolution and short analysis time. It can be used for various types of material analysis because there is no limitation on color development reagents.

4. Cell Viability Assay

Cell Viability Assay is a test method using the reduction of MTT Tetrazolium, a yellow water-soluble substrate, to Formazan Crystal, a blue-violet water-insoluble substrate, by the dehydrogenase action of mitochondria in cells. In order to test the safety of the material and confirm the cell proliferative ability, the material was treated with skin keratinocytes (HaCaT), fibroblast, and dermal papilla cells (HDP, Human Dermal Papilla Cell), and the cell proliferative ability test was conducted.

5. Wound Healing Assay

The cell regeneration inducing effect of the fermented product-derived peptide was verified through the Wound Healing Assay, which tests the cell recovery ability through the growth of skin cells. The middle of the human skin keratinocyte cell line grown to 100% fill in a 6-well plate was uniformly wounded using a tip, and then washed twice with PBS. The experimental time was based on a total of 12 and 24 hours, and the degree of recovery of the wound during each time was confirmed through image equipment, and this was quantitatively confirmed through Image J program.

6. Collagen synthesis ability test

It was cultured for 24 hours at 37°C in a 5% CO ₂ humidified incubator with 10% FBS and DMEM culture medium. Change the medium to Serum-free medium and incubate for 24 hours by treating with TJEs, TJFs, and PEP 1 μg/ml. Collagen synthesis in the cell supernatant was quantified using the Collagen Type 1 c-Peptide ELISA Kit (TKARA, BioInc.) according to the manufacturer's protocol.

7. qPCR

The human epithelial cell line cultured for 1 day was treated with the extract, and nucleotide was separated by adding RiboEX RNA Purification Solution to the cells treated with the extract. After adding chloroform and isopropanol and centrifuging through a refrigerated centrifuge, only RNA is separated. After quantification of extracted RNA, cDNA was synthesized through Reverse Transcription PCR. After amplifying a specific gene related to skin barrier loss in Real Time PCR using a specific primer, the expression level of the specific gene compared to GAPDH was quantitatively analyzed.

[Test result]

1. Comparison of material changes before and after fermentation

As a result of analyzing substances before and after fermentation and substances by solvent fractions, it was confirmed that the size of substances after fermentation was relatively small and evenly distributed compared to before fermentation. This means that it can penetrate the skin more effectively when used as an external agent such as the skin. In addition, when checking each solvent, many substances were identified mainly in the water fraction and the buthanol fraction, confirming that the active ingredient was highly likely to be water-soluble (Fig. 2).

2. Cell viability test results

Cell proliferative ability tests were conducted on fibroblast cells, keratinocytes (HaCaT cells), and dermal papilla cells (HDP Cell) for the extract and fermented product. The results are shown in Fig. As in 3, cytotoxicity was not shown for all cells, and it was confirmed that the viability was increased in a concentration dependent manner. In particular, it was found that the cell proliferation ability was better in the fermented product than in the fermented product. In particular, since the viability in dermal papilla cells indicates that the material has a hair growth effect, it was confirmed that the hair growth effect of the fermented chrysanthemum extract and fermented product was confirmed. As a result, it can be confirmed that both the chrysanthemum extract and the fermented product are safe and effective materials for the skin.

3. Skin regeneration test results

The wound healing effect confirmation test was conducted through the Wound Healing Assay in skin keratinocytes (HaCaT Cell) for the extract and the fermented product. As a result, compared to Vitamin-C used as a control group, it was confirmed that the effect was equivalent or higher. In particular, at a concentration of 300 μg/ml, it showed a far superior effect than Vitamin-C, confirming that the regeneration effect was very excellent when used on the skin (Fig. 4~5).

4. Collagen Synthesis Test Result

In order to confirm the anti-wrinkle effect of sanguk, a test was conducted on the ability to synthesize collagen. As a result of confirmation, it showed excellent efficacy compared to the control, and in particular, at a concentration of 300 μg / ml or more, it showed better efficacy than the positive control group, and more excellent efficacy was confirmed in fermented sanguk than in sanguk extract.

5. qPCR test results

IGF (Insulin-like Growth Factor) is a factor that promotes hair growth by promoting the proliferation of dermal papilla cells. At a concentration of 100 μg/ml, sanguk extract had no effective effect compared to the control, but sanguk fermented product was 8 times more effective than sanguk extract. It showed a double expression increase, and at a concentration of 500 μg/ml, a 35-fold increase in the expression level was confirmed, indicating that the fermented sanguk extract has a far superior effect on hair growth than the sanguk extract.

VEGF (Vascular Endotherial Growth Factor) is a new blood vessel growth factor known to promote hair growth through improvement of blood circulation. As a result of qPCR, in the case of fermented sanguk, compared to sanguk extract, the expression increased about 2 times at 100 μg / ml and about 4 times at 500 μg / ml, and it was confirmed that there is a hair growth effect by improving blood circulation.

As examined above, the fermented chrysanthemum extract and chrysanthemum fermented product according to the present invention promote cell proliferation without toxicity to cells, and have excellent skin regeneration effects, confirming their potential as cosmetic materials. It can be applied to the field of anti-aging, and it was confirmed that it has excellent efficacy as a material for hair growth by increasing the activity of dermal papilla cells and the expression of IGF and VEGF.

Claims (5)

A cosmetic composition for preventing hair loss and for hair growth, containing chrysanthemum extract or a fermented product thereof as an active ingredient. According to claim 1,
Chrysanthemum chrysanthemum extract is a cosmetic composition for preventing hair loss and growing hair, characterized in that obtained through hot water extraction of 100 ~ 150 ℃. According to claim 1,
A cosmetic composition for preventing hair loss and growing hair, characterized in that the fermented product of chrysanthemum extract is obtained through complex fermentation of two strains, Bacillus methylotrophicus and Bacillus subtilis. delete delete