KR102149223B1 - Cosmetic composition for skin whitening containing the fermented complex natural extracts - Google Patents

Abstract

The present invention relates to a cosmetic composition for skin whitening comprising a fermented complex natural extract. The present invention provides a cosmetic composition for skin whitening containing a fermented complex natural extract obtained by sequentially fermenting mountain mulberry, mistletoe, inula flower, Weeping Forsythia, and poncirus with Lactobacillus sakei and Aspergillus niger and carrying out an extraction. The fermented complex natural extract shows has an excellent tyrosinase expression inhibitory activity and an excellent melanin production inhibitory activity, and thus can be useful as a skin-whitening cosmetics.

Description

Cosmetic composition for skin whitening containing the fermented complex natural extracts}

The present invention relates to a cosmetic composition containing a natural complex fermentation extract, specifically, mountain mulberry, mistletoe, sunbokhwa, Yeongyo, and fat to Lactobacillus sakei ( Lactobacillus sakei ) and Aspergillus niger ( Aspergillus niger ) It relates to a technology for producing an extract having excellent skin whitening activity by sequential fermentation extraction and using it as a cosmetic composition for skin whitening.

It is everyone's constant desire to have white and fair skin. In general, the color of human skin, hair, and eyes is determined by melanin carotene and hemoglobin. Human skin color is determined by several factors, such as the distribution of active blood vessels of melanocytes, which make melanocytes, the thickness of the skin and the presence or absence of pigments inside and outside the body such as carotenoids and bilirubin. The most important is melanin, a black pigment produced by the action of enzymes. The formation of melanin pigment is influenced by genetic factors, physiological factors related to hormone secretion and stress, and environmental factors such as ultraviolet irradiation. Melanin exists in the skin and has an important function to protect the body from ultraviolet rays, but it is known that overproduction of melanin not only forms spots or freckles, but also promotes skin aging and plays an important role in causing skin cancer.

The main function of melanin is to protect the skin from damage by removing these harmful radicals. Therefore, a high amount of melanin means that you have an effective countermeasure system that can protect your skin from physical or chemical toxins. Factors that promote the production of melanin include estrogen or prostagladin, as well as sunlight and ultraviolet rays. Melanin biosynthesis is the continuous oxidation of specific enzymes such as tyrosinase, tyrosinase-related protein-1 (TRP-1), and dopachrome tautomerase (DCT) from the melanocytes of melanocytes in the epidermal basal layer. It occurs by a reaction, and among them, an experiment for inhibiting tyrosinase activity, which is a major enzyme in melanin synthesis, is being adopted in the early stages of the development of melanin polymer inhibitors. However, besides the oxidation reaction by tyrosinase, among the factors affecting the melanin production pathway in the skin, there are advanced glycation end products (AGEs) reactions or non-enzymatic glycation reactions. Glycation is a non-enzymatic reaction between sugar and protein, and reacts with the carbonyl group of the reducing sugar and the free amino group of the protein, lysine or arginine, to form the initial glycosylation product, the schiff base. It is a reaction that forms and forms irreversible final glycation end products (AGEs), which are browning compounds, through a series of reactions such as rearrangement and cross-linking.

This is because lysine or arginine, which is a free amino group of protein, and the carbonyl group of reducing sugar react to form a schiff base, an initial glycation product, and the compounds formed at this time continuously undergo a series of complex reactions such as condensation, repotentialization, oxidation, cleavage, and cyclization. It is a reaction that makes a compound (melanoidine). When these AGEs are produced and accumulated, melanin pigments are produced and accumulated in the skin. The production of melanin is synthesized in an organelle called melanosomes in melanocytes in the basal layer. When AGEs are produced, the receptor for AGE (RAGE) acts on the receptor site. After binding to RAGE, AGE promotes Microphthalmia associated transcription factor (MITF), an important transcriptional regulator in the process of melanin synthesis, promotes the expression of the enzyme tyrosinase, which plays the most important role in melanin production, and finally, melanin Is created. The synthesized melanin pigment is delivered to adjacent cells, keratinocytes, through dendrites of melanocytes, and the delivered melanin pigments are distributed in various parts of the skin through the keratinocytes.

Ascorbic acid, kojic acid, arbutin, hydroquinone, glutathione, or derivatives thereof to treat or alleviate skin pigmentation symptoms such as spots, freckles or pigmentation, and excessive melanin pigmentation caused by UV exposure. Substances with tyrosinase inhibitory activity have been formulated and used in cosmetics and pharmaceuticals, but their use is restricted due to insufficient whitening effect, safety issues for the skin, formulation and stability issues when blending cosmetics.

The inventors of the present invention were searching for natural products with skin whitening activity, and a compound extracted by sequentially fermenting mountain mulberry, mistletoe, sunbok flower, yeonkyo and fatty fruit with Lactobacillus sakei and Aspergillus niger . The present invention was completed by confirming that the fermented extract exhibits excellent skin whitening activity.

(001) Korean Patent Registration No. 10-0731613 (2007.06.22) (002) Republic of Korea Patent Publication No. 10-2019-0009164 (2019.01.28)

An object of the present invention is to provide a cosmetic composition for skin whitening showing excellent skin whitening activity by containing as an active ingredient a complex fermentation extract of mountain mulberry, mistletoe, sunbokhwa, yeonkyo and fat.

In addition, another object of the present invention is to provide a method for producing a complex fermented extract of mountain mulberry, mistletoe, sunbokhwa, yeonkyo and fat.

In order to achieve the above object, according to the present invention, a complex fermented extract extracted by sequential fermentation with Lactobacillus sakei and Aspergillus niger from mountain mulberry, mistletoe, sunbok flower, Yeongyo and fat A cosmetic composition for skin whitening containing is provided.

The complex fermentation extract is a weight ratio of 1-5:1-5:1-5:1-5:1-5, more preferably 1-2:1 It is prepared by extracting after mixing at a weight ratio of ~2:1~2:1~2:1~2.

The complex fermentation extract extracted by sequential fermentation of mountain mulberry, mistletoe, sunbok flower, yeongyo and fat with Lactobacillus sakei and Aspergillus niger is 0.001 to 20 weight based on the total weight of the cosmetic composition % Is included.

The cosmetic composition is skin lotion, skin softener, skin toner, astringent, lotion, milk lotion, moisture lotion, nutrition lotion, massage cream, nutrition cream, moisture cream, hand cream, essence, pack, soap, shampoo, cleansing foam, cleansing It may be made of a formulation of lotion, cleansing cream, body lotion, body cleanser, emulsion, press powder, loose powder, or eye shadow.

According to the present invention in order to achieve the above other object, (A) preparing and sterilizing a mixture of mountain mulberry, mistletoe, sunbokhwa, Yeongyo and fat; (B) inoculating the mixture with Lactobacillus sakei and then culturing to prepare a first fermented product; (C) preparing a second fermented product by inoculating and culturing the first fermented product with Aspergillus niger ; And (D) there is provided a method for producing a complex fermentation extract of mountain mulberry, mistletoe, sunbokhwa, Yeongyo and fat extract comprising the step of extracting by adding a solvent to the secondary fermentation product.

At this time, as the solvent, at least one selected from the group consisting of water, anhydrous or hydrous alcohol having 1 to 4 carbon atoms, ethyl acetate, acetone, glycerin, ethylene glycol, propylene glycol and butylene glycol may be used.

The complex fermented extract extracted by sequential fermentation of mountain mulberry, mistletoe, sunbok flower, lotus root, and fat with Lactobacillus sakei and Aspergillus niger is safe and has excellent skin whitening activity by its synergy As it shows, it can be usefully used as a skin whitening cosmetic.

1 is a graph showing the inhibitory effect of tyrosinase activity of a complex fermented extract according to an embodiment of the present invention.
Figure 2 is a graph showing the melanin production inhibitory effect of the complex fermented extract according to an embodiment of the present invention.

Hereinafter, the present invention will be described in more detail.

Morus bombycis is a deciduous small tree of the Morus family Morus, a dicotyledonous plant. It grows in mountains or on the banks of rice fields. It is 7-8m in height and about 50cm in diameter. The bark is grayish brown. Small branches with or without fine hairs gradually become blackish brown. The leaves are egg-shaped or broad egg-shaped, 8-15cm long and 4-8cm wide. There are irregular and sharp serrations on the edge, hairs a little on the back vein, and the end is long like a tail. Stipules fall early, petioles have fine hairs, and the length is 5-25mm. It is used for tools, carving, and boiled water, the leaves are used for silkworm feed, and the bark is used for medicinal or papermaking. In this regard, Korean Patent Registration No. 10-1722615 discloses a skin moisturizing cosmetic composition containing a mountain mulberry extract, and Korean Patent No. 10-0731613 discloses a whitening and yellowing cosmetic composition containing a mountain mulberry stem heartwood extract. The composition is disclosed.

Mistletoe ( Viscum album ) is an evergreen parasitic shrub of the Mistletoe family, a dicotyledonous plant. It is a semi-parasitic plant that makes chlorophyll through photosynthesis by itself, and has green leaves in all four seasons. The flower of mistletoe is yellow in March and blooms at the tip of the branch, and the fruit is round and ripens in light yellow in October. Mistletoe, which mainly lives in the highlands along the Baekdu-daegan, is a plant that survives winter, so it is valuable because it cannot be harvested unless it is in a high area. Mistletoe has chlorophyll, so it can assimilate carbon by itself to make nutrients, and it blooms flowers and bears beautiful fruit even without contact with soil throughout life. In oriental medicine, stems and leaves are used for molesting, pyeongboje, toothache, and low back pain.

Seonbokhwa (旋覆花) is the dried flowers of the asteraceae plant, Geumbulcho. Inula britannica var. japonica grows in wetlands and propagates while the rhizomes are extended. It is 30~60cm high, has hairs all over, and the stem stands straight. Leaves are alternate, no petiole, long oval or lanceolate with fine serrations. Leaves are 5 to 10 cm long and 1 to 3 cm wide, and the bottom is narrowed to wrap the stem and have hairs on both sides. Flowers bloom yellow in July-September, and the whole hangs on the ends of the main stem and branches, and the whole hangs in the form of a corymbula (揀房狀). The guns are hemispherical and the guns are arranged in 5 rows. Snowflakes are 16-19mm long and 1.5-2mm wide. The achene has hairs along with 10 ridges, and the tube hair is about 5mm long. Young shoots are edible as herbs or soup. Seonbokhwa (旋覆花) of herbal medicine, dried this flower, has the effect of geodam, jinhae, geonwi, and soothing. Outposts and roots are also called Geumbulcho and Geumbulchogeun, respectively.

Yeongyo (連翹) refers to the dried fruit of Forsythia viridissima Lindley or Forsythia suspensa Vahl of the ash family. It is a capsule of ovate or long ovate, with sharp ends, and the stem of the fruit remains at the base. The outer surface is light brown or dark brown, with small pale gray ridges scattered here, with two longitudinal grooves. The well-opened one is split along the longitudinal groove and the end is twisted. The inside of the split skin is yellowish brown, and there is a partition wall in the middle. The seed has elongated oval wings. As a pharmacological action, antibacterial action, anti-inflammatory action, blood pressure lowering, hemostasis action, liver treatment action, antipyretic action, dust and diuretic action have been reported. Major chemical components include forsythol, sterol compounds, saponin, and oleanolic acid.

Jisil (枳實) is the dried under-ripe fruit of the mountain plant, Poncirus trifoliata Rafin. Pharmacological experiments revealed uterine contraction, gastrointestinal peristaltic activity, blood pressure elevation, cardiovascular function, and diuretic activity. In Oriental Medicine, food (食滯), digestive disorders, abdominal bloating, constipation, dysentery, uterine ptosis, stomach and sewage, and prolapse are written. In this regard, Korean Patent Publication No. 10-2019-0009164 discloses a skin soothing cosmetic composition containing a fermented fat extract as an active ingredient, and Korean Patent Publication No. 10-2008-0024589 discloses a fat extract as an active ingredient. A skin astringent cosmetic composition containing is disclosed.

According to the present invention, a cosmetic composition for skin whitening containing a complex fermentation extract extracted by sequentially fermenting mountain mulberry, mistletoe, sunbokhwa, yeonkyo and fat with Lactobacillus sakei and Aspergillus niger Is provided. The present invention is a complex fermentation extract extracted by sequential fermentation of mountain mulberry, mistletoe, sunbokhwa, yeonkyo and fat with Lactobacillus sakei and Aspergillus niger , more excellent whitening by its synergy It is characterized in that it exhibits activity.

The extract as an active ingredient is prepared by extraction with at least one extraction solvent selected from the group consisting of water, anhydrous or hydrous alcohol having 1 to 4 carbon atoms, ethyl acetate, acetone, glycerin, ethylene glycol, propylene glycol, and butylene glycol. will be.

In the present invention, mountain mulberry, mistletoe, sunbokhwa, Yeongyo and Jisil are used after being cleaned with purified water, and dried according to the extraction method and minced or made into powder. In addition, the extract of the present invention may be a concentrate obtained by concentrating the extract extracted with an extraction solvent under reduced pressure, a freeze-dried powder of the concentrate, or a powder dried by spray-drying the concentrate.

In the preparation of the extract of the present invention, the mountain mulberry is used at least one selected from the group consisting of stems and leaves, and mistletoe outpost is used.

Mountain mulberry tree, mistletoe, seonbokhwa, Yeongyo and Jisil complex fermentation extract of the present invention comprises the steps of (A) sterilizing by preparing a mountain mulberry, mistletoe, sunbokhwa, Yeongyo and Jisil mixture; (B) inoculating the mixture with Lactobacillus sakei and then culturing to prepare a first fermented product; (C) preparing a second fermented product by inoculating and culturing the first fermented product with Aspergillus niger ; And (D) extracting by adding a solvent to the secondary fermentation product.

In this case, the mixture is a weight ratio of 1-5:1-5:1-5:1-5:1-5, and more preferably 1-2:1 of mountain mulberry, mistletoe, sunbokhwa, Yeongyo and Jisil, respectively. It is made by mixing at a weight ratio of ~2:1~2:1~2:1~2. Most preferably, it exhibits the best whitening activity when it is mixed in the same weight ratio.

According to one embodiment of the present invention, after sterilizing the mixture, Lactobacillus sakei is inoculated with 1.6 × 10 ⁹ cfu and cultured for 2 to 3 days to prepare a first fermented product. Subsequently, 1.0 × 10 ⁴ cfu of Aspergillus niger was inoculated to the first fermented product and cultured for 3 to 5 days to prepare a second fermented product.

Then, a solvent is added to the secondary fermented product, extracted according to a conventional method, filtered, and concentrated to prepare a complex fermented extract. As the extraction solvent, at least one selected from the group consisting of water, anhydrous or hydrous alcohol having 1 to 4 carbon atoms, ethyl acetate, acetone, glycerin, ethylene glycol, propylene glycol and butylene glycol may be used.

The mountain mulberry tree, mistletoe, sunbokhwa, Yeongyo and Jisil complex fermentation extracts, respectively, the mountain mulberry, mistletoe, sunbokhwa, Yeongyo and Jisil, respectively, and Lactobacillus sakei Each fermented extract may be prepared by sequential fermentation extraction with Aspergillus niger , and then may be prepared by mixing them.

It was confirmed that the extracts of fermented mountain mulberry, mistletoe, sunbokhwa, Yeongyo, and Jisil complex of the present invention exhibited more excellent whitening activity by synergistic action according to their mixing (Test Example).

Mountain mulberry, mistletoe, sunbok flower, Yeongyo and branch fruit with Lactobacillus sakei The complex fermentation extract extracted by sequential fermentation with Aspergillus niger is preferably contained in an amount of 0.001 to 20% by weight based on the total weight of the cosmetic composition.

The cosmetic composition is skin lotion, skin softener, skin toner, astringent, lotion, milk lotion, moisture lotion, nutrition lotion, massage cream, nutrition cream, moisture cream, hand cream, essence, pack, soap, shampoo, cleansing foam, cleansing It may be made of a formulation of lotion, cleansing cream, body lotion, body cleanser, emulsion, press powder, loose powder, or eye shadow.

[Example]

Hereinafter, the present invention will be described in detail through the following examples and test examples, but the present invention is not limited by these examples.

Preparation Example 1: Preparation of mountain mulberry extract

The washed, dried, pulverized mountain mulberry leaf and stem mixture was sterilized after adding 150% water to 100 g by weight. Lactobacillus sakei diluted with a 50% dilution of the weight of the original product was inoculated with 1.6 × 10 ⁹ cfu, cultured for 2 days, and then sterilized. Aspergillus niger diluted with a dilution of 50% by weight of the raw material to the sterilized primary fermentation product was inoculated with 1.0 × 10 ⁴ cfu, cultured for 3 days, and then sterilized. After adding 70% ethanol of 10 times the mass of the raw material to the sterilized secondary fermentation product, it was immersed and extracted at room temperature for 1 day.The filtered filtrate was collected, concentrated under reduced pressure, and dried to obtain the extracted powder in a yield of 7.6% of the original product. Got it.

Preparation Example 2: Preparation of mistletoe extract

Washed, dried, and pulverized mistletoe outpost 100 g was prepared in the same manner as in Preparation Example 1 to obtain an extract powder in a yield of 19.5% compared to the original product.

Preparation Example 3: Preparation of Sunbokhwa extract

Washed, dried, and pulverized 100 g of predicament was prepared in the same manner as in Preparation Example 1 to obtain an extract powder in a yield of 11.6% compared to the original product.

Preparation Example 4: Preparation of Yeongyo extract

Washed, dried, and pulverized 100 g of bristle was subjected to the same method as in Preparation Example 1 to obtain an extract powder with a yield of 21.4% compared to the original.

Preparation Example 5: Preparation of fat extract

Washed, dried, and pulverized 100 g of fat was obtained in the same manner as in Preparation Example 1 to obtain an extract powder with a yield of 26.5% compared to the original.

Examples 1 to 6: Preparation of mountain mulberry, mistletoe, sunbokhwa, Yeongyo and fat fermented extract

Washed, dried, pulverized mountain mulberry leaf and stem mixture, mistletoe outpost, sunbokhwa, yeonggyo and fat were mixed in the weight ratio of Table 1 to prepare a mixture 500g, and 150% of water by weight was added to the mixture. And then sterilized. Lactobacillus sakei diluted with a 50% dilution of the weight of the original product was inoculated with 1.6 × 10 ⁹ cfu, cultured for 2 days, and then sterilized. Aspergillus niger diluted with a dilution of 50% by weight of the raw material to the sterilized primary fermentation product was inoculated with 1.0 × 10 ⁴ cfu, cultured for 3 days, and then sterilized. To the sterilized secondary fermentation product, 70% ethanol, 10 times the weight of the raw material was added, followed by immersion, extraction at room temperature for 1 day, the filtered filtrate was collected, concentrated under reduced pressure, and dried to obtain an extract powder.

Extract (weight ratio) Example 1 Example 2 Example 3 Example 4 Example 5 Example 6 Mountain mulberry One 2 2 One 2 One mistletoe One One 2 One One One Boat One One One One 2 2 Yeongyo One One One 2 One 2 Jisil One One One 2 One One

Test Example 1: Evaluation of inhibition of tyrosinase activity in cells

The ability of the extracts of Preparation Example and Example 1 to inhibit intracellular tyrosinase activity was confirmed. To confirm this, it was tested as follows.

Mouse melanoma cells, B16F10 cells, were cultured in DMEM medium containing 10% Fetal bovine serum (FBS), 2% HEPES, and 1% Antibiotic-Antimycotic at 37° C. and 5% CO ₂ . Remove the cultured B16F10 cells with 1×rypsin-EDTA, spread 1×10 ⁵ cells per well on a 6-well plate, and incubate for 24 hours. Five kinds of complex fermentation extracts (Example 1) of Hwa, Yeongyo and Jisil were treated in a medium containing 1 μM α-MSH and cultured. On the fifth day, the medium was removed, washed with DPBS, and then a hemolysis buffer was added and the cells were crushed. After centrifugation at 4° C. and 13,000 rpm for 20 minutes, the supernatant was taken, and then 50 μl was added to a 96-well plate. Then, 50 μl of 2 mg/ml L-DOPA was added and reacted at 37° C. for 1 hour, and the absorbance was measured at 475 nm using an absorbance meter to evaluate the ability to inhibit tyrosinase activity.

The results are shown in FIG. 1.

As can be seen in Figure 1, compared to the extracts of the preparation example, mountain mulberry, mistletoe, sunbokhwa, Yeongyo and fat were extracted by sequential fermentation with Lactobacillus sakei and Aspergillus niger It was confirmed that the complex fermentation extract of Example 1 exhibited a more excellent inhibitory effect on intracellular tyrosinase activity.

Test Example 2: Confirmation of inhibition of melanogenesis

The ability of the extracts of Preparation Example and Example 1 to inhibit melanin production was confirmed. To confirm this, it was tested as follows.

Mouse melanoma cells, B16F10 cells, were cultured in DMEM medium containing 10% Fetal bovine serum (FBS), 2% HEPES, and 1% Antibiotic-Antimycotic at 37° C. and 5% CO ₂ . The cultured B16F10 cells were detached with 1× trypsin-EDTA, spread on a 6-well plate at 1×10 ⁵ cells per well, and incubated for 24 hours, and then on the 2nd and 3rd days, extracts of Preparation Example and mountain mulberry, mistletoe, Five kinds of complex fermented extracts (Example 1) of Seonbokhwa, Yeongyo and Jisil were treated in a medium containing 1 μM α-MSH and cultured. On the fifth day, the medium was removed, washed with DPBS, harvested with a scraper, and then dried at 70°C. After dissolving melanin in 1N NaOH solution at 70° C. for 30 minutes, absorbance was measured at 490 nm using an absorbance meter to evaluate the effect of inhibiting melanin formation.

The results are shown in FIG. 2.

As can be seen in Figure 2, compared with the extracts of the preparation example, it was confirmed that the five kinds of complex fermentation extracts of mountain mulberry, mistletoe, sunbokhwa, yeonkyo and fat in Example 1 exhibited more excellent melanin production inhibitory effect.

Formulation Example 1: Preparation of serum containing complex fermentation extract

A serum containing the complex fermentation extract (Example 1) was prepared according to a conventional method with the composition and contents shown in Table 2 below.

Raw material Content (% by weight) Complex fermentation extract (Example 1) 2.0 Beeswax 1.0 Polysorbate 60 1.5 Sorbitan sesquioleate 0.5 Mineral oil 10.0 Sorbitan Stearate 0.5 Glycerin monostearate 1.0 Stearic acid 1.5 Glyceryl Stearate/Pig-400 Stearate 1.0 Propylene glycol 3.0 Carboxypolymer 0.1 Triethanolamine 0.1 Phenoxyethanol Appropriate amount Purified water To 100

Formulation Example 2: Preparation of cream containing complex fermentation extract

A cream containing the complex fermentation extract (Example 1) was prepared according to a conventional method in the composition and content of Table 3 below.

Raw material Content (% by weight) Complex fermentation extract (Example 1) 2.0 Shea Butter 2.0 Polysorbate 60 1.5 Sorbitan sesquioleate 0.8 Mineral oil 10.0 Dimethicone 3.0 Sorbitan Stearate 0.5 Glycerin monostearate 1.0 Cetearyl alcohol 2.0 Glyceryl Stearate/Pig-400 Stearate 1.0 Propylene glycol 3.0 Carboxypolymer 0.25 Triethanolamine 0.25 Phenoxyethanol Appropriate amount Purified water To 100

Test Example 4: Confirmation of formulation stability

For the formulations prepared in Formulation Examples 1 and 2, put in an opaque glassware container in an opaque glassware container in a room maintained at room temperature (25°C), refrigerated (4°C) and constant temperature (50°C) and stored at a constant temperature (50°C) for 12 weeks. And observation (discoloration, discoloration and separation), and stability was confirmed. The results are shown in Table 4 below.

Temperature condition Stability check (discoloration, discoloration and separation) Formulation Example 1 Formulation Example 2 Room temperature (25℃) 0 0 Refrigeration (4℃) 0 0 Constant temperature (50℃) 0 0

<Formulation stability grade>

0: No change 1: Subtle change 2: Change 3: Extreme change

As shown in the table above, it was confirmed that both formulations Examples 1 and 2 were stable without discoloration and discoloration and segregation under the temperature conditions of 25°C, 4°C and 50°C.

Claims (6)

For skin whitening containing a complex fermentation extract extracted by sequential fermentation with Lactobacillus sakei and Aspergillus niger after mixing mountain mulberry, mistletoe, sunbok flower, yeonkyo and fat in the same weight ratio Cosmetic composition. delete The cosmetic composition for skin whitening according to claim 1, wherein the complex fermentation extract is contained in an amount of 0.001 to 20% by weight based on the total weight of the cosmetic composition. delete delete delete

Images