KR102761562B1 - Cosmetic composition or pharmaceutial composition comprising protease, and production method thereof - Google Patents

Abstract

The present invention relates to a fermented product obtained by fermenting a mixture of three or more selected from the group consisting of mugwort, soybeans, tea tree, purslane, birch bark, and evening primrose with a strain producing a protein-decomposing enzyme, or a cosmetic composition or a skin external application composition comprising a protein-decomposing enzyme obtained from the fermented product, and a method for producing the same, thereby providing a cosmetic composition and a skin external application composition which are environmentally friendly and minimize skin irritation, while being very stable and capable of exhibiting an excellent exfoliating effect.

Description

{Cosmetic composition or pharmaceutical composition comprising protease, and production method thereof}

The present invention relates to a cosmetic composition, a pharmaceutical composition containing a protein-decomposing enzyme, and a method for producing the same.

Generally, the skin is divided into three parts from the outside of the skin: the epidermis, the dermis, and the subcutaneous fat layer. The epidermis is further divided into the stratum corneum, the granular layer, the spinous layer, and the basal layer. The epidermal cells in the basal layer differentiate as they rise to the upper part of the skin and finally reach the stratum corneum. The epidermal cells that reach the stratum corneum lose their nuclei and are filled with an insoluble protein called keratin, turning them into dead cells.

Although keratinocytes are continuously produced one after another, they maintain a certain thickness because the old keratinocytes in the outermost layer are shed. This process in which new cells rise to the stratum corneum and become dead cells and fall off is called turnover. In the case of normal skin, the stratum corneum is composed of 15 to 20 layers, and it takes 15 to 20 days for them to be completely separated (British J. of Dermatology, 86, 14-19, 1974; K.M.Halprin, Hyangjanghoeji, 12(4), 265-271, 1988), so it can be said that about one layer of keratinocytes falls off per day.

If this keratin layer is not normally exfoliated and remains on the skin surface, the keratin layer becomes thicker and the complexion becomes dull and dark. In addition, impurities remaining on the skin surface or in the hair follicles are oxidized or decomposed by oxygen or microorganisms, causing skin troubles such as inflammation. The main component of these impurities is keratin, which remains on the skin surface even after the cells die.

The above keratin has a large molecular weight, is insoluble, and is not easily broken down, so it is not easily removed with a detergent alone, but it can be effectively removed by breaking it down into small pieces using enzymes. Using a stable protein-decomposing enzyme like this can remove the keratin layer attached to the skin surface, making the skin smoother.

Most of the keratinases developed so far are obtained from microorganisms selected from animal sources such as chicken feathers or poultry waste, and in most cases, they are unstable to heat, unstable to surfactants, and unstable to heat, acids, bases, and metal ions, so they are not very useful for formulating cosmetic compositions or skin external preparation compositions. In addition, there is an increasing need for the use of natural substances to reduce skin irritation. Therefore, in order to develop eco-friendly cosmetic raw materials, it is necessary to develop plant- or soil-derived keratinase, and it is desirable to develop a technology that can be applied to cosmetics or skin external preparation compositions that are more eco-friendly, minimize skin irritation, and exhibit excellent effects.

Republic of Korea Patent Publication No. 10-2014-0055397 Republic of Korea registered patent 10-2394644

In order to solve the problems described above, the present invention aims to provide a cosmetic or skin external application composition that is environmentally friendly, minimizes skin irritation, and is very stable and exhibits an excellent exfoliating effect against various surfactants, acids, bases, temperature, metal ions, etc. used in cosmetics.

In order to solve the above-described problem, the present invention provides a fermented product obtained by fermenting a mixture of three or more selected from the group consisting of mugwort, soybeans, tea tree, purslane, birch bark, and evening primrose with a strain producing a protein-decomposing enzyme, or a cosmetic composition comprising a protein-decomposing enzyme obtained from the fermented product.

According to one embodiment of the present invention, the strain producing the protein-decomposing enzyme of the cosmetic composition is at least one strain selected from Bacillus cloudi, Lactobacillus plantarum, and a transformant introduced with a protein-decomposing enzyme gene.

According to one embodiment of the present invention, the protein decomposing enzyme of the cosmetic composition is keratinase.

According to one embodiment of the present invention, the cosmetic composition further comprises at least one selected from alpha hydroxy acid (AHA), beta hydroxy acid (BHA), lipo hydroxy acid (LHA), and poly hydroxy acid (PHA).

According to one embodiment of the present invention, the cosmetic composition is used for preventing dead skin cells, removing dead skin cells, promoting dead skin cell exfoliation, alleviating acne, preventing atopic dermatitis, alleviating atopic dermatitis, preventing skin trouble, alleviating skin trouble, improving skin moisturizing, or improving skin tone.

The present invention also provides a fermented product obtained by fermenting a mixture of three or more selected from the group consisting of mugwort, soybeans, tea tree, purslane, birch bark, and evening primrose with a strain producing a protein-decomposing enzyme, or a pharmaceutical composition for external skin use for removing dead skin cells or preventing, alleviating, or treating atopic dermatitis, comprising a protein-decomposing enzyme obtained from the fermented product.

According to one embodiment of the present invention, the strain producing the protein-decomposing enzyme of the pharmaceutical composition is at least one strain selected from Bacillus cloudi, Lactobacillus plantarum, and a transformant introduced with a protein-decomposing enzyme gene.

According to one embodiment of the present invention, the proteolytic enzyme of the pharmaceutical composition is keratinase.

According to one embodiment of the present invention, the pharmaceutical composition is for preventing dead skin cells, removing dead skin cells, promoting dead skin cell exfoliation, alleviating acne, treating acne, preventing atopic dermatitis, alleviating atopic dermatitis, treating atopic dermatitis, preventing skin trouble, alleviating skin trouble, and treating skin trouble.

According to one embodiment of the present invention, the pharmaceutical composition further comprises at least one selected from alpha hydroxy acid (AHA), beta hydroxy acid (BHA), lipo hydroxy acid (LHA), and poly hydroxy acid (PHA).

The present invention also provides a method for producing a cosmetic composition, comprising a step of fermenting a mixture of three or more selected from the group consisting of mugwort, soybeans, tea tree, purslane, birch bark, and evening primrose with a strain producing a protein-decomposing enzyme.

According to one embodiment of the present invention, the method for producing the cosmetic composition further includes a step of purifying and obtaining a protein decomposing enzyme from a fermented product obtained through the fermentation step.

The present invention also provides a method for producing a pharmaceutical composition for external skin application for removing dead skin cells or preventing, alleviating or treating atopic dermatitis, comprising a step of fermenting a mixture of three or more selected from the group consisting of mugwort, soybeans, tea tree, purslane, birch bark and evening primrose with a strain producing a protein-decomposing enzyme.

According to one embodiment of the present invention, the method for preparing the pharmaceutical composition further comprises a step of purifying and obtaining a protein decomposing enzyme from a fermented product obtained through the fermentation step.

According to the present invention as described above, a cosmetic composition and a skin external application composition are provided which are environmentally friendly and minimize skin irritation, while being very stable and exhibiting an excellent exfoliating effect against various surfactants, acids, bases, temperatures, metal ions, etc. used in cosmetics.

Through this, the skin barrier function is restored through the exfoliation improvement effect without skin irritation, improving skin troubles and maximizing functionality.

Figure 1 shows the results of an experiment on the thermal stability of the present invention.
Figure 2 shows the results of the DHA test experiment of the present invention. (Melanin index)
Figure 3 is a photograph showing changes in skin observed after applying the present invention to the skin for two weeks.

Hereinafter, the present invention will be described in detail.

The present invention provides a fermented product obtained by fermenting a mixture of three or more selected from the group consisting of mugwort, tea tree, rock soybean, purslane, birch bark, and evening primrose with a strain producing a protein-decomposing enzyme, or a cosmetic composition comprising a protein-decomposing enzyme obtained from the fermented product.

The above-mentioned herb can use at least one of the whole plant, leaves, and roots of the herb. Specifically, the leaves of the herb can be used.

The above tea tree can use one or more of the leaves, stems, bark, and roots of the tea tree. Specifically, tea tree leaves can be used.

The above stone soybean can use one or more of the stone soybean leaves, stems, roots, and seeds. Specifically, stone soybean seeds can be used.

The above herb can be used as one or more of the whole plant, leaves, and roots of the herb. Specifically, the whole plant of the herb can be used.

The above birch bark is the bark of a birch tree trunk.

The above evening primrose can be made using one or more of the evening primrose leaves, stems, roots, seeds, and flowers. Specifically, evening primrose seeds can be used.

The above fermented product is a fermented product obtained by fermenting a mixture of three or more selected from the group consisting of the above-mentioned Houttuynia cordata, tea tree, rock soybean, centella asiatica, birch bark, and evening primrose with a strain producing a protein-decomposing enzyme, and includes all fermented products obtained therefrom. In addition, the fermented product can also be obtained by mixing fermented products obtained by fermenting three or more plants selected from the group consisting of the above-mentioned Houttuynia cordata, tea tree, rock soybean, centella asiatica, birch bark, and evening primrose with a strain producing a protein-decomposing enzyme.

The above fermented product can be obtained by inoculating an appropriate strain into each or a mixture of three or more plants selected from the group consisting of the above-mentioned mugwort, tea tree, rock soybean, purslane, birch bark, and evening primrose, or their crushed products, supernatants of their crushed products, dried products, dried product powder, extracts, fractions, etc., and culturing them.

For example, the fermented solution of a strain culture obtained by aerobic fermentation of a mixture extract; a supernatant obtained by centrifuging and then filtering the fermented solution of a strain culture of the mixture extract; an extracted fraction obtained by using a fractional extraction solvent such as propanol, butanol, ethyl acetate, ethyl ether, chloroform, dichloromethane, and hexane on the fermented solution of a mixture extract; etc., and the fermented solution, supernatant, diluted solution, concentrate, or dried product of the fraction described above may be mentioned, but are not limited thereto.

The above extract may be obtained using a conventional extraction and separation method. At this time, the extraction solvent of the mixture extract includes both a polar solvent-soluble extract and a non-polar solvent-soluble extract. For example, the mixture extract may be an extract of one or more extraction solvents selected from purified water, methanol, ethanol, butanol, glycerin, ethyl acetate, ethyl ether, chloroform, dichloromethane, and hexane, and specifically, may be an extract of only a polar solvent such as purified water, methanol, ethanol, butanol, and glycerin, or a mixed solvent of the polar solvent and a non-polar solvent such as ethyl acetate, ethyl ether, chloroform, dichloromethane, and hexane.

At this time, the extraction temperature is preferably 15 to 45°C, and the extraction time is preferably 5 to 48 hours. If the extraction temperature and extraction time are exceeded, the extraction efficiency may decrease or the components may change. The extraction method is not particularly limited, and examples thereof include hot water extraction, cold extraction, ultrasonic extraction, and reflux extraction. After obtaining the extract using the solvent above, a liquid may be obtained by cold-drying, heating, and filtering at room temperature using a conventional method known in the art, or the solvent may be additionally evaporated, spray-dried, or freeze-dried.

The above fermented product includes all of the obtained fermentation products and may include a fermented solution, a supernatant, a dried product, a dried product powder, an extract, a fraction, etc.

The composition of the present invention has an excellent effect in decomposing keratin, a main component of skin keratin, when applied to the skin, and is also suitable for removing skin keratin as it causes very little irritation to the skin as a natural ingredient.

In one embodiment of the present invention, the fermented product may include all of mugwort, tea tree, soybean, purslane, birch bark and evening primrose fermented by a strain producing a protein-decomposing enzyme, or a protein-decomposing enzyme obtained from the fermented product. Specifically, it may be a mixture of mugwort, tea tree and soybean.

The fermented product obtained by fermenting the above mixture or the protein decomposing enzyme obtained from the fermented product is not inhibited by a surfactant compared to conventional keratinase, maintains activity under high pH conditions (stable at pH 4.5 to 11.5), and exhibits a stable effect not only at low temperatures (20 to 30°C) but also at high temperatures (55 to 65°C). In particular, when the keratinase-producing strain is cultured in a medium containing the extract of the present invention, compared to the case where the keratinase-producing strain is cultured in a general medium, the activity of the keratinase is excellent and the protein decomposition ability also increases significantly. In addition, it has the advantage of not being degraded by heavy metals such as cadmium, copper, manganese, and magnesium, and maintaining activity even in the presence of oxidizing and reducing agents. In addition, it maintains enzyme activity even in salt, and has the advantage of being stable even in a salt solution of 10% or more.

In one embodiment of the present invention, the fermented product or the protein decomposing enzyme obtained from the fermented product may be included in an amount of 0.001 to 5 wt% based on the total weight of the composition, and specifically, may be included in an amount of 0.005 to 2 wt%, or 0.01 to 1 wt%.

According to one embodiment of the present invention, the strain producing the protein-decomposing enzyme of the cosmetic composition may be at least one strain selected from Bacillus cloudi, Lactobacillus plantarum, and a transformant introduced with a protein-decomposing enzyme gene. Specifically, it may be Bacillus clausii I-52 (Accession No.: KCTC 10277BP), Lactobacillus plantarum APsulloc 331261 (Accession No.: KCCM11179P), Lactobacillus plantarum APsulloc 331263 (Accession No.: KCCM11180P), Lactobacillus plantarum APsulloc 331266 (Accession No.: KCCM11181P), or Lactobacillus plantarum Apsulloc 331269 (Accession No.: KCCM11182P). Among the transformants introduced with a protein-decomposing enzyme gene, there is a transformant introduced with the expression vector pHPS-BCAP isolated from Bacillus clausii I-52 (Accession Number: KCTC 10277BP).

According to one embodiment of the present invention, the protein decomposing enzyme of the cosmetic composition may be keratinase.

According to one embodiment of the present invention, the cosmetic composition may further include at least one selected from alpha hydroxy acid (AHA), beta hydroxy acid (BHA), lipo hydroxy acid (LHA), and polyhydroxy acid (PHA). The polyhydroxy acid may be at least one selected from the group consisting of gluconolactone, lactobionic acid, and maltobionic acid.

In addition to the above-described effective ingredients, the cosmetic composition of the invention may further contain conventional auxiliary agents and carriers, such as antioxidants, stabilizers, solubilizers, vitamins, pigments, fragrances, etc., which are commonly used in cosmetic compositions. For example, the cosmetic composition may further contain auxiliary ingredients, such as glycerin, butylene glycol, polyoxyethylene hydrogenated castor oil, tocopheryl acetate, citric acid, panthenol, squalane, sodium citrate, and allantoin. The blending amount of the above-described ingredients can be easily selected by those skilled in the art within a range that does not impair the purpose and effect of the present invention, and the blending amount may be about 0.01 to 5 wt%, specifically 0.01 to 3 wt%, based on the total weight of the composition.

Since the cosmetic composition of the present invention is basically applied to the skin, it can be manufactured into any formulation that is commonly manufactured with reference to cosmetic compositions in the art. For example, it can be formulated into a solution, suspension, emulsion, paste, gel, cream, lotion, powder, soap, surfactant-containing cleansing, oil, powder foundation, emulsion foundation, wax foundation, and spray, but is not limited thereto. More specifically, it can be manufactured into the formulation of a flexible toner, a nourishing toner, a nourishing cream, a massage cream, an essence, an eye cream, a cleansing cream, a cleansing foam, cleansing water, a mask pack, a spray, or a powder.

When the formulation of the present invention is a paste, cream or gel, the carrier component may include animal oil, vegetable oil, wax, paraffin, starch, tragacanth, cellulose derivatives, polyethylene glycol, silicone, bentonite, silica, talc, zinc oxide, etc.

When the formulation of the present invention is a powder or spray, the carrier component may include lactose, talc, silica, aluminum hydroxide, calcium silicate, polyamide powder, etc., and particularly, when it is a spray, it may additionally include a propellant such as chlorofluorohydrocarbon, propane/butane, dimethyl ether, etc.

When the formulation of the present invention is a solution or emulsion, the carrier component may include a solvent, a solubilizer, an emulsifier, etc., and specifically, water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butyl glycol oil, glycerol aliphatic ester, polyethylene glycol, fatty acid ester of sorbitan, etc.

When the formulation of the present invention is a suspension, the carrier component may include a liquid diluent such as water, ethanol, or propylene glycol; a suspending agent such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester, or polyoxyethylene sorbitan ester; microcrystalline cellulose, aluminum metahydroxide, bentonite, agar, or tragacanth.

When the formulation of the present invention is a surfactant-containing cleansing agent, the carrier component may include fatty alcohol sulfate, fatty alcohol ether sulfate, sulfosuccinic acid monoester, isethionate, imidazolinium derivative, methyl taurate, sarcosinate, fatty acid amide ether sulfate, alkyl amidobetaine, fatty alcohol, fatty acid glyceride, fatty acid diethanolamide, vegetable oil, lanolin derivative, ethoxylated glycerol fatty acid ester, etc.

According to one embodiment of the present invention, the cosmetic composition is used for preventing dead skin cells, removing dead skin cells, promoting dead skin cell exfoliation, alleviating acne, preventing atopic dermatitis, alleviating atopic dermatitis, preventing skin trouble, alleviating skin trouble, improving skin moisturizing, or improving skin tone.

The term "keratin" in the present invention is a type of fibrous structural protein and is the main structural material that constitutes the epithelial cells of skin, hair, and nails. It varies greatly in size, is charged, and is insoluble, and its most important function is to enhance the ability of cells to withstand trauma. In addition, it is responsible for functions such as cell signaling and cell proliferation regulation. It is representatively present in the epidermal area of the skin, especially in the skeleton of keratinocytes, and is present in the inner root sheath and hair shaft of hair. In addition, it is present in the cornea of the eye, the upper part of the basal layer of the oral mucosa, and the epithelial cells of the intestine.

The present invention also provides a fermented product obtained by fermenting a mixture of three or more selected from the group consisting of mugwort, tea tree, rock soybean, purslane, birch bark, and evening primrose with a strain producing a protein-decomposing enzyme, or a pharmaceutical composition for external skin application for removing dead skin cells or preventing, alleviating, or treating atopic dermatitis, comprising a protein-decomposing enzyme obtained from the fermented product.

According to one embodiment of the present invention, the strain producing the protein-decomposing enzyme of the pharmaceutical composition may be at least one strain selected from Bacillus cloudi, Lactobacillus plantarum, and a transformant introduced with a protein-decomposing enzyme gene.

According to one embodiment of the present invention, the protein decomposing enzyme of the pharmaceutical composition may be keratinase.

According to one embodiment of the present invention, the pharmaceutical composition may be used for preventing keratin, removing keratin, promoting keratin exfoliation, alleviating acne, treating acne, preventing atopic dermatitis, alleviating atopic dermatitis, treating atopic dermatitis, preventing skin trouble, alleviating skin trouble, or treating skin trouble.

According to one embodiment of the present invention, the pharmaceutical composition may further include at least one selected from alpha hydroxy acid (AHA), beta hydroxy acid (BHA), lipo hydroxy acid (LHA), and poly hydroxy acid (PHA).

The pharmaceutical composition may be provided in any formulation suitable for topical application. For example, it may be a solution, suspension, emulsion, gel, patch or spray for external use, but is not limited thereto. The formulation may be easily prepared according to a conventional method in the art, and may appropriately use surfactants, excipients, wetting agents, emulsifying promoters, suspending agents, salts or buffers for osmotic pressure control, coloring agents, flavoring agents, stabilizers, preservatives, preservatives or other commonly used auxiliary agents.

The effective ingredient of the pharmaceutical composition according to one embodiment of the present invention will vary depending on the age, sex, weight, pathological condition and its severity, the route of administration, or the judgment of the prescriber of the subject. Determination of the application amount based on these factors is within the level of those skilled in the art, and the daily use dose thereof may be, for example, 0.1 mg/kg/day to 5000 mg/kg/day, more specifically, 50 mg/kg/day to 500 mg/kg/day, but is not limited thereto.

The present invention also provides a method for producing a cosmetic composition, comprising a step of fermenting a mixture of three or more selected from the group consisting of mugwort, tea tree, rock soybean, purslane, birch bark, and evening primrose with a strain producing a protein-decomposing enzyme.

The above fermentation step can be obtained by inoculating a strain into a mixture of mugwort, tea tree, soybean, purslane, birch bark and evening primrose, or by inoculating and cultivating an extract of these mixtures with a strain. The extract can be prepared using a solvent suitable for a person skilled in the art, and for example, water or a mixed solvent of water and an organic solvent can be used. The organic solvent that can be used is

According to one embodiment of the present invention, the method for producing the cosmetic composition may further include a step of purifying and obtaining a protein decomposing enzyme from a fermented product obtained through the fermentation step.

The method for obtaining the above-mentioned purified product is carried out by methods known to those skilled in the art, for example, centrifugation, filtration, ultrafiltration and dialysis. Preferably, ultrafiltration and dialysis are carried out using a polyethersulfone membrane that retains molecules having a molecular weight exceeding 10,000 Da.

The present invention also provides a method for producing a pharmaceutical composition for external skin application for removing dead skin cells or preventing, alleviating or treating atopic dermatitis, comprising a step of fermenting a mixture of three or more selected from the group consisting of mugwort, tea tree, rock soybean, purslane, birch bark and evening primrose with a strain producing a protein-decomposing enzyme.

According to one embodiment of the present invention, the method for preparing the pharmaceutical composition may further include a step of purifying and obtaining a protein decomposing enzyme from a fermented product obtained through the fermentation step.

Hereinafter, embodiments of the present invention will be described in detail with reference to the attached drawings.

[Example 1]

After washing and preparing the leaves of mugwort, tea tree leaves, soybean seeds, purslane whole plant, birch bark, and evening primrose seeds, they were extracted with hot water at 60℃ for 1 hour. The obtained hot water extract was filtered through a 100 mesh screen. Bacillus cloudi I-52 (Accession No.: KCTC 10277BP) was inoculated into the culture medium and primary cultured at 30℃ for 24 hours. The primary culture was mixed with the extract obtained above and secondary cultured (30℃, 24 hours). It was centrifuged, filtered (0.45 ㎛), purified, and concentrated.

[Example 2]

It was prepared in the same manner as Example 1, except that the extracts of mugwort leaves, tea tree leaves, and soybean seeds were mixed and cultured.

[Example 3]

The composition of Example 1 obtained above was prepared by mixing salicylic acid (0.45% and 1%, respectively, based on the total weight of the composition).

[Comparative example]

It was prepared in the same manner as in Example 1, except that the extracts of mugwort leaves, tea tree leaves, soybean seeds, purslane whole plant, birch bark, and evening primrose seeds were mixed and cultured. That is, Bacillus cloudi I-52 (Accession No.: KCTC 10277BP) was inoculated into the culture medium and cultured primarily at 30°C for 24 hours, and then cultured secondary at 30°C for 24 hours. The mixture was centrifuged, filtered (0.45 μm), purified, and concentrated.

[Experimental Example 1] Measurement of protein decomposition ability

The protein (casein) decomposition ability of the keratinase obtained in Examples 1 and 2 was tested at each concentration. Papain (Control 1) and Comparative Example (Control 2) were used as a control group.

The zone of inhibition was measured. When a paper disc impregnated with a sample (control or Example 1) is placed on a medium consisting of 0.5% casein and 1% agar, the sample diffuses onto the medium and the protein is decomposed. Therefore, the protein degradation ability is determined by the area of the zone formed around the sample. The wider the zone, the better the degradation ability.

Specifically, Example 1, Example 2, Control Group 1 (Papain), and Control Group 2 (Comparative Example) were treated at 0.001%, 0.01%, 0.1%, and 1%, respectively, and then observed after culturing at 37°C for 24 hours.

The experimental results are shown in Table 1. It was confirmed that Example 1 of the present invention had casein decomposition ability that was about 3 times better than that of Control Group 1 (papain).

Sample concentration Example 1 Example 2 Control group 1 (papain) Control group 2 (comparative example) 0.001% 0.2 mm 0.15 mm 0 mm 0.1 mm 0.01% 4 mm 3 mm 0 mm 2 mm 0.10% 8 mm 6 mm 2 mm 4 mm 1% 12 mm 9 mm 4 mm 6 mm

[Experimental Example 2] Thermal Stability Evaluation

The thermal stability of the keratinase obtained in Example 1 was evaluated. Papain (Control 1) and Comparative Example (Control 2) were used as a control group.

Keratinase activity was analyzed using keratin azure (Sigma Aldrich) as a substrate. 1.0 ml of Example 1, Control Group 1, and Control Group 2 were each added to 1.0 ml of 50 mM Tris-HCl buffer solution in which 5 g of keratin azure powder (pH 8.0) was suspended. After centrifugation at 1500 rpm for 30 minutes, the supernatant was separated, and the absorbance at 595 nm was measured using a spectrophotometer. After incubating them at 37°C and 60°C for 60 minutes, 2 ml of TCA was added. After centrifugation, the keratinase activity value was determined using a spectrophotometer.

The experimental results are shown in Table 2 and Fig. 1. At a temperature of 60°C, both Example 1 and the control group showed relatively stable results, but at 37°C, which is the temperature of human skin, Example 1 of the present invention was found to maintain stability that was up to 4 times higher than that of the control group 1 compared to papain.

　 Example 1 Control group 1 (papain) Control group 2 (comparative example) temperature 37℃ 60℃ 37℃ 60℃ 37℃ 60℃ Keratinase U/g 1600 825 400 675 1000 760

[Experimental Example 3] DHA Test

A DHA test was conducted on 10 men and women in their 20s to 40s.

Specifically, a 1.5 X 1.5 cm area was designated on the subject's forearm, artificial pigmentation was induced with a 10% DHA solution, and Example 3 was applied to the skin twice a day to observe the degree of skin turnover for two weeks. A 0.45% salicylic acid solution was applied as a control group.

The results of measuring the melanin index (M.I.) using a mexameter are shown in Table 3 and Fig. 2. In addition, the results of observing skin changes for two weeks are shown in Fig. 3.

Day Control (salicylic acid) Example 1 Example 3 1 0 0 0 4 3 2 6 7 7 5 17 11 15 7 27 14 17 8 27

Salicylic acid (BHA, control), which is well known as a chemical peeling agent, has an excellent peeling effect, but is known to irritate the skin when used in high concentrations. Through this experiment, it was confirmed that the extract of the present invention and salicylic acid have a synergistic effect on exfoliation when used together. In other words, it is possible to implement a better peeling effect with less skin irritation.

[Manufacturing example]

A cosmetic composition was prepared by mixing the ingredients in the following amounts.

Ingredient name Content (%) water 91.5 Eoseongcho leaf extract 1.0 Tea tree leaf extract 1.0 Soybean Seed Extract 1.0 Whole plant extract of Centella asiatica 1.0 Birch bark extract 1.0 Evening primrose seed extract 1.0 Example 1 0.5 1,2-Hexanediol 2.0 Total 100

Hereinafter, while the embodiments of the present invention have been described with reference to the attached drawings, those skilled in the art will understand that the present invention can be implemented in other specific forms without changing the technical spirit or essential characteristics thereof. Therefore, the present invention is not limited to the embodiments disclosed below, but can be implemented in various different forms, and the present embodiments are provided only to make the disclosure of the present invention complete and to fully inform those skilled in the art of the scope of the present invention, and the present invention is defined only by the scope of the claims.

Claims (14)

A cosmetic composition comprising a fermented product obtained by fermenting three or more mixtures selected from the group consisting of an extract of mugwort leaves, an extract of tea tree leaves, an extract of soybean seeds, an extract of whole purslane, an extract of birch bark, and an extract of evening primrose seeds with a strain producing a protein-decomposing enzyme, or a protein-decomposing enzyme obtained from the fermented product,
The above mixture contains extracts of mugwort leaf, tea tree leaf, and soybean seed.
A cosmetic composition wherein the strain producing the above protein-decomposing enzyme is a Bacillus cloudi strain.
delete In the first paragraph,
A cosmetic composition wherein the above protein-decomposing enzyme is keratinase.
In the first paragraph,
The above cosmetic composition further comprises at least one selected from alpha hydroxy acid (AHA), beta hydroxy acid (BHA), lipo hydroxy acid (LHA), and poly hydroxy acid (PHA).
In the first paragraph,
The above cosmetic composition is a cosmetic composition for preventing dead skin cells, removing dead skin cells, or promoting dead skin cell exfoliation.
A fermented product obtained by fermenting a mixture of three or more selected from the group consisting of an extract of mugwort leaves, an extract of tea tree leaves, an extract of soybean seeds, an extract of whole purslane, an extract of birch bark, and an extract of evening primrose seeds with a strain producing a proteolytic enzyme, or a pharmaceutical composition for external skin application for exfoliation comprising a proteolytic enzyme obtained from the fermented product,
The above mixture contains extracts of mugwort leaf, tea tree leaf, and soybean seed.
A pharmaceutical composition for external skin application, wherein the strain producing the above protein-decomposing enzyme is a Bacillus cloudi strain.
delete In Article 6,
A pharmaceutical composition for external use in the skin, wherein the protein-decomposing enzyme is keratinase.
In Article 6,
The above pharmaceutical composition is a pharmaceutical composition for external use on the skin for preventing keratin, removing keratin, or promoting keratin exfoliation.
In Article 6,
The pharmaceutical composition for external use in the skin further comprises at least one selected from alpha hydroxy acid (AHA), beta hydroxy acid (BHA), lipo hydroxy acid (LHA), and poly hydroxy acid (PHA).
A method for producing a cosmetic composition comprising a step of fermenting a mixture of three or more selected from the group consisting of an extract of mugwort leaves, an extract of tea tree leaves, an extract of soybean seeds, an extract of the whole plant of Centella asiatica, an extract of birch bark, and an extract of evening primrose seeds, with a strain producing a protein-decomposing enzyme,
The above mixture contains extracts of mugwort leaf, tea tree leaf, and soybean seed.
A method for producing a cosmetic composition, wherein the strain producing the above protein-decomposing enzyme is a Bacillus cloudi strain.
In Article 11,
A method for producing a cosmetic composition, wherein the above manufacturing method further comprises a step of purifying and obtaining a protein decomposing enzyme from a fermented product obtained through the above fermentation step.
A method for producing a pharmaceutical composition for external application of skin for exfoliation, comprising the step of fermenting three or more mixtures selected from the group consisting of an extract of mugwort leaves, an extract of tea tree leaves, an extract of soybean seed, an extract of whole purslane, an extract of birch bark, and an extract of evening primrose seeds, with a strain producing a protein-decomposing enzyme,
The above mixture contains extracts of mugwort leaf, tea tree leaf, and soybean seed.
A method for producing a pharmaceutical composition for external skin application for exfoliation, wherein the strain producing the above protein-decomposing enzyme is a Bacillus cloudi strain.
In Article 13,
A method for producing a pharmaceutical composition for external skin application for exfoliation, wherein the above-mentioned manufacturing method further comprises a step of purifying and obtaining a protein-decomposing enzyme from a fermented product obtained through the above-mentioned fermentation step.