KR20190063071A - Cosmetic materials composition having inhibitor of skin photo-aging and regeneration of skin wound-cell, Cosmetic materials using the same and Manufacturing method thereof - Google Patents

Abstract

The present invention relates to a cosmetic composition for inhibiting skin photoaging and for regenerating skin damaged cells and a cosmetic composition for inhibiting skin photoaging and regenerating skin damaged cells manufactured by using the same. More specifically, the present invention relates to an invention to provide a cosmetic for inhibiting skin photoaging and regenerating skin damaged cells using a bio peptone fermented product, a method for manufacturing the same, and a skin external agent such as functional cosmetics using the same.

Description

TECHNICAL FIELD The present invention relates to a cosmetic composition for inhibiting skin photo-aging and regenerating skin damaged cells, a cosmetic composition for regenerating skin damaged cells and a method for producing the same, the same and manufacturing method thereof}

The present invention relates to a cosmetic composition having suppressed skin photoaging and regenerating effect on damaged cells of skin, a composition for use in the preparation thereof, and a method for preparing the same, and more specifically, it relates to a cosmetic composition which inhibits photoaging using a biop peptone fermentation product, / RTI > and / or preventing skin cells damaged by aging, and regenerating skin cells damaged by photoaging or the like.

The main source of protein is the rising consumption of popular foods due to nutritional value and various functions scientifically revealed. Recently, it has been known that soybeans not only have a therapeutic effect on cardiovascular diseases, diabetes, cancer, overweight, and obesity but also improve skin such as promoting collagen production, inhibiting melanin production, inhibiting skin aging, and antiinflammation . Soybean is cultivated all over the world and it is a representative vegetable protein source which not only makes it easy to receive the raw materials but also contains 20-40% of the protein in the building. In addition, since it is a plant material that has secured safety within the body through its long history, it has been widely utilized in various physiological activities and functional studies.

A biological active peptide (BPs) is a small molecular weight amino acid sequence having a physiological activity derived from a protein. The amino acid sequence contained in the precursor protein is largely inactive, but the small peptides generated through the degradation process have various biological actions. Protein degradation is a necessary process for the production of bioactive peptides, and the physiological activity of the peptides produced by degradation of the proteins varies depending on the type of original protein source, type of enzyme, and production conditions. In addition, the physiologically active peptide produced by the degradation is not only improved in penetration into the skin with low molecular weight but also increased in solubility in water, so that it is very easy to apply to various cosmetic materials.

The skin is an organ directly exposed to the external environment, and it always comes in direct contact with the ultraviolet rays such as sunlight. Ultraviolet rays are classified into three types according to the wavelength. UVA (320-400 nm), UVB (280-320 nm), and UVC (200-280 nm) are short wavelengths. About 90% of the ultraviolet radiation reaching the Earth's surface is UVA, and UVB is mostly absorbed in the atmosphere and only reaches about 10% of the Earth's surface. UVC does not reach the surface and does not affect the skin.

The minimum erythema dose for a minimum amount of light of a specific wavelength light is cause redness to the skin (minimal erythema dose, MED) as to when the minimum erythema dose skin in Asians in the case of the ^{UVA 50 ~ 70 J / cm 2} , UVB 50 ~ 70 mJ / cm ² to be. The wavelength of the light beam is inversely proportional to the energy, so even though the absolute amount of UVB has at least a strong energy, it can affect the gene of the cell. Actual UVB can cause erythema with only 1/1000 of UVA. The minimum erythema of UVB is 20 ~ 80 mJ / cm ² in hairless mouse, which is widely used in skin barrier studies, although it varies from report to report. Also, the longer the wavelength, the deeper the penetration into the skin. Although UVB differs depending on the thickness of the skin, it mainly affects the epidermis and upper dermis, and UVA can affect the lower dermis. In other words, both UVA and UVB cause skin damage, but UVA does not directly damage the DNA of skin cells, whereas UVB directly acts on the DNA of cells, resulting in burns, photoaging, skin cancer and the like.

Exposure to UVB causes oxidative stress to the skin due to an increase in reactive oxygen species (ROS). This leads to increased secretion of inflammatory cytokines from epidermal keratinocytes and induction of matrix metalloproteinases (MMPs) in the dermal fibroblast cell line, leading to the destruction of collagen, a major component of the extracellular matrix (ECM). Even at extremely low amounts of UVB exposure, MMP can be activated, which ultimately promotes skin aging, such as wrinkles.

In addition, ultraviolet light also affects the biosynthesis of the melanin pigment. Melanin is produced through a stepwise enzymatic reaction in the melanosome of the melanocyte, the melanocyte, located in the basal layer of the epidermis. Melanin promotes biosynthesis as a mechanism against skin damage caused by exposure to ultraviolet rays. Melanin pigment plays a positive role in protecting the skin. However, excessive production such as ultraviolet rays causes excessive production of spots, freckles and skin spots The toxicity of the melanin precursors can lead to cell death and skin cancer. Tyrosinase is known as a major enzyme involved in melanin biosynthesis. When tyrosinase activity is inhibited, overexpression of melanin pigment due to exposure to ultraviolet light can be suppressed.

In addition, as a melanogenesis inhibitor, there has been studied a technique of mixing together plant extracts such as wheat germ, soybean, and hops extract together with a cultured product of licorice and / or extract (Japanese Patent Publication No. JP-A-150-15017) there is a problem.

The present invention also relates to a method for producing an anti-aging cosmetic composition, such as skin whitening, which comprises extracting a specific component, alittiamine, by fermenting it as a strain to Saccharomyces cerevisiae using a plant containing a vitamin B1-containing wheat germ, malt and allysine as a culture component The technique used (Korea Publication No. 2010-0084209) has been studied.

Various raw materials that help skin whitening are developed, but there are problems such as adverse effects and long-term stability. In the case of materials such as existing melanin production inhibitors and cosmetic compositions, the skin absorption rate is low and new Raw materials and / or products of natural materials are required.

In order to solve the above-mentioned problems, the present invention relates to a cosmetic composition for inhibiting skin photoaging and regenerating skin damaged cells, which comprises: a biop peptone fermentation product; Alpha-amino acids; sugars; Acetylglucosamine; And citric acid.

In one preferred embodiment of the present invention, the biop peptone fermentation product may include a biop peptone fermentation product or a biop peptone fermentation product.

In one preferred embodiment of the present invention, when the fermented product is a fermented product of biopeptone, the fermented product of biopeptone may contain 86 to 97.5% by weight of fermented product, 2.0 to 7% by weight of alpha-amino acid, 0.3 to 5% by weight of saccharides, To 1 wt% and citric acid 0.1 to 2 wt%.

In one preferred embodiment of the present invention, when the fermented product of the biopeptone is a fermented product of biopeptone, it is preferable that the fermented product is a fermented product of 40 to 60 wt% of the biopeptone fermentation product, 25 to 40 wt% of the alpha-amino acid, 10 to 20 wt% 0.1 to 2 wt% and citric acid 1 to 5 wt%.

As a preferred embodiment of the present invention, the fermented product of the biopeptone is a fermented product obtained by fermenting a peptone-digested protein digestion product with lactic acid bacteria, and contains a low molecular peptide having a weight average molecular weight of 200 to 500 in an amount of 100 wt% (including other unavoidable impurities) can do.

In one preferred embodiment of the present invention, the low-molecular peptide may include at least one selected from a dipeptide, a tripeptide, and a tetrapeptide.

In one preferred embodiment of the present invention, the lactic acid bacteria used in the fermentation may include a strain of Lactobacillus sp.

As a preferred embodiment of the present invention, The peptone-degraded protein degradation product can be produced by using pepsin as a soybean containing at least one selected from soybean, seaweed, seomyeotei, black tea, kidney bean, fence bean, fowl bean, mung bean, Or may be peptonized.

In one embodiment of the present invention, the Lactobacillus genus strains Lactobacillus ramno suspension (Lactobacillus rhamnosus), Lactobacillus casei (Lactobacillus Casei) and Lactobacillus flat Lanthanum (Lactobacillus plantarum), Lactobacillus brevis (Lactobacillus brevis) , Lactobacillus acidophilus , and the like.

In one preferred embodiment of the present invention, the alpha-amino acid of the composition is selected from the group consisting of isoleucine, leucine, lysine, methionine, phenylalanine, threonine, tryptophan ), Valine, glutamic acid, and acetylcysteine (N-acetylcysteine).

In one preferred embodiment of the present invention, the saccharide in the composition may include at least one selected from glucose, galactose and xylose.

Another object of the present invention is to provide a cosmetic composition for regenerating skin-damaged skin and regenerating skin-damaged cells prepared using the composition described above, which comprises the cosmetic composition and the phospholipid mixture in various forms as described above.

In one preferred embodiment of the present invention, the cosmetic composition may contain 40 to 80% by weight of the mixture and 20 to 60% by weight of the phospholipid mixture.

In one preferred embodiment of the present invention, the phospholipid mixture may contain 1 to 10 parts by weight of ceramides and 10 to 100 parts by weight of a solvent, based on 100 parts by weight of lecithin.

In a preferred embodiment of the present invention, the solvent is caprylic / capric triglyceride caprylic / capric / stearic triglyceride and caprylic / capric / Caprylic / Capric / Myristic / Stearic Triglyceride may be included.

As one preferred embodiment of the present invention, the cosmetic for skin photoaging inhibition and regeneration of skin damaged cells according to the present invention comprises a mixture of the above cosmetic composition; And a phospholipid coating film for wrapping a mixture of the cosmetic composition.

Another object of the present invention is to provide a method for producing the liposomal cosmetic using the above-described composition, comprising the steps of: preparing a solution of a ceramide solution in which ceramide is dissolved in a solvent; Mixing the ceramide solution with lecithin and stirring at low speed to prepare a solution containing the phospholipid mixture; Adding a mixture of the cosmetic composition to the solution containing the phospholipid mixture, followed by slow stirring to produce a liposomal suspension; And filtering the liposome to recover the liposome.

As a preferred embodiment of the present invention, the low-speed stirring of the second and third stages may be performed at a stirring speed of 1,000 to 2,000 rpm at 10 to 30 ° C for 15 to 30 minutes.

In one preferred embodiment of the present invention, the interior of the liposome in step 4 includes a mixture of cosmetic compositions, and the liposome surface may be formed of a phospholipid coating film that wraps a mixture of the cosmetic composition.

In a preferred embodiment of the present invention, the bio-peptone fermentation product of the composition of the cosmetic composition of the third step includes a step 3-1 of producing a culture medium containing a peptized protein degradation product; Step 3-2 of inoculating the culture medium with the lactic acid bacteria; A step 3-3 of culturing the inoculated lactic acid bacteria in the second step to ferment the peptone-digested protein degradation product to produce a low molecular weight peptide culture; And a step (3-4) of removing the cells and insoluble components from the low molecular weight peptide culture to obtain a fermentation broth of the biopeptide.

In a preferred embodiment of the present invention, the method may further include a step 3-5 of drying and pulverizing the fermentation broth of the biopeptide of the third to fourth steps to obtain a fermentation powder of the biopeptide.

In one preferred embodiment of the present invention, the first step of the peptonized protein degradation product may be a peptone-modified soybean protein degradation product.

In one preferred embodiment of the present invention, in the preparation of the fermentation broth of the present invention, the culture medium of the first step contains at least one selected from the group consisting of glucose, sucrose, maltose, fructose, lactose, xylose, galactose and arabinose Carbon sources contained; And at least one inorganic element compound selected from magnesium sulfate, manganese sulfate, zinc sulfate, cooper sulfate, sodium acetate, potassium phosphate and calcium chloride.

In a preferred embodiment of the present invention, the inoculation of the second step in the preparation of the fermentation broth of the present invention is characterized in that the lactic acid bacterium is inoculated into the medium at a concentration of 1 × 10 ⁴ to 1 × 10 ⁷ cfu / ml .

In a preferred embodiment of the present invention, the fermentation in the third step may be performed at 25 to 35 for 12 to 72 hours during the preparation of the fermented biopgetone fermentation broth.

Still another object of the present invention is to provide a cosmetic composition for inhibiting skin photoaging and regenerating skin damaged cells or an external preparation for skin using the cosmetic composition, wherein the external preparation for skin may be cosmetic, ointment or ampoule.

In one preferred embodiment of the present invention, the cosmetic may be an emulsion type, a lotion type, a cream type, a lotion type, an emulsion type, an essence type, a pack type, a gel type, or a mist type cosmetic.

In one preferred embodiment of the present invention, the external preparation for skin of the present invention may contain the cosmetic composition or the cosmetic composition at a concentration of 50 μg / mL to 20,000 μg / mL.

In another preferred embodiment of the present invention, when the external preparation for skin is a cosmetic product and the cosmetic product is a cream or lotion type, the cosmetic composition or the cosmetic composition, glycerin, butylene glycol, di Propylene glycol, carboxyl vinyl polymer, polysorbate, hydrogenated castor oil, glyceryl stearate, cetearyl alcohol, cetyl ethylhexanoate and licorice extract.

In another preferred embodiment of the present invention, when the external preparation for skin is a cosmetic product and the cosmetic product is an emulsion type, a lotion type, an emulsion type, an ampule type or an essence type, Cosmetics, dipropylene glycol, 1,3-butylene glycol, polysorbate, dipotassium glycyrrhizate, allantoin, panthenol and glycerin.

The cosmetic for skin photoaging inhibition and regeneration of skin damaged cells according to the present invention is superior in skin absorption rate to the case of using the biopeptone fermented product itself as a component for external application for skin and the like and also has a skin damaging and / It is excellent in regeneration effect of cells and is excellent in compatibility with other compositions used in the preparation of external preparations for skin, so that external preparations for skin can be easily manufactured in various types such as cosmetics, ampoules and ointments.

FIG. 1 a is a schematic process diagram for producing a biop peptone fermentation broth as a skin-damaging cell regeneration component of skin photoaging inhibition of the present invention. FIG. 1 b is a graph showing the results of liposomalization of the liposome- Which is a schematic diagram of manufacturing.
2A is a schematic view of a liposome-type cosmetic composition of the present invention, and FIG. 2B is an optical microscope photograph taken before and after liposomes in the manufacture of a skin photoinhibition inhibitor.
3A to 3B are measurement results of the biopeptone fermentation product prepared in Example 1, as a result of HLPC measurement performed in Experimental Example 1. FIG. FIG. 3A shows the results from 0 to 60 minutes, and FIG. 3B shows the results from 0 to 25 minutes.
Each of (1) to (5) in FIG. 4 is a result of MS spectral measurement performed in Experimental Example 1, which is a result of confirming the molecular weight of the portion shown in FIG. 3 (b).
5A and 5B show the cytotoxicity confirmation test results of Experimental Example 2. FIG.
Figs. 6 and 7 show the measurement results of the cell protection effect by UVB irradiation in Experimental Example 3. Fig.
Fig. 8 shows the result of measuring collagen type synthesis promoting effect in Experimental Example 4-1. Fig.
FIG. 9 shows the results of measuring collagen type synthesis promoting effect of cells damaged by UVB irradiation in Experimental Example 4-2.
Fig. 10 shows the results of tyrosinase inhibitory effect measurement in Experimental Example 5. Fig.
Figs. 11A and 11B show the measurement results of inhibitory effect on melanin pigment formation in Experimental Example 6. Fig.
12A and 12B show the results of the antioxidative effect measurement in Experimental Example 7.

As used herein, the term [biop peptone fermentation product] refers to a fermentation product obtained by fermenting protein nucleic acid of peptone-modified soybean peptone by pepsin, and unless otherwise specified, the biop peptone fermentation product and the biop peptone fermentation product It is meant to include. And, [protein degradation product] means the protein degradation product of soybean unless otherwise noted. The peptone-degraded protein degradation product before fermentation Refers to a mixture of an oligopeptide and a polypeptide having various molecular weights and the fermented product of the fermented biopeptone has a weighted average of a di-peptide, a tri-peptide, and a tetra-peptide. And a peptide having a low molecular weight of 200 to 500 in molecular weight.

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

The cosmetic composition for inhibiting skin photoaging and regenerating skin damaged cells according to the present invention comprises a biopeptone fermentation product comprising a biopeptone fermentation broth or a biopeptone fermentation powder; Alpha-amino acids; sugars; Acetylglucosamine; And citric acid.

The biop peptone fermentation product will be described as follows.

The fermentation broth of the three-step biopeptone may be prepared through a process shown schematically in FIG. 1A. Specifically, Step 3-1 of producing a culture medium containing a peptone-digested protein degradation product; Step 3-2 of inoculating the culture medium with the lactic acid bacteria; A step 3-3 of culturing the inoculated lactic acid bacteria in the second step to ferment the peptone-digested protein degradation product to produce a low molecular weight peptide culture; And a step (3-4) of removing the cells and the insoluble component from the low-molecular peptide culture to obtain a fermentation broth of the biopeptone.

In addition, the step (3-5) of drying and pulverizing the fermented product of the biopeptone may be further followed by pulverization, followed by liquefaction.

In the preparation of the biopeptone fermentation product, the culture medium of Step 3-1 is a carbon source containing at least one selected from the group consisting of glucose, sucrose, maltose, fructose, lactose, xylose, galactose and arabinose; And one or more inorganic element compounds selected from magnesium sulfate, manganese sulfate, zinc sulfate, cooper sulfate, sodium acetate, potassium phosphate and calcium chloride.

In addition, in the production of the biopeptone fermentation product, in the step 3-2, the lactic acid bacteria are inoculated at a concentration of 1 × 10 ⁴ to 1 × 10 ⁷ cfu / ml, preferably 1 × 10 ⁵ to 5 × 10 ⁶ cfu / ml And then inoculating the culture medium with the lactic acid bacteria. If the amount of lactic acid bacteria inoculated is less than 1 x 10 < ⁴ > cfu / ml, the fermentation time may be too long. Even if the inoculated amount of lactic acid bacteria is more than 1 x 10 < ⁷ > cfu / It is not economical since it does not increase anymore.

In the fermentation of the bioepitone fermentation product, the fermentation in the step 3-3 is preferably performed at 25 to 35 for 12 to 72 hours, preferably 24 to 48 hours. If the fermentation time is less than 12 hours, the degradation time of the peptone-modified soybean protein degradation product may be insufficient, so that the content of the low molecular weight peptide may be too small. If the fermentation time exceeds 72 hours, the economical efficiency may deteriorate. It is better to do it.

The biop peptone fermented product thus prepared may include a low molecular weight biopeptide having a weight average of 200 to 500, and the low molecular weight peptide may include at least one selected from a peptide, a tripeptide, and a tetrapeptide.

The biop peptone fermented product may be characterized in that the peptone-degraded protein degradation product is fermented with lactic acid bacteria. The peptone-degraded protein decomposition product may be selected from the group consisting of soybean, seabream, seomotei, black beef, kidney beans, fried beans, Soybean, soybean, soybean, soybean, soybean, soybean, soybean, soybean, soybean, soybean, soybean, soybean, soybeans, soybeans, soybeans, soybeans and soybeans are preferably peptized by using pepsin .

In addition, the lactic acid bacteria used in the fermentation are Lactobacillus ramno suspension (Lactobacillus rhamnosus), Lactobacillus casei (Lactobacillus Casei) and Lactobacillus flat Lanthanum (Lactobacillus plantarum), Lactobacillus brevis (Lactobacillus brevis), Lactobacillus ash FIG filler switch (Lactobacillus acidophilus) can use a one Lactobacillus (Lactobacillus) sp or more species selected from, preferably, can be used Lactobacillus ramno suspension and Lactobacillus casei Lactobacillus sp 1 or more species selected from the group consisting of.

The cosmetic composition for inhibiting skin photoaging and regenerating skin damaged cells according to the present invention is characterized in that when the biopeptone fermentation product is a biopeptone fermentation broth, the biop peptone fermentation liquid contains 86 to 97.5% by weight, alpha-amino acid is 2 to 7% Preferably 0.1 to 2% by weight of acetylglucosamine, 0.05 to 1% by weight of acetylglucosamine and 0.1 to 2% by weight of citric acid, preferably 92 to 97% by weight of the biopeptone fermentation broth, 2.0 to 4% by weight of alpha-amino acid, 0.1 to 0.5% by weight of acetylglucosamine and 0.1 to 0.5% by weight of citric acid.

In the cosmetic composition for inhibiting skin photoaging and regenerating skin damaged cells according to the present invention, when the biop peptone fermentation product is a biopeptone fermentation powder, it is preferable that the biop peptone fermentation powder contains 40 to 60 wt%, alpha-amino acid 25 to 40 wt% Preferably from 10 to 20% by weight, from 0.1 to 2% by weight of acetylglucosamine and from 1 to 5% by weight of citric acid, preferably from 45 to 55% by weight of the biop peptone fermentation powder, from 27 to 36% by weight of the alpha- To 18% by weight, acetylglucosamine 0.5-1.5% by weight and citric acid 1 - 3.5% by weight.

Among the components of the composition of the present invention, the alpha-amino acid constitutes a protein and constitutes collagen, increases skin metabolism, and effectively moisturizes the skin. The use amount thereof may be so small that the synthesis of the protein may not be exerted, and if it is used in excess of the above range, the protein may be precipitated.

The saccharides of the composition of the present invention act as a skin moisturizing agent. When the total weight of the composition is less than the above-mentioned range, there is a problem that the amount of the saccharide used is so small that the moisturizing effect is lowered. , There may be a problem of causing stickiness.

Among the components of the composition of the present invention, the acetylglucosamine acts as a skin moisturizer. When the total weight of the composition is less than the above range, there is a problem that the amount of the acetylglucosamine used is so small that the moisturizing effect is lowered. If used, there may be a problem with solubility.

In the composition of the present invention, the citric acid serves as an acidity controlling agent, a neutralizing agent and a preservative. When the total weight of the composition is less than the above range, the amount of the citric acid to be used is small, In case of exceeding the range, there may be a problem that irritation occurs due to the effect of acidity.

The present invention also relates to cosmetic compositions for skin photoaging suppression and regeneration of skin damaged cells (hereinafter referred to as "cosmetics") used in external preparations for skin prepared using the above-described skin photoaging inhibiting and cosmetic composition for regenerating skin damaged cells will be. The cosmetic of the present invention may be a mixture of the cosmetic compositions described above, or may be a liposomized mixture of cosmetic compositions as described below.

More specifically, the cosmetic of the present invention is a liposomized formulation as shown in the schematic diagram of FIG. 2, comprising a biop peptone fermentation broth; And a phospholipid coating film for wrapping the biop peptone fermentation broth.

The phospholipid coating layer may have a single layer or multiple layers of phospholipids formed from liposome and ceramide solution. At this time, the ceramide solution is a solution in which ceramide is dissolved in a solvent.

The cosmetic composition of the present invention preferably contains 40 to 80% by weight of the biop peptone fermentation broth and 20 to 60% by weight of the phospholipid mixture, preferably 55 to 75% by weight of the biop peptone fermentation broth and 25 to 45% by weight of the phospholipid mixture, 60 to 72.5% by weight and 27.5 to 40% by weight of the phospholipid mixture. If the content of the phospholipid mixture is less than 20% by weight, the amount of the phospholipid coating film may be too small to sufficiently form a phospholipid coating film. If the content of the phospholipid mixture exceeds 60% by weight, The skin absorption rate of the cosmetic may be lowered, and there may be a problem that impurities may remain in the production of cosmetics.

Wherein the phospholipid mixture comprises lecithin, ceramide, and a solvent, wherein the content of lecithin is 1 to 10 parts by weight and the solvent is 10 to 100 parts by weight, preferably 100 to 100 parts by weight of lecithin, 4 to 8.5 parts by weight of a solvent and 30 to 85 parts by weight of a solvent, more preferably 5 to 8 parts by weight of a ceramide and 35 to 80 parts by weight of a solvent, based on 100 parts by weight of lecithin.

If the content of ceramide is less than 1 part by weight with respect to 100 parts by weight of lecithin, it is difficult to expect an effect of improving the skin barrier. If the content of ceramide is more than 10 parts by weight, the problem that the ceramide does not sufficiently dissolve due to low solubility of ceramide Therefore, it is preferable to use within the above range.

The solvent serves to dissolve the ceramide to increase compatibility with lecithin and to prevent water evaporation. The solvent is used in an amount of less than 10 parts by weight based on 100 parts by weight of lecithin and the amount thereof is too small to sufficiently dissolve the ceramide There may be a problem that the amount of the solvent used is more than 100 parts by weight and the viscosity of the phospholipid mixture is excessively lowered due to excessive use, which may interfere with the formation of the phospholipid coating film. The solvent may include caprylic / capric triglyceride caprylic / capric / stearic triglyceride and caprylic / capric / myristic / And one or more selected from the group consisting of Caprylic / Capric / Myristic / Stearic Triglyceride, or a mixture of two or more thereof.

The cosmetic composition of the present invention may be prepared by a process comprising the steps of: (1) preparing a solution of a ceramide solution in which a ceramide is dissolved in a solvent, Mixing the ceramide solution with lecithin and stirring at low speed to prepare a solution containing the phospholipid mixture; Adding a mixture of the cosmetic composition to the phospholipid mixture-containing solution, and then stirring at a low speed to prepare a liposomal suspension; And filtering the liposome to recover the liposome.

The composition and use amount of the ceramide, solvent, lecithin, and the biopeptone fermentation solution used in the manufacturing process are the same as those described above.

The low-speed stirring of the second and third stages may be carried out at a stirring speed of 1,000 to 2,000 rpm at 10 to 30 ° C for 15 to 30 minutes, preferably at a temperature of 15 to 25 ° C at 1,200 to 1,600 rpm It is preferable to carry out stirring for 20 minutes to 30 minutes at a stirring speed. At this time, if the temperature is less than 10 ° C, the temperature is too low to effectively form the phospholipid coating film, and if the temperature exceeds 40 ° C, the formed phospholipid coating film may be loosened. If the agitation speed is less than 1,000 rpm, the agitation speed may be too slow to sufficiently mix the biop peptone fermentation product and the ceramide solution. If the agitation speed exceeds 2,000 rpm, the agitation speed may become too fast. There may be a problem with unwinding.

Then, the liposomization was carried out through three-stage low-speed agitation, and an optical microscope photograph before and after the low-speed agitation was shown in FIG. 2B.

The skin photoinhibition inhibitor of the present invention, which is manufactured through the above steps, comprises a mixture of a cosmetic composition containing a biopeptone fermentation product in a liposome, and the liposome surface forms a phospholipid coating film for wrapping the mixture (See FIG. 2).

The cosmetic preparation of the present invention produced by the above-mentioned method has little skin cytotoxicity, promotes the synthesis of collagen type I in cells damaged by ultraviolet rays, has a tyrosinase inhibiting effect, Is excellent.

The cosmetic of the present invention can be applied as a skin external preparation such as cosmetics, ointments or ampoules of various formulations.

The cosmetic may be a cosmetic such as an emulsion type, a lotion type, a cream type, a lotion type, an emulsion type, an essence type, a pack type, a gel type, or a mist type.

In addition, the external preparation for skin of the present invention may be administered at a concentration of 50 μg / mL to 20,000 μg / mL, preferably 400 μg / mL to 10,000 μg / mL, more preferably 500 μg / mL to 5,000 μM ml, more preferably at a concentration of 450 μg / ml to 3,500 μg / ml. The skin photoinhibition inhibitor of the present invention has a very high skin absorbing ability, and the same effect can be obtained by using only 40 to 70% of the amount of the non-liposomized biop peptone fermentation liquid (or the biop peptone fermentation product) There is a number.

The present invention is not limited to this, but specific examples thereof include a skin photoinhibition agent, a glycerin, a butylene glycol, a dipropylene glycol, a carboxyvinyl The mixture may be prepared by mixing a polymer, polysorbate, hydrogenated castor oil, glyceryl stearate, cetearyl alcohol, cetyl ethylhexanoate and licorice extract, and further mixed with purified water, flavor, preservative and the like.

More specifically, mixing a carboxyvinyl polymer, purified water, glycerin, butylene glycol and dipropylene glycol to prepare a first mixture; Preparing a second mixture by mixing polysorbate, hydrogenated castor oil, glyceryl stearate, cetearyl alcohol, and cetyl ethylhexanoate; The first mixture and the second mixture are heated to 70 ° C to 90 ° C, preferably 75 ° C to 85 ° C, and then the warmed first mixture and the second mixture are mixed and stirred to prepare a third mixture step; And lowering the temperature to 35 ° C to 50 ° C, preferably 38 ° C to 45 ° C while stirring the third mixture, and mixing the skin photoinhibition inhibitor, the licorice extract and the additive, Type or lotion type external preparation for skin.

At this time, the glycerin serves as a skin moisturizer, and 600 to 1,600 parts by weight, preferably 800 to 1,400 parts by weight, of glycerin may be used per 100 parts by weight of the skin photoinhibition inhibitor. If the amount of glycerin used is less than 600 parts by weight, there may be a problem of deteriorating the moisturizing effect. If the glycerin is used in excess of 1,400 parts by weight, there may be a problem of causing stickiness.

The butylene glycol serves as a skin moisturizing agent, and 900 to 2,000 parts by weight, preferably 1,350 to 1,900 parts by weight, of butylene glycol may be used relative to 100 parts by weight of the skin photoinhibition agent. In this case, If the amount is less than 900 parts by weight, there may be a problem of deteriorating the moisturizing effect. If the amount is more than 2,000 parts by weight, there is a possibility that the formulation may become unstable.

The dipropylene glycol serves to moisturize the skin, and 1,400 to 3,000 parts by weight, preferably 1,700 to 2,200 parts by weight of dipropylene glycol may be used relative to 100 parts by weight of the skin photoinhibition agent. In this case, If the amount of glycol is less than 1,400 parts by weight, there may be a problem of reducing the moisturizing effect. If the amount of glycol is more than 3,000 parts by weight, there may be uneconomical problems.

The carboxyl vinyl polymer serves as an increasing agent. The carboxyl vinyl polymer may be used in an amount of 50 to 200 parts by weight, preferably 100 to 180 parts by weight, based on 100 parts by weight of the skin photoinhibition agent, If the amount of the vinyl polymer is less than 50 parts by weight, the viscosity may be lowered. If the amount of the vinyl polymer is more than 200 parts by weight, there may be a problem that it causes abrasion (not absorbed by the skin) It is good to do.

The polysorbate acts as a solubilizing agent, and 80 to 300 parts by weight, preferably 120 to 260 parts by weight, of polysorbate may be used relative to 100 parts by weight of the skin photoinhibition agent. In this case, If the amount is less than 80 parts by weight, there may be a problem of separating the formulations. If the amount is more than 300 parts by weight, there may be a problem of stickiness and foaming during application of the skin.

The hardened castor oil serves as a solubilizing agent. The hardened castor oil may be used in an amount of 900 to 1,800 parts by weight, preferably 1,350 to 1,700 parts by weight, based on 100 parts by weight of the skin photoinhibition agent. If the amount is less than 900 parts by weight, there may be a problem of separating the formulations. If the amount is more than 1,800 parts by weight, there may be a problem of foaming during application.

The glyceryl stearate serves as an emulsifier, and glyceryl stearate may be used in an amount of 140-280 parts by weight, preferably 160-260 parts by weight, based on 100 parts by weight of the skin photoinhibition agent. In this case, If the amount of the used amount is less than 140 parts by weight, there may be a problem that the emulsion stability is lowered. If the amount is more than 280 parts by weight, there is a problem that the emulsion stability is lowered.

The cetearyl alcohol serves as an emulsifier. The cetearyl alcohol may be used in an amount of 250 to 550 parts by weight, preferably 300 to 450 parts by weight, based on 100 parts by weight of the skin photoinhibition agent. If the amount of the alcohol is less than 250 parts by weight, the emulsion stability may be deteriorated. If the amount of the alcohol is used in excess of 550 parts by weight, the emulsion stability may be deteriorated.

The cetyl ethylhexanoate serves as an oil component to improve the sensation of use, and 700 to 1,600 parts by weight, preferably 900 to 1,450 parts by weight, of cetyl ethylhexanoate can be used relative to 100 parts by weight of the skin photoinhibition agent If the amount of cetylethylhexanoate used is less than 700 parts by weight, there may be a problem that the feeling of use is lowered. If the amount of cetylethylhexanoate is more than 1,600 parts by weight, there may be a problem of casting.

The licorice extract has a role of skin moisturizing and anti-inflammation, and 15 to 80 parts by weight, preferably 20 to 60 parts by weight, of the licorice extract may be used per 100 parts by weight of the skin photoinhibition agent. In this case, If the amount is less than 15 parts by weight, the skin may not be able to function as a moisturizing and anti-inflammatory agent. If it is used in excess of 80 parts by weight, skin irritation may be caused.

The purified water serves as a solvent and a viscosity control agent. The purified water may be used in a controlled amount depending on the type of external preparation for skin, preferably 20,000 to 35,000 parts by weight based on 100 parts by weight of the skin photoinhibition agent.

When the cosmetic of the present invention is manufactured by an emulsion type, a lotion type, an emulsion type, an ampule type, an essence type or the like, it is preferable to use the skin photoinhibition agent, dipropylene glycol, 1,3 -Butylene glycol, polysorbate, dipotassium glycyrrhizate, allantoin, panthenol, and glycerin, and may further be prepared by mixing purified water, flavor, preservative and the like. And, this type of external preparation for skin may include a skin photoinhibition agent, dipropylene glycol, 1,3-butylene glycol, polysorbate, dipotassium glycyrrhizate, allantoin, panthenol and glycerin.

The dipropylene glycol serves as a skin moisturizing agent. Dipropylene glycol may be used in an amount of 1,500 to 3,000 parts by weight, preferably 2,000 to 2,800 parts by weight, based on 100 parts by weight of the skin photoinhibition agent. In this case, If the amount is less than 1,500 parts by weight, there may be a problem of deteriorating the moisturizing effect. If the amount is more than 3,000 parts by weight, it may be insoluble.

The 1,3-butylene glycol serves as a skin moisturizing agent, and 1,500 to 2,500 parts by weight, preferably 1,800 to 2,400 parts by weight, of 1,3-butylene glycol can be used relative to 100 parts by weight of the skin photoinhibition agent If the amount of 1,3-butylene glycol used is less than 1,500 parts by weight, there may be a problem of reducing the moisturizing effect. If the amount of 1,3-butylene glycol is more than 2,500 parts by weight, there is a possibility that the formulation may become unstable. It is good to use.

The polysolvate serves as a solubilizing agent, and 600 to 1,250 parts by weight, preferably 700 to 1,100 parts by weight, of polysorbate may be used relative to 100 parts by weight of the skin photoinhibition agent. In this case, If the amount is less than 600 parts by weight, there may be a problem of separating the formulations. If the content is more than 1,250 parts by weight, there may be a problem of stickiness and foaming during application of the skin.

The dipotassium glycyrrhizate has a role of relieving irritation, and 120 to 250 parts by weight, preferably 140 to 220 parts by weight, of dipotassium glycyrrhizate can be used relative to 100 parts by weight of the skin photoinhibition inhibitor. If the amount of dipotassium glycyrrhizinate is less than 120 parts by weight, there may be a problem that it does not play a role of stimulation relaxation. If it exceeds 250 parts by weight, there may be a problem of breaking the stability of the formulation. It is good to use.

The allantoin has a role of relieving irritation, and it is possible to use 90 to 280 parts by weight, preferably 140 to 220 parts by weight, of allantoin relative to 100 parts by weight of the skin photoinhibition inhibitor, wherein the amount of allantoin used is less than 90 parts by weight There may be a problem that it does not play a role of stimulation relaxation. If it exceeds 280 parts by weight, there is a problem of breaking the stability of the formulation.

The panthenol serves to supply nutrients to the skin with a stimulant and vitamin B5. The panthenol may be used in an amount of 80 to 300 parts by weight, preferably 120 to 200 parts by weight, based on 100 parts by weight of the skin photoinhibition agent, If the amount of the allantoin used is less than 80 parts by weight, there may be a problem that the irritation is relieved and the skin nutrient supply does not play a role. If the amount is more than 250 parts by weight, the stability of the formulation may be broken. It is good to use.

The glycerin serves as a skin moisturizing agent. The glycerin may be used in an amount of 650 to 1,550 parts by weight, preferably 800 to 1,250 parts by weight, based on 100 parts by weight of the skin photoinhibition agent. When the amount of the allantoin used is less than 650 parts by weight There may be a problem that skin moisturization may be deteriorated, and when it is used in excess of 1,550 parts by weight, there may be a problem of causing stickiness, so it is preferable to use within the above range

Hereinafter, the present invention will be described in more detail with reference to the following examples. However, the following examples should not be construed as limiting the scope of the present invention, and should be construed to facilitate understanding of the present invention.

[Example]

Preparation Example 1: Preparation of a biopeptone fermentation broth

Peptonized protein degradation products were purchased from Fluka (Sigma-Aldrich, St. Quentin-Fallavier, France). 500 g of soybean peptone, which is a peptone-degraded protein degradation product, was added to the components and the contents of the following Table 1, and the mixture was sterilized by heating at 121 占 폚 for 15 minutes. Lactobacillus rhamnosus was inoculated as lactic acid bacteria at an initial bacterial count of 1 x 10 ⁶ cfu / ml, and cultured and fermented at 30 ° C for 48 hours with stirring at 50 rpm. A peptone fermentation culture was obtained.

Next, the fermentation broth obtained from the above procedure was heat treated to a final temperature of 95 ° C and centrifuged (3,000 rpm, 15 min) to remove the cells and insoluble components, thereby preparing a biopeptone fermentation broth.

No. Ingredients Content (w / w) One Soybean peptone 5.000% 2 Glucose 0.500% 3 Potassium phosphate 0.2500% 4 Sodium acetate 0.120% 5 Ammonium citrate (Dimmonium citrate) 0.120% 6 Magnesium sulfate (MgSO ₄ ) 0.012% 7 Manganese sulfate (MnSO ₄₎ 0.003% 8 Zinc sulfate (ZnSO ₄ ) 0.001% 9 Copper sulfate (CuSO ₄ ) 0.001% 10 Purified water 93.993% Sum 100,000%

Preparation Example  2: Preparation of biop peptone fermentation powders

About 45 g of the lyophilized biopeptide fermentation product of the biopeptone fermentation broth prepared in Preparation Example 1 was obtained.

Experimental Example  1: high performance liquid chromatography ( HPLC ) And mass spectrometry (MS) measurements Through bio  Molecular weight confirmation test of peptone fermentation broth

To confirm the molecular weight of the biopeptone fermentation broth of Preparation Example 1, High performance liquid chromatography and MS spectra were measured.

Accela UHPLC (manufacturer: Thermo Scientific) was used for high performance liquid chromatography measurement. The lyophilized biopeptone fermentation product was dissolved in 0.1% formic acid, and the temperature was determined as RT (Room Temperature). The column used was Water BEH C18 (2.1 × 100 mL, 1.7 μm) and the flow rate was maintained at 300 μL per minute. The sample analysis time was up to 60 minutes. The measurement results from 0 to 60 minutes are shown in FIG. 3A, and the measurement results from 0 to 25 minutes are shown in FIG. 3B.

MS spectra were also measured using an LTQ Orbitrap XL mass spectrometer (manufactured by Thermo Scientific). Five peaks having large areas among the single components separated by liquid chromatography were arbitrarily selected, and the molecular weight was confirmed by MS spectrum. The results are shown in FIG.

4 shows data obtained by measuring the molecular weights of the peaks in the portions indicated by (1) to (5) in FIG. 3B. From the results of (1) to (5) in FIG. 4, it can be seen that most of the peaks have a weight average molecular weight of 100 to 500 or less It is possible to estimate and / or confirm that the peptide is a peptide composed of 2 to 4 amino acids and has a weight average molecular weight of 200 to 300.

Example  1: skin Photoaging  For inhibition and skin damaged cell renewal Cosmetics  Preparation of composition

The mixture of the biop peptone fermentation broth prepared in Preparation Example 1, 3.2 wt% of alpha-amino acid, 1.5 wt% of saccharides, 0.1 wt% of acetyl glucosamine and 0.2 wt% of citric acid was mixed and stirred to prepare a skin photoaging A mixture of cosmetic compositions for inhibition and skin damaged cell regeneration was prepared.

In this case, the alpha-amino acid was prepared by mixing isoleucine, leucine, lysine, methionine, phenylalanine, threonine, tryptophan, valine, glutamic acid and acetylcysteine. The saccharides were prepared by mixing glucose, galactose and xylose Respectively.

Example  2

50% by weight of the biop peptone fermentation powder prepared in Preparation Example 2, 32% by weight of alpha-amino acid, 15% by weight of saccharides, 1% by weight of acetylglucosamine and 2% by weight of citric acid were mixed and stirred, A mixture of cosmetic compositions for inhibiting skin photoaging and for regenerating skin damaged cells was prepared.

Comparative Example

As a comparative example, a biopeptone fermentation broth of Preparation Example 1 was prepared.

Category (% by weight) Example 1 Example 2 Comparative Example Biopeptone fermentation broth 95 - 100 Biop peptone fermentation powder - 50 - Alpha-amino acid Isoleucine 0.30 3 - Leucine 0.30 3 - Lysine 0.50 5 - Methionine 0.20 2 - Phenylalanine 0.20 2 - Threonine 0.30 3 - Tryptophan 0.20 2 - Balin 0.20 2 - Glutamic acid 0.50 5 - Acetylcysteine 0.50 5 - sugars Glucose 0.50 5 - Galactose 0.50 5 - Xylose 0.50 5 - Acetylglucosamine 0.10 One - Citric acid 0.20 2 - Sum 100.00 100.00 100.00

Experimental Example  2: In vitro cytotoxicity test

(1) Human dermal fibroblasts, human dermal fibroblasts, were purchased from Innoprot (Bizkaia, Spain) and cultured in DMEM (Dulbecco's modified Eagle's medium containing 10% FBS (Fetal bovine serum, Welgene, Daegu, Korea) , Welgene, Daegu, Korea) at 37 ° C in a 5% CO ₂ cell incubator. The culture medium was changed every 3 days. When the number of cells grew by more than 80%, the cells were transplanted and the cells were maintained in a stable state for 2 weeks.

(2) WST-1 assay was carried out to confirm that the fibroblasts of Example 1 and Comparative Example were toxic. Cells were seeded in a 96-well plate at a density of 1 × 10 ⁴ cells / well in a volume of 200 μL per well and cultured in a 5% CO ₂ cell incubator at 37 ° C. for 24 hours. DMEM containing no FBS The medium and skin photoinhibition agent were treated for 24 hours. To each well, 20 μL of WST-1 solution (Cell Proliferation Reagent WST-1, Cat No. 05 015 944 001, Roche) was added to make 10% of the whole, followed by incubation at 37 ° C in a 5% CO ₂ cell incubator for 2 hours Lt; / RTI >

Absorbance was measured at 450 nm and 620 nm using a VersaMax microplate reader (Molecular Devices, Sunnyvale, CA) for the measurement of absorbance, and the survival ratio of the cells was calculated based on the control group. The results are shown in FIGS. 5A and 5B . Statistical analysis was performed at the p <0.05 level using SPSS 12.0K (SPSS Inc., Chicago, IL, USA) and the t- test And Duncan's multiple range test.

In FIGS. 5A and 5B, the control group was obtained by culturing only the DMEM medium without the treatment of the EGF and the biopeptone fermentation product. The EGF was obtained from EGF (Epidermal Growth of Sigma aldrich), which is known as a growth factor promoting epithelial cell proliferation, Factor human, Cat. No. E9644) at 20 μg / mL. 5A is a graph showing the results of measurement of the concentration (100 μg / mL to 2,000 μg / mL) of the comparative example (fermented biopeptone).

As shown in FIG. 5A, no cytotoxicity was observed in the entire concentration range of 0 to 2,000 μg / mL, and cell proliferation was actively observed. In particular, the cell proliferation effect was very high in the vicinity of 300 to 1,000 μg / mL, and the highest cell proliferation effect was observed particularly in the vicinity of 600 to 900 μg / mL.

(2) FIG. 5 (b) shows the results of measuring the cell proliferation effect after using the biopeptone fermented product having the best cell proliferation effect of 800 μg / ml in Examples and Comparative Examples.

Referring to FIG. 5B, it can be seen that the cell proliferation results of the examples are somewhat higher than those of the comparative example using only the biopeptone fermentation product.

Experimental Example  3: In UVB irradiation  Measurement of cell protection effect

(1) The human dermal fibroblasts of the human dermal fibroblasts of Experimental Example 2 were cultured in a 60 mm cell culture dish. Then, the culture medium was removed before irradiation with ultraviolet light, ) And the cosmetic composition of Example 1 and the comparative example were added at a concentration of 800 μg / mL. UVB was irradiated at 312 m with 50 mJ / cm ² intensity using a UV lamp (VL-6LM, Vilber Lourmat, France).

Immediately after irradiation with ultraviolet light, the cells were replaced with serum-free cell culture medium and cultured in a 5% CO ₂ cell incubator at 37 ° C. In the control group, the whole process except the ultraviolet irradiation process was treated in the same manner. Statistical analysis was performed at the p <0.05 level using SPSS 12.0K (SPSS Inc., Chicago, IL, USA) and the t- test And Duncan's multiple range test.

6, the cell viability was decreased to 48% in the control group 2 irradiated with UVB compared to the control group not irradiated with UVB, whereas the comparative example and Example 1 showed a relatively high cell survival rate when compared with the control group 2 And in particular, Example 1 showed relatively better cell viability than the Comparative Example. From these results, it can be confirmed that the cosmetic composition of the present invention inhibits cell death due to UVB and effectively protects skin from ultraviolet rays

FIG. 7 shows photographs taken before and after the experiment.

Experimental Example  4-1: Collagen type (Collagen type) Measurement of synthesis promoting effect

Collagen, which accounts for about 70-80% of the dry weight of skin epithelial cells, is known to be a major factor in skin elasticity and wrinkles, along with hyaluronic acid and elastin. Generally, the activity of MMP-1 (Matrix metalloproteinase-1), which is one of the collagen type synthesis and degradation enzymes, is generally balanced in the skin. However, in aging skin tissue, synthesis of collagen type is decreased and enzyme of MMP- It is known that the activity is increased. Therefore, the promoting effect of collagen synthesis is an important factor for reducing elasticity and preventing wrinkles.

In addition to the cytoprotective effect by the UVB irradiation of the cosmetic composition for restraining photoaging and skin damaged cell regeneration, the additional effect of promoting collagen synthesis was measured as described below.

Experiments were conducted using a PIP (Procollagen Type C-peptide) EIA kit (Cat no. MK101, Takara Bio) to confirm the promoting effect of type 1 collagen biosynthesis on the cosmetic composition.

The human dermal fibroblast, which was the same human dermal fibroblast cell line used in Experimental Example 2, was inoculated at 6 × 10 ⁵ cells / well on a 6-well plate and cultured in a 5% CO ₂ cell incubator at 37 ° C. After 24 hours, the cosmetic composition of the present invention of Example 1 was treated with DMEM (Dulbeco's Modified Eagle's Media) -free medium containing no serum (FBS) for each concentration and cultured at 37 ° C under 5% CO ₂ for 24 hours. The culture broth was recovered.

Next, 10 占 퐇 of the Antibody-POD Conjugate Solution was treated per well in the Anti-PIP Monoclonal Antibody Plate, and 20 占 퐇 of the recovered cell culture medium was added and reacted for 3 hours in a 5% CO ₂ cell incubator at 37 占 폚. After washing the wells with PBS 4 times, the reaction was stopped by treating 100 μL of the substrate solution (TMBZ, 3,3 ', 5,5'-tetramethylbenzene), and a VERSAmax microplate reader (Molecular Devices, Sunnyvale, CA ) Was used to measure the absorbance at 450 nm and the synthesized collagen (PIP) was calculated based on the standard curve. Statistical analysis was performed at the p <0.05 level using SPSS 12.0K (SPSS Inc., Chicago, IL, USA) and the t- test And Duncan's multiple range test.

In FIG. 8, the control group was obtained by culturing the EGF and biop peptone fermentation product without DMEM medium alone. EGF was cultured in EGF (Epidermal Growth Factor human, Sigma aldrich), which is known as a growth factor promoting epithelial cell proliferation, Cat. No. E9644) at 20 μg / mL. And, Comparative Example and Example 1 are the results of measurement at 800 μg / mL.

8, it can be confirmed that the cosmetic composition of Example 1 has the best collagen synthesis promoting effect. In particular, at a concentration of 800 μg / mL or more, the cosmetic composition of the present invention exhibits an effect of promoting collagen synthesis, which is close to 250% as compared with the control group, and thus the cosmetic composition of the present invention can increase skin elasticity or effectively prevent wrinkles .

Experimental Example  4-2: In UVB irradiation  Collagen type (collagen type) of damaged cells

(One) The Since the cosmetic composition for suppressing photoaging and skin damage cell regeneration not only inhibits UVB-induced cell damage but also promotes excellent collagen synthesis in a human-derived fibroblast cell, Whether the collagen synthesis is promoted is experimentally conducted using a skin photoinhibition inhibitor.

After irradiation with UVB in the same manner as in Experimental Example 3, collagen synthesis promotion was measured in the same manner as in Experimental Example 4-1, and the composition of Example 1 was used. The results are shown in FIG.

9, the cell survival rate was decreased to 40% in the control group 2 irradiated with 50 mJ / cm ² UVB compared with the control group not irradiated with UVB. In contrast, the cell survival rate in the group irradiated with UVB was significantly higher than that in the group irradiated with the control (800 μg / mL) and Example 1 (800 μg / mL) % Cell survival rate was increased. Therefore, it is considered that the cosmetic composition of the present invention can prevent the photoaging of the skin due to ultraviolet rays, thereby suppressing the generation of wrinkles of the skin to the utmost.

Experimental Example  5: tyrosinase enzyme ( Tyrosinase ) Inhibition effect measurement

Tyrosinase is an enzyme involved in the initial synthesis step in the human melanin biosynthetic pathway. When melanocytes are exposed to stress caused by external or internal factors such as ultraviolet rays, drugs, and α-MSH, tyrosine, an amino acid, catalyzes tyrosinase to form L-DOPA It forms a melanin pigment through the pathway of dopaquinone. Therefore, in order to confirm the inhibitory effect of tyrosinase acting as an important catalyst in the melanin biosynthesis, the tyrosinase enzyme inhibitory effect involved in the melanin biosynthesis step of the cosmetic composition of the present invention was studied.

Melanocyte B16F10 (human epidermal melanocytes), a melanocyte-forming cell line, was purchased from Innoprot (Bizkaia, Spain) and cultured in Dulbecco's modified Eagle's medium containing 10% FBS (Fetal bovine serum, Welgene, Daegu, Korea) , Welgene, Daegu, Korea) at 37 ° C in a 5% CO ₂ cell incubator. The culture medium was changed every 3 days. When the number of cells was more than 80%, the cells were transplanted and the cells were maintained in a stable state for 2 weeks.

Next, 550 μL of distilled water (DW), 100 μL of 100 U tyrosine oxidase (Mushroom tyrosinase), 50 μL of 0.1 M KPB (potassium phosphate buffer), 50 μL of the melanocyte cell and the cosmetic composition of the present invention of Example 1 2,000 μg / mL), and pre-incubated at 37 ° C for 5 minutes. Then, 200 μL of 10 mM L-DOPA as a substrate was added and reacted at 37 ° C. for 10 to 20 minutes for enzyme activity. Next, the reaction solution was transferred to a 96-well plate and the absorbance at 475 nm was measured using a VersaMax microplate reader (Molecular Devices, Sunnyvale, Calif.), And the inhibition of tyrosinase enzyme activity was measured in comparison with the control group. Control group 1 was treated with 1X PBS (Dulbecco's Phosphate Buffer Saline from Gibco Co., Cat. No. 21300-025) instead of Example 1, and control group 2 was treated with arbutin (100 μg / mL) . The measurement results thereof are shown in Fig.

10, it was confirmed that the inhibitory effect of tyrosinase activity was observed in both the control group 2, the comparative example (800 μg / mL) and the example 1 (800 μg / mL). It was confirmed that Example 1 effectively inhibited tyrosine activity from tyrosine by inhibiting tyrosinase activity relatively more than Comparative Example.

Experimental Example  6: From Melanocyte (Melanocyte)  Melanin pigment inhibitory effect

(1) In order to measure the inhibitory effect of the cosmetic composition of the present invention on melanin pigment formation in melanocytes, Melanocyte B16F10 (Human Epidermal Melanocytes) was divided into 6 well plates at 2 × 10 ⁵ cells / CO ₂ . After 24 hours, 0.1 mM IBMX (3-isobutyl-1-methylxanthine) was treated with the sample and incubated in a 5% CO ₂ cell incubator at 37 ° C for 48 hours to induce intracellular melanin production. Arbutin / mL), Comparative Example (800 μg / mL) and Example 1 (800 μg / mL) were treated with 800 μg / mL. The cells were washed with PBS, centrifuged using a cell scraper, centrifuged, and 800 μL of 1N-NaOH was added to the pellet obtained by removing the supernatant, which was then reacted in a thermostat at 80 ° C to dissolve the cells . Next, the absorbance was measured at 405 nm using a VersaMax microplate reader (Molecular Devices, Sunnyvale, Calif.), And a small amount of melanin was calculated in comparison with the control group. The results are shown in FIG. 11A. In addition, a photograph of melanin production observed at a magnification of 100 at a microscope (phase contrast microscope) is shown in FIG. 11B. Here, the control group 1 was not using any of IBMX, arbutin and the cosmetic composition of the present invention, and the control group 2 was treated with IBMX.

11A, the melanin pigment was increased to about 146% of the control group 1 in IBMX-added cells of the control group 2, whereas the addition of the comparative example (800 μg / mL) of IBMX to the control group 1 resulted in a production rate of melanin pigment of 117% Respectively. In contrast, when IBMX and Example 1 (800 μg / mL) were added together, the production rate of melanin pigment was about 105% lower than that of the Comparative Example.

Experimental Example  7: DPPH  Antioxidant effect by assay

DPPH assay experiments were conducted to measure the antioxidant effect of the cosmetic composition of the present invention. DPPH powder was diluted with ethanol and dissolved at 0.2 mM. The DPPH (1,1-diphenyl-2-picrylhydrazyl) solution and the cosmetic composition of Example 1 were reacted at a weight ratio of 1: 1 for 30 minutes, and the mixture was transferred to a VERSAmax microplate reader , Sunnyvale, CA), and the radical scavenging activity (IC50 value) was calculated on the basis of the control group. The results are shown in FIGS. 12A and 12B. Also, the DPPH (1,1-diphenyl-2-picrylhydrazyl) solution and the comparative biopeptone fermentation product were reacted for 30 minutes at a weight ratio of 1: 1, and the radical scavenging activity was measured in the same manner. FIG. 12A shows the results of measurement of free radical scavenging ability (%) by concentration of ascorbic acid, and FIG. 12B shows the results of measurement of radical scavenging ability of the cosmetic composition of Comparative Example and Example 1 by concentration.

As shown in Table 12b, the free radical scavenging ability was increased in a concentration-dependent manner. Compared with ascorbic acid, the radical scavenging ability was lowered when the same amount was applied, but it was confirmed that the radical scavenging ability was excellent. It was confirmed that Example 1 had a relatively superior radical scavenging ability than the Comparative Example.

Example  3: Liposome  Type Cosmetic  Produce

Lecithin was added to the beaker, and then 57 parts by weight of the ceramide solution was mixed with 100 parts by weight of lecithin, followed by stirring at 20 to 21 DEG C for 15 minutes at a stirring speed of 1,350 to 1,400 rpm to prepare a phospholipid mixture. At this time, the ceramide solution was prepared by adding 7 parts by weight of ceramide to 50 parts by weight of caprylic / capric triglyceride based on 100 parts by weight of lecithin.

Subsequently, 35.7% by weight of the phospholipid mixture and 64.3% by weight of the cosmetic composition prepared in Example 1 were mixed and stirred at 20 to 21 ° C for 15 minutes at a stirring speed of 1,350 to 1,400 rpm for 15 minutes to prepare a liposome suspension 1b).

Next, the liposome formed by leaving the liposome suspension was immersed, and the liposome was recovered by filtering the liposome suspension to prepare a cosmetic composition for restricting skin photoaging and regenerating skin damaged cells.

Example  2 ~ Example  5 and Comparative Example  1-4

The skin photoinhibition inhibitor was prepared in the same manner as in Example 1, except that the composition of the skin photoinhibition inhibitor was prepared as shown in Table 3 below.

division Example 1 Phospholipid mixture content lecithin Ceramide menstruum Example 3 64.3 wt% 35.7 wt% 100
Weight portion 7
Weight portion 50
Weight portion

Manufacturing example  1: Functionality Ampoule  Produce

Using the liposome type cosmetics of Example 3, ampoules having the components shown in Table 4 below were prepared. More specifically, dipropylene glycol, 1,3-butylene glycol, dipotassium glycyrrhizate, allantoin, panthenol, and glycerin were added to purified water and mixed well to prepare a first mixture. Polysorbate 80 was added thereto, followed by solubilization, followed by addition of a preservative and mixing to prepare a second mixture. Next, a preservative, a flavoring agent, and a fermentation product of biopeptone were added to the second mixture and stirred well to prepare an ampoule-type functional skin external preparation containing a skin photoinhibition agent at a dose of about 2,630 μg / mL. Ampoule production was possible at room temperature without warming, so the product was prepared at 26 ℃.

division ingredient Manufacturing ratio
(Parts by weight) One The liposome type cosmetic of Example 3 100 2 Dipropylene glycol 2,500 3 1,3-butylene glycol 2,200 4 Polysorbate 80 950 5 Dipotassium glycyrrhizate 200 6 Allantoin 190 7 Panthenol 150 8 glycerin 900 9 Purified water 30,500 10 antiseptic 150 11 Spices 120

Manufacturing example  2: Preparation of functional external preparation (cream)

Cream type external preparations having the components shown in Table 5 below were prepared using the liposome type cosmetics of Example 3 above. More specifically, purified water was added to a carboxyvinyl polymer and mixed to prepare a carboxyvinyl polymer. After that, glycerin, butylene glycol, and dipropylene glycol were added to prepare a first mixture. Further, a second mixture was prepared by adding Polysorbate 80, hydrogenated castor oil, glyceryl stearate, cetearyl alcohol, and cetyl ethylhexanoate. Then, the first and second mixtures were heated to 80 DEG C, and then mixed and stirred to prepare a third mixture. Next, the third mixture was evenly poured, the temperature was lowered to 40 ° C, and preservatives, flavors, biop peptone fermented product and licorice extract were added thereto, followed by stirring and mixing to obtain a fermented product containing about 2,560 μg / One cream type functional external preparation was prepared.

division ingredient Manufacturing ratio
(Parts by weight) One The liposome type cosmetic of Example 3 100 2 glycerin 1,200 3 Butylene glycol 1,800 4 Dipropylene glycol 1,900 5 Carboxyl vinyl polymer 150 6 Polysorbate 80 180 7 Hardened castor oil 1500 8 Glyceryl stearate 200 9 Cetearyl alcohol 380 10 Cetyl ethyl hexanoate 1,200 11 Licorice extract 45 12 Purified water 30,000 13 antiseptic 180 14 Spices 170

Through the above Examples and Experimental Examples, it was confirmed that the skin photoinhibition inhibitor of the present invention is excellent in the inhibition of melanin pigment formation through promotion of collagen synthesis and inhibition of tyrosinase in cells damaged by skin UVB and the like, , Ampoules, ointments, and the like.

Claims (15)

A fermented product of a biopeptone comprising a fermented product of fermented biopeptone or fermented product of fermented biopeptone; Alpha-amino acids; sugars; Acetylglucosamine; And citric acid. The cosmetic composition for inhibiting skin photoaging and regenerating skin damaged cells. The pharmaceutical composition according to claim 1, which comprises 86 to 97.5% by weight of the biopeptone fermentation broth, 2.0 to 7% by weight of alpha-amino acid, 0.3 to 5% by weight of saccharides, 0.05 to 1% by weight of acetylglucosamine, and 0.1 to 2% A cosmetic composition for inhibiting skin photoaging and regenerating skin damaged cells. The method of claim 1, wherein the biop peptone fermentation powder comprises 40 to 60 wt% of alpha-amino acid, 25 to 40 wt% of saccharide, 0.1 to 2 wt% of acetylglucosamine, and 1 to 5 wt% Wherein the cosmetic composition is a cosmetic composition for inhibiting skin photoaging and regenerating skin damaged cells. The fermented product according to claim 1, wherein the biopeptone fermented product is a fermented product obtained by fermenting a peptoned protein degradation product with a lactic acid bacterium,
Wherein the biop peptone fermented product comprises 100 wt% of a low molecular weight peptide having a weight average molecular weight of 200 to 500 (including other unavoidable impurities). 5. The method according to claim 4, wherein the lactic acid bacterium comprises at least one selected from the group consisting of Lactobacillus rhamnosus, Lactobacillus plantarum and Lactobacillus brevis. A cosmetic composition for regenerating skin damaged cells. [Claim 5] The cosmetic composition according to claim 4, wherein the peptone-modified protein degradation product is peptone-modified soybean, at least one selected from soybean, seomyeitae, kidney bean and mung bean. . A cosmetic for inhibiting skin photoaging and regenerating skin damaged cells, which comprises a mixture of the composition of any one of claims 1 to 6. The cosmetic according to claim 7, wherein the mixture contains 40 to 80% by weight of the mixture and 20 to 60% by weight of a phospholipid mixture. [9] The cosmetic according to claim 8, wherein the phospholipid mixture comprises 1 to 10 parts by weight of ceramides and 10 to 100 parts by weight of a solvent, based on 100 parts by weight of lecithin. A mixture of cosmetic compositions according to any one of claims 1 to 6; And
And a liposome comprising a phospholipid coating film for wrapping a mixture of the cosmetic composition on the skin. A step of preparing a ceramide solution in which ceramide is dissolved in a solvent;
Mixing the ceramide solution with lecithin and stirring at low speed to prepare a solution containing the phospholipid mixture;
Adding a mixture of the cosmetic composition of any one of claims 1 to 6 to the phospholipid mixture-containing solution, and then slowly stirring to prepare a liposomal suspension; And
And recovering the liposome by filtration. The method of manufacturing a cosmetic for regenerating skin damaged cells and inhibiting skin photoaging. 12. The method of claim 11, wherein the interior of the liposome comprises a mixture of a cosmetic composition, and the surface of the liposome is formed of a phospholipid coating film for wrapping the mixture of the cosmetic composition. A method for producing a cosmetic for reproduction. 12. The method according to claim 11, wherein the low-speed stirring of the second and third stages is performed at a stirring speed of 1,000 to 2,000 rpm at 10 to 30 DEG C for 15 to 30 minutes. Wherein the cosmetic material is a cosmetic. A composition for external application for skin, which comprises a mixture of the composition of any one of claims 1 to 6. 15. The external preparation for skin according to claim 14, wherein the external preparation for skin is cosmetic, ointment or ampoule.

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