KR102155489B1 - A composition comprising a mixture of nucleic acid fragments and uses thereof - Google Patents

Abstract

It relates to compositions comprising a mixture of nucleic acid fragments and to use thereof for improving skin conditions. According to the composition according to one aspect, since it has excellent UV protection, cell regeneration, skin barrier strengthening, skin moisturization, and antioxidant effects, it can be applied to various fields such as cosmetics, foods, pharmaceutical compositions for improvement, prevention or treatment of skin conditions. .

Description

A composition comprising a mixture of nucleic acid fragments and uses thereof

It relates to compositions comprising a mixture of nucleic acid fragments and to use thereof for improving skin conditions.

Ribonucleic acid (RNA) is a type of nucleic acid that forms nucleotides based on ribose, unlike deoxyribonucleic acid (DNA), which has deoxyribose as a base. A single helix has a long twisted structure, and it forms a complementary base pair inside the molecule, so that it has a partial double helix structure. Unlike deoxyribonucleic acid, uracil instead of thymin makes a base pair with adenine, ribonucleic acid acts in the process of protein synthesis, and some viruses use ribonucleic acid instead of deoxyribonucleic acid as a genetic material. I have it. Ribonucleic acid is a ribosomal ribonucleic acid (rRNA) that composes ribosomes according to function and structure, messenger ribonucleic acid (mRNA) that has genetic information on protein synthesis received from deoxyribonucleic acid, and corresponds to the codon of messenger ribonucleic acid. It can be divided into transporting ribonucleic acid (tRNA) and the like that deliver amino acids.

Such nucleic acids are components of the human body and are not toxic when applied to the skin, and thus have excellent safety, so it is important to develop a composition for using the nucleic acids as cosmetics, skin external preparations, foods, or pharmaceuticals for improving skin conditions.

One aspect provides a cosmetic composition for improving skin condition, or a composition for external application, including a mixture of nucleic acid fragments.

Another aspect provides a pharmaceutical composition for preventing or treating a skin condition comprising a mixture of nucleic acid fragments.

One aspect provides a cosmetic composition for improving skin conditions comprising a mixture of nucleic acid fragments as an active ingredient.

Another aspect provides a composition for external application for skin for improving skin conditions comprising a mixture of nucleic acid fragments.

The mixture of nucleic acid fragments according to an aspect has excellent UV protection, cell regeneration, antioxidant, and skin moisturizing effects, and thus can be usefully used to improve skin conditions.

The term "improving skin condition" may include one or more of skin wound improvement, dermatitis improvement, skin regeneration, skin barrier strengthening, skin moisturizing, antioxidant, and combinations thereof.

The term "strengthening the skin barrier" may refer to any action that enhances the function of the skin barrier, which is located on the outermost side of the skin and prevents loss of moisture and nutrition.

The term "skin regeneration" may refer to any action that supplements the part of the skin or promotes proliferation of skin cells when a part of the skin is lost.

The term "included as an active ingredient" means that a mixture of nucleic acid fragments is added to the composition of the present invention to the extent that it can exhibit skin improvement effects, and various components are added as subcomponents for drug delivery and stabilization. It is meant to include being formulated.

The term "improvement" can mean any action that at least reduces the severity of a parameter, such as a symptom, associated with remission or treatment of the condition.

The term "prevention" refers to any action that improves the skin condition by administering a composition containing the nucleic acid fragment mixture to become closer to a normal control group.

The term "treatment" refers to any action in which the skin condition is improved or advantageously changed to the same or similar to that of a normal control group by administration of a composition comprising the nucleic acid fragment mixture.

The "nucleic acid fragment" may refer to a form of a fragment having a reduced molecular weight of a nucleic acid, which is a biopolymer. The "mixture of nucleic acid fragments" may refer to an unspecified form in which nucleic acid fragments are mixed.

The mixture of nucleic acid fragments is a nucleic acid polymer, and because it is a biological component, there is no allergic reaction or rejection reaction in the body, and the main component is made of DNA or RNA having a negative charge, so it is hydrophilic, so it is very easy to apply industrially, and thus pharmaceutical compositions and cosmetics It can be used in the composition.

In one embodiment, the nucleic acid may be DNA.

The nucleic acid fragment is about 10 kDa to 2000 kDa, about 10 kDa to 1900 kDa, about 10 kDa to 1800 kDa, about 10 kDa to 1700 kDa, about 15 kDa to 1700 kDa, about 15 kDa to 1650 kDa, about 15.5 to 1650 kDa, it may have a molecular weight of about 16 to 1650 kDa.

The GC content of the mixture of nucleic acid fragments may be about 35% to 40%, about 36 to 39%, and about 37% to 38%. The GC content of the mixture of nucleic acid fragments may be measured according to a method known in the art, and may be calculated by calculating the average of the nucleic acids in the mixture of nucleic acid fragments.

The nucleic acid fragment may be derived from yeast (Saccharomyces cerevisiae), virus, or bacteria. Preferably, the nucleic acid fragment may be derived from yeast.

The nucleic acid fragment and the mixture of nucleic acid fragments are extracted with saline [J. Biol. Chem., 203, 167(1953)] or by treatment with an organic solvent or a surfactant such as SDS [J. Chem. Soc., 1129(1947); J. Biol. Chem., 190, 165(1951)], known in the art, such as a method of sedimenting and separating nucleic acid complexes through several steps, such as heat treatment in aqueous solution obtained after treatment, centrifugation to obtain a supernatant, and then lowering the pH again. Can be separated according to the usual method.

The composition may be one that increases the expression of filaggrin or claudin. The main function of keratinocytes, which mainly forms the stratum basal of the skin's epidermis, is to form a barrier against damage from outside the skin. In normal skin, keratinocytes protect the skin from moisture loss and various environmental invasions by expressing structural proteins important for maintaining skin barrier functions such as keratin or filaggrin through the differentiation process. The differentiation process of keratinocytes proceeds by protease and protease inhibitors present in the epidermis of the skin. Through the normal differentiation process of the skin, profilaggrin is synthesized in the granular layer of the epidermis, and proteolytic enzymes act on the synthesized propylraggrin to produce filaggrin. Filaggrin is again decomposed into free amino acids, and the decomposed free amino acids are converted into important pyrolidone carboxylic acid as a natural moisturizing factor (NMF), which is then converted into the stratum corneum. It functions to keep moisture. Claudin is a family of proteins, which is the most important component of 　close joint 　zonulae occludentes　 together with occludin　. Tight junction establishes a barrier between cells that controls the flow of molecules in the intercellular space between epithelial cells and cells. It has four transmembrane domains with N- and C-terminals in the cytoplasm.

The composition increases the expression of genes and/or proteins of claudin and/or filaggrin in human keratinocytes, thereby providing an excellent skin moisturizing effect.

The composition can provide excellent skin regeneration effect by promoting cell growth of human fibroblasts.

The composition can provide excellent skin regeneration effects by promoting cell proliferation of stimulated or damaged human fibroblasts.

The composition can improve the epidermal layer of the skin by improving the lipid biosynthesis effect of human keratinocytes, thereby providing excellent skin barrier strengthening and moisturizing effects.

The composition can provide excellent antioxidant effects by inhibiting the generation of active oxygen in human fibroblasts.

The composition may exhibit an excellent UV blocking effect.

The composition according to an embodiment may contain the mixture in an effective amount or as an active ingredient. The effective amount can be appropriately selected according to the individual. The severity of the disease or condition, the age, weight, health, sex, sensitivity of the individual to the mixture, the time of administration, the route of administration and the rate of excretion, the duration of administration, factors including other compositions used in combination with or concurrently with the composition, and others. It can be determined according to factors well known in the field of physiology or medicine.

The composition for preventing, improving, or treating a skin condition according to an embodiment may further include a cosmetically, food, or pharmaceutically acceptable excipient or carrier. The carrier may be an excipient, a disintegrant, a binder, a lubricant, or a combination thereof. The excipient may be microcrystalline cellulose, lactose, low-substituted hydroxycellulose, or a combination thereof. The disintegrant may be sodium starch glycolate, anhydrous calcium monohydrogen phosphate, or a combination thereof. The binder may be polyvinylpyrrolidone, low-substituted hydroxypropylcellulose, hydroxypropylcellulose, or a combination thereof. The lubricant may be magnesium stearate, silicon dioxide, talc, or a combination thereof.

The composition may be formulated as a parenteral dosage form. The parenteral dosage form may be an injection or an external preparation for skin. The external preparation for skin may be a cream, gel, ointment, skin emulsifier, skin suspension, transdermal delivery patch, drug-containing bandage, lotion, or a combination thereof.

The above skin external preparations are ingredients commonly used in external preparations for skin such as cosmetics and pharmaceuticals, such as aqueous ingredients, oily ingredients, powder ingredients, alcohols, moisturizers, thickeners, ultraviolet absorbers, whitening agents, preservatives, antioxidants, surfactants, fragrances. , Colorants, various skin nutrients, etc. can be appropriately formulated as needed.

The external preparations for the skin include metal sequestering agents such as disodium edetate, trisodium edetate, sodium citrate, sodium polyphosphate, sodium metaphosphate, and gluconic acid, caffeine, tannin, bellapamil, licorice extract, glavidine, and caline. Hot water extract of fruit, various herbal medicines, drugs such as tocopherol acetate, glytilithic acid, tranexamic acid and derivatives or salts thereof, vitamin C, ascorbic acid magnesium phosphate, ascorbic acid glucoside, arbutin, kojic acid, glucose, fructose, Sugars such as trehalose can also be appropriately blended.

The composition according to an embodiment may be a cosmetic composition. In this case, the mixture is a lotion (skin lotion), skin softener, skin toner, astringent, lotion, milk lotion, moisture lotion, nourishing lotion, massage cream, nourishing cream, moisturizing cream, hand cream, foundation, essence, nourishing essence, pack , Soap, cleansing foam, cleansing lotion, cleansing cream, body lotion, body cleanser, suspension, gel, powder, paste, mask pack, or sheet or aerosol composition. In addition, it can be applied in various properties such as liquid, cream, paste, and solid. Compositions of these formulations can be prepared according to methods conventional in the art. The cosmetic composition may further contain preservatives, stabilizers, surfactants, solubilizers, moisturizers, emollients, ultraviolet absorbers, preservatives, disinfectants, antioxidants, pH adjusters, organic and inorganic pigments, fragrances, cooling agents or limiting agents. have. The blending amount of the additional ingredients such as the moisturizing agent can be easily selected by a person skilled in the art within a range that does not impair the object and effect of the present invention, and the blending amount is 0.001 to 5% by weight, or 0.01 to 5% by weight based on the total weight of the composition. It can be %.

The composition according to one embodiment may be a food composition. In this case, it may be formulated into a conventional health functional food formulation known in the art. The food composition may be prepared in general formulations such as, for example, powders, granules, tablets, pills, capsules, suspensions, emulsions, syrups, needles, liquids, and extracts, and meat, sausages, breads, chocolates, candies, snacks , Confectionery, pizza, ramen, other noodles, gums, jelly, dairy products including ice cream, various soups, beverages, tea, drinks, alcoholic beverages, and may be prepared in any health food form such as vitamin complex. For the formulation of the health food, a food pharmaceutically acceptable carrier or additive may be used, and any carrier or additive known to be usable in the art may be used to prepare a formulation to be prepared. As the additive, used in various nutrients, vitamins, electrolytes, flavoring agents, coloring agents, pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohols, carbonated beverages It may contain a carbonation agent and the like. In addition, it may contain flesh for the production of natural fruit juice, fruit juice beverage and vegetable beverage.

These additive components may be used independently or in combination, and the proportion of the additive may be 0.001 to 5% by weight, specifically 0.01 to 5% by weight, based on the total weight of the composition.

The content of the mixture in the food composition may be appropriately determined depending on the purpose of use (prevention or improvement). In general, it may be contained in an amount of 0.01 to 15% by weight of the total food weight, and when prepared as a beverage, it may be contained in an amount of 0.02 to 10 g, specifically 0.3 to 1 g, based on 100 mL.

The beverage may further contain ingredients other than the mixture of the nucleic acid fragments, and may further contain various flavoring agents or natural carbohydrates that are commonly used in beverages. The natural carbohydrates include monosaccharides (e.g., glucose, fructose, etc.), disaccharides (e.g., maltose, sucrose, etc.), polysaccharides (e.g., dextrin, cyclodextrin, etc.), and conventional sugars such as xylitol, sorbitol, erythritol, etc. Sugar alcohol of may be contained. In addition, natural flavoring agents (eg, taumatin, stevia extract, etc.) and synthetic flavoring agents (eg, saccharin, aspartame, etc.) may be included as flavoring agents. The ratio of the natural carbohydrate may be generally contained in about 1 to 20 g, specifically about 5 to 12 g per 100 mL of beverage.

The composition according to one embodiment may be a pharmaceutical composition. Specifically, the pharmaceutical composition may be a pharmaceutical composition for preventing or treating a skin condition.

The skin condition may be a dermatitis or a skin wound.

Another aspect provides a method of improving a skin condition in a subject comprising administering the composition to the subject. The composition is the same as described above.

Specifically, the method may be a skin condition of an individual, such as skin wound improvement, dermatitis improvement, skin regeneration, skin barrier strengthening, skin moisturizing and antioxidant methods.

The terms “administering”, “introducing” and “transplanting” are used interchangeably and one embodiment into a subject by a method or route that results in at least partial localization of a composition to a desired site according to one embodiment. It may mean the arrangement of the composition according to. It can be administered by any suitable route that delivers at least a portion of the mixture of nucleic acid fragments of the composition according to one embodiment to a desired location in a living subject.

Administration may be administered by a method known in the art. Administration can be administered directly to a subject by any means, for example by routes such as intravenous, intramuscular, oral, transdermal, mucosal, intranasal, intratracheal or subcutaneous administration. I can. The administration can be administered systemically or locally.

The subject may be a mammal, for example a human, cow, horse, pig, dog, sheep, goat, or cat. The subject may be a subject in need of skin soothing, eg, anti-inflammatory, atopic dermatitis treatment, skin moisturizing, or erythema reduction effect.

The administration is 0.1 mg to 1,000 mg per individual per day of the nucleic acid fragment mixture, for example, 0.1 mg to 500 mg, 0.1 mg to 100 mg, 0.1 mg to 50 mg, 0.1 mg to 25 mg, 1 mg to 1,000 mg , 1 mg to 500 mg, 1 mg to 100 mg, 1 mg to 50 mg, 1 mg to 25 mg, 5 mg to 1,000 mg, 5 mg to 500 mg, 5 mg to 100 mg, 5 mg to 50 mg, 5 mg To 25 mg, 10 mg to 1,000 mg, 10 mg to 500 mg, 10 mg to 100 mg, 10 mg to 50 mg, or 10 mg to 25 mg may be administered. However, the dosage may be variously prescribed according to factors such as formulation method, administration mode, patient's age, weight, sex, pathological condition, food, administration time, route of administration, excretion rate, and response sensitivity. Taking these factors into account, the dosage can be appropriately adjusted. The number of administration can be once a day or two or more times within the range of clinically acceptable side effects, and the administration site can be administered at one or two or more locations, daily or at intervals of 2 to 5 days. The number of days of administration can be administered from 1 to 30 days per treatment. If necessary, the same treatment can be repeated after an appropriate time period. For animals other than humans, the same dose per kg as human or, for example, the volume ratio (e.g., average value) of the target animal and the human organ (heart, etc.) One amount can be administered.

According to the composition according to one aspect, since it has excellent UV protection, cell regeneration, skin barrier strengthening, skin moisturization, and antioxidant effects, it can be applied to various fields such as cosmetics, foods, pharmaceutical compositions for improvement, prevention or treatment of skin conditions. .

1 is before UVB irradiation to the nucleic acid fragment mixture of Example 1 1% (a), 3% (b), 5% (c), 10% (d) and the nucleic acid fragment mixture (e) of Comparative Example 1 It is the result of measuring the absorbance (% transmission) after ,.
Figure 2 is a graph showing the results of measuring the cell growth rate of the nucleic acid fragment mixture of Example 1 50, 100, 500, 1000 ppm, FBS 1% addition group and control group.
3 is a photograph of cell proliferation of the nucleic acid fragment mixture of Example 1, the nucleic acid fragment mixture of Comparative Example 1, the FBS 1% addition group, and the control group after scraping human fibroblasts.
4 is a graph measuring the relative mRNA levels of claudin and filaggrin, which are moisturizing factors.
5 is a result of photographing lipid-stained keratinocytes.
6 is a graph showing the results of measuring active oxygen in human fibroblasts after UVB irradiation.

Hereinafter, the present invention will be described in more detail through examples. However, these examples are for illustrative purposes only, and the scope of the present invention is not limited to these examples.

Example 1: Preparation of nucleic acid fragment mixture

A mixture of yeast-derived DNA fragments was obtained from Moachem Co., Ltd. (Korea). Yeast (Saccharomyces cerevisiae) was cultured with shaking in YPD (Yeast extract-Peptone-Dxtrose, Difco) at 25 to 30°C, 200 rpm, and aerobic conditions for 48 to 72 hours. The cultured fermentation broth was heat-treated at 100° C. for 5 to 10 minutes to dissolve, and then centrifuged at 5000 to 8000 rpm for 30 to 60 minutes to obtain a supernatant. DNA was precipitated by adding equal amounts of ethanol and an organic solvent to the supernatant. As a result of checking the GC content according to a known method, it was found that adenosine and thymine were 64.2%, and guanosine and cytosine were 37.99%.

Comparative Example 1: Preparation of nucleic acid fragment mixture

A mixture of DNA fragments derived from wheat (Triticum 　 aestivum (vulgare)) was obtained from Moachem Co., Ltd. (Korea). As a result of checking the GC content according to a known general method, it was found that adenosine and thymine were 54.5%, and guanosine and cytosine were 45.5%.

Experimental Example 1: UVB shielding effect confirmation

The UVB shielding effect of the mixture of nucleic acid fragments of Example 1 and Comparative Example 1 was confirmed as follows. After dispensing 50 μl of the Example 1 composition (1, 3, 5, 10%) into a glass plate, it was dried overnight in a desiccator. UVB having a wavelength of 312 nm was irradiated at 160 J/cm ² for 24 hours, and then dissolved sufficiently by mixing in 20 ml of tertiary distilled water. The absorbance was measured before and after UVB irradiation.

1 is before UVB irradiation to the nucleic acid fragment mixture of Example 1 1% (a), 3% (b), 5% (c), 10% (d) and the nucleic acid fragment mixture (e) of Comparative Example 1 It is the result of measuring the absorbance (% transmission) after ,.

As shown in FIG. 1, the difference in absorbance before and after UVB irradiation increased in a concentration-dependent manner when the nucleic acid fragment mixture of Example 1 was treated. In addition, when treating the nucleic acid fragment mixture of Comparative Example 1 used as a control, there was no difference in absorbance before and after UVB irradiation.

Therefore, it was confirmed that the nucleic acid fragment mixture of Example 1 has excellent UVB shielding effect, and this effect increases depending on the concentration of the mixture.

Experimental Example 2: Cell regeneration effect evaluation

2.1. Cell growth assessment

In order to evaluate the cell regeneration effect of the nucleic acid fragment mixtures of Example 1 and Comparative Example 1, the effect on cell growth was confirmed. Human fibroblast Hs68 cells (purchased from Korea Cell Line Bank) were inoculated in a 24-well cell culture dish at a density of 1x10 ⁵ and cultured in a 37°C, 5% CO ₂ incubator for 24 hours. 50, 100, 500, and 1000 ppm of the compositions of Example 1 and Comparative Example 1 were added to the cultured human fibroblasts and cultured for 24 hours. As a positive control, an FBS-added group cultured by adding 1% Fetal bovine serum (FBS) was used, and as a control, cells to which no nucleic acid fragment composition was added were used. Cell growth rate was measured as a ratio to the control.

Figure 2 is a graph showing the results of measuring the cell growth rate of the nucleic acid fragment mixture of Example 1 50, 100, 500, 1000 ppm, FBS 1% addition group and control group.

As shown in Figure 2, the composition of Example 1 was confirmed the effect of increasing the growth rate of about 20% compared to the control. Therefore, it can be seen that the skin cell proliferation effect of the composition of Example 1 is excellent.

2.2. Cell proliferation assessment

In order to evaluate the cell regeneration effect of the nucleic acid fragment mixtures of Example 1 and Comparative Example 1, the effect on proliferation after wounding the cells was confirmed. Human fibroblasts (Hs68 cells (purchased from Korea Cell Line Bank) were inoculated in a 6-well cell culture dish at a density of 4x10 ⁵ and cultured for 24 hours in an incubator at 37° C. and 5% CO ₂ .) Using a cell scraper After scraping the center of the cultured cells, 100 ppm of the composition of Example 1 and Comparative Example 1 were added and cultured for 24 hours. As a positive control, FBS (Fetal bovine serum) 1% was added and cultured FBS was added. A group was used, and as a control, cells to which the nucleic acid fragment composition was not added were used.

3 is a photograph of cell proliferation of the nucleic acid fragment mixture of Example 1, the nucleic acid fragment mixture of Comparative Example 1, the FBS 1% addition group, and the control group after scraping off human fibroblasts.

As shown in Fig. 3, it was confirmed that the composition of Example 1 significantly proliferated skin cells compared to the composition of Example 1.

Accordingly, it was confirmed that the composition of Example 1 has an effect of increasing cell growth and improving proliferation of damaged cells, and thus has excellent cell regeneration effect.

Experimental Example 3: Checking the moisturizing effect

In order to evaluate the skin moisturizing effect of the nucleic acid fragment mixture of Example 1 and Comparative Example 1, the following experiment was performed. Human keratinocytes HaCaT cells (purchased from Korea Cell Line Bank) were inoculated in a 6-well cell culture dish at a density of 4x10 ⁵ and cultured in a 37°C, 5% CO ₂ incubator for 24 hours. Thereafter, 100 ppm of the compositions of Example 1 and Comparative Example 1 were added and then cultured for an additional 24 hours, and the cells were recovered, and 1 ml of trizol (RNA iso, DAKARA, Japan) was added to isolate RNA. After RNA was quantified using Nanodrop 2000 (Thermo, USA), cDNA was synthesized by reacting at 42°C for 55 minutes and 70°C for 15 minutes (Reverse Transcriptase Mix, ELPIS biotech, Korea). Using the synthesized cDNA as a template, a real-time PCR machine using a mixture to which cybergreen (SYBR Green supermix, Applied Biosystems, USA) was added to the target genes claudin and filaggrin One Plus, Applied Biosystems, USA) carried out real-time polymerase chain reaction. Real-time PCR reaction conditions were 94 ℃, after polymerase activation for 5 minutes, 95 ℃ 30 seconds, 54 ℃ 1 minute, 72 ℃ 1 minute under the conditions of 40 cycles were performed under the conditions of the primer sequences are shown in Table 1 below. The expression level of the gene was finally analyzed through correction for the β-actin gene.

Primer name Explanation order Claudin Forward direction 5'-GCTCTAGAATTCCGAGCGAGTCATGGCCAACGC-3' Reverse 5'-GCTCTAGAATTCTCACACGTAGTCTTTCCCGCT-3' Pillar Green Forward direction 5'-AGTGCACTCAGGGGGCTCACA-3' Reverse 5'-CCGGCTTGGCCGTAATGTGT-3' β-actin Forward direction 5'-GGCCATCTCTTGCTCGAAGT-3' Reverse 5'-GACACCTTCAACACCCCAGC-3'

4 is a graph measuring the relative mRNA levels of claudin and filaggrin, which are moisturizing factors.

As shown in FIG. 4, it was confirmed that the gene expression of claudin was increased by about 60% and filaggrin by about 31% compared to the group treated with the composition of Comparative Example 1 when the composition of Example 1 was treated.

Therefore, it can be seen that the nucleic acid fragment mixture according to an aspect has excellent skin moisturizing effect by increasing the expression of claudin and filaggrin.

Experimental Example 5: Lipid Biosynthesis Effect

In order to evaluate the lipid biosynthesis effect of the nucleic acid fragment mixture of Example 1 and Comparative Example 1, the following experiment was performed. Human keratinocytes were inoculated in a 6-well cell culture dish at a density of 4x10 ⁵ and then cultured in a 37°C, 5% CO ₂ incubator for 24 hours. At this time, 10 ppm of the composition of Example 1 was added. Cells to which nothing was added were used as a negative control, and a group to which rosiglitazone was added was used as a positive control. The cultured cells were stained with Nile red and then fluorescence was measured.

5 is a result of photographing lipid-stained keratinocytes.

As shown in FIG. 5, it was confirmed that the composition of Example 1 had a larger amount of lipid observed compared to the composition of Comparative Example 1.

Therefore, it was confirmed that the nucleic acid fragment mixture according to an aspect has excellent effects of promoting lipid biosynthesis, thereby enhancing skin barrier and moisturizing skin.

Experimental Example 6: Confirmation of antioxidant effect

In order to evaluate the antioxidant effect of the nucleic acid fragment mixture of Example 1 and Comparative Example 1, the following experiment was performed. Human fibroblasts were inoculated in a 6-well cell culture dish at a density of 5x10 ⁵ , and then cultured in a 37° C., 5% CO ₂ incubator for 24 hours, and then UVB generating reactive oxygen (ROS) was irradiated. After 24 hours of irradiation, DCF-DA treatment was performed to measure fluorescence. During cell culture, the composition of Example 1 was treated with 50, 125, and 250 ppm as an experimental group. A cell group cultured without any treatment was used as a negative control, and a group treated with 50 ppm of trolox was used as a positive control.

6 is a graph showing the results of measuring active oxygen in human fibroblasts after UVB irradiation.

As shown in FIG. 6, it was confirmed that the active oxygen increased by UVB decreased when the composition of Example 1 was treated, and significantly increased to about 49, 41, and 43% at 50, 125, and 250 ppm treatments, respectively. In addition, it was confirmed that such an increase effect was superior to the comparative example 1 composition-treated group.

Therefore, it was confirmed that the nucleic acid fragment mixture according to an aspect has excellent effect of reducing ROS, and thus has excellent antioxidant effect.

According to the above results, since the nucleic acid fragment mixture of one aspect has excellent antioxidant, skin moisturizing, skin barrier strengthening, cell regeneration, and wound healing effects, it can be usefully used in a composition for improving skin. In addition, it was confirmed that the effect was different when compared to the nucleic acid fragment mixture of one aspect and the nucleic acid fragment mixture having a different GC content.

<110> COSMAX, INC. <120> A composition comprising a mixture of nucleic acid fragments and uses thereof <130> PN125371 <160> 6 <170> KoPatentIn 3.0 <210> 1 <211> 33 <212> DNA <213> Artificial Sequence <220> <223> claudin forward primer <400> 1 gctctagaat tccgagcgag tcatggccaa cgc 33 <210> 2 <211> 33 <212> DNA <213> Artificial Sequence <220> <223> claudin reverse primer <400> 2 gctctagaat tctcacacgt agtctttccc gct 33 <210> 3 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> filaggrin forward primer <400> 3 agtgcactca gggggctcac a 21 <210> 4 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> filaggrin reverse primer <400> 4 ccggcttggc cgtaatgtgt 20 <210> 5 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> beta actin forward primer <400> 5 ggccatctct tgctcgaagt 20 <210> 6 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> beta actin reverse primer <400> 6 gacaccttca acaccccagc 20

Claims (10)

A cosmetic composition for sun protection comprising a mixture of nucleic acid fragments, wherein the mixture has a GC content of 35% to 40%, the nucleic acid is DNA, and the mixture is 3% by weight based on the total weight of the cosmetic composition Cosmetic composition that contains to 10% by weight. The cosmetic composition of claim 1, wherein the nucleic acid fragment has a molecular weight of 10 kDa to 2000 kDa. The cosmetic composition of claim 1, wherein the mixture has a GC content of 37% to 38%. The cosmetic composition of claim 1, wherein the nucleic acid fragment is derived from yeast. delete The cosmetic composition of claim 1, wherein the cosmetic composition further exhibits an effect of improving one or more skin conditions selected from the group consisting of skin wound improvement, dermatitis improvement, skin regeneration, skin barrier strengthening, skin moisturizing and antioxidant. delete A composition for external skin for sun protection comprising a mixture of nucleic acid fragments, wherein the GC content of the mixture is 35% to 40%, the nucleic acid is DNA, and the mixture is 3 based on the total weight of the composition for external application for skin. The composition for external application for skin that is contained in a weight% to 10% by weight. The composition for external application for skin according to claim 8, wherein the composition for external application for skin further exhibits an effect of improving one or more skin conditions selected from the group consisting of skin wound improvement, dermatitis improvement, skin regeneration, skin barrier strengthening, skin moisturizing and antioxidant. delete

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