JP5025254B2 - Cosmetic preparations and cosmetics for improving the skin groove density - Google Patents

Description

  The present invention relates to a cosmetic formulation that promotes the regeneration or activation of human skin (skin) cells and improves the crevice density (texture), and a cosmetic containing the cosmetic formulation.

Rough skin and stains include effects of hormonal balance due to aging, stimulation of ultraviolet rays from sunlight, adverse effects on the epidermis due to increased peroxide concentration in the body, excessive secretion of sebum, decreased blood flow to the epidermis, nutrition It is thought to be caused by various things such as badness and mental stress.
The fact that rough skin and wrinkles are a serious concern for people's beauty has recently appeared in an increase in the number of cosmetic products for men as well as for women. Because the face is a big factor that gives a first impression to others, many people want to have glossy and firm skin (beautiful skin) regardless of age or gender, therefore rough skin and wrinkles The general public's interest in cosmetics that improve the quality of the cosmetics is increasing.

  In conventional cosmetics, many attempts have been made to enhance the moisturizing effect of the skin by adding a moisturizing agent such as glycerin or an oil component to make the skin glossy. Recently, cosmetics containing L-ascorbic acid and its derivatives, which have the function of suppressing melanin production and promoting collagen production, have been introduced with the intention of having a beautiful skin effect.

Meanwhile, in recent years, health foods using deoxyribonucleic acid (DNA), ribonucleic acid (RNA) or nucleoprotein as raw materials or active ingredients have been provided, reflecting the increasing public interest in health. In order to make high molecular weight nucleoprotein water-soluble and easily digestible, it has also been proposed to reduce the molecular weight (Patent Document 1).
Although deoxyribonucleic acid (DNA), ribonucleic acid (RNA) or nucleoprotein is known to have an anti-aging effect, attempts to apply it to cosmetics have not been made sufficiently.
JP 2004-16143 A

Causes of skin troubles such as rough skin and wrinkles are various, such as facial skin aging, poor blood circulation, the influence of ultraviolet rays, stress, etc., but the decrease in skin cell metabolism and the renewal function of new cells directly Cause.
Conventional cosmetics do not work by acting on the metabolism mechanism of skin cells, but merely pursue effects on the surface of the skin. For this reason, the effect until it can be said sufficient with respect to rough skin, wrinkles, etc. is not acquired.
Moreover, in the method of applying the above-mentioned L-ascorbic acid and its derivative, since it is not stable and the ultraviolet-ray inflammation prevention effect is inadequate, the content component of the said compound is calculated | required directly as an active ingredient of cosmetics. It is difficult.
In other words, there is a demand for a product with a high skin beautifying effect that enhances the regenerative and activating ability of human skin cells to promote the regeneration of new skin cells and prevents rough skin and wrinkles. The effect is still not enough.
Therefore, it has been desired to develop a new cosmetic product that activates the cells themselves to make the skin alive.

As a result of earnest research to obtain a new cosmetic product that improves the texture of human skin, the present inventors have obtained a deoxyoligonucleotide, deoxynucleotide obtained by enzymatic or hydrolytic treatment of nucleoprotein and / or DNA. Excellent cell regeneration for degradation products containing mononucleotides or oligopeptides, or degradation products containing oligonucleotides or mononucleotides obtained by enzymatic degradation or hydrolysis of RNA, or mixtures thereof And found that it has a cell activation effect, which leads to an improvement effect of the skin groove density, an improvement effect of the moisture retention ability of the skin and a balance of sebum amount, and an effect of preventing wrinkles, etc. Was completed.

That is, the cosmetic compounding agent according to the present invention is a fraction having a molecular weight of 1000 to 3000 obtained by reducing the molecular weight of RNA by enzymatic degradation or hydrolysis, and 20 to 20 oligonucleotides having a chain length of 2 to 12. It is a cosmetic preparation for improving the crevice density containing RNA degradation products containing up to 50% (except for a preparation containing β-alanine).
The RNA degradation product preferably contains 41.1% of an oligonucleotide having a chain length of 2 to 12.
Alternatively, the cosmetic compounding agent for improving the crevice density according to the present invention has a molecular weight of 1,000 to 1,000 obtained by further reducing the molecular weight of nucleoprotein and DNA by enzymatic degradation or hydrolysis in addition to the RNA degradation product. The fraction of 3000 may contain nucleoprotein degradation products and DNA degradation products containing 20-50% deoxyoligonucleotides with chain lengths of 2-12.

In a preferred embodiment of the cosmetic compounding agent of the present invention, the nucleoprotein and DNA are preferably obtained from fish larvae such as salmon, salmon, salmon and salmon.
The RNA is obtained from yeast, preferably from yeast selected from the group consisting of beer yeast, torula yeast, milk yeast and baker's yeast.

  Furthermore, the cosmetic product according to the present invention is characterized by containing a cosmetic compounding agent for improving the skin groove density.

The cosmetic compounding agent of the present invention is a degradation product containing 20 to 50% of a fraction having a molecular weight of 1000 to 3000 obtained by reducing the molecular weight of nucleoprotein and / or DNA or RNA by enzymatic degradation or hydrolysis. A deoxyoligonucleotide, deoxymononucleotide, oligopeptide, oligonucleotide, mononucleotide separated from the degradation product, or a mixture of at least two selected from the degradation product or the compound as an active ingredient is there.
Since the deoxyoligonucleotides and the like have a relatively small molecular weight, they are easily absorbed percutaneously, and when absorbed percutaneously, they have a function of enhancing the cell regenerative ability or cell activation ability of the skin. Therefore, when applied to the epidermis of the face, it is effective in improving the skin groove density (texture), moisture and sebum, and further improving wrinkles.

  In addition, since the cosmetic of the present invention comprises the above-mentioned cosmetic compounding agent, when applied to the epidermis of the face, the cosmetic compounding agent has excellent skin roughness, wrinkles and the like, and has a skin beautifying effect.

  The deoxyoligonucleotide or deoxymononucleotide used as a refined product as an active ingredient of the cosmetic compounding agent of the present invention is obtained by subjecting DNA to an enzymatic degradation treatment or hydrolysis treatment, and the oligonucleotide or mononucleotide is an enzyme that transforms RNA into an enzyme The oligopeptide can be obtained by subjecting a nucleoprotein to an enzymatic degradation treatment or a hydrolysis treatment, respectively.

  DNA and nucleoprotein can be obtained, for example, by extracting and purifying from fish larvae. Examples of the fish include salmon, salmon, salmon and salmon, and salmon is particularly preferable.

Hereinafter, DNA will be described in more detail.
The DNA that is the raw material for producing the cosmetic compounding agent of the present invention may have various forms, for example, double-stranded, single-stranded or circular DNA. The source of DNA is various organisms such as animals, plants, and microorganisms. Fish, particularly sharks, sharks, sharks, and shark testes (shirako), which are wastes from marine processing, contain a lot of DNA, but they have not been effectively used as resources, and many of them have been discarded. Therefore, it is desirable to use these testis-derived DNA also from the viewpoint of waste recycling. In addition, DNA obtained from thymus of mammals and birds, for example, cows, pigs, chickens and the like can be used. In addition, synthetic DNA can also be used.

In addition, in order to obtain DNA from a fish white child, the extraction / purification method described in JP-A-2005-245394 can be used.
Specifically, firstly, the larvae of the fish are crushed, the crushed fish larvae are subjected to a proteolytic enzyme (protease) treatment under conditions where DNA is not degraded, and the enzyme-treated solution is filtered. The filtrate is dialyzed using a hollow fiber membrane having a molecular weight cut off of 2,000 to 1,000,000 to remove decomposed proteins and ions and concentrate double-stranded DNA. Further, the dialysis solution is precipitated as a double-stranded DNA salt, or the solution is concentrated, and the precipitate or concentrate is recovered.
A powdered DNA salt obtained by drying the DNA salt obtained by the above method can be used as a raw material for producing the cosmetic compounding agent of the present invention.

  RNA can be obtained by extracting and purifying from a yeast selected from the group consisting of beer yeast, torula yeast, milk yeast and baker's yeast.

  The enzyme that processes DNA and RNA is, for example, a nuclease, and particularly, a nuclease derived from blue mold.

  Oligopeptides can be obtained by hydrolyzing nucleoprotein (nuclear protein) contained in fish larvae with protease and nuclease.

  The protease is mainly composed of trypsin. Trypsin is a serine protease having high specificity and selectively hydrolyzes peptide bonds on the carboxyl side of arginine and lysine, and is therefore suitable for hydrolysis of protamine containing a large amount of arginine. The protease can also contain other proteases such as chymotrypsin in addition to trypsin. An example of a good protease is a protease manufactured by Novozymes Japan.

The nuclease hydrolyzes the 3 ′, 5′-phosphodiester bonds of deoxyribonucleic acid (DNA) and ribonucleic acid (RNA) to produce 5 ′-(deoxy) nucleotides for oligobody polymerization. Although there is no restriction | limiting in particular about the property of this nuclease, It is preferable to provide a certain amount of thermal stability. Such a nuclease is commercially available from, for example, Amano Enzyme Co., Ltd., Sigma Co., etc.

What is important in the hydrolysis treatment of DNA, RNA and nucleoprotein using the nuclease is the temperature at which the reaction is carried out. The reaction temperature must be in the range of 60-75 ° C, most preferably 70 ° C. When the reaction is carried out at a temperature lower than the temperature range, DNA, RNA
In addition, the molecular weight of the nucleoprotein does not sufficiently progress, and the degradation product does not become water-soluble. On the other hand, when the reaction is performed at a temperature higher than the above temperature range, the reduction of the molecular weight proceeds excessively and the excellent effect of the nucleoprotein (nucleoprotein) may be lost.

As described above, DNA, RNA, and nucleoprotein are treated by hydrolysis using nuclease at 60 to 75 ° C., so that a fraction having a molecular weight of 1000 to 3000 is contained in an amount of 20 to 50%, and the molecular weight is 1000 or less. The molecular weight of the fraction is usually higher than that of the fraction having a molecular weight of 1000 to 3000, for example, 30 to 50%, thereby improving water solubility and transdermal absorbability. Combined DNA, RNA and nucleoprotein degradation products can be produced.
Nucleoprotein degradation products and DNA degradation products obtained by the above treatment contain deoxyoligonucleotides, deoxymononucleotides and oligopeptides that have been reduced in molecular weight as the main part or most of the reduced molecular weight. Insufficient deoxynucleotides and the like are contained as a very small portion.
In addition, RNA degradation products obtained by the same treatment contain oligonucleotides and mononucleotides as major parts or most parts, and nucleotides with insufficient molecular weight are contained in very small parts. .
Therefore, the nucleotides (deoxyoligonucleotides, deoxymononucleotides, oligonucleotides, mononucleotides) contained in these degradation products are almost all or most of them are mono- or complete ones that do not inherently have a helical chain. It is composed of a chain oligo body and an oligo body having only a part of a double helix structure. In other words, even if it is assumed that the above-described reduction in molecular weight has not progressed sufficiently, the double helix ratio of nucleotides contained in the degradation product does not exceed 20%.
In order to obtain an oligopeptide by hydrolyzing a nucleoprotein, a protease treatment and a nuclease treatment are required to obtain an oligonucleotide. Preferably, treatment with a protease is performed first, followed by nuclease treatment. It is desirable from the viewpoint of work convenience and the quality of the final product obtained.

The cosmetic preparation of the present invention has, as its active ingredient, a degradation product obtained by enzymatic degradation or hydrolysis of nucleoprotein and / or DNA, or an enzymatic degradation or hydrolysis treatment of RNA. The resulting degradation product can be used as such (without purification). In the decomposition product, for example, an amino acid or the like may be contained.
In addition, as an active ingredient of the cosmetic compounding agent of the present invention, the following compound separated and purified from nucleoprotein and / or DNA or RNA degradation products using conventional separation means and / or purification means: Deoxyoligonucleotide , Deoxymononucleotides, oligopeptides, oligonucleotides and mononucleotides can be used.
At this time, deoxyoligonucleotides and oligonucleotide strands contained in the degradation products of nucleoprotein, DNA and RNA, or separated / purified from the degradation products using conventional separation means and / or purification means The length is preferably 2-12.

The decomposition product and the compound may be used alone or in combination of at least two of them.
When the decomposition product or the compound is used in combination, the mixing ratio can be appropriately selected.
At this time, the degradation product and the compound include at least one of the deoxyoligonucleotide having a chain length of 2 to 12 or the oligonucleotide, and the deoxyoligonucleotide having a chain length of 2 to 12 and the compound. It is particularly preferable that the total content of oligonucleotides is 20% or more based on the total amount of the degradation products and the compounds.

The concentration of the active ingredient (the decomposition product and / or the compound) in the cosmetic preparation of the present invention can be appropriately selected.
Moreover, you may use the said decomposition product and the said compound further combining with the conventional additive component of the cosmetic compounding agent in a predetermined ratio.

The cosmetic of the present invention contains the cosmetic compounding agent.
The dosage form that can be taken by the cosmetic of the present invention can be appropriately selected as long as it is a dosage form applicable to the skin. Preferably, the cosmetic is in a form that is directly applied to the skin, such as lotion, emulsion, cream, gel, jelly, essence, pack, mask, foundation. These cosmetics can be used not only on the skin of the face but also on the entire body such as the neck and limbs.

  The cosmetics of the present invention can be blended with known ingredients that can be blended into existing cosmetics in addition to the cosmetic compounding agents. For example, a fragrance | flavor, a moisturizer, etc. can be mix | blended individually or in combination.

In the cosmetic of the present invention, vitamin E or a derivative thereof, for example, vitamin E acetate; a vasodilator such as an acetylcholine derivative; a skin function enhancer such as cephalanthin; glycyrrhetinic acid or a derivative thereof; estradiol, estrone, etc. Female hormone agents; amino acids such as serine, methionine, arginine; vitamin A, vitamin B ₁ ,
Vitamins such as vitamin B ₆ , biotin, pantothenic acid or derivatives thereof can be used alone or in combination.

  Furthermore, in the cosmetics of the present invention, additives that are usually used for external preparations for skin such as cosmetics and pharmaceuticals, for example, oils, preservatives, surfactants, dispersion stabilizers, thickeners, wetting are optionally used. An agent, an ultraviolet absorber, an antioxidant, a pH adjuster, purified water, alcohol and the like can be used alone or in combination.

In the following examples and comparative examples, the present invention will be described in detail and in detail. The following examples are for illustrative purposes only and are not intended to limit the technical scope of the present invention.
The compounding amount of each component in the following examples and comparative examples is mass% with respect to the total amount. In addition, the amount of prototype 1 to prototype 3 (each containing a degradation product obtained by enzymatic degradation of DNA salt, DNA and nucleoprotein, or RNA) used in the examples is the solid content Show.

1. Production of (Deoxy) Oligonucleotide Limited digestion was performed on DNA derived from coconut bud using a nuclease [for example, enzyme preparation nuclease “Amano” (manufactured by Amano Enzyme)] approved as a food additive. The produced deoxymononucleotide and deoxyoligonucleotide were analyzed with an electrophoresis apparatus to determine the optimum conditions.
Specifically, powdered DNA-Na salt is added as raw material to warm water adjusted to around 65 ° C., stirred, and further heated to 70 ° C., and nuclease is 0.05 to 0.25% based on the raw material. An appropriate amount was added within the range of 3 to react for 3 hours. Next, after heating at 85 ° C. for 10 minutes to inactivate the nuclease, the mixture was centrifuged, and spray drying was applied to the supernatant to obtain a dry powder (degradation product) containing deoxyoligonucleotide.

2. Analysis of (deoxy) oligonucleotide A powdery DNA-Na salt derived from white coconut was added as raw material to warm water adjusted to around 65 ° C.,
After stirring, the mixture was further heated to 70 ° C., 0.05% of the enzyme preparation nuclease “Amano” (manufactured by Amano Enzyme) was added to the raw material and reacted for 3 hours to obtain a decomposition product. Next, the nuclease was inactivated by heating at 85 ° C. for 10 minutes, and then the decomposition product was analyzed by HPLC.
FIG. 1 shows an analysis example of deoxyoligonucleotides as degradation products by HPLC (high performance liquid chromatography). In FIG. 1, 5′-deoxymononucleotide and 3′-deoxymononucleotide are eluted up to peak 20, and the subsequent relatively large peak, that is, peak 26 and later, can be regarded as absorption of deoxyoligonucleotide. Further, after the peak 41, it can be regarded as absorption of a decomposition product having a molecular weight exceeding 3000. For this reason, as a result of calculating from the peak intensities of the peaks 26 to 41, it was found that in this example, a fraction of 31% deoxyoligonucleotide (molecular weight 1000 to 3000) was contained with respect to the entire decomposition product. .

3. Manufacture of cosmetics using (deoxy) oligonucleotide A dry powder (DNA salt degradation product) containing deoxyoligonucleotide obtained by the above-described manufacturing method is used as prototype 1, and using this prototype 1, the cosmetic of the present invention Was manufactured as follows. As a control, a cosmetic product containing no prototype 1 was produced. These compositions are summarized in Table 1 below.
[Example 1]
Purified water is added to 95% ethanol, and polyoxyethylene (25 mol) hydrogenated castor oil ether, glycerin and propylene glycol are added and stirred. I got cosmetics.
[Comparative Example 1]
A cosmetic product of Comparative Example 1 was obtained in the same manner as in Example 1 except that the same amount of glycerin was used instead of Prototype Product 1.

4). Evaluation of skin condition using skin image analysis system For subjects in their 30s and 50s, the cosmetic product of Example 1 and the cosmetic product of Comparative Example 1 were placed on the left and right halves of the face, about 1 g / time / day after bathing. , Respectively.
Using an In-Forward Robo Skin Analyzer CS50 system set, the skin condition of the applied part before the test (initial value) and after 4 weeks was evaluated.
4-1. Evaluation of water content and sebum content The states of water content and sebum content were measured using an oil moisture sensor. The sensor was applied to the left and right cheeks, and the measurement results were evaluated by the following measurement method and evaluation criteria.
・ Oil content (the amount of sebum secreted on the skin surface)
The oil component rolled by pressing the sensor part is measured by the refractive index and evaluated by the area. The state where the entire surface was covered with oil was evaluated as “oil saturated” (100), and the state where no oil was present was evaluated as “oil free (0)”.
・ Moisture (water content of living body surface)
The capacitance of the skin is measured by pressing the sensor part and measured as a value representing moisture. The state where the physiological saline itself was measured was evaluated as “saturated state (100)”, and the state where no water was present was evaluated as “zero state (0)”.
4-2. Evaluation of skin groove density (skin texture) The state of skin groove density on the left and right cheeks was photographed using a microscope, and the photographed images were analyzed and evaluated with a Robo Skin Analyzer.
In the analysis method, the dark part in the photographed image (converted into a monochrome image) is a skin groove, the bright part is a skin hill, the light and dark part is processed and binarized, and a regular triangle texture model with a side of 0.4 mm. Compared with (referring to the condition of the skin under the chin), the model image is taken as 100 and the photographed image is evaluated.
4-3. Evaluation of wrinkles The condition of the wrinkles under the eyes was evaluated by analyzing the skin image measured in the full face photography box.
In the photographed image (converted to a monochrome image), a portion where the density was extremely steep and dark was detected as a wrinkle under the eyes, and the number of wrinkles was counted.

The evaluation of the test is shown in the following Table 2 (subject: 30s) and Table 3 (subject: 50s), and the skin condition change (photographed image) of each subject is shown in FIG. 2 and FIG.
Moreover, the figure (graph) which described the said test result and the average value of each age is shown in FIG.4 and FIG.5.

As shown in Table 2, Table 3, FIG. 4 and FIG. 5, the results of Example 1 showed improvement in water content, sebum content, and crevice density in subjects in their 30s and 50s. The amount of water (30's and 50's) and skin groove density (30's) that were below the average before the start of the test recovered to the average after 4 weeks. The number of wrinkles also decreased to the average value or close to the average value after 4 weeks.
Moreover, it has confirmed visually also from the picked-up image shown in FIG.2 and FIG.3 that the skin groove density (texture) was improved.
On the other hand, in Comparative Example 1, due to the moisturizing effect of the blended glycerin and the like, the result was somewhat improved or comparable to that before the start of the test (initial value), but improved compared to Example 1 above. As a result, there was a big difference in the effect.

5. Production and analysis of DNA and nucleoprotein degradation product (prototype 2) or RNA degradation product (prototype 3) Obtained by enzymatic degradation of DNA and nucleoprotein produced by Nissei Bio Inc. as active ingredients Prototype 2 (containing deoxyoligonucleotides, deoxymononucleotides and oligopeptides) containing degradation products, and obtained by enzymatic digestion of RNA produced by Nissei Bio Inc. as an active ingredient The product containing the resulting degradation product was designated as prototype 3 (containing oligonucleotide and mononucleotide).
The manufacturing method and analysis results of Prototype 2 and Prototype 3 are as follows.

5-1. Production method and analysis of Prototype 2 (DNA and nucleoprotein degradation products) Prototype 2 is a degradation product obtained by reducing the molecular weight of DNA by enzymatic degradation using the same production method as in Prototype 1. And an equal amount of degradation products obtained by reducing the molecular weight of nucleoprotein by enzymatic degradation. The detail of the manufacturing method of each decomposition product is as follows.
<DNA degradation product>
[Production method]
Production was carried out in the same manner as in “1. Production of (deoxy) oligonucleotide” above to obtain a DNA degradation product.
[Structure and composition]
Analysis is performed in the same manner as in “2. Analysis of (deoxy) oligonucleotide” above, and a fraction of 31% deoxyoligonucleotide (molecular weight 1000 to 3000) is contained in the entire degradation product. I found out.
<Nucleoprotein degradation products>
[Production method]
Nucleoprotein derived from white cocoon (Nissei Bio Co., Ltd.) is added to water as a raw material, heated and stirred at 50 ° C., and then the enzyme protease “PTN” (manufactured by Novozymes Japan Co., Ltd.) 0.065% is added and reacted for 4 hours, and further heated and stirred at 70 ° C. Subsequently, 0.1% of the enzyme preparation nuclease “Amano” (manufactured by Amano Enzyme) was added and reacted for 3 hours. Next, the nuclease was inactivated by heating at 85 ° C. for 10 minutes, followed by centrifugation and applying a spray drying method to obtain a dry powder (decomposition product) containing deoxyoligonucleotide.
[Structure and composition]
The obtained decomposition product was analyzed by HPLC.
FIG. 6 shows an analysis example of deoxyoligonucleotide as a decomposition product by HPLC (high performance liquid chromatography). In FIG. 6, peaks (peak 1 to peak 4) within a retention time of 19 minutes to 24 minutes can be regarded as oligonucleotide absorption. For this reason, as a result of calculating from the peak intensities of peaks 1 to 4, in this example, a fraction of 33.4% deoxyoligonucleotide (molecular weight 1000 to 3000) is included in the entire decomposition product. all right.

5-2. Production method and analysis of Prototype 3 (RNA degradation product) Prototype 3 is a degradation method obtained by reducing the molecular weight of RNA instead of raw DNA by enzymatic degradation using the same production method as in Prototype 1. Details of the product are as follows. <RNA degradation product>
[Production method]
Yeast-derived RNA (Nissei Bio Co., Ltd.) is added as raw material to warm water adjusted to around 70 ° C, stirred, and further heated to 70 ° C. The enzyme preparation nuclease "Amano" (Amano Enzyme) is added to the raw material. 0.05%) was added and reacted for 3 hours. Next, the nuclease was inactivated by heating at 85 ° C. for 10 minutes, followed by centrifugation and applying a spray drying method to obtain a dry powder (decomposition product) containing the oligonucleotide.
[Structure and composition]
The obtained decomposition product was analyzed by HPLC.
FIG. 7 shows an analysis example of oligonucleotides of degradation products by HPLC (high performance liquid chromatography). In FIG. 7, a peak (peak 2 to peak 5) within a retention time of 13 minutes to 24 minutes can be regarded as oligonucleotide absorption. For this reason, as a result of calculating from the peak intensities of peaks 2 to 5, it was found that in this example, a fraction of 41.1% of oligonucleotide (molecular weight 1000 to 3000) was contained with respect to the entire degradation product. It was.

6). Manufacture of various cosmetics Prototype 2 (deoxyoligonucleotide, deoxymononucleotide and oligopeptide) containing degradation products obtained by enzymatic degradation of DNA and nucleoprotein produced by Nissei Bio Inc. as active ingredients And an active ingredient containing a degradation product obtained by enzymatic degradation of RNA produced by Nissei Bio Inc. Stuff).
Using the obtained prototype 2 and prototype 3, the cosmetic of the present invention was produced as follows. Moreover, the cosmetics of the comparative example which does not contain the prototype 2 and the prototype 3 were manufactured as a control. The compositions of these cosmetics are summarized in Tables 4 to 7 below.
In addition, the acquisition source of the product used for the following cosmetics is shown in parentheses.

6-1. Gel-like cosmetic [Example 2 (Examples 2-1 to 2-3)]
While stirring in purified water, Pemulen TR-1 (Nikko Chemicals Co., Ltd.) was gradually added and sufficiently dispersed, and then Ultraz-10 (Nikko Chemicals Co., Ltd.) was stirred and dispersed, and then concentrated glycerin was added to the gel. A substrate was used. Separately, Resinol WS-50 (Nikko Chemicals Co., Ltd.), Jojoba Oil, Squalane, Real Curve SR-1000 (LIALCARB: Nikko Chemicals Co., Ltd.), AMITER MA-HD (Nihon Emulsion Co., Ltd.) and Rosehip Oil are heated. Dissolved and mixed to obtain an oily component. Separately, Ceralipid W-2 (Nikko Chemicals Co., Ltd.), 1,3-butylene glycol (BG), dipropylene glycol (DPG) and 2-phenoxyethanol were added and superheated with stirring (300 rpm, maintained at 80 ° C. for 5 minutes). Then, after adding purified water and stirring sufficiently, the oil components were combined and mixed thoroughly to obtain a blended component stock solution.
The gel base material and the blended component stock solution were mixed, ethanol was added thereto, 18% sodium hydroxide was added, and the mixture was neutralized and stirred. Each of the prototype 2, mixed with purified water, and a mixture of equal amounts of the prototype 2 and the prototype 3 (prototype 4) was dissolved in the gels of Examples 2-1 to 2-3. A cosmetic product was prepared.
[Comparative Example 2]
In the same manner as in Example 2-1, Example 2-2, and Example 2-3 except that 2% by mass of concentrated glycerin was added instead of prototype 2 and prototype 3, and 7% by mass in total was added. A gel cosmetic product of Comparative Example 2 was prepared.

6-2. Emulsion cosmetics [Example 3 (Examples 3-1 to 3-3)]
Concentrated glycerin was added to purified water and heated to 70 ° C. (aqueous phase). Separately, monooleate and glycerol monostearate were added to cetyl alcohol, beeswax, petrolatum, squalane and dimethylpolysiloxane and heated to 70 ° C. This was added to the previously prepared aqueous phase and pre-emulsified. Furthermore, the quince seed extract and ethanol were added and stirred, and the emulsified particles were made uniform with a homomixer.
When this emulsion liquid reached 40 ° C., each of Prototype 2, Prototype 3, and an equal mixture of Prototype 2 and Prototype 3 (Prototype 4) mixed with purified water was dissolved. -1 to 3-3 milky cosmetics were prepared.
[Comparative Example 3]
The emulsion of Comparative Example 3 was prepared in the same manner as in Examples 3-1 to 3-3, except that 2% by mass of concentrated glycerin was added instead of Prototype 2 and Prototype 3, and a total of 10% by mass was added. A cosmetic product was prepared.

6-3. Lotion cosmetics [Example 4 (Examples 4-1 to 4-3)]
1,3-butylene glycol, concentrated glycerin, a buffering agent and a browning inhibitor were dissolved in purified water at room temperature. Also, each of Prototype 2, Prototype 3, and Equal Mix of Prototype 2 and Prototype 3 (Prototype 4) mixed with oleyl alcohol, sorbitan monolaurate and purified water in ethanol is dissolved and mixed. The mixture was stirred to prepare lotion-like cosmetics of Examples 4-1 to 4-3.
[Comparative Example 4]
The lotion form of Comparative Example 4 was prepared in the same manner as in Examples 4-1 to 4-3 except that 2% by mass of concentrated glycerin was added instead of Prototype 2 and Prototype 3 and a total of 6% by mass was added. A cosmetic product was prepared.

6-4. Creamy cosmetic [Example 5 (Examples 5-1 to 5-3)]
1,3-Butylene glycol, pentylene glycol, concentrated glycerin, and polyquaternium were sequentially dissolved in purified water and heated to 80 ° C. to obtain an aqueous phase. On the other hand, polyglyceryl stearate, stearyl alcohol, behenyl alcohol, batyl alcohol, cetyl palmitate, glyceryl stearate, Crodaran SWL (Croda Japan Co., Ltd.), isopropyl palmitate, squalane, octyldodecyl myristate, macadamia nut oil, trioctanoin Dimethicone was mixed and heated and stirred at 85 ° C. to obtain an oil phase. The oil phase was poured into the aqueous phase, stirred (300 rpm), and emulsified with a high viscosity homomixer (4000 rpm).
When this emulsified substance reaches 40 ° C., trial water 2, sodium citrate and purified water mixed with purified water, prototype 3, and a mixture of prototype 2 and prototype 3 in equal amounts (prototype) Each of 4) was dissolved to prepare creamy cosmetics of Examples 5-1 to 5-3. [Comparative Example 5]
Instead of Prototype 2 and Prototype 3, 2% by mass of concentrated glycerin was added, and a total of 7% by mass was added, and the cream of Comparative Example 5 was used in the same manner as Example 5-1 to Example 5-3. A cosmetic product was prepared.

7). Performance test of cosmetics (gel, milky lotion, lotion, cream) The cosmetics of Examples 2 to 5 and Comparative Examples 2 to 5 were evaluated as follows. The results are summarized in Tables 4 to 7.
7-1. State of skin groove density (texture) For the subjects of middle-aged and elderly women, the cosmetics of Examples 2 to 5 and the cosmetics of Comparative Examples 2 to 5 on the left and right half of the face, about 1 g / time / day after bathing, respectively. Applied.
Using the above-mentioned Inforward Robo Skin Analyzer CS50 system set, the skin crevice density of the applied part after 4 weeks was evaluated as described above. The test results were determined according to the following criteria.
++: The numerical value of the skin groove density of the applied part increased by 10% or more compared with the comparative example (high effect)
+: The numerical value of the skin groove density of the coated part was increased by 5 to 10% compared to the comparative example (effective)
±: The numerical value of the skin groove density of the applied part increased by 0 to 5% compared to the comparative example (somewhat effective)
-: The numerical value of the skin groove density of the coated part was equal to or less than that of the comparative example (invalid)

7-2. Skin roughness improvement test 10 middle-aged and elderly women with skin roughness in the lower limbs were tested, and the cosmetics of Examples 2 to 5 and Comparative Examples 2 to 5 were each placed at different locations on the left and right lower limbs at about 1 g / It was applied once / day.
The test results were judged from the viewpoints of the peeled state and moisture state of the skin, using the following judgment criteria, depending on the state of the skin at the place where the test was applied and one month later.
7-2-1. Criteria for skin peeling and judgment of effectiveness The state of skin peeling before and after the test was judged according to the following criteria.
0: No peeling 1: Mild peeling 2: Moderate peeling 3: Peeling severity Based on the above judgment, the result after the test was improved by one level from the level before the test was regarded as “slightly effective” and improved by two or more levels. Was “valid”, and those with the same level or worsened were “invalid”.
7-2-2. Criteria for determination of moisture state The moisture state (moisturizing property) of the skin after one month was measured with a moisturizer, and determined according to the following criteria.
<Judgment criteria for moisture status>
++: The moisturizing property of the applied part increased by 5% or more from before the test (high effect)
+: The moisturizing property of the applied part increased by 2 to 5% (effective) from before the test.
±: Moisturizing property of the applied part increased by 0 to 2% from before the test (slightly effective)
−: The moisture retention of the applied part was equal to or less than that before the test (invalid)

7-3. Evaluation test by trial The evaluation test of the cosmetics of Examples 2 to 5 and the cosmetics of Comparative Examples 2 to 5 was conducted in a single-blind manner for middle-aged women. The number of examinees for each cosmetic product was 10. The evaluation was based on a questionnaire for the examinees, and answers were obtained before use and after 1 month of use for two items of “smoothness” and “fineness of texture”.
Based on the obtained answers, the test results were determined according to the following criteria.
++: Improved compared to before use (effective)
+: Slightly improved compared to before use (somewhat effective)
±: Less than or equal to before use (invalid)

As shown in Tables 4 to 7, the cosmetics of Examples 2 to 5 containing prototype 2, prototype 3, or prototype 4 (an equivalent mixture of prototype 2 and prototype 3) are prototypes 2 to Compared with the cosmetics of Comparative Examples 2 to 5 not containing 4, the state of the skin groove density was remarkably improved.
In addition, the cosmetics of Examples 2 to 5 obtained results that the effectiveness was almost remarkable in the rough skin improvement test, and the test results that the improvement effect was remarkably recognized also in the evaluation test by trial use were obtained. .
On the other hand, the cosmetics of Comparative Examples 2 to 5 showed almost no improvement effect in both the rough skin improvement test and the evaluation test by trial use, and the effectiveness of the cosmetics including the prototypes 2 to 4 became clear.

  In Examples 1 to 5 described above, prototype 1 (DNA salt degradation product), prototype 2 (DNA and nucleoprotein degradation product), prototype 3 (RNA degradation product), and prototype as active ingredients. 4 (mixture of Prototype 2 and Prototype 3) was used, but instead of Prototypes 1 to 4, deoxyoligonucleotides, deoxymononucleotides, oligopeptides, oligonucleotides or mononucleotides separated and purified from them Even when these were used alone or in combination, the same effects as in the case of using prototypes 1 to 4 were recognized.

  That is, from this example, a preparation containing deoxyoligonucleotide or the like (at least one of deoxyoligonucleotide, deoxymononucleotide, oligopeptide, oligonucleotide, mononucleotide) has a skin condition (water content, sebum amount, skin groove). It was found to have an effect of improving density and wrinkles). In particular, it can be said that the test result that the crevice density is improved is a result supporting that deoxyoligonucleotide or the like has cell regeneration or cell activation ability.

Examples of prescription of cosmetics using the prototypes 1 to 4 are shown as Examples 6 to 8 below. The “prototype (class)” described here is any one selected from prototypes 1 to 3 alone or an equivalent mixture of prototype 2 and prototype 3 (prototype 4). It shall represent meaning.
In any of the examples, it was confirmed that the skin water content and sebum content were improved, and the condition of the crevice density was improved, and the effect of reducing wrinkles was observed.
It was found to have an effect of improving the smoothness and fineness of the skin.

Example 6: Cold cream
<Manufacturing method>
Prototype (s) and soap powder were added to purified water and dissolved by heating and kept at 70 ° C. (aqueous phase). The other components were mixed, heated and melted and kept at 70 ° C. (oil phase). The oil phase was gradually added to the aqueous phase with stirring, and after completion, the mixture was uniformly emulsified with a homomixer. After emulsification, the mixture was cooled to 30 ° C. with good stirring to prepare a cold cream.

Example 7: Clay pack
<Manufacturing method>
1,3-butylene glycol, glycerin, PEG-20 cetyl glucose sesquistearate, xanthan gum, preservative, and purified water were mixed and stirred to make the whole uniform.
While stirring this with a homomixer, the prototype (s) and kaolin were added to make the whole uniform, and ethanol was further added and stirred and mixed to prepare a clay pack.

Example 8: Foundation
<Manufacturing method>
Glycerin oleate, triglyceryl diisostearate, cyclohexanedioctyl, cetyl alcohol, microcrystalline wax, liquid paraffin, trimethylsiloxysilicate solution, talc and iron oxide were dissolved by heating and mixed (A). Glycerin, magnesium sulfate and purified water were mixed and heated, (A) was added and mixed, and then cooled. A prototype (s) was dissolved in purified water and added, and then a fungicide and a fragrance were added to prepare a base foundation.

FIG. 1 is a diagram showing an analysis example of deoxyoligonucleotide by HPLC of a nuclease-treated product (degradation product) of DNA. FIG. 2 is a photographed image showing changes in the skin condition of subjects in their 30s before the test and four weeks after the start of the test. FIG. 3 is a photographed image showing changes in the skin condition of subjects in their 50s before the test and four weeks after the start of the test. FIG. 4 is a diagram showing the test results of the moisture amount and sebum amount and the average values for each age. FIG. 5 is a diagram showing the results of skin groove density and wrinkle tests and the average values for each age. FIG. 6 is a diagram showing an analysis example of deoxyoligonucleotide by HPLC of a nucleoprotein nuclease-treated product (degradation product). FIG. 7 is a diagram showing an analysis example of oligonucleotides by HPLC of RNA nuclease-treated products (degradation products).

Claims (6)

A fraction with a molecular weight of 1000 to 3000 obtained by reducing the molecular weight of RNA by enzymatic degradation or hydrolysis, and containing an RNA degradation product containing 20 to 50% of an oligonucleotide having a chain length of 2 to 12 A cosmetic preparation for improving the crevice density (except for a preparation containing β-alanine). The cosmetic preparation for improving the crevice density according to claim 1, wherein the RNA degradation product contains 41.1% of an oligonucleotide having a chain length of 2 to 12. The cosmetic preparation according to claim 1 or 2, wherein the RNA degradation product is obtained from a yeast selected from the group consisting of beer yeast, torula yeast, milk yeast and baker's yeast. In addition to the RNA degradation product, a deoxyoligomer having a molecular weight of 1000 to 3000 obtained by lowering the molecular weight of nucleoprotein and DNA by enzymatic degradation or hydrolysis, and having a chain length of 2 to 12 The cosmetic preparation according to claim 1, comprising a nucleoprotein degradation product containing 20 to 50% nucleotides and a DNA degradation product. 5. The cosmetic preparation according to claim 4, wherein the nucleoprotein and DNA are obtained from a fish larva selected from the group consisting of salmon, salmon, salmon and salmon. A cosmetic for improving the skin groove density, comprising the cosmetic compounding agent according to any one of claims 1 to 5.