JP2007291062A - Compounding agent for base cosmetic, and base cosmetic - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a compounding agent for a base cosmetic and the base cosmetic. <P>SOLUTION: This compounding agent for the base cosmetic contains at least 2 kinds of mixtures selected from a nucleoprotein and/or the decomposition product by an enzyme or hydrolysate of a DNA or RNA, a deoxyoligonucleotide, a deoxymononucleotide, an oligopeptide, an oligonucleotide or a mononucleotide isolated from the decomposition products, or the above decomposition products or compounds as active ingredients, and the base cosmetic containing the compounding agent is also provided. Since the active ingredients such as the deoxyoligonucleotide, etc., have relatively small molecular weights, they are easily absorbed percutaneously, and have a cell-activating effect and bloodstream-promoting effect on being absorbed percutaneously, and on applying the cosmetic on the epidermis of a face surface, they exhibit excellent preventing effect of skin roughness, spots, freckles, wrinkles, etc., to show a beautiful skin-whitening effect. <P>COPYRIGHT: (C)2008,JPO&INPIT

Description

  The present invention relates to a basic cosmetic preparation and a basic cosmetic, and more particularly, a nucleoprotein and / or DNA or RNA enzymatic degradation product or hydrolysis product having a cell activation effect and a blood circulation promoting effect, and the degradation product. A basic cosmetic formulation comprising as an active ingredient a deoxyoligonucleotide, deoxymononucleotide, oligopeptide, oligonucleotide, mononucleotide, or at least two mixtures selected from the degradation products or the compounds separated from The present invention relates to a basic cosmetic containing the basic cosmetic compounding agent.

Rough skin, spots, freckles, wrinkles, etc. are caused by the effects of the balance of female hormones 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, blood to the epidermis It is thought to be caused by various things such as decreased flow, malnutrition, and mental stress.
The fact that rough skin, spots, freckles, wrinkles, and the like are serious cosmetic concerns of people has recently appeared in an increase in the number of basic cosmetic products for men as well as for women. Because the face is a big factor that gives a first impression to others, there are many people who want to have glossy and firm skin (beautiful skin) or white and clear skin (whitening) regardless of age or gender. Therefore, there is a growing public interest in basic cosmetics that improve rough skin, spots, freckles, wrinkles and the like.

  In conventional cosmetics that are in the category of “basic 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 derivatives thereof having a function of suppressing melanin production and hydroquinone derivatives have been introduced with the intention of whitening the skin.

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 basic cosmetics have not been made sufficiently.
JP 2004-16143 A

Causes of skin troubles such as rough skin, spots, freckles, wrinkles, etc. are various factors such as facial skin aging, poor blood circulation, influence of ultraviolet rays, stress, etc., but skin cell metabolism decreases and new cell regeneration functions Falling is a direct cause.
Conventional basic cosmetics do not work by acting on the metabolism mechanism of skin cells to exert their functions, 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, a spot, freckles, wrinkles, whitening, etc. is not acquired.
In addition, the above-described L-ascorbic acid and its derivatives, which have a function of suppressing melanin production, are not stable and have insufficient ultraviolet inflammation prevention effects, and hydroquinone derivatives have a safety problem. Therefore, it is difficult to directly obtain the melanin biosynthesis inhibitory effect and melanin bleaching action of the above compounds as active ingredients in cosmetics.
That is, a product with a high skin whitening effect that promotes the regeneration of new cells by cell activation and blood circulation promotion and prevents rough skin, spots, freckles, wrinkles, etc. is desired, but the effect in conventional basic cosmetics is still sufficient. Not.
Therefore, there has been a demand for the development of a new basic cosmetic product that activates the cells themselves to make the skin alive.

  As a result of earnest research to obtain a new basic cosmetic having an effect of preventing rough skin, spots, freckles, wrinkles and the like, and a skin whitening effect, the present inventors have carried out an enzymatic degradation treatment or a hydrolysis treatment of nucleoprotein and / or DNA. Degradation products containing deoxyoligonucleotides, deoxymononucleotides or oligopeptides, or degradation products containing oligonucleotides or mononucleotides obtained by enzymatic degradation or hydrolysis of RNA Alternatively, the present inventors have found that a mixture thereof has an excellent effect of preventing rough skin, spots, freckles, wrinkles and the like and a skin whitening effect, thereby completing the present invention.

That is, the basic cosmetic compounding agent of the present invention is
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 by enzymatic degradation or hydrolysis, or deoxy separated from the degradation product An oligonucleotide, deoxymononucleotide or oligopeptide, or
A degradation product containing 20 to 50% of a fraction having a molecular weight of 1000 to 3000 obtained by degrading RNA by enzymatic degradation or hydrolysis, or an oligonucleotide or mononucleotide separated from the degradation product, Or
A mixture of at least two selected from the group consisting of the nucleoprotein and / or DNA degradation product, the RNA degradation product, deoxyoligonucleotide, deoxymononucleotide, oligopeptide, oligonucleotide and mononucleotide;
Is contained as an active ingredient.

In a preferred embodiment of the combination product for basic cosmetics of the present invention, the nucleoprotein and DNA are preferably obtained from fish larvae such as salmon, 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 basic cosmetic of the present invention is characterized by containing the above basic cosmetic compounding agent.
The basic cosmetic is preferably selected from the group consisting of lotion, milky lotion, cream, gel, jelly, essence, lip balm, pack or mask.

  The basic cosmetic compound of the present invention contains 20 to 50% of a fraction having a molecular weight of 1,000 to 3,000 obtained by reducing the molecular weight by enzymatic degradation or hydrolysis of nucleoprotein and / or DNA or RNA. , Deoxyoligonucleotides, deoxymononucleotides, oligopeptides, oligonucleotides and mononucleotides separated from the degradation products, or at least two mixtures selected from the degradation products or the compounds as active ingredients It is. Since the deoxyoligonucleotides and the like have a relatively small molecular weight, they are easily absorbed transdermally, and they have cell activation and blood circulation promoting effects when absorbed transdermally. Therefore, when applied to the epidermis of the face, excellent skin roughening, spots, freckles, wrinkles and the like are prevented, and a skin whitening effect is achieved.

Further, since the basic cosmetic composition of the present invention comprises the above-mentioned basic cosmetic compounding agent, when applied to the epidermis of the face, the basic cosmetic compounding agent has excellent rough skin, spots, freckles, wrinkles, etc. Provides a whitening effect.

  Deoxyoligonucleotide or deoxymononucleotide, which is also used as a refined product as an active ingredient of the basic cosmetic compounding agent of the present invention, enzymatically decomposes or hydrolyzes DNA, and oligonucleotide or mononucleotide enzymatically degrades RNA 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, salmon, and salmon, and salmon is particularly preferable.

Hereinafter, DNA will be described in more detail.
The DNA which is the raw material for producing the basic cosmetic compounding agent of the present invention may be in 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, which are wastes from marine processing, especially salmon, salmon, salmon, salmon and salmon testes (white larvae) contain a lot of DNA, but they have not been effectively used as resources and have been discarded in the past. . Therefore, it is desirable to use DNA derived from these testes from the viewpoint of recycling waste. Moreover, DNA obtained from thymus of mammals and birds, for example, pigs, chickens, etc. 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 fish larva is roughly crushed, and the crushed fish larva is treated with a proteolytic enzyme (protease) 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 DNA. Furthermore, it precipitates as a DNA salt from the solution which performed the dialysis process, or concentrates a solution, and collect | recovers these precipitates or concentrates.
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 basic cosmetic compounding agent of the present invention.
It is not limited to this method to obtain DNA from fish larvae, and a known method may be used.

  RNA is obtained from yeast, and preferably can be obtained by extraction from a yeast selected from the group consisting of beer yeast, torula yeast, milk yeast and baker's yeast and purification.

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

  The oligopeptide can be obtained by hydrolyzing a nucleoprotein (nuclear protein) contained in fish larvae with a protease.

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. As a good protease, a protease manufactured by Novozymes Japan K.K. (former Novo Nordisk Bio Industry Co., Ltd.) can be mentioned.

  The nuclease hydrolyzes the 3 ', 5'-phosphodiester bonds of deoxyribonucleic acid (DNA) and ribonucleic acid (RNA) to generate 5'-(deoxy) nucleotides for oligopolymerization. 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. (former Amano Pharmaceutical 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 this temperature range, the molecular weight of DNA, RNA and nucleoprotein does not sufficiently proceed, 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.

As an active ingredient of the formulation for basic cosmetics of the present invention, a degradation product obtained by enzymatic degradation or hydrolysis of nucleoprotein and / or DNA, or obtained by enzymatic degradation or hydrolysis 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 basic cosmetic preparation of the present invention, the following compounds separated from nucleoprotein and / or DNA or RNA degradation products using conventional separation means and / or purification means: deoxyoligonucleotide, deoxy Mononucleotides, 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 these compounds or at least two kinds may be mixed and used. 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.
Moreover, you may use the said decomposition product and the said compound in combination with the conventional additive component of the basic cosmetic compounding agent in a predetermined ratio.

  The concentration of the active ingredient (the decomposition product and / or the compound) in the formulation for basic cosmetics of the present invention can be appropriately selected.

Moreover, the basic cosmetic of the present invention contains the above-mentioned compounding agent for basic cosmetics.
The dosage form that can be taken by the basic cosmetic of the present invention can be appropriately selected as long as it is a dosage form applicable to the skin. Preferably, the basic cosmetic is a lotion, emulsion, cream, gel, jelly, essence, lip balm, pack or mask.

  In the basic cosmetic of the present invention, in addition to the basic cosmetic compounding agent, known components that can be mixed into existing basic cosmetics can be blended. For example, a fragrance | flavor, a moisturizer, etc. can be mix | blended individually or in combination.

The basic cosmetics of the present invention include vitamin E or a derivative thereof such as vitamin E acetate; a vasodilator such as an acetylcholine derivative; a skin function-enhancing agent such as cephalanthin; glycyrrhetinic acid or a derivative thereof; Female hormone agents such as: amino acids such as serine, methionine, and arginine; vitamins such as vitamin A, vitamin B ₁ , vitamin B ₆ , biotin, pantothenic acid, and derivatives thereof can be used alone or in combination.

  Furthermore, in the basic cosmetics of the present invention, if necessary, additives usually used for external preparations for skin such as cosmetics and pharmaceuticals, such as oils, preservatives, surfactants, dispersion stabilizers, thickeners, Wetting agents, ultraviolet absorbers, antioxidants, pH adjusters, purified water, alcohols, 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 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, DNA and nucleoprotein, or RNA) used in the examples is shown as a solid content. .

1. Manufacture of (deoxy) oligonucleotides Restricted by using a nuclease [for example, the enzyme preparation nuclease “Amano” (Amano Enzyme (former Amano Pharmaceutical Co., Ltd.)), which is approved as a food additive, for DNA derived from silkworm Decomposition was performed. 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) oligonucleotides Powdered DNA-Na salt derived from white coconut was added as raw material to warm water adjusted to around 65 ° C., stirred, and further heated to 70 ° C. Amano ”(Amano Enzyme (former Amano Pharmaceutical Co., Ltd.)) was added at 0.05% 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 basic cosmetics using (deoxy) oligonucleotide A dry powder (decomposition product) containing deoxyoligonucleotide obtained by the above production method is designated as prototype 1, and using this prototype 1, basic cosmetic of the present invention Was manufactured as follows. As a control, a basic 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. A basic cosmetic was obtained.
[Comparative Example 1]
A basic cosmetic 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 1.

4). Blood flow measurement test Each 10 μL of the basic cosmetic of Example 1 and the basic cosmetic of Comparative Example 1 were applied to the human upper arm, and the blood flow was measured with a laser Doppler meter one hour after the application. The test results were determined according to the following criteria.
++: The blood flow increased significantly compared to the comparative example (high effect)
+: Blood flow increased compared to the comparative example (effective)
±: The blood flow increased slightly compared to the comparative example (somewhat effective)
−: The blood flow was equal to or less than that of the comparative example (invalid)
The results are summarized in Table 1 below.

5). Skin roughness improvement test 10 middle-aged and elderly women with skin roughness in the lower limbs were tested, and the basic cosmetic of Example 1 and the basic cosmetic of Comparative Example 1 were each placed at different locations on the left and right lower limbs about 1 g / time after bathing. / Applied for 1 day.
Depending on the skin condition of the applied part before and 1 month after the test, using the following criteria,
The test results were judged from the viewpoints of the peeled state and moisture state of the skin.
5-1. 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”.
5-2. 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)
The results are summarized in Table 1 below.

6). Evaluation test by trial The evaluation test of the basic cosmetic of Example 1 and the basic cosmetic of Comparative Example 1 was conducted in a single-blind manner for middle-aged women. The number of examinees for each cosmetic product was 10. The evaluation is a questionnaire for the examinees. Five items, “smoothness”, “fineness of texture”, “whitening condition”, “elasticity of skin” and “improvement of freckles” are used before and 1 month. The answer was obtained after use.
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)
The results are summarized in Table 1 below.

7). 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.

7-1. 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. 2 shows an analysis example of deoxyoligonucleotide as a decomposition product by HPLC (high performance liquid chromatography). In FIG. 2, peaks within a retention time of 19 minutes to 24 minutes (peak 1 to peak 4) 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.

7-2. 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. 3 shows an analysis example of oligonucleotides of degradation products by HPLC (High Performance Liquid Chromatography). In FIG. 3, peaks within a retention time of 13 minutes to 24 minutes (peak 2 to peak 5) 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.

8). Production of various basic cosmetics Using the prototype 2 (DNA and nucleoprotein degradation products) and the prototype 3 (RNA degradation products) obtained by the above-described method, the basic cosmetics of the present invention were produced as follows. . Moreover, the basic 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 2 to 5 below.
The source of products used for the following basic cosmetics is shown in parentheses.

8-1. Gel-like basic cosmetics 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. Into this, each of prototype 2, mixed with purified water, prototype 3, and equivalent mixture of prototype 2 and prototype 3 (prototype 4) was dissolved, and Example 2-1, Example 2-2, and implementation were performed. The basic cosmetic product of Example 2-3 was obtained.
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 basic cosmetic product of Comparative Example 2 was obtained.

8-2. Emulsion basic cosmetic Example 3 (Examples 3-1 to 3-3):
Concentrated glycerin was added to purified water and the mixture was heated to 70 ° C. 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 preliminarily 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, mixed with purified water, prototype 3, and equivalent mixture of prototype 2 and prototype 3 (prototype 4) was dissolved and stirred. Deaeration, filtration, and cooling were performed to obtain the emulsion-type basic cosmetics of Example 3-1, Example 3-2, and Example 3-3.
Comparative Example 3:
A method similar to Example 3-1, Example 3-2 and Example 3-3, except that 2% by mass of concentrated glycerin was added in place of prototype 2 and prototype 3, and a total of 10% by mass was added. Thus, a basic cosmetic product of Comparative Example 3 was obtained.

8-3. Lotion-like basic cosmetics (lotion)
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. Each of prototype 2, prototype 3, and equivalent mixture of prototype 2 and prototype 3 (prototype 4) mixed with oleyl alcohol, sorbitan monolaurate and purified water in ethanol was dissolved. The mixture was stirred and the lotion-shaped basic cosmetics of Example 4-1, Example 4-2, and Example 4-3 were obtained.
Comparative Example 4:
A method similar to Example 4-1, Example 4-2, and Example 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. Thus, a basic cosmetic product of Comparative Example 4 was obtained.

8-4. Creamy basic cosmetics 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., prototype 2, sample 3 and equivalent mixture of prototype 2 and prototype 3 in which purified water is mixed with sodium hydroxide, sodium citrate and purified water (prototype 4) ) Were mixed and stirred to obtain creamy basic cosmetics of Example 5-1, Example 5-2, and Example 5-3.
Comparative Example 5:
A method similar to Example 5-1, Example 5-2, and Example 5-3, except that 2% by mass of concentrated glycerin was added instead of prototype 2 and prototype 3, and a total of 7% by mass was added. Thus, a creamy basic cosmetic product of Comparative Example 5 was obtained.

9. Performance test of basic cosmetics (gels, emulsions, lotions, creams) For the basic cosmetics of Examples 2 to 5 and Comparative Examples 2 to 5, the blood flow measurement test, the rough skin improvement test, and the evaluation tests by trial are performed, respectively. It carried out on the same conditions as Example 1 and Comparative Example 1.
The results are summarized in Tables 2 to 5.

From the results shown in Table 1 and Tables 2 to 5, the following was found.
(1) Evaluation of the cosmetic blood flow measurement test of Example 1 or Example 2 to Example 5 is ++ [the blood flow is extremely increased compared to the comparative example (highly effective)], and Examples 1 to It is clear that the active ingredient of the cosmetic product of Example 5 penetrates from human skin and significantly increases blood flow.
(2) In the cosmetic rough skin improvement test of Example 1 or Examples 2 to 5, all cosmetics including prototype 1, prototype 2, and prototype 4 containing prototype 2 are effective. The effectiveness of the cosmetics including Prototype 3 was confirmed. In addition, the improvement of rough skin was hardly recognized in the left lower limb which was not applied as a control, and the effectiveness of the cosmetics including Prototype 1 to Prototype 4 became clear.
(3) In the test results shown in Table 1, the evaluation test based on the trial use of cosmetics including the trial product 1 (DNA degradation product) is particularly “smoothness”, “fineness of texture”, “elasticity of the skin” It was an evaluation result that a remarkable improvement effect was recognized.
(4) In the comparison of the test results for the same cosmetic products shown in Tables 2 to 5, the evaluation test by trial use of the cosmetic products including the prototypes 2 to 4 is the prototype 2> prototype in any cosmetic form. 4> Prototype 3> Excellent in order of comparison.
In addition, each cosmetic product including prototype 2 to prototype 4 was evaluated as having a remarkable improvement effect, particularly in terms of “smoothness”, “fineness of texture”, and “elasticity of skin”. .
(5) On the other hand, in the evaluation tests by trial in Example 1 or Examples 2 to 5, “whitening effect” and “improvement of freckles” were observed in some test subjects, but the metabolism of skin, etc. As a result, the results indicate that longer use is required.

  In Examples 1 to 5, as an active ingredient, prototype 1 (DNA salt degradation product), prototype 2 (DNA and nucleoprotein degradation product), prototype 3 (RNA degradation product) and prototype 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, deoxyoligonucleotides, deoxymononucleotides, oligopeptides, oligonucleotides or monocosmetics containing basic cosmetics and basic cosmetics containing the basic cosmetics have increased blood flow and rough skin. It was recognized that it had an improving effect, as well as an effect of improving the smoothness of the skin, the fineness of the skin, and the elasticity of the skin.

Examples of prescription of basic cosmetics using the prototypes 1 to 4 are shown as Examples 6 to 8. Note that 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). The meaning of
In any of the examples, it was confirmed that the blood flow rate was increased, the rough skin was improved, and the skin smoothness, fine texture, and skin elasticity were improved.

＜製造方法＞
上記配合成分のうち香料以外の成分を混合、溶解した後、香料を加えて撹拌混合した後、型に流し込み冷却してリップクリームを調製した。 Example 6: Lip Balm

<Manufacturing method>
After mixing and dissolving components other than the fragrance among the above blended components, the fragrance was added and stirred and mixed, then poured into a mold and cooled to prepare a lip balm.

＜製造方法＞
精製水に試作品（類）、石鹸粉末を加え加熱溶解して７０℃に保った（水相）。他の成分を混合し、加熱融解して７０℃に保った（油相）。水相に油相を撹拌しながら徐々に加え、終了後、ホモミキサーで均一に乳化し、乳化後よく撹拌しながら３０℃まで冷却し、
コールドクリームを調製した。 Example 7: 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). Gradually add the oil phase to the aqueous phase with stirring, and after completion, emulsify uniformly with a homomixer, cool to 30 ° C. with good stirring after emulsification,
Cold cream was prepared.

＜製造方法＞
１，３−ブチレングリコール、グリセリン、セスキステアリン酸ＰＥＧ−２０セチルグルコース、キサンタンガム、防腐剤、精製水を混合・撹拌して全体を均一とした。
これをホモミキサーで撹拌しながら、試作品（類）及びカオリンを加え、全体を均一とし、さらにエタノールを加えて撹拌・混合し、クレイパックを調製した。 Example 8: 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.

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 diagram showing an analysis example of deoxyoligonucleotide by HPLC of a nucleoprotein nuclease-treated product (degradation product). FIG. 3 is a diagram showing an analysis example of an oligonucleotide by HPLC of a nuclease-treated product (degradation product) of RNA.

Claims (5)

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 by enzymatic degradation or hydrolysis, or deoxy separated from the degradation product An oligonucleotide, deoxymononucleotide or oligopeptide, or
A degradation product containing 20 to 50% of a fraction having a molecular weight of 1000 to 3000 obtained by degrading RNA by enzymatic degradation or hydrolysis, or an oligonucleotide or mononucleotide separated from the degradation product, Or
A mixture of at least two selected from the group consisting of the nucleoprotein and / or DNA degradation product, the RNA degradation product, deoxyoligonucleotide, deoxymononucleotide, oligopeptide, oligonucleotide and mononucleotide;
A basic cosmetic preparation, characterized in that it contains as an active ingredient. 2. The basic cosmetic preparation according to claim 1, wherein the nucleoprotein and DNA are obtained from fish larvae such as salmon, salmon, salmon, salmon and salmon. The basic cosmetic preparation according to claim 1, wherein the RNA is obtained from a yeast selected from the group consisting of beer yeast, torula yeast, milk yeast and baker's yeast. A basic cosmetic comprising the basic cosmetic compounding agent according to claim 1. The basic cosmetic according to claim 4, wherein the basic cosmetic is selected from the group consisting of lotion, milky lotion, cream, gel, jelly, essence, lip balm, pack or mask.

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