JPH04187618A - Skin external preparation - Google Patents

Abstract

PURPOSE:To obtain a skin external preparation having excellently beautifying effects on skin, propagating effects on cells, stability to metal ion, etc., and safety, comprising an amino acid derivative of a kojic acid and/or a peptide derivative of the kojic acid. CONSTITUTION:A base cosmetic, a make-up cosmetic, a face washing agent, soap, a bathing agent, etc., is blended with 0.001-30wt.% one or more of an amino acid derivative of a kojic acid and a peptide derivative of the kojic acid shown by the formula (R is amino-protecting amino acid or amino- protecting peptide) as an active ingredient to prepare the objective substance. The substance is made into the form such as solution, emulsion, ointment, oil, wax, gel, sol, powder or spray. The compound shown by the formula has 10-20 times as high inhibitory effects on tyrosinase activity as kojic acid alone does.

Description

DETAILED DESCRIPTION OF THE INVENTION <Industrial Application Field> The present invention relates to an external skin preparation. More specifically, the present invention relates to a safe external preparation for skin, which has properties such as skin whitening effect, cell proliferation effect, and stability against metal ions and the like.

<Prior Art and Background of the Invention> Kojic acid, kojic acid fatty acid esters, kojic acid ethers, and certain other kojic acid derivatives have been shown to have a skin whitening effect, that is, a tyrosinase activity inhibiting effect (melanin formation inhibiting effect, It is known to have the effect of preventing burns. Based on these actions and effects, kojic acid and the above-mentioned kojic acid derivatives have been used in various skin-whitening cosmetics, skin-whitening foods, and the like.

Examples of the conventional uses include the following. For example, first, conventional examples of using kojic acid alone or an isomer of kojic acid alone as an active ingredient for whitening effects, etc.
Japanese Patent Publication No. 61-010447, Japanese Patent Publication No. 53-3538, and Japanese Patent Publication No. 62-059084).

Second, one or two fatty acid esters (monoesters and diesters) of kojic acid are used as active ingredients for whitening effects, etc.
As a conventional example of using seeds or more, there is
No. 022152, Special Publication No. 60-7961, Special Publication No. 1983
-9722, Japanese Patent Publication No. 01-319473, Special Publication No. 6
1-060801, etc.

Thirdly, there are conventional examples of selectively using kojic acid and various derivatives of kojic acid, and these can be further classified into the following two types. That is, A] Active ingredients such as kojic acid, kojic acid salts, kojic acid fatty acid esters (including monoesters and diesters), kojic acid ethers, kojic acid derivatives such as kojic acid phosphate esters, and When one or more of the other kojic acid derivatives (that is, the fatty acid esters, ethers, kojic acid derivatives other than the phosphoric acid esters, etc.) are selectively used.

Conventional examples that are considered to belong to this section include:
JP 60-10005, JP 61-60802, JP 60-137253, JP 60-2330
No. 71, JP-A-61-057574, JP-A-61-1
No. 97506, JP-A-61-289086, JP-A-6
No. 3-277670, etc. are recognized.

B] Kojic acid by selectively using one or more of the salts of kojic acid and kojic acid derivatives, and also using other substances unrelated to kojic acid or kojic acid derivatives. Or when aiming for a synergistic effect of various effects such as the whitening effect of kojic acid derivatives.

Conventional examples that are considered to belong to this section include:
Japanese Patent Publication No. 62-3820, Japanese Patent Publication No. 64-83008,
JP-A-64-083009, JP-A-64-08301
No. 0, JP 64-083011, JP 01-09
No. 6109, JP-A-01-100112, JP-A-01
-100113, JP 01-100114, JP 01-100114, JP 01-121205, JP 01-121206, JP 01-1212
No. 07, JP 02-028105, JP 02-02-1
No. 08614, etc. are recognized.

However, as suggested in the conventional example above, the physiological activities of known kojic acid and kojic acid derivatives, such as whitening effects (tyrosinase activity inhibition effect), are not sufficient by themselves. By using appropriate substances in addition to derivatives, various effects can be improved and
We are trying to improve. In addition, the presence or absence of physiological activity of various substances,
At present, it is difficult to determine the success or failure of the improvement in physiological activity based solely on the chemical structure of various substances, and it is currently impossible to judge this without conducting specific comparative tests.

Furthermore, in the presence of metal ions, kojic acid forms a type of chelate compound, resulting in significant coloring, which is extremely inconvenient when used in cosmetics, pharmaceuticals, etc. In order to prevent this, it was necessary to take various additional measures.

<Problems to be Solved by the Invention> As a result of intensive research aimed at developing a substance that is a kojic acid derivative and has a stronger tyrosinase activity inhibiting effect than conventional substances, the inventor discovered that an amino acid or peptide and kojic acid Both the amine-protected amino acid derivative of kojic acid and the amine-protected peptide derivative of kojic acid produced in the process of synthesizing the ester compound of Moreover, ■ Both the amino-protected amino acid derivatives of kojic acid and the amino-protected peptide derivatives of kojic acid are harmless, and ■ The amino-protected amino acid derivatives of kojic acid and the amino-protected peptide derivatives of kojic acid are stable against metal ions. The present invention was completed based on the discovery that they each have excellent physiological activity and properties, such as being able to prevent coloring of products, etc.

Note that the amino-protected amino acid derivatives of kojic acid and the amine-protected peptide derivatives of kojic acid are naturally included in the respective superordinate concepts of amino acid derivatives of kojic acid and peptide derivatives of kojic acid. However, when it is necessary to clearly distinguish between various substances, in this specification, as necessary, various substances with amino-protecting groups will be referred to as "amino-protected amino acid derivatives of kojic acid". ”, “Amino-protected peptide derivatives of kojic acid”
On the other hand, various substances obtained by removing the amino-protecting group from the various substances with the above-mentioned amino-protecting group by a conventional method are referred to as ``-y-mi/acid Fa derivative of kojic acid'' and ``peptide of kojic acid.'' referred to as "derivatives".

The present invention focuses on the biological activity of the amino-protected amino acid derivatives of kojic acid and the amino-protected peptide derivatives of kojic acid, and has developed amino-protected amino acid derivatives of kojic acid and amino-protected peptide derivatives of kojic acid without using other substances in combination as in the past. Simply containing one or more substances selected from the group of substances consisting of amino-protected peptide derivatives of acids has the effect of inhibiting tyrosinase activity and cell proliferation, and can reduce rough skin caused by sunburn and dead skin cells. It prevents dark skin, age spots, freckles, skin aging, etc. It also speeds up the turnover (metabolism) of epidermal cells, removes aging and keratinized skin, and gives elasticity, tension, and luster to the skin. The object of the present invention is to provide an external skin preparation suitable for sun care cosmetics and pharmaceuticals in particular.

<Means for Solving the Problems> In order to achieve the above object, in this invention, as described in the claims, “-form [■] (N (wherein R is an amino-protected amino acid or an amine 1. An external preparation for skin, characterized in that it contains as an active ingredient one or more substances selected from the group of amino acid derivatives of kojic acid (protected peptide) and peptide derivatives of kojic acid. We decided to configure J.

Amine-protected amino acid derivatives of kojic acid that can be used in the present invention include pyranyl pyrane of kojic acid (5-hydroxy-2-hydroxymethyl-γ-pyrone, 5-hydroxy-2-hydroxymethyl-4H-bilan-4-one). The hydroxyl group of the hydroxymethyl group present at the 2-position hydroxymethyl group in the ring structure (that is, the hydroxyl group bonded to the 7-position carbon element) and the carboxyl group of the amino-protected amino acid It can be freely selected and used from amino-protected amino acid derivatives of kojic acid formed by ester bonding. There are no particular limitations on the amino acids introduced into the kojic acid. In principle, all amino acids, that is, all amino acids can be introduced into kojic acid, regardless of the type of amino acid or the optical isomer such as D-type, L-type, or racemic form. Among them, for example, glycine (hereinafter referred to as rcty J or "G").

Alanine (hereinafter referred to as "rA1a" or "A").

Valine (hereinafter referred to as rVal J or ``V''), leucine (hereinafter referred to as ``rLeu'' or ``L''), isoleucine (hereinafter referred to as ``■le'' or ``river''), serine (
(hereinafter referred to as "rser" or "S").

Threonine (hereinafter referred to as "rThe" or "T"), cysteine (hereinafter referred to as "rCys" or "rcu"), methionine (hereinafter referred to as "rMet J" or "M"), proline (hereinafter referred to as "rPro" or "rpJ"), aspartic acid (hereinafter referred to as [^ SL] J or “D”)
, asparagine (hereinafter referred to as rAsn or “N”)
, glutamic acid (hereinafter referred to as rG1u J or "E"), glutamine (hereinafter referred to as rGln J or "Q"), histidine (hereinafter referred to as rHis) or rl (J), lysine (hereinafter referred to as rLys J or "K"),
Arginine (hereinafter referred to as rArg or “R”),
Phenylalanine (hereinafter referred to as rPhe J or r)-J), tyrosine (hereinafter referred to as rTyr J or "Y"), tryptophan (hereinafter referred to as rTrp j or rw")
Particularly preferred are amino-protected amino acid derivatives of kojic acid in which a biogenic amino acid is introduced into kojic acid.

Further, the amino-protected peptide derivative of kojic acid that can be used in the present invention is similar to the above-mentioned "amino-protected amino acid derivative of kojic acid", and the hydroxyl group ( In other words, 7-
It is possible to freely select and use amino-protected peptide derivatives of kojic acid, which are formed by an ester bond between a hydroxyl group (bonded to the carbon atom at position 1) and a carboxyl group of an amino-protected peptide. Furthermore, the peptide introduced as an amino-protected peptide derivative of kojic acid is not particularly limited. In principle all common peptides can be used. That is, not only relatively low-molecular peptides such as dipeptides, tripeptides, and tetrapeptides, but also oligopeptides consisting of about 2 to 10 amino acids and polypeptides consisting of about 10 to 100 amino acids can be used. Furthermore, protein decomposition products can also be used.

Among these, kojic acid and low molecular weight peptides (oligopeptides) (2 to 5 amino acids) are preferred.

Furthermore, the amino-protecting groups that can be used in the group of effective substances according to the present invention, that is, the group of amino-protected amino acid derivatives of kojic acid and the amino-protected peptide derivatives of kojic acid, include all known amino-protecting groups that are commonly used in peptide synthesis. Amino protecting groups are applicable. For example, the applicable amino protecting group is a carbobenzoxy group (hereinafter “
t-butyloxycarbonyl group (hereinafter referred to as rBoc-J), p-toluenesulfonyl (hereinafter referred to as [Tos-J), triphenylmethyl (hereinafter referred to as ``T
rt-), other amino protecting groups can be used.

As a matter of course, various substance groups obtained by removing the various amino-protecting groups from the various effective substances according to the present invention, that is, the [amino-protected amino acid derivatives of kojic acid] and the "amino-protected peptide derivatives of kojic acid" (
Similar to the various amino-protected derivatives of kojic acid (amino acid derivatives of kojic acid and peptide derivatives of kojic acid), they also have excellent physiological activities (tyrosinase activity inhibition, cell proliferation, etc.) and properties (stability against metal ions, etc.). sex)
etc. exist.

In addition, in order to remove various amino protecting groups from these amino-protected amino acid derivatives of kojic acid and amino-protected peptide derivatives of kojic acid, for example, catalytic reduction (20° C.
This can be achieved by known treatment methods depending on the various amino protecting groups described above, such as normal pressure).

By the way, as demonstrated in the Examples below, amino-protected amino acid derivatives of kojic acid and amino-protected peptide derivatives of kojic acid are about 10 to 20 times more effective in suppressing tyrosinase activity than kojic acid alone. found. That is, it is suggested that amino acid derivatives of kojic acid and peptide derivatives of kojic acid have extremely superior skin whitening effects, sunburn prevention effects, and melanin production suppressing effects compared to kojic acid alone.

Furthermore, the amino-protected amino acid derivatives of kojic acid and the amino-protected peptide derivatives of kojic acid according to the present invention have cell proliferation effects (human healthy skin cells-XX-male).
It was found that it has an excellent effect on cell proliferation (cell proliferation).

In other words, while kojic acid alone does not have the cell proliferation effect, amino-protected amino acid derivatives of kojic acid and amino-protected peptide derivatives of kojic acid exhibit cell proliferation effects when added to the medium at 0.05mM. It was found that the increase was approximately 12-32%.

The amino-protected amino acid derivatives of kojic acid and the amino-protected peptide derivatives of kojic acid used in the external skin preparation of this invention are non-toxic, and no problems were observed when either substance was absorbed through the skin.

The skin external preparation according to the present invention contains, as an active ingredient, one or more substances selected from the group consisting of the amino-protected amino acid m8 form of kojic acid and the amino-protected peptide derivative of kojic acid. It can be provided by blending an effective amount. A skin external preparation can usually be provided by incorporating the active ingredient in an amount of about 0.001 to 30% by weight (hereinafter referred to as i%), preferably about 0.05 to 10% by weight. In addition, in the skin external preparation according to the present invention, when blending one or more substances selected from the substance group of the amine-protected amino acid derivatives of kojic acid and the amino-protected peptide derivatives of kojic acid, Even if a small amount of unreacted substances, reaction intermediates, etc. that could not be completely removed during the production and purification process of the protected amino acid derivative and the amine-protected peptide derivative of kojic acid are present, there is no negative effect on the efficacy of the external skin preparation according to the present invention. I was not able to admit.

Product forms of external skin preparations to which this invention can be applied include creams, milky lotions, lotions, packs, facial cleansers9 soaps, various basic cosmetics such as bath additives, foundations, various make-up products such as Habo-ko, etc. It can be applied to a wide range of cosmetics, etc.

The various skin external preparations can be applied in various forms such as solutions, emulsions, ointments, oils, waxes, gels, sol, powders (paratars), sprays (aerosols), and the like.

By the way, the method for manufacturing and purifying the amino-protected amino acid derivative of kojic acid and the amine-protected peptide derivative of kojic acid used in the external skin preparation according to the present invention is a known method. That is, using a carbohendioxy group (hereinafter referred to as "Z") as an amino protecting group, kojic acid (hereinafter r
an amino-protected amino acid derivative of r Z -11e -0-Kojiam e'
The case of synthesizing will be explained as an example.

First, p~dimethylsulfonylphenol methyl sulfate (HODMSP) was dissolved in acetonitrile,
Add Z-Ice-OH to this and stir. Further dicyclohexylcarbodiimide (DCC) is added at 0°C. The reaction mixture was gently stirred at O''C for 2 hours and continued stirring at room temperature for 2 hours. After filtering off the precipitate, the solvent was distilled off using an evaporator and Z-11e-
While obtaining ODMSP, the above Z-11 in which kojic acid was dissolved in water and triethylamine) was dissolved in acetonitrile.
Add to e-ODMSP. The reaction mixture is then stirred at room temperature for 8 hours. After removing acetonitrile using an evaporator, the pH of the mixture was adjusted to 1, and Z -11e -〇-
Kojiate (an amino-protected amino acid derivative of kojic acid) is extracted with ethyl acetate. Zlle-0-Koji by recrystallization from ethyl acetate ether
Purify ate.

<Effect> The skin external preparation according to the present invention (skin external preparation containing one or more substances selected from the group of substances consisting of amino-protected amino acid derivatives of kojic acid and amino-protected peptide derivatives of kojic acid) has a synergistic effect. Based on the above-mentioned tyrosinase activity inhibiting effect (whitening effect/melanin formation inhibiting effect), the whitening effect is far superior to that of conventional kojic acid (approximately 10 to 20 times more effective) without adding any other substances to achieve this effect. The same whitening effect can be achieved with an addition amount of about 1/10 to 1/20 of the concentration), and the whitening effect can be further enhanced by accelerating skin turnover based on the cell proliferation effect. A skin external preparation can be provided.

Furthermore, since the skin external preparation according to the present invention has a slow cell proliferation effect as mentioned above, it speeds up the turnover (metabolism) of epithelial cells, thereby reducing keratinized and aged skin. It has the effect of removing it and leaving your skin fresh and beautiful. On the other hand, since the skin external preparation according to the present invention has an excellent cell proliferation effect, it promotes the production of Mucopoly II and collagen in dermal cells, thereby increasing the elasticity and tension of the skin. Based on these effects, the skin becomes even more fresh.

With the discovery of these effects, we have created an excellent external skin preparation that has not existed before, by containing one or more substances selected from the substance group of amino-protected amino acid derivatives of kojic acid and amino-protected peptide derivatives of kojic acid. This means that we can provide the following.

<Example> Next, according to Examples, (1) Examples of preparation and purification of amino acid derivatives of kojic acid and peptide derivatives of kojic acid, (2)
Example of a test for measuring the inhibition rate of tyrosinase activity of kojic acid and the various derivatives of kojic acid, (3) Example of a test for measuring the cell proliferation effect of kojic acid and the various derivatives of kojic acid, (4)
Examples of stability tests against iron ions of kojic acid and the various derivatives of kojic acid, (5) application formulation examples of the various derivatives of kojic acid, etc. are illustrated.

It goes without saying that the present invention is not limited to these examples.

(1) Examples of preparation and purification of amino acid derivatives of kojic acid and peptide derivatives of kojic acid: ■Amine-protected amino acid derivative of kojic acid (carbobenzoxyisoleucine-kojic acid WI form: Z-11e-0
-Kojiate) purification example.

p-dimethylsulfonylphenol methyl sulfate (HODMSP) (5.30g, 20m
mol) to 80rn1. dissolved in acetonitrile,
Z-11e-OH (5.30g, 20m mol)
Add and stir. Furthermore, dicyclohexylcarbodiimide (DCC) (4.12 g, 20 mmol) was o
Add at 'c. The reaction mixture is stirred gently for 2 hours at 0°C. Further, stirring is continued for 2 hours at room temperature. After removing the precipitate by filtration, the solvent was distilled off using an evaporator and
-1ie-ODMSP is obtained.

On the other hand, Z -11e -ODMS P in which kojic acid (2,84 L 20 mmol) was dissolved in water and triethylamine (2,80 d) and dissolved in acetonitrile 40 ml.
Add to. The reaction mixture is then stirred for 8 hours. After removing acetonitrile using an evaporator, the pH of the mixture was adjusted to 1.
and extract Z-11e-0-Kojiate with ethyl acetate.

The Z-1ie-0-Kojiate is purified by recrystallization from ethyl acetate ether to obtain white crystals.

Similarly, as amino-protected amino acid derivatives of kojic acid and amino-protected peptide derivatives of kojic acid,
A total of 6 types were obtained as shown below and used as samples for subsequent tyrosinase activity suppression effect tests, cell proliferation effect tests, etc.

◎Amino-protected amino acid derivatives of kojic acid 1) Carbobenzoxyleucine-kojic acid derivatives (r Z -Le
2) Carbobenzoxyisoleucine-kojic acid derivative (hereinafter referred to as r Z -11e -0-Kojiate J) [The IR of this derivative is shown in Figure 2, and the analysis results of "C-NMR and 'H-NMR are shown in Table 2."

] 3) Carbobenzoxyalanine-kojic acid derivative (hereinafter referred to as 'Z Ala-○-Kojiate') [IR of this derivative is shown in Figure 3, +3C-NMR and 'H
-NMR analysis results are shown in Table 3. ] 4) Carbobenzoxyparin-kojic acid m1 body (hereinafter '
Z Vat -〇-Kojiate J) [IR of this derivative is shown in Figure 4, "'C-NMR and 'H
-NMR analysis results are shown in Table 4. ] ◎ Amino-protected peptide derivative of kojic acid 5) Carbobenzoxyphenylalanine-glycine-kojic acid derivative (hereinafter referred to as rZ -Phe -Gly -0-Kojiat
e J) [The IR of this derivative is shown in Figure 5, and the analysis results of I''C-NMR and 'H-NMR are shown in Table 5.] 6) Carpobenzoxyleucine-alanine-kojic acid derivative ( Below 'Z Leu-Ala-〇-KojiateJ
) (The IR of this derivative is shown in Figure 6, "C-NMR and '
The results of H-NMR analysis are shown in Table 6. ] [Margins below this page] Table 1: Amine-protected amino acid derivatives of kojic acid (Z −
Physical properties of Leu , -0-Koj 1ate ).

Table 2: Amino-protected amino acid derivatives of kojic acid (Z-1
Physical properties of 1e-〇-Kojiate).

Table 3: Amino-protected amino acid derivatives of kojic acid (Z
Physical properties of Ala O-Kojiate).

Table 4: Amino-protected amino acid derivatives of kojic acid (Z −
Physical properties of Val-0-Koj1ate).

Table 5: Amine-protected peptide derivatives of kojic acid (Z −
Physical properties of Phe-Gly-〇-Kojiate).

Table 6: Amine-protected peptide derivatives of kojic acid (Z −
Physical properties of Leu-Ala-0-Kojiate).

(2) Test to measure the inhibitory effect on tyrosinase activity of kojic acid, amino-protected amino acid derivatives of kojic acid, and amino-protected peptide derivatives of kojic acid: (a) Test sample and control: Among the test samples, a commercially available product was used for kojic acid. However, for the other samples, each kojic acid derivative prepared by the method (1) above was used. That is, ■Control: 1-1) As a negative control (blank), a 5% aqueous solution of dimethyl sulfoxide (hereinafter referred to as rDMSOJ) is used.

Measurement of tyrosinase activity inhibition rate of kojic acid and test sample The test results were determined by measuring the tyrosinase activity inhibition rate (%) based on this negative control.

1-2) Kojic acid (hereinafter referred to as "Koji J") commercially available product: purity = 99% or more, melting point -152 to 155'C, molecular weight = 142°. Used as a positive control.

■Amino-protected amino acid derivatives of kojic acid 2-1) Carbobenzoxyleucine-kojic acid derivatives (hereinafter referred to as “Z-Le
u-0-Kojiate") 2-2) Carbobenzoxyisoleucine-kojic acid derivative (hereinafter referred to as "Z-1
1e-0-Koj 1ate J) 2-3) Carbohenzoxyalanine-kojic acid derivative (hereinafter referred to as rZ-A
2-4) Carbobenzoxyvaline-kojic acid derivative (hereinafter referred to as rZ-Val
-0-Kojiate'') ■ Amino-protected peptide derivatives of kojic acid 3-1) Carbobenzoxyphenylalanine-glycine-kojic acid derivatives (r Z-P
he-Gly--O-Koj 1ate J) 3-2) Carbobenzoxyleucine-alanine-kojic acid derivative (hereinafter referred to as 'Z-Leu-Ala-0-Koj 1
(b) Measuring method: When measuring the tyrosinase activity inhibition rate of the sample samples (1) amino-protected amino acid derivatives of kojic acid and (2) amino-protected peptide derivatives of kojic acid, first the poorly water-soluble sample was Dissolve in DMSO and then dilute with water to make DMS.
(Adjust so that the temperature is 14 degrees for a 5% DMSo aqueous solution.

Regarding the measurement of the tyrosinase activity inhibition rate of each sample, 0.4mM of each sample was used as the standard, and 1/1°115, 1/1 of this concentration (0.4m11) was measured.
lo, a concentration of 1/100 (i.e., Q, 4mM, 0
．． The inhibition rate of tyrosinase activity of each diluted sample solution at each concentration of 0.08 mM, 0.04 mM, and 0.004 mM is measured.

Add L-tyrosine solution (concentration: 0.3 mg) to each test tube.
/d) 1st grade and pine kilvain buffer (Mcllv)
aine's Buffer 5 solution
) (pH 6, 8), add 0.9 mA each of the sample diluted test solution and DMSo 5% aqueous solution for blank test to each test tube, and place it in a constant temperature water bath at 37 °C. Add 0.1 d of the reaction solution (tyrosinase solution: 1 mg/m・Matsukilhain buffer) to the incubated solution for 10 minutes, stir, and immediately set each reaction solution on a spectrophotometer at 475 nm. Measure the absorbance over time. (The absorbance value at each time point is indicated with a subscript 0 for immediately after addition of the tyrosinase solution, and a subscript X for after incubation for X minutes after addition.) The tyrosinase activity inhibition rate is calculated by substituting the absorbance value into the following 0 formula.In addition, in calculating the tyrosinase activity inhibition rate in this invention, 15 minutes after adding the reaction solution (tyrosinase solution)
Use the absorbance after minutes.

The reason for setting the time 15 minutes after the addition of the reaction solution is to ensure that the color development of the reaction solution is sufficiently stable and does not fade, and is the most suitable time for absorbance measurement to measure the inhibition rate of tyrosinase activity. This is because there are two time periods.

Tyrosinase activity inhibition rate (Bx-Bo) 1----------・----------■Formula Bo Absorbance value after 0 minutes BX Absorbance value after χ minutes of Ni blank solution Ao: Absorbance value of the test solution after 0 minutes AX: Absorbance value of the test solution after X minutes Note that each of the above absorbance values is measured based on the amount of dopachrome (precursor of melanin) produced.

(C) Example of test results: Table 7 shows example test results of the tyrosinase activity inhibition rate for each sample. Furthermore, Table 8 shows the measurement results of the tyrosinase activity inhibition rate of various amino-protected amino acids (carbobenzoxy amino acids) alone, and Table 9 shows the measurement results of the tyrosinase activity inhibition rate of various amino acids alone.

[Margins below this page] (d) Discussion: From the results in Table 7, for example, 0.04mM of kojic acid
showed a tyrosinase activity inhibition rate of 57% *2), while the amino-protected peptide of kojic acid = S form (Z--Leu
-Ala-Koj 1ate) shows a tyrosinase activity inhibition rate of 52% (*2), so the amino-protected peptide derivative of kojic acid exhibits a tyrosinase activity inhibition rate about 10 times that of kojic acid. It suggests that. In addition, the tyrosinase activity inhibitory effect of amino-protected amino acid derivatives of kojic acid was similarly observed (*2).
By comparing the inhibition rate of tyrosinase activity of kojic acid and the measured concentration of each, it was found that 0.08mM of kojic acid showed an inhibition rate of tyrosinase activity of about 80% *1).On the other hand, the amino-protected amino acid derivative of kojic acid (Z -11e4oj
0.004m? The fact that I exhibits a tyrosinase activity inhibition rate of about 81% (*1) suggests that the amino-protected amino acid derivative of kojic acid exhibits a tyrosinase activity inhibition rate about 20 times that of kojic acid.

Note that Table 8 shows the measurement results of the tyrosinase activity inhibition rate of several types of amino-protected amino acids, and Table 9 shows the measurement results of the tyrosinase activity inhibition rate of representative single amino acids. Furthermore, the results in Tables 8 and 9 suggest that Z-amino acids and single amino acids (excluding cysteine) have no tyrosinase activity inhibitory effect, or that the effect is incomparably small. ing.

Note: Amino protecting group = carbobenzoxy group] ((rz-
(abbreviated as ")") (3) Test example of cell proliferation action/effect of kojic acid, amino-protected amino acid derivatives of kojic acid, and amino-protected peptide derivatives of kojic acid: This measurement of cell proliferation action/effect was carried out on XX-male cells ( This is done by examining the culture proliferation of healthy human skin cells.

(a) Sample sample: The same sample used in the above (2) test for measuring the inhibitory effect on tyrosinase activity is used.

(b) Test basic medium: Eagle's MEM medium containing 5% tallow serum is used as the test basic medium used for cell proliferation measurement.

(C) Method for preparing the sample solution: Amino-protected amino acid derivatives of kojic acid and amino-protected peptide derivatives of kojic acid are hardly soluble in water, but DM
The kojic acid, amino-protected amino acid derivatives of kojic acid, and kojic acid have been shown to be soluble in SO and that an aqueous solution of DMSO of 0.5% or less does not adversely affect the growth of the XX-male cells. The amino-protected peptide derivative of was first dissolved in a small amount of DMSO, and then dissolved in the basic test medium to make DMSOO, 5% MEM medium, and the final concentration of each test sample was 0.0.
Each test sample contained 0.5% D to give a concentration of 5mM.
Prepare MSOMEM medium.

(d) Experimental method: Eagle's MEM medium containing 5% tallow serum is used as the basic medium.

8. XX-male cells were placed in a 4 or 5 Cφ plastic petri dish containing 1 d of the above basal medium. Seed 10' OX and culture for 1 day at 37°C and 5% carbon dioxide.

After 1 day, discard the medium and add 1 portion of the MEM medium containing each sample.
d and cultured for 4 days at 37°C and 5% carbon dioxide.

(e) Measurement of proliferation of cultured cells: The cell proliferation effect is measured by measuring the proliferation of cells after culturing.

As a specific method, after culturing the cells described in (d) above for 4 days, the cells are fixed with a 10% formalin solution, and
．． 05% naphthol blue black (9% acetic acid, 0
．． Stain the cells for 30 min with 1M sodium acetate solution).

After that, the plate was thoroughly washed with water, and after drying, the dye was eluted with a 0.05N caustic soda aqueous solution, and the absorbance at 630 nm was measured, and a quantitative curve showing the correlation between the previously measured absorbance and the number of cells was used to determine the growth rate. Determine the number of cells.

(f) Results: Table 10 shows examples of test results for measuring cell proliferation effects of kojic acid, amino-protected amino acid derivatives of kojic acid, and amino-protected peptide derivatives of kojic acid.

[Margins below this page]

From the results in Table 10 above, kojic acid has no cell proliferation activity, whereas amino-protected amino acid derivatives of kojic acid and amino-protected peptide derivatives of kojic acid have significant cell proliferation activity. It suggests that.

(4) Examples of metal ion stability tests of kojic acid, amino-protected amino acid derivatives of kojic acid, and amino-protected peptide derivatives of kojic acid: (a) Test sample: The test sample was tested in the above (2) Tyrosinase activity inhibition effect measurement test. , (3) As in the cell proliferation activity measurement test, kojic acid,
These are amine-protected amino acid derivatives of kojic acid and amino-protected peptide derivatives of kojic acid.

(c) Measuring method: An aqueous solution of 0.5 mM of the sample was prepared. On the other hand, a ferric chloride solution (concentration: 0.IM) was prepared, and the concentration of the ferric chloride solution was 511 IM for each sample (10d).
500n of the reaction solution 30 minutes after addition.
The absorbance at m was measured. (The λnmx of the reaction solution of kojic acid and ferric chloride is 500 nm, and the absorbance value of 0.5 mM of ferric chloride against the negative control (water) at 500 nm is 0.022.) Table 11 shows examples of stability test results for iron ions of kojic acid, amino-protected amino acid derivatives of kojic acid, and amino-protected peptide derivatives of kojic acid.

[Margins below this page]

[Margins below this page] From the results in Table 11, the amino-protected amino acid derivatives of kojic acid and the amino-protected peptide derivatives of kojic acid have a coloring degree of about 1/4 to 1 of that of kojic acid in the presence of ferric chloride ions. /2, which suggests that it has an excellent coloring prevention effect, and suggests that product control by coloring is extremely advantageous compared to kojic acid when blended into a product. The reason for this great anti-coloring effect is not clear, but the introduction of amino acids or peptides into kojic acid causes changes in the electron distribution state within the molecule, causing chelation with metal ions such as iron. It is thought that the formation of is inhibited.

(5) Example as a skin external preparation (prescription example) Note that this prescription example is merely an example, and it goes without saying that the present invention is not limited to the examples described below.

(a) Cream (wt%
)・Amino-protected amino acid derivative of kojic acid−・−1−−
−・・−・・−1,0 ・Stearic acid−−−−−−−−−−1−−−−−−
−−−−−−−−−−−−−−−−−−−2,0・stearyl alcohol−1−−−−−−−−−−−−
---7,0・Setanolooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooong
−−1−・・−・−=−・−・−−−−−−−4,
0.Squalane---------1------
−−−−−−−−−−−−・−・−−−−−−・10.
0.Polyoxyethylene cetyl ether (25E, O,)
−1−−−−−−−・・−3,0・Lipophilic glycerin monostearate・・−2,0・Preservative/antioxidant−−
−−１・・・−−−−−−１−−−・−・・−−−−−
1. Appropriate amount/Fragrance: 1.
−−−−・−−1−−−−−・・・・−−−−−−−−
・−・−・−・−Amount・Glycerin−−−−−−1−
−−−−−−−−−−−−−−・・−・−−−−−・−
-----------5,0・Purified water---------・・
−・・・・−・−−−１−−−−−−−・・・−
・・・Remaining amount (prescription) Heat the oil phase to 80°C and dissolve it. Meanwhile, the aqueous phase was
Heat to °C to dissolve. Add the oil phase to the water phase and stir well. Cool while stirring, add fragrance at 40°C, and take out at 30°C to make the product (b) Emulsion
(wt%) Amine-protected peptide derivative of kojic acid -1
−・・1.0・Setanol・−−m−−−−・・
−−−−−−m−・−・〜・・−・・−・−・・−・−
・-1,0.Stearic acid-1-----・-1--
−−−−−−−−・−・・・・・〜−−−−・・−−−
−−2,0・Squalane・−・−・−・−−−−−−
−−−１−−・・・・−・−・−１１１１・−10,0
・Polyoxyethylene monooleate (10E, O,)-・-・-1
, 5. Preservative/Antioxidant・・・・・・・−・−−−111−−−・−・−・−Adequate amount・Carboxyvinyl polymer−−−−−−−−−−−・・・0.2・Glycerin−・−・−・−・・・・−・・−−−−−1−・・・
・・−−−−−−・・−5,0・Potassium hydroxide−−−・・・・・・−−−−−−−・−・−・・−−
-----・0. ethanol・−−−・−・−・
−・−−−−−・・−・・−・・・・−−−−1・−7
,0・Purified water−・−・−・−−−−1−−−−−−−・
・−・・・−・−・−・−・−・・−・〜・・・・Remaining amount/Fragrance−−−−−−m−−−・・−・−−m−
−−−・−・・−１−−−−・−−１−−−−−・
-------Appropriate amount (prescription) Heat the oil phase to 80°C and dissolve.

Heat the aqueous phase to 80°C and dissolve.

Add the oil phase to the water phase and stir well. Stir while cooling, add ethanol and fragrance at 40°C, and take out at 35°C to prepare the product.

(C) Lotion (weight%)
・Polyoxyethylene sorbitan monolaurate (20E, O,) ------1・---・
2.0 Amino-protected amino acid derivative of kojic acid--
−−−−−−−−−−−−・−・・−・0.2・Ethanol−−−−−−−−−−・・・・−・−・
−−−−−−−−−m−・−・−・7.0・Preservatives/antioxidants・・・−・−−−−−−−・−・・
・−Appropriate amount・Fragrance・−・・−・・−・−1−−−−−・−
・−・−・−−−−−−−−−−−−−−−−−−−−
・−・−・Appropriate amount・Glycerin−・−・−・・・−−−−
1--
−5,0・Purified water・−−1−−・・・・・・・・・
−−−−−−−−−−1−・−・−・・・・−・−−−
-----------1 Remaining amount (prescription) Mix glycerin and purified water well.

Stir and mix the other ingredients well and add the water phase. The mixture is filtered to obtain a product.

(d) Pack (weight%
)・Amino-protected peptide derivative of kojic acid・・・・−
・−−−−−−−−−0, 5・Olive oil・・−−
−−−−−・−・−・・−１−−〜−１−−−−−−
−−−−−・−・−・5.0・Titanium oxide−・・・−・
−・−・−・−・−−１−−−−−・−・・・・・−・−
−−−8,0・Kaolin−・・−・−・・−・・−・
−・−・−・−・・−・−・・・・・・−・7.0・Preservatives・Antioxidants・−−−−・−・−・・−・−・−・−
...Appropriate amount of vinyl acetate resin emulsion------
--15.0, polyvinyl alcohol ---
−・・−・−10,0・Glycerin −・−・・・・−−
−−−−・・−１−−−−−−・・・・−・−・・−・
−・5.0・Ethanol−・−−1−−−−・・−−−
−１・・・−・−１−−−・・−・−−m−−−−−・
−5,0・Fragrance−−−−−−−−−・・・・−−−1−
−−−・−１−−−・＝・・−−−−−−−−・−−−
−−−・−・−・−Appropriate amount・Purified water・−・−・−・−
−−−−−−・・・・−・−−−−m−−・−・−−−
-1...--Remaining amount (prescription) Heat the whole to 80°C and stir sufficiently to uniformly disperse the resin and powder raw material. Cool with stirring and add flavoring at 40"C. Remove at 35°C to prepare product.

(e) Bath oil (wt%)
・Amino-protected amino acid derivative of kojic acid---1・
------0,5-polyoxyethylene lauryl ether-----io,.

・Coconut oil fatty acid polyoxyethylene glycerin--25,
0.Isopropyl myristate-・・・・・
20.0・Squalane・−−−−−−−−−
−−−・−−−−−−−−m−・−・−・・・・−・
Remaining amount/Fragrance・−・−・・−・−・−・−−−−−1−・
−・−・・−−１−−・−−１−−−・・−１−
-----Appropriate amount (prescription) Stir and mix thoroughly to make the product.

(e) Foundation (weight%
)・Stearic acid-11−−−−−−−−−−−−−
−−−−−−Unicurved surface upper−−−−5,0・Lipophilic glycerin monostearate−・−・−−−−2,5・Cetostearyl alcohol−−−−−−・−・−・
-1,0 Propylene glycol monolaurate -------3,0
・Squalane・−・〜・・−−−−1・−−−−−1・−−−−−−−1−1−−7,0・Isopropyl myristate−・−・−・−8,0・Amino protected amino acid derivative of kojic acid−−−−111・−−
---1---・-1,0・Preservative/Antioxidant----
−−−・−−−−1−−−−−−−−−−・−Appropriate amount・Purified water・−−−−−−・−−−−−〜・−・−−
−−11−−−−−−−−−−−−−−−・−−−
--1 remaining amount・Triethanolamine・・・−−1−−・−−−1,2・Glycerin−−
−−１−〜−−・−・・−・・−１−−−−・・・−・
−・−3,0・Titanium oxide−−−−・・−・−・−
−−−−１−−−−・・−−−−−−−−１・−m−−
−−・・8.0・Kaolin −・−・−−−−−・−・・
−・〜−−−−−・・・・−−−−−1・−・・−−−
−−5,0・talc ・−・−・−・−・・−・−1−
−−・−・・−−１−−−−・−−−−−１−・・−２
,0・Bentonite−・・−・−・−−−−−・−−−
1---------------・・--------...1.0・
Fragrance-1--------・--・-m--------・・・
−・−−−1−−−−−−−−・・−・−・−Appropriate amount (prescription) Use part of the oil phase for the powder raw material (let it blend in).

Meanwhile, heat the oil phase to 80° C. and stir to dissolve.

Heat the aqueous phase to 80°C and add the oil phase to it. Cool while stirring. At 40°C add the fragrance and after stirring take out at 35°C to make the product.

(6) Cleansing F (weight%)
・Stearic acid・−・−・−・−・−・−・−・−−−
−−m−−−−・・−14,0・Palmitic acid・−・
−・−・・−・・−１１１−−−−−・・−・・−・−
10,0・Myristic acid・・・・・・−・−−1−−
−−・・−・−・−・・−・ 12.0・Oleyl alcohol・−・・−・・−・−・−−−1−−−・2
．． Amino-protected amino acid derivative of 0.kojic acid------
-1----------
--1----Appropriate amount, glycerin, --m----・--
−−−11−−−−−−−−−−−−−−−−−m−
---・18.0・Potassium hydroxide--1---・-・
−−−−−m−−−−・−−−1−・−・・−6,0・
Purified water -1------・--- −・・−−−１−−−・−−−１−−・−−１−
−−−−−・−・−−−〜−−−・−・−・−−1−−
----Appropriate amount (prescription) Heat and dissolve the oil phase to 70°C.

Meanwhile, the aqueous phase is heated to 70°C and dissolved.

Gradually add the oil phase to the water phase and cool while stirring well. Add fragrance at 50°C, stir until 30°C, and take out the product.

<Effects of the Invention> (a) The amino-protected amino acid derivatives of kojic acid and the amino-protected peptide derivatives of kojic acid, which are blended as active ingredients in this invention, have an extremely excellent tyrosinase activity inhibiting effect (approximately 10 to 20% higher than that of kojic acid alone).
Since it has double the tyrosinase activity inhibitory effect), it is possible to provide external skin preparations such as cosmetics and pharmaceuticals that have a whitening effect.

(b) Since the amino-protected amino acid derivatives of kojic acid and the amino-protected peptide derivatives of kojic acid, which are blended as active ingredients in this invention, have extremely excellent cell proliferation effects, they promote skin turnover, thereby promoting the above-mentioned ( Effects that synergistically enhance the whitening effect of a)
In addition to being effective, it is absorbed into the skin and produces mucopolysaccharides and collagen in the dermal cells, thereby increasing the elasticity of the skin, improving the firmness of the skin, and making the skin fresh and beautiful.

(C) Since the amino-protected amino acid derivatives of kojic acid and the amino-protected peptide derivatives of kojic acid that are blended as active ingredients in this invention have extremely high stability against metal ions, these substances can be used in cosmetics, pharmaceuticals, etc. When blended into a product, it is possible to provide an extremely stable product without causing product changes such as coloring or discoloration, or loss of commercial value, as with conventional kojic acid-containing products.

(d) The amino-protected amino acid derivatives of kojic acid and the amino-protected peptide derivatives of kojic acid that are blended as active ingredients in this invention are kojic acid, which is harmless to humans and animals, and kojic acid, which is also harmless to humans and animals. Amino-protected amino acids.

Since they are derivatives with amino-protected peptides, they are harmless to humans and animals, and are ideal for inclusion in cosmetics and pharmaceuticals.

etc., it has an outstanding effect of achieving the purpose of the invention.

[Brief explanation of drawings]

・Figure 1 shows the amino-protected amino acid derivative of kojic acid (Z-
Infrared absorption spectrum of kojic acid (Leu-0-Kojiate), Figure 2 shows the amino-protected amino acid derivative of kojic acid (Z
-Tie-0-Koj 1ate), ・Figure 3 is the infrared absorption spectrum of the amino-protected amino acid derivative of kojic acid (Z-Ala-0-Koj 1ate), ・Figure 4 is the amine-protected kojic acid Infrared absorption spectrum of amino acid derivative (Z-Val-0-Koj 1ate), Figure 5 is infrared absorption spectrum of amine-protected peptide derivative of kojic acid (Z-Phe-Gly-Kojiate), Figure 6 is kojic acid. amino-protected peptide derivative (Z-
FIG. 2 is a spectrum diagram showing an infrared absorption spectrum of Leu-A1a4ojiate). Patent Applicant: Tomo Tomo Kunimasa Attorney: Patent Attorney Takashi Komatsu Procedural Amendment (Ho) April 2, 2001 1 Case Description 1990 Patent Application No. 3194
No. 62 No. 2 Title of the invention Skin external preparation 3 Relationship with the case of the person making the amendment Patent applicant address 3-15 Kamitsumuro, Takatsuki City, Osaka 569 +06-302 Name Tomoharu Kunimasa 4 Agent 6 Subject of amendment (1) Drawings (Figures 1 to 6) 7 Contents of amendments

Claims (1)

[Claims] (1) General formula [I] ▲Mathematical formulas, chemical formulas, tables, etc.▼[I] (In the formula, R is an amino-protected amino acid or an amino-protected peptide) Amino acid derivatives of kojic acid and peptide derivatives of kojic acid An external skin preparation characterized by containing one or more substances selected from various substance groups as an active ingredient.