JP2001316221A - Antiaging agent and cosmetic - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain an antiaging agent and cosmetic including effective liquid plant extract(s) as active ingredient which has (have) collagenase inhibitory action, lastase inhibitory action and Maillard reaction inhibitory action. SOLUTION: The excellent antiaging agent and cosmetic are respectively obtained by being based on collagenase inhibitory action, elastase inhibitory action and Maillard reaction inhibitory action observed in various kinds of extracts from respective plants. The above mentioned collagenase inhibitor, elastase inhibitor and Maillard reaction inhibitor are natural products contained in various kinds of extracts from respective plants and so safe, stable to heat, having little side effect and being capable of developing applications in wide and several technological fields including not only cosmetic field on the above mentioned cosmetics but also pharmaceuticals, foods and the like, thus demonstrating remarkable effect to achieve the purpose of the invention.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

The present invention relates to a collagenase inhibitor, an elastase inhibitor and a Maillard reaction inhibitor selected from various extract groups of each plant, and one or more of these inhibitors as an active ingredient And a cosmetic. In particular, the cosmetic according to the present invention is mainly intended to provide an anti-aging cosmetic such as skin, but the field of application of the present invention is not limited to the cosmetics field of the cosmetic, It can be widely applied to various technical fields such as food and food.

[0002]

2. Description of the Related Art It is a fate for humans to age physiologically with aging, but in recent years, physiological aging has been caused by deterioration of environmental conditions, changes in lifestyles, and increased stress accompanying the complication of social life. Needless to say, the factors that are promoted are increasing. Above all, the skin is a tissue that is constantly exposed to ultraviolet rays in addition to this physiological aging, and is considered to be a tissue that is liable to cause so-called photoaging.

[0003] Specifically, skin aging refers to the reduction or cross-linking of collagen (collagenous fibrils) and elastin (elastic fibers) in the skin extracellular matrix, the reduction of mucopolysaccharides such as hyaluronic acid, DNA damage by ultraviolet rays, etc. Is recognized as a phenomenon that promotes the formation of "sag" and "wrinkles" on the skin.

Among collagens (collagen fibers) occupying 70 to 80% of the dry weight of the dermal extracellular matrix, type I collagen, which greatly contributes to skin “stiffness”, is exposed to ultraviolet rays and aging. , Are denatured and degraded by overexpression of type I collagenase which is a type collagen degrading enzyme. Therefore, inhibiting the activity of type I collagenase is important in that it gives the skin a firmness and prevents aging of the skin.

[0005] In addition, the skin dermis extracellular matrix 2
Elastin (elastic fiber), which accounts for ~ 4%, forms crosslinks with each other and contributes to the elasticity of tissues. However, it is said that it is denatured and degraded by overexpression of elastin, an elastin-degrading enzyme, due to exposure to ultraviolet light and aging. . Therefore, inhibiting the activity of elastase is important in giving skin "elasticity" and preventing skin aging.

[0006] In general, aging means that various functions of the body deteriorate, and various physiological functions in a living body include:
In particular, proteins contribute significantly. In other words, aging is considered to be a state in which the protein is unable to perform its original function due to some kind of disorder given to this protein, and one of the disorders is non-enzymatic saccharification of protein. . This is a reaction that impairs the original function of the protein due to the binding of the reducing sugar and the amino acid or protein, and is called a Maillard reaction (browning reaction).

As dysfunction due to protein saccharification, “sags” and “wrinkles” appear on the skin of a person in contact with the outside world with aging, and “stiffness” and “gloss” of the skin decrease. Come. This is largely due to changes in the extracellular matrix of the skin dermis, and collagen (collagenous fibrils) due to glycation of proteins by the Maillard reaction.
And elastin (elastic fibers) cause crosslinking.
In particular, the cross-linking between collagen fibers associated with protein saccharification is necessary to increase mechanical strength at the beginning,
It is said that gradually increasing unnecessary cross-linking causes a decrease in skin elasticity, resulting in "sags" and "wrinkles". Therefore, inhibiting the Maillard reaction is important in that it gives the skin “tension” and “elasticity” and prevents aging of the skin.

As described above, the occurrence of “sag” and “wrinkle” due to a decrease in skin elasticity, which is seen as one of the skin aging symptoms, is caused by the generation of collagen (collagenous fibrils) and elastin (elastic fibers) in the skin extracellular matrix. It is thought to be due to the decrease or excessive progression of the formation of cross-links, and it is possible to directly add soluble collagen or elastin to the external preparation for skin or to activate fibroblasts that synthesize matrix components such as collagen. Formulation is also being considered.

However, soluble collagen or the like is a protein having a large molecular weight, and has problems in that it does not reach the site of action and stability because it is difficult to be absorbed transdermally. Care must also be taken to maintain quality such as prevention of decay and denaturation. Furthermore, there is a concern that after percutaneous absorption, it may show antigenicity, and it is hard to say that it is suitable for blending into an external preparation for skin. On the other hand, various fibroblast activators have been studied, but those with excellent percutaneous absorption, action effects, and stability, and with less adverse effects on the skin such as skin irritation and sensitization are still in use. It is slight. Also, the addition of an ultraviolet ray protective agent or an active oxygen scavenger is not sufficient to prevent a decrease in skin elasticity not caused by ultraviolet rays or active oxygen species.

[0010]

The present invention has been studied for the purpose of searching for a substance having an effect of preventing skin aging from natural products (plants). The present invention has been completed based on the finding of a substance having an elastase inhibitory effect and a substance having a Maillard reaction inhibiting effect.

Therefore, the present invention provides a stable and safe collagenase inhibitor, elastase inhibitor, and Maillard reaction inhibitor, which has no side effects selected from various extract groups of each plant, and furthermore, one of these inhibitors It is intended to provide an anti-aging agent and a cosmetic, characterized by containing one or more species as active ingredients, and each of a collagenase inhibitor, an elastase inhibitor and a Maillard reaction inhibitor according to the present invention. The active ingredient is
The cosmetics can be widely used not only in the cosmetics field but also in technical fields such as pharmaceuticals and foods.

[0012]

Means for Solving the Problems In other words, the present inventors have selected from a group of various extracts extracted from a plant consisting of cherry, red scape, blueberry, bearberry, whiteberry, chickweed, scorpion, henna and lalea. Collagenase inhibitor, elastase inhibitor and Maillard reaction inhibitor, further provided by providing an anti-aging agent and cosmetics characterized by containing one or more of these inhibitors as an active ingredient, The present invention has been completed. Hereinafter, the process leading to the present invention will be described.

[0013]

BEST MODE FOR CARRYING OUT THE INVENTION A collagenase inhibitor, an elastase inhibitor and a Maillard reaction inhibitor according to the present invention contain, as an active ingredient, one or more selected from various extract groups of each plant. The anti-aging agent and the cosmetic according to the present invention contain one or more selected from these inhibitors as an active ingredient.

[0014] The various extract groups of each plant include flowers of each plant,
After drying one or more parts (hereinafter referred to as "the original substance") of the whole plant or its leaves, branches, bark, roots, etc., or pulverizing without drying, water and / or methanol,
Use lower alcohols or lower alcohol aqueous solutions such as ethanol and propanol, polyhydric alcohols such as propylene glycol, 1,3-butylene glycol and glycerin or polyhydric alcohol aqueous solutions singly and / or in any combination of two or more solvents be able to.

The extract of each plant with lower alcohol can be obtained using a Soxhlet extractor. For other extracts, various extracts can be obtained by immersing a predetermined amount of the drug substance in a predetermined amount of solvent under predetermined conditions. And various plant extracts, lower alcohols, various organic solvents or organic solvent aqueous extracts, respectively, the lower alcohol, the organic solvent is distilled off and concentrated under reduced pressure,
With respect to the water extract, the extract can be concentrated under reduced pressure and then freeze-dried to obtain each extract. In addition, specific preparation examples of various plant extracts will be described in detail in Examples.

A collagenase inhibitor according to the present invention,
The amount (content) of various active ingredients (various extracts of each plant) in the elastase inhibitor and the Maillard reaction inhibitor depends on the type of the active ingredient and / or a combination thereof, as well as the purpose, mode, and form of use of the active ingredient. There is no particular limitation because it can be varied according to the number of uses and the like. In principle, it is sufficient that an effective amount be present, but in general, 0.000.00 for each composition of collagenase inhibitor, elastase inhibitor and Maillard reaction inhibitor.
01 to 100% by weight, preferably 1 to 10% by weight can be used.
Furthermore, although the active ingredient (various extracts of each plant) according to the present invention can exert its function and effect even by one kind, an excellent synergistic effect can be obtained by appropriately combining two or more kinds of active ingredients. Can be played. Of course, one or two of the collagenase inhibitor, elastase inhibitor and Maillard reaction inhibitor according to the present invention
An anti-aging agent containing at least one species as an active ingredient can also exhibit an excellent synergistic effect when used in combination with various known anti-aging agents.

[0017] The collagenase inhibitory effect of the various extracts of each plant according to the present invention was measured by the following method.

The collagenase inhibitory effect was measured using purified type I collagenase (0.25 unit / ml). That is, the denaturation temperature of collagen degraded by collagenase is reduced by about 10 ° C. to about 35 ° C., and furthermore, using only the collagenase activity measurement method utilizing the fact that only the denatured collagen molecular chains are soluble in ethanol, the presence or absence of an active ingredient is determined. Was searched. The specific measurement method and the method of calculating the collagenase inhibition rate (%) will be described in detail in Examples.

The collagenase inhibitor according to the present invention has industrial utility from the following viewpoints. That is, 70-80% of the dry weight of the dermal extracellular matrix
Among collagen (collagen fibrils) that occupy, type I collagen, which is greatly involved in skin "hardness", is denatured and decomposed by overexpression of type I collagen-degrading enzyme type I collagenase due to exposure to ultraviolet light and aging. It has been done. Therefore, inhibiting the activity of type I collagenase is important in that it gives the skin a firmness and prevents aging of the skin. Further, the collagenase inhibitory effect according to the present invention is present in various extracts of each plant, and among them, it has an extremely excellent collagenase inhibitory effect and has stable properties. In particular, various extracts of each plant are composites, some of which have an active oxygen scavenging effect in addition to the collagenase inhibitory effect, and have a remarkable property that they have excellent stability to heat and the like, and are useful. It is possible to provide a novel anti-aging agent and cosmetics. Furthermore, the collagenase inhibitor according to the present invention can be used in industrial fields such as pharmaceuticals and foods.

That is, the collagenase inhibitor according to the present invention was selected from a group of various extracts extracted from a plant consisting of cherry, red keiten, blueberry, bearberry, whiteberry, chickweed, turkey, henna and lalea. It is characterized by containing one or more kinds as an active ingredient.

The elastase inhibitory effect of the various extracts of each plant according to the present invention was measured by the following method.

The elastase inhibitory effect was measured using normal human fibroblasts. That is, Suc- (Ala) ₃ -p-Nitro
Using an elastase activity measurement method using anilide as a substrate,
Each was searched for the presence or absence of an active ingredient. The specific measurement method and the method of calculating the elastase activity inhibition rate (%) will be described in detail in Examples.

The elastase inhibitor according to the present invention has industrial utility from the following viewpoints. In other words, elastic fiber elastin (elastic fiber), which accounts for 2 to 4% of the dermal extracellular matrix, forms crosslinks with each other and contributes to the elasticity of tissues. However, elastase, an elastin-degrading enzyme, is exposed to ultraviolet light and aging. It is said to be denatured and degraded by overexpression of. Therefore, inhibiting the activity of elastase gives the skin "elasticity",
It is important in preventing skin aging. Further, the elastase inhibitory effect according to the present invention is present in various extracts of each plant, and among them, it has an extremely excellent elastase inhibitory effect and has stable properties. In particular, various extracts of each plant are composites, some of which have an active oxygen scavenging effect in addition to the elastase inhibitory effect, and have a remarkable property of being excellent in stability against heat and useful. It is possible to provide a novel anti-aging agent and cosmetics. Furthermore, the elastase inhibitor according to the present invention can be used in industrial fields such as pharmaceuticals and foods.

That is, the elastase inhibitor according to the present invention was selected from a variety of extracts extracted from a plant consisting of cherry, red keiten, blueberry, bearberry, whiteberry, chickweed, scotch, henna and lalea. It is characterized by containing one or more kinds as an active ingredient.

The measurement of the Maillard reaction inhibitory effect of various extracts of each plant according to the present invention was performed by the following method.

The Maillard reaction inhibitory effect was determined by comparing the inhibitory activity with aminoguanidine, which is known as a drug that inhibits this reaction, to quantify the inhibitory activity of this reaction, and to search for the presence or absence of an active ingredient, respectively. The specific measurement method and the method for calculating the Maillard reaction inhibition rate (%) will be described in detail in Examples.

The Maillard reaction inhibitor according to the present invention has industrial utility from the following viewpoints. In other words, as dysfunction due to protein saccharification in the Maillard reaction, sagging and wrinkles are seen on the skin of people who come into contact with the outside world with aging, and `` stiffness '' and `` glossy '' of the skin decrease Come. This is because changes in the extracellular matrix of the dermis have a great influence, and collagen (collagenous fibrils) and elastin (elastic fibers) crosslink due to glycation of proteins by the Maillard reaction. In particular, cross-linking between collagen fibers due to protein saccharification is necessary at the beginning to increase mechanical strength, but gradually increases unnecessary cross-linking, causing a decrease in skin elasticity and causing sagging and wrinkles. " Therefore, inhibiting the Maillard reaction
It is important in giving the skin "tension" and "elasticity" and preventing skin aging. Further, the Maillard reaction inhibitory effect according to the present invention is present in various extracts of each plant, and among them, it has an extremely excellent Maillard reaction inhibitory effect and has stable properties. In particular,
Various extracts of each plant are composites, some of which have an active oxygen scavenging effect other than the Maillard reaction inhibitory effect, etc. An anti-aging agent can be provided. Furthermore, the Maillard reaction inhibitor according to the present invention can be used in industrial fields such as pharmaceutical products and foods.

That is, the Maillard reaction inhibitor according to the present invention is selected from a group of various extracts extracted from a plant consisting of cherry, red vine, blueberry, bearberry, whiteberry, chickweed, squid, henna and lalea. It is characterized by containing one or more kinds as active ingredients.

The amounts of the various active ingredients according to the present invention in the cosmetics may be varied according to the kind of the active ingredients and / or the combination thereof, the purpose, the mode of the cosmetics, the form of use of the cosmetics and the like. Is not particularly limited. In principle, it suffices that an effective amount be present, but generally 0.0001 to 100% by weight, preferably 0.01 to 10% by weight, especially 0.1 to 10% by weight, of the cosmetic composition is available.
1-5.0% by weight is optimal. In particular, in the case of a cosmetic prepared at the time of use of the powder, the collagenase inhibitor, the elastase inhibitor and the Maillard reaction inhibitor according to the present invention are used in a high blending ratio including 100% by weight.

The application range of the cosmetic according to the present invention is:
There is no particular limitation. In other words, the present invention can be applied to all cosmetics that can utilize the respective effects according to the effects of the active ingredient of the present invention.

For example, one or more of the various active ingredients according to the present invention may be blended with various cosmetic bases and the like to prepare various basic cosmetics such as creams, emulsions, lotions, packs, face wash, etc. Various makeup products such as foundation, blusher, lipstick, white powder, etc., various hair cosmetics such as hair washes, hair tonics, shampoos, rinses, soaps, beautiful nails,
Widely applicable to other cosmetics such as cologne. In addition, the above-mentioned various aspects of cosmetics can be applied in various aspects such as solutions, emulsions, ointments, oils, waxes, sols, gels, powders, and sprays.

(Effects) The various extracts of each plant according to the present invention are all composites, and as described above, the unique effects of each plant extract besides the collagenase inhibitory effect, elastase inhibitory effect and Maillard reaction inhibitory effect. As a result, an active oxygen eliminating effect can be expected. Further, it has excellent heat stability, and has an excellent property that it can provide a highly safe anti-aging agent.

Further, as described above, various extracts of each plant according to the present invention are stable without side effects,
Moreover, it is possible to provide a cosmetic containing one or more of a safe collagenase inhibitor, an elastase inhibitor and a Maillard reaction inhibitor as an active ingredient, and thus it is possible to achieve prevention of skin aging.

[0034]

EXAMPLES Next, the present invention will be described in more detail with reference to examples, but the present invention is not limited to these examples. In addition, the part in an Example shows a weight part unless there is particular notice.

1. Preparation Examples of Various Plant Extracts (1) Preparation Example 1 (Water Extract) 2-5 g of the original substance of each plant is put in a thimble filter paper, immersed in 50-100 ml of ion-exchanged water, and heated and extracted at 60 ° C. for 8 hours. To obtain a solution. This operation was repeated three times, and all the liquids were combined and freeze-dried to obtain a water extract of each plant.

(2) Preparation Example 2 (Various Lower Alcohol Extracts) After extracting various lower alcohols as extraction solvents with a Soxhlet extractor for 8 hours, the solvent was distilled off, and the extract was powdered to obtain each plant. Various lower alcohol extracts were obtained.

(3) Preparation Example 3 (Extracts of Various Organic Solvents or Aqueous Solutions of Organic Solvents) In the extraction operation of the water extract, various organic solvents or aqueous solutions of organic solvents were used instead of water. All extracts were combined, the solvent was distilled off as much as possible, concentrated, and then lyophilized to obtain extracts of various organic solvents or aqueous organic solvent of each plant.

[0038] 2. First, the following (1) 0.25 unit / ml purified type I collagenase solution, (2) 1 mg / ml fluorescein isothiocyanate (hereinafter referred to as "FITC") labeled type I collagen solution,
(3) Prepare an enzyme reaction terminator / extract. (1) 0.25 unit / ml purified type I collagenase solution : purified human type I collagenase derived from normal human dermal fibroblasts is 0.1 M tris (hydroxymethyl) aminomethane (hereinafter simply abbreviated as "tris")-hydrochloric acid buffer (pH 7) .5, 0.4M NaCl, 10
(including CaCl ₂ , 0.04% NaN ₃ ) to make a 0.25 unit / ml solution. (2) 1 mg / ml FITC-labeled type I collagen solution : FITC-labeled type I collagen is prepared into a 1 mg / ml solution with 0.01 M acetic acid aqueous solution. (3) Enzyme reaction stopping agent / extract solution : O-phenanthroline is dissolved in a 50% by volume aqueous ethanol solution, and adjusted to a 40 mM solution with 0.5 M Tris-HCl buffer (pH 9.5, containing 0.2 M NaCl).

0.1 mg / ml sample solution (various extracts of each plant
(A 50% by volume ethanol aqueous solution) was comparatively evaluated for its inhibitory effect on collagenase. In addition, as blank
A 50% by volume aqueous ethanol solution was used for the test.

The “enzyme / sample sample (S)” was prepared by adding 0.25 unit / ml purified type I collagenase solution and 1 mg / m
l Add a FITC-labeled type I collagen solution, incubate at 35 ° C for 2 hours, then add a reaction terminator / extract to the reaction solution, treat at 35 ° C for 30 minutes, and contain collagen degraded by ethanol precipitation. Obtain the supernatant. The fluorescence intensity (Es) of the collagen contained in the supernatant at an excitation wavelength of Ex = 495 nm and a fluorescence wavelength of Em = 520 nm is measured using a fluorescence spectrophotometer. Also, `` Enzyme sample (B) ''
The same operation as in the measurement of the above “enzyme / sample sample” was performed on a blank instead of each sample solution, and Ex = 495
The fluorescence intensity (Eb) at nm, Em = 520 nm is measured.

On the other hand, in the “enzyme-free sample specimen (SB)”, 0.25 unit / ml purified type I collagenase solution and 1 mg / ml FITC-labeled type I collagen solution were added to each sample solution, and then the reaction was stopped. After adding the agent / extract and treating at 35 ° C. for 30 minutes, a collagen-containing supernatant decomposed by the ethanol precipitation method is obtained. The fluorescence intensity of collagen contained in this supernatant was measured by a fluorescence spectrophotometer, Ex = 495 nm, Em = 520.
The fluorescence intensity (Esb) at nm is measured. In addition, the “enzyme-free sample (BB)” was obtained by performing the same operation as in the measurement of the above “enzyme-free sample sample” on a blank instead of each sample solution to obtain fluorescence at Ex = 495 nm and Em = 520 nm. Measure the strength (Ebb). The collagenase inhibitory effect of various extracts of each plant according to the present invention was expressed by calculating the collagenase inhibitory rate (%) according to Equation 1. The results are shown in Table 1.

[0042]

(Equation 1)

[0043]

[Table 1]

From these results, collagenase inhibitory effects are observed in various extracts of various plants according to the present invention.

[0045] 3. First, the following (1) HAM'S F-12 medium containing 15% by volume of FBS,
Prepare (2) cell lysate and (3) substrate buffer. (1) HAM'S F-12 medium containing 15% by volume FBS : HAM'S F-12
To 106.4 g of a culture medium (manufactured by Dainippon Pharmaceutical Co., Ltd.) was added 1 liter of distilled water, and a final concentration of 15% by volume of FBS (fetal calf serum), 0.12% by weight of sodium bicarbonate, 100 mg / l of streptomycin sulfate and 70 mg / l of benzylpenicillin potassium were added. Prepare by adding. (2) Cell lysate : 0.5% by volume Triton X-100 aqueous solution. (3) Substrate buffer : 1.2 mM Suc- (Ala) ₃ -p-Nitroanilid
e, Buffer prepared by adjusting 100 mM Tris aqueous solution containing 1 mM PMSF to pH 8.0 with hydrochloric acid at room temperature.

0.1 mg / ml sample solution (various extracts of each plant
(50% by volume ethanol aqueous solution) was comparatively evaluated for its inhibitory effect on elastase. In addition, as blank
A 50% by volume aqueous ethanol solution was used for the test.

Normal human dermal fibroblast cell line (HFSKF-II)
Was adjusted to 1 × 10 ⁵ cells / ml in HAM'S F-12 medium containing 15% by volume of FBS, and seeded at 200 μl on a 96-well microplate.
The cells were cultured at 37 ° C. under 5% carbon dioxide and saturated steam.

After 24 hours, the culture was aspirated off and PBS (-)
After washing the cells twice, a cell lysis solution was added, and the cells were lysed by leaving at room temperature for 30 minutes, and this was used as an enzyme solution.

In a 96-well microplate, add 25 μl of the enzyme solution.
After adding 50 μl of the substrate buffer solution to each sample solution and further adding 25 μl of each sample solution and stirring, the plate reader: measured the absorbance (Esb) before each reaction at 405 nm and the absorbance (Ebb) of the blank before the reaction, Then, the reaction was carried out at 37 ° C. for 2 hours.
, The absorbance after each reaction (Es) and the blank after the reaction (Eb) were measured. The elastase inhibitory effect of the various extracts of each plant according to the present invention was expressed by calculating the elastase inhibition rate (%) according to Equation 2. Table 2 shows the results
It was shown to.

[0050]

(Equation 2)

[0051]

[Table 2]

From these results, various extracts of various plants according to the present invention have an elastase inhibitory effect.

[0053] 4. Method for Measuring Maillard Reaction Inhibitory Effect First, the following (1) 100 mM sodium hydrogen phosphate buffer,
(2) 2M glucose solution, (3) 4 mg / ml bovine serum albumin solution, (4) 100 mM aminoguanidine solution, (5) 100
% (W / v) trichloroacetic acid solution and (6) an alkaline phosphate buffer (-) are prepared.

(1) 100 mM sodium hydrogen phosphate buffer
(PH 7.4): the _{Na 2 HP0 4 · 12H 2 0} 3.6g dissolved in distilled water, NaH
It was prepared in ₂ P0 ₄ at pH7.4 to a total volume of 100 ml. (2) 2M glucose solution : Dissolve 18 g of glucose in distilled water to make a total volume of 50 ml. (3) 4 mg / ml bovine serum albumin solution : 4 mg / ml aqueous solution of bovine serum albumin (prepared before use). (4) 100 mM aminoguanidine solution : 13.2 mg / ml aqueous solution of aminoguanidine sulfate. (5) 100% (w / v) trichloroacetic acid solution : Dissolve 100 g of trichloroacetic acid in distilled water to make 100 ml. (6) Alkaline phosphate buffer (-) : A 0.25N aqueous sodium hydroxide solution is diluted 40-fold with a phosphate buffer (-).

0.1 mg / ml sample solution (50% of various extracts of each plant)
Volume% aqueous ethanol solution), the inhibitory effect on the Maillard reaction was comparatively evaluated. In addition, as a positive control
A 50% by volume aqueous ethanol solution was used for the test.

Apparent inhibition rate In 50 μl of each sample solution, 100
mM sodium hydrogen phosphate buffer (final concentration 50 mM) 250μ
l, 2M glucose solution (final concentration 200mM) 50μl, 4mg / ml
100 μl of bovine serum albumin solution (final concentration 800 μg / ml)
Add 50 μl of distilled water and react at 60 ° C. for 30 hours. After the reaction is completed, remove the tube from the heat block, place in a refrigerator at 4 ° C., cool, stir by vortex, and add 100 μl of the reacted solution to a new Eppendorf tube. there,
Add 10 μl of 100% (w / v) trichloroacetic acid solution. Again, vortex and then centrifuge in the cold (4 ° C, 15000
(rpm, 4 minutes), remove the supernatant by suction, and dissolve with 400 μl of alkaline phosphate buffer (-). Transfer 200 μl to a 96-well microplate for fluorescence measurement, measure the fluorescence intensity with a fluorescence plate reader: Ex = 360 nm, Em = 460 nm, and calculate the apparent inhibition rate (%) by Equation 3.

[0057]

(Equation 3)

Quenching Effect The amount of each sample solution itself that absorbs fluorescence is measured. Mix 10 μl of each sample solution with 100 μl of the positive control reaction solution. Take 100 μl into a new Eppendorf tube and put 10
After adding 10 μl of 0% (w / v) trichloroacetic acid solution, stirring by vortex, and then performing centrifugation under cooling (4 ° C., 15000 rpm, 4 minutes), the supernatant is removed by suction, and the alkaline phosphate buffer solution is removed.
Dissolve in (-) 400 μl. 200 μl was transferred to a 96-well microplate for fluorescence measurement, and a fluorescence plate reader: Ex = 360 nm,
Measure fluorescence intensity at Em = 460nm and quench effect (%)
Is calculated by Equation 4.

[0059]

(Equation 4)

True Inhibition Rate The Maillard reaction inhibitory effect of various extracts of each plant according to the present invention is calculated from the apparent inhibition rate (%) and the quenching effect (%) by the following equation (5). Calculated and expressed. The results are shown in Table 3.

[0061]

(Equation 5)

[0062]

[Table 3]

From these results, various extracts of various plants according to the present invention have a Maillard reaction inhibitory effect.

Next, a cosmetic according to the present invention was prepared using an anti-aging agent having a collagenase inhibitor, elastase inhibitor, and Maillard reaction inhibitory effects according to the present invention. The mixing ratio is part by weight.

[0065]

[Formulation Example 1] Lotion

[Production method] The above-mentioned raw materials are added to purified water and mixed uniformly.

[0067]

[Formulation Example 2] Cosmetic cream

[Production Method] The raw materials for the aqueous phase are mixed and heated to 70 ° C. to form an aqueous phase. On the other hand, the raw materials of the oil phase are mixed and dissolved by heating to 70 ° C. to obtain the oil phase. This oil phase is added to the above-mentioned aqueous phase to perform preliminary emulsification, homogenize uniformly with a homomixer, and cool to 30 ° C. to obtain a cosmetic cream.

[0069]

[Formulation Example 3] Cosmetic emulsion

[Production Method] The raw materials for the aqueous phase are mixed and heated to 70 ° C. to form an aqueous phase. Meanwhile, mix other ingredients,
Heat and melt to 70 ° C to make oil phase. The oil phase is added to the above-mentioned aqueous phase and emulsified, and cooled to 30 ° C. to obtain a cosmetic emulsion.

[0071]

[Formulation Example 4] Packing agent

[Production Method] The raw materials of the aqueous phase are mixed and made uniform. Further, other raw materials are mixed and stirred until the mixture becomes uniform to obtain a pack.

[0073]

[Formulation Example 5] Creamy foundation

[Production Method] A part of the oil phase and the powder are put on a three-roll mill, and the remaining oil phase is added and dissolved by heating, and kept at 80 ° C. Next, the aqueous phase dissolved by heating is gradually added, and the mixture is emulsified at 80 ° C. and cooled to room temperature while stirring to obtain a creamy foundation.

[0075]

The present invention can provide an excellent anti-aging agent and cosmetic based on the collagenase inhibitory effect, elastase inhibitory effect, and Maillard reaction inhibitory effect of various extracts of each plant. Moreover, since the collagenase inhibitor, elastase inhibitor and Maillard reaction inhibitor are natural products contained in various extracts of each plant, they are safe, stable in heat, etc., have few side effects, and have the cosmetics of the cosmetic. The present invention can exert remarkable effects to achieve the object of the invention, such as opening up various fields such as pharmaceuticals and foods as well as various fields.

[Brief description of the drawings]

 None

──────────────────────────────────────────────────続 き Continued on the front page (51) Int.Cl. ⁷ Identification symbol FI Theme coat ゛ (Reference) A61K 7/021 A61K 7/021 35/78 35/78 CNW A61P 43/00 107 A61P 43/00 107 111 111 C12N 9/99 C12N 9/99 F term (reference) 4B018 MD61 ME10 MF01 4C083 AA082 AA111 AA112 AA122 AB032 AB242 AB432 AB442 AC012 AC022 AC102 AC122 AC182 AC352 AC422 AC432 AC442 AD092 AD112 BB51 CC01 CC04 CC05 CC07 CC12 DD23 DD23 EE12 FF01 FF05 4C088 AB12 AB40 AB44 AB52 AC01 BA09 BA10 CA06 MA02 MA22 MA28 MA63 NA14 ZA89 ZC20

Claims (4)

[Claims] 1. An active ingredient comprising one or more active ingredients selected from a group of various extracts extracted from a plant consisting of cherry, red scape, blueberry, bearberry, whiteberry, chickweed, himauikyo, henna and lalea. A collagenase inhibitor characterized by containing 2. An active ingredient comprising one or more selected from a variety of extracts extracted from a plant consisting of cherry, red scape, blueberry, bearberry, whiteberry, chickweed, chickweed, henna, lalea. An elastase inhibitor characterized by containing 3. An active ingredient comprising one or more selected from a variety of extracts extracted from a plant consisting of cherry, red scape, blueberry, bearberry, whiteberry, chickweed, chickweed, henna and lalea. A Maillard reaction inhibitor characterized by containing 4. An active ingredient comprising one or more selected from the collagenase inhibitors, elastase inhibitors and Maillard reaction inhibitors according to claim 1, 2 and 3. Anti-aging agents and cosmetics.