JP2014221740A - Cosmetic composition - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide an external composition for skin having an excellent function independently or by combining an extract derived from multiple herbal medicines having a biologically active effect.SOLUTION: Provided are an external composition for skin which contains herbal medicine extracts obtained from one or more kinds selected from Pruni cortex, Magnolia bark, Glechoma hederacea subsp. grandis and pine needles, and an external composition for skin which contains herbal medicine extracts containing a pine-needle extract and also containing one or more kinds of herbal medicine extracts selected from Glechoma hederacea subsp. grandis and Pruni cortex, and Pruni cortex. The composition has a biologically active effect as a Maillard reaction inhibitor, an involucrin production promoter, an antioxidant, an elastase inhibitor, a hyaluronidase inhibitor, and a fibroblast activator, and is useful as cosmetics or medicated cosmetics.

Description

  The present invention relates to a cosmetic composition. Specifically, the present invention relates to a herbal extract having various actions useful for prevention and improvement of various symptoms that occur in daily life, and a composition for external use with the skin.

Attention has been focused on the physiological activity of plant-derived natural products such as herbal extracts, and attempts have been made to blend them into cosmetics.
Cherry bark is a herbal medicine that uses bark such as Yamazakura of the Rosaceae family, and the main components are flavonoids and flavonoid glycosides. As a pharmacological effect by ingestion, detoxification, antipyretic, antitussive, expectorant, draining action, astringent action, and the like are known, and the topical skin preparation is used for the treatment of eczema and urticaria. As the use for cosmetics, attempts to blend into whitening cosmetics and hair nourishing cosmetics are described (Patent Documents 1 to 3). Moreover, utilization as a hyaluronic acid degradation inhibitor has also been reported, and attempts have been made to blend it into cosmetics in anticipation of the effect of preventing skin roughness, wrinkles, and roughness (Patent Document 4).

  Koboku is a herbal medicine made from the bark of the magnolia magnolia genus. The main components are essential oil components, sesquiterpenes, phenylpropanoids, and alkaloids. As pharmacological effects by oral intake, antibacterial action, antispasmodic action, healthy stomach action and the like are known.

  Examples of the use for external preparations or cosmetics include whitening cosmetics (Patent Documents 5 to 7, 14, 16), acne skin improvement agents (Patent Document 8), antioxidants (Patent Document 9), photoaging protection agents ( Patent Literature 10), elastase activity inhibitor (Patent Literature 11), histamine release inhibitor (Patent Literature 12), collagenase activity inhibitor (Patent Literature 15), and preadipocyte differentiation inducer (Patent Literature 17). It has been reported. In addition, an attempt has been made to blend into cosmetics in anticipation of the effect of preventing skin roughness and maintaining the gloss and elasticity of the skin, using a solvent extract of the related flower of the genus Otamatamaki as an antioxidant (patented) Reference 13).

  Kakidoshi is a herbal medicine that uses the whole plant of the medicinal plant of Lamiaceae Kakidooshi, also known as ream, grass, and turf. The main ingredients are sesquiterpenes, flavonoids, saponins, tannins. Although it is native to Japan, it has long been used as a folk medicine (medical herb) in Europe. As a pharmacological effect by ingestion, tonicity, convergence, fever, diuresis, hypoglycemic action and the like are known.

  As for use in cosmetics, it has been reported that a monitor test was conducted with an extract of ethanol, propylene glycol or purified water in anticipation of a moisturizing effect and an improvement effect on the skin. It has not been done (Patent Document 18).

  Matsuba is a herbal medicine using Pinus densiflora leaves, and the main components are essential oil components, waxy components, and pine ani. As a pharmacological effect by ingestion, blood flow promoting action, expectorant / analgesic / healthy stomach action and the like are known.

  As cosmetic use, hair nourishing makeup containing matsuya extract, red pine needle extract and red pine root extract skin moisturizing cosmetic composition (Patent Document 19), pine fresh leaves alcohol extract (Patent Document 20) is described.

  Natural products derived from plants such as herbal medicines are guaranteed to be safe because they have been used orally for many years by humans. When used on human skin as a raw material for cosmetics, quasi drugs or pharmaceuticals, Is also safe. However, when natural products are used alone, many of them have weak physiological activity compared to synthetic products. In addition, cosmetics are expected to have multiple effects on a single product, such as preventing spots, wrinkles and rough skin, maintaining moisture and flexibility, adjusting the texture of the skin, and giving a beam, gloss and smoothness. Many. In addition, recently, “short-time cosmetics” that can make up quickly and easily have become popular, and there is an ever-increasing demand for a single product to have multiple effects such as serum, emulsion, and makeup base. .

Patent No. 2799603 Japanese Patent No. 29602558 Japanese Patent No. 2871863 JP-A-10-130162 Japanese Patent No. 3592918 Patent 3696271 JP-A-8-92055 JP2005-8572 JP 2002-154922 A JP 2002-104924 JP 2000-178168 A JP 2000-169383 A Japanese Patent No. 3268965 Japanese Patent No. 2957334 JP 2003-146876 A 7-91179 JP2002-138045 Patent No. 4057170 Special table 2012-519165 JP-A-2-164811

  The present applicants have been studying the use of a substance derived from a natural product as a raw material for cosmetics (Japanese Patent No. 4795475, “Agent containing ume extract”, etc.). And this time, when the extract obtained from a specific some crude drug was used for skin, it discovered various effects useful for prevention and improvement of various symptoms which occur with daily life. It is an object of the present invention to provide an external composition for skin having an excellent function by blending such extracts alone or in combination.

  The present inventors diligently investigated the physiological activity of extracts obtained from various herbal medicines. In addition to the high Maillard reaction inhibitory action and the involucrin production promoting action, each of the herbal extract of cherry bark, Koboku, Kakidoshi, and Matsuba The inventors have found a plurality of previously unknown actions such as an oxidizing action, an elastase activity inhibitory action, a hyaluronidase activity inhibitory action, a fibroblast activation action, etc., and completed the present invention.

That is, the present invention provides an external composition for skin containing a herbal extract obtained from one or more selected from cherry bark, Kobak, Kakitsu, and Matsuba.
A preferred embodiment of the external composition for skin of the present invention containing two or more kinds of herbal extracts is one or more extracts selected from Kakitsu, Kobaku, and cherry bark in addition to the pine needle extract. It is a composition for external use containing the skin. Another preferred embodiment is an external composition for skin comprising an extract obtained from pine needles and cherry bark. Another preferred embodiment is an external composition for skin comprising an extract obtained from pine needles and fences. Yet another preferred embodiment is an external composition for skin comprising an extract obtained from pine needles and Kompak. Yet another preferred embodiment is an external composition for skin comprising an extract obtained from Matsuba, Kobaku, and Kakitsu. Yet another preferred embodiment is an external composition for skin comprising an extract obtained from pine needles, Kobak, Kakitsu, and cherry bark.

The composition for external use of the skin of the present invention contains a substance derived from another natural product in addition to a herbal extract obtained from one or two or more selected from cherry bark, Kobak, Kakitsu, matsuba. You may go out.
The present invention also provides a Maillard reaction inhibitor containing a cherry bark extract as an active ingredient.

Moreover, this invention provides the Maillard reaction inhibitor which contains a pine needle extract as an active ingredient.
Moreover, this invention provides the Maillard reaction inhibitor which contains a magnolia extract as an active ingredient.

The present invention also provides an external composition for skin containing any of the Maillard reaction inhibitors described above alone or in combination.
The present invention also provides an involucrin production promoter containing a pine needle extract as an active ingredient.

The present invention also provides an involucrin production promoter containing a cherry bark extract as an active ingredient.
Moreover, this invention provides the involucrin production promoter which contains a mackerel extract as an active ingredient.

The present invention also provides an involucrin production promoter containing a fence extract as an active ingredient.
The present invention also provides a composition for external use on skin containing any of the above involucrin production promoters alone or in combination.

The present invention also provides an antioxidant containing a cherry bark extract as an active ingredient.
Moreover, this invention provides the antioxidant which contains a magnolia extract as an active ingredient.
Moreover, this invention provides the antioxidant which contains a pine needle extract as an active ingredient.

The present invention also provides an external composition for skin containing any one of the above antioxidants alone or in combination.
The present invention also provides an elastase activity inhibitor containing a cherry bark extract as an active ingredient.

The present invention also provides an elastase activity inhibitor containing a pine needle extract as an active ingredient.
The present invention also provides an external composition for skin containing any of the above elastase activity inhibitors alone or in combination.

The present invention also provides a hyaluronidase activity inhibitor containing a cherry bark extract as an active ingredient.
The present invention also provides a hyaluronidase activity inhibitor containing a pine needle extract as an active ingredient.

The present invention also provides an external composition for skin containing any one of the above hyaluronidase activity inhibitors alone or in combination.
The present invention also provides a fibroblast activator containing a cherry bark extract as an active ingredient.

Moreover, this invention provides the fibroblast activator which contains a fence extract as an active ingredient.
The present invention also provides an external composition for skin containing any of the fibroblast activators described above alone or in combination.

  The above-mentioned external composition for skin is particularly useful as a cosmetic or medicated cosmetic.

  The external composition for skin according to the present invention contains an active ingredient having an antioxidant action, an anti-aging action, an anti-inflammatory action, and an action for maintaining a healthy structure of the epidermis and dermis. Excellent in preventing and improving skin barrier function deterioration, texture roughness and rough skin caused by ultraviolet rays and the like.

The migration result of the Maillard reaction product when the sample is allowed to act at 4.0% is shown. The migration result of the Maillard reaction product when the sample is allowed to act at 2.0% is shown. Continuation of FIG.

  Cherry bark (Ohhi), Koboku, Kakidoshi (Oki-doshi), and Matsuba (Matsuba) used in the present invention are all so-called herbal medicines (Japanese pharmacopoeia or externally listed products, or Food).

Cherry bark refers to dried bark of the Rosaceae Yamazakura.
Koboku refers to the dried bark of the magnoliaceae honoki.
Kakidoshi refers to dried whole plant of Lamiaceae oyster.

Matsuba refers to the dried leaves of Pinus densiflora.
Herbal extracts of cherry bark, Kobak, Kakidori, and Matsuba are obtained by drying the extract obtained by using each of the above-mentioned materials as an extraction raw material, a diluted or concentrated solution of the extract, and the extract. A dried product, or any of these roughly purified products or purified products is included.

  There are no particular limitations on the method of preparing the extract of each of the herbal medicines of cherry bark, Kobaku, Kakitsu, and Matsuba, and it can be obtained from each of the above materials by an extraction method that is generally used for plant extraction. As an example of the preparation method used in the present invention, for example, extraction can be performed from room temperature to warm using an appropriate solvent. As a solvent used for extraction, water, a hydrophilic organic solvent, etc. can be used individually by 1 type or the mixed solvent which combined 2 or more types, for example.

  Examples of the water that can be used as the extraction solvent include purified water, distilled water, pure water, tap water, well water, mineral water, and the like, and those obtained by subjecting them to various treatments. Examples of the treatment applied to water include heating, sterilization, filtration, ion exchange, adjustment of osmotic pressure, buffering, and the like. The water that can be used as the extraction solvent includes physiological saline, phosphate buffer, phosphate buffered saline, and the like.

  Examples of the hydrophilic organic solvent include lower alcohols (methanol, ethanol, 1-propanol, 2-propanol, 1-butanol, 2-butanol, etc.), polyhydric alcohols (glycerin, propylene glycol, 1,3- Butylene glycol, 1,3-propanediol, etc.), ketones (acetone, methyl ketone, etc.), ethers (diethyl ether, tetrahydrofuran, etc.), and a mixed solvent of these hydrophilic organic solvents and water can be used. it can. Examples of preferred extraction solvents are ethanol, butylene glycol or a mixed solvent of 1,3-propanediol and water.

It is not necessary to employ a special extraction method for obtaining a crude drug extract from each of the extraction raw materials, and extraction can be performed using an arbitrary extraction device at room temperature or under reflux heating.
For example, when performing extraction at room temperature, each extraction raw material is put in a treatment tank filled with the extraction solvent so that the amount of the extraction raw material becomes 1.5 to 10% (mass ratio), and further stirred as necessary. On the other hand, the extract can be obtained by standing for 1 to 3 days to elute soluble components and then filtering to remove solids. In order to obtain a diluted or concentrated solution of the extract, a dried product of the extract, or a crude purified product or purified product thereof, the obtained extract is diluted, concentrated, dried, purified, etc. according to a conventional method. Processing may be performed.

  Further, the extract can be purified for the purpose of decolorization, deodorization, etc. within a range that does not cause a decrease in its physiological activity, but there is no practical problem even if it is not purified. Such purification can be performed by, for example, activated carbon treatment, adsorption resin treatment, ion exchange resin treatment, and the like. There is no restriction | limiting in particular as a means for deodorizing, The existing technique used for the same objective can be applied. One typical method is to use activated carbon.

  The extract can be formulated in accordance with a conventional method, and the formulated product is Maillard reaction inhibitor, involucrin production promoter, antioxidant, elastase activity inhibitor, hyaluronidase activity inhibitor, fibroblast It may be used as an activator.

  Furthermore, the Maillard reaction inhibitor, involucrin production promoter, antioxidant, elastase activity inhibitor, hyaluronidase activity inhibitor, and fibroblast activator of the present invention can be used in combination with other compositions. As such a composition, an external composition for skin is preferable.

  The external composition for skin of the present invention can be produced according to a conventional method. In addition, the composition for external use of the skin of the present invention is not limited to general cosmetics, but skins of quasi drugs (for example, medicinal cosmetics or other quasi drugs), designated quasi drugs, external drugs, etc. Including external preparations, the dosage form can be arbitrarily selected according to the purpose.

  In the composition for external use of the skin of the present invention, in addition to the Maillard reaction inhibitor, the involucrin production promoter, the antioxidant, the elastase activity inhibitor, the hyaluronidase activity inhibitor, the fibroblast activator, other ingredients to be formulated include cosmetics Alternatively, any drug that can be normally used in each drug can be used, and may be appropriately selected depending on the efficacy and effect.

  In the composition or agent of the present invention, the content of each herbal extract described above is not particularly limited as long as the desired effect is exhibited, but was obtained by the method described in Example 1 of the present invention. As an amount corresponding to the solvent extract, usually about 0.01 mass (w / w)% (hereinafter, expressed as%) or more can be blended. The content range of the Spruce extract is 0.01% to 50%, preferably 0.1% to 10%, more preferably 0.3% to 4%. The content range of the extract is from 0.01% to 50%, preferably from 0.02% to 10%, more preferably from 0.05% to 1%. The content range of the oyster extract is 0.01% to 50%, preferably 0.1% to 10%, more preferably 0.2% to 2%. The content range of the pine nut extract is 0.01% to 50%, preferably 0.1% to 10%, more preferably 0.5% to 1.5%. When referring to the content of the herbal extract in the present invention, it means the amount corresponding to the extract obtained by the method described in Example 1 of this specification, unless otherwise specified.

  The composition or agent of the present invention contains a substance derived from another natural product in addition to a herbal extract obtained from one or two or more selected from cherry bark, Kobak, Kaki-dori, and Matsuba. May be included. Examples of such substances include: Ume extract described in Japanese Patent No. 4795475, for example, ume root extract and ume fruit water. Steam-distilled water derived from plants described in JP 2012-116761, for example, steam distillation of one or more kinds of plants selected from passion fruit, Shimamikan, banana, Atemoya, plum, tankan, lemon lime, ginger water. Deep ocean water described in JP-A-2000-290159. Deep sea water concentrate described in JP-A-2007-217356. Toki extract described in Japanese Patent No. 3544505. Seawater-derived yeast culture described in JP2009-029788. Plum juice fermented liquid described in Japanese Patent No. 2563662. Yunohana and / or Yunohana extract or oxidized product described in Japanese Patent No. 3967357. However, substances derived from natural products that can be contained in the composition or agent of the present invention are not limited to these, and any other substance can be used.

  In the composition or agent of the present invention, fats, waxes, hydrocarbons, fatty acids, alcohols, esters, which are components used in ordinary compositions or agents for external use, as long as the desired effects are not impaired. , Amino acids, vitamins, surfactants, pH adjusters, preservatives, fragrances, moisturizers, powders, UV absorbers, thickeners, pigments, antioxidants, whitening agents, anti-inflammatory agents, anti-wrinkle agents In addition, components such as a rough skin improving agent, an acne agent, alkalis, a chelating agent, and a metal sequestering agent may be blended. In addition, ingredients such as sodium sulfate, sodium hydrogen carbonate, calcium carbonate, potassium chloride, oily ingredients, emulsifiers, succinic acid, herbal medicines, inorganic pigments, fragrances, and pigments, which are components used in ordinary bath additives, You can also.

  The composition of the present invention can also be prepared into compositions of various dosage forms such as a solid agent, a semisolid agent, and a liquid agent depending on the purpose of use. The composition of the present invention can be in any form of cosmetics, quasi drugs, and pharmaceuticals.

The form as a pharmaceutical can be a plaster, an ointment, a poultice, a liniment, a lotion, for example.
The form as a cosmetic is not particularly limited, but for example, a facial cleanser, a cleansing agent, a lotion, a cosmetic liquid, a pack, a massage cream, an emulsion, a moisture cream, a lip balm, etc. as a skin care cosmetic, a foundation, a white powder as a makeup cosmetic Lipsticks, cheeks, eye shadows, body care cosmetics such as soap, liquid cleaning agents, bath preparations, etc. hair care cosmetics such as shampoos, rinses, hair treatments, hair conditioners, hair tonics, hair restorers, scalp treatments, etc. it can. Various cosmetic ingredients generally used in cosmetics can be appropriately blended in the cosmetic.

  When the composition of the present invention is in the form of a milky lotion, cream, lotion or serum, the preferred content of each herbal extract is about 0.05% to 10%, more preferably about 0.1 to 5%, more preferably about 0.2-2%. When the composition of the present invention is in the form of cleansing oil, it is effective with a smaller blending amount than other forms of cosmetics, and the preferred content of each herbal extract is 0.005% to 0.05%, for example, about 0.01 %.

  Cosmetics or medicinal cosmetics containing the Maillard reaction inhibitor, involucrin production promoter, antioxidant, elastase activity inhibitor, hyaluronidase activity inhibitor, fibroblast activator of the present invention are reduced skin roughness and barrier function, Since it is excellent in the effect of preventing and improving the roughness of the texture, cosmetics in the form for external use that appeal for such an effect are suitable.

  In the present invention, the “Maillard reaction” refers to a reaction in which a brown substance (melanoidin) is generated from a sugar and a protein by an aminocarbonyl reaction, unless otherwise specified. The Maillard reaction is usually a non-enzymatic reaction in which an Amadori rearrangement product (Amadori compound) is generated after a reducing sugar reacts with an amino group of a protein to form a Schiff base. Then, through complex reactions such as oxidation, dehydration, condensation, intramolecular and intermolecular cross-linking, the mixture gradually becomes yellowish brown and AGEs (advanced glycation endproducts) that are late stage products of the Maillard reaction are produced.

  The Maillard reaction that occurs in vivo is considered to be involved in the formation of brown spots on the skin found in diabetic patients and the like. AGE is included in the pigment that causes brown spots. In vivo Maillard reaction has also been shown to be associated with lifestyle-related diseases, diseases accelerated by aging (eg, diabetic complications, Alzheimer's disease, arteriosclerosis). AGE is a generic name for various structures produced from the Amadori rearrangement product through complex reactions such as oxidation, dehydration, and condensation, including carboxymethyllysine, pyralin, pentosidine, croslin, imidazolone, etc. A polymer of protein dimer or higher is also known as AGE. It is thought that wrinkles, dullness, sagging, etc. are recognized by cross-linking and denaturation of dermal constituent proteins such as collagen and elastin as the Maillard reaction proceeds in the skin.

  The presence and / or degree of Maillard reaction suppression ability can be confirmed and measured by a person skilled in the art as appropriate using known means, for example, inhibition of production of Amadori rearrangement products or inhibition of AGE production as an index. it can. For detailed conditions, reference can be made to the Examples section of this specification. In the present invention, when the presence and / or degree of Maillard reaction suppression ability is mentioned, it is based on the method described in the examples of the present specification, unless otherwise specified.

  In the present invention, “involucrin production” means production of involucrin by human keratinocytes (human skin epidermal keratinocytes), unless otherwise specified. Involucrin is one of the precursor proteins involved in the formation of the peripheral zone of the stratum corneum and plays an important role in the barrier function of the skin epidermis. The stratum corneum is composed of various substances such as keratin and lipid, covers the living body, and retains moisture and protects against intruders. The surrounding zone is an insoluble structure that lines the cell membrane of keratinocytes, and involcrine made from keratinocytes in the spinous layer is cross-linked by loricrin made in the granular layer and enzymes such as transglutaminase. Arise. A part of involucrin binds to ceramide and participates in the formation of an intercellular lipid lamellar structure. The effect of normalizing the barrier function of the skin epidermis can be expected by promoting the involucrin production of keratinocytes.

  The presence and / or extent of the ability to promote involucrin production can be appropriately confirmed and measured by those skilled in the art using known means. For detailed conditions, reference can be made to the Examples section of this specification.

  “Antioxidation” refers to suppressing active oxygen (not generating, erasing, suppressing action). The presence and / or degree of the antioxidant ability of the test substance can be confirmed and measured as appropriate by those skilled in the art using known means. For example, DPPH (1,1-diphenyl-2-picrylhydrazyl) radical scavenging, superoxide dismutase (SOD) -like action, hydrogen peroxide (H2O2) scavenging, TRAP (Total Radical-trapping Antioxidant Parameter), FRAP (Ferric Reducing Ability of Plasma), β-carotene fading, etc. can be used as indicators.

  In the present invention, the presence or absence and / or degree of antioxidant ability refers to the method (DPPH radical scavenging effect test) described in the examples of the present specification, unless otherwise specified. For detailed conditions for the test, reference can be made to the Examples section of this specification.

  The presence of the DPPH radical scavenging effect indicates the possibility of removing unpaired electrons from radicals made from oxygen. That is, radicals made from oxygen do not oxidize other substances in the cell, and an effect of suppressing diseases and aging caused by active oxygen can be expected.

  In the present invention, “elastase” refers to a protease having elastin as a typical substrate, unless otherwise specified. Elastin is a kind of a hard protein that is rich in elasticity and constitutes connective tissues, tendons, aortic skin, cervical cord and the like of higher animals. There are many crosslinks between peptide chains constituting this protein, which brings about elasticity.

  In the dermis, elastin binds the reticulated collagen and maintains the firmness of the skin together with the collagen. Therefore, when elastin is reduced by aging, ultraviolet rays, active oxygen, stress, elastin degrading enzyme (elastase), it causes aging such as wrinkles and sagging. From this, it can be expected that an ingredient that inhibits elastase has an effect of restoring or maintaining the firmness of the skin and, as a result, keeping the skin youthful.

  The presence and / or extent of the ability to inhibit elastase activity can be appropriately confirmed and measured by those skilled in the art using known means. For detailed conditions, reference can be made to the Examples section of this specification. In the present invention, the presence and / or degree of inhibition of elastase activity is based on the method described in the examples of the present specification, unless otherwise specified.

  “Hyaluronidase” is an enzyme that lowers the molecular weight of hyaluronic acid and hydrolyzes the N-acetyl-D-hexosaminide bond of hyaluronic acid. Hyaluronic acid is a kind of glycosaminoglycan having a repeating structure of only disaccharides of O-β-D-glucuronosyl (1 → 3) -N-acetyl-β-D-glucosaminyl (1 → 4) units. Hyaluronic acid is abundant in connective tissues such as animal joint fluid and dermis surface, and is involved in joint lubrication and skin softening. Hyaluronidase is present in the skin and is thought to play a part in cellular activities in the reconstruction of connective tissue by degrading hyaluronic acid. Therefore, an inhibitor against hyaluronidase that degrades hyaluronic acid in connective tissue is expected to have an effect of improving the state of the skin in which the degradation of hyaluronic acid is enhanced. It is also considered that hyaluronidase is activated during inflammation, destroys the structure of tissues, and enhances the permeability of inflammatory cells. It is also involved in the release of histamine from mast cells, and is thought to be involved in the development of type I allergies. Therefore, it is possible to predict the anti-inflammatory action or anti-allergic action of the test substance by measuring the hyaluronidase activity inhibitory effect.

  The presence and / or extent of the ability to inhibit hyaluronidase activity can be confirmed and measured as appropriate by those skilled in the art using known means. For detailed conditions, reference can be made to the Examples section of this specification. In the present invention, the presence and / or degree of inhibition of hyaluronidase activity is based on the method described in the examples of the present specification, unless otherwise specified.

  In the present invention, “fibroblast activation (activity, effect)” means that the proliferation ability of normal human dermal fibroblasts (NHDF) can be enhanced. The presence / absence / degree of cell proliferation can be determined by counting the number of living cells. Various methods are known for determining the number of viable cells.

  The presence or absence and / or degree of the ability to activate fibroblasts can be appropriately confirmed and measured by those skilled in the art using known means. For detailed conditions, reference can be made to the Examples section of this specification. In the present invention, the presence or absence and / or degree of fibroblast activation activity is based on the method described in the examples of the present specification, unless otherwise specified.

  Fibroblasts are present in the dermis and produce components such as collagen, elastin, and hyaluronic acid. As the age increases, the growth rate of the cells decreases and the amount of components produced from the cells decreases, and the elasticity of the skin is lost, so-called aging symptoms such as sagging and wrinkles appear. Fibroblast activation can be said to be a means corresponding to such an aging phenomenon.

[Example 1: Production example of sample for evaluation]
Preparation of Spruce Extract: 3 % (w / w) of cherry bark in 30% (w / w) mixed solvent of purified water and butylene glycol or 50% (w / w) mixed solvent of purified water and ethanol. It was immersed so as to become w) and extracted at 25 ° C. for 3 days. After extraction, aseptic filtration was performed with a 0.45 μm filter to obtain a spruce extract.

Preparation of Kokuboku extract Kobaku was added to 3% (w / w) of 30% (w / w) mixed solvent of purified water and butylene glycol, or 50% (w / w) mixed solvent of purified water and ethanol. It was immersed so as to become w) and extracted at 25 ° C. for 3 days. After extraction, aseptic filtration was performed with a 0.45 μm filter to obtain a kumoku extract.

Preparation <br/> fence through the ground ivy extract, 30% (w / w) mixed solvent of purified water and butylene glycol or 50% of the purified water and ethanol (w / w) mixed solvent, or purified water and ethanol, Was immersed in a 95% (w / w) mixed solvent or purified water at 1% (w / w) and extracted at 25 ° C. for 1 day. After extraction, aseptic filtration was performed with a 0.45 μm filter to obtain a kakidoshi extract.

Preparation of Matsuba extract The pine needle was mixed with 30% (w / w) mixed solvent of purified water and butylene glycol, or 5% (w / w) in 50% (w / w) mixed solvent of purified water and ethanol. And extraction was performed at 25 ° C. for 3 days. After extraction, aseptic filtration was performed with a 0.45 μm filter to obtain a Matsuba extract.

  In addition, these extract solutions were also subjected to deodorization treatment. After the extraction liquid was prepared in the same manner as above, activated carbon was added so as to be 5% (w / w), and the mixture was shaken at 110 rpm at room temperature for 3 hours for deodorization treatment. After shaking, aseptic filtration was performed with a filter in the same manner to obtain a Matsuba extract from which activated carbon had been removed.

[Example 2: Evaluation of extract]
Test method:
(1) Elastase activity inhibitory effect test (derived from pancreas)
Add 50 μL of evaluation sample prepared with purified water to 96 well plate, 35 μL of 0.1 M Tris-HCl buffer, 100 μL of 1 mM Suc-Ala-Ala-Ala-pNA, 15 μL of 0.25 U / mL elastase, 37 ° C., After reacting for 30 minutes, the absorbance at 405 nm was measured with a microplate reader. Using purified water as a control, the inhibition rate was calculated using the following formula.

Elastase activity inhibition rate (%) = {1- (A−B) / (C−D)} × 100
A: Absorbance when sample is added, B: Absorbance when sample is added and elastase is not added
C: Absorbance when purified water is added, D: Absorbance when purified water is added and elastase is not added
(2) Elastase activity inhibitory effect test (derived from fibroblasts)
Human dermal fibroblasts were seeded in a 96-well microplate at a concentration of 20 × 10 ⁴ cells / mL at 100 μL / well. The seeding medium was cultured in a CO ₂ incubator (5% CO ₂ , 37 ° C.) for 24 hours using a DMEM medium containing 5% fetal bovine serum (FBS). After 24 hours, 0.5% Triton X-100 / purified water was added to prepare a crude enzyme solution. Add 50 μL of the evaluation sample to the crude enzyme solution and 150 μL of Suc-Ala-Ala-Ala-pNA adjusted to 2 mM with 0.1 M Tris-HCl buffer, react at 40 ° C. for 2 hours, and then absorb the absorbance at 405 nm with a microplate reader. It was measured. The inhibition rate was calculated using the following formula as a purified water control.

Elastase activity inhibition rate (%) = {1- (A−B) / (C−D)} × 100
A: Absorbance when sample is added, B: Absorbance when sample is added and elastase is not added
C: Absorbance when purified water is added, D: Absorbance when purified water is added and elastase is not added
(3) DPPH radical scavenging effect test
Microplate reader: 15 μL of evaluation sample prepared with purified water in 96 well plate, 15 μL of 0.2 M MES buffer 60 μL, 50 μL of 95% ethanol, 25 μL of 0.6 mM DPPH radical, reacted at 30 ° C. for 30 minutes The absorbance at 540 nm was measured. The elimination rate was calculated using the following formula with purified water as a control.

DPPH radical scavenging rate (%) = {1- (A−B) / (C−D)} × 100
A: Absorbance when sample is added, B: Absorbance when sample is added and DPPH radical is not added
C: Absorbance when purified water is added, D: Absorbance when purified water is added and DPPH radical is not added
(4) Hyaluronidase activity inhibitory effect test
Test sample prepared with purified water in 96-well plate 12 μL, 400 U / mL hyaluronidase 12 μL was mixed, incubated at 40 ° C. for 20 minutes, 0.1 mg / mL Compound 48/80 12 μL was added, and reaction was carried out at 40 ° C. for 20 minutes Then, 12 mg of 1 mg / mL potassium hyaluronate was added and reacted at 40 ° C. for 40 minutes.

  Thereafter, 12 μL of 0.4N NaOH was added, the reaction was stopped by cooling with ice water for 5 minutes, 12 μL of 0.8 M potassium borate solution was added, heated for 3 minutes with boiling water, and then cooled with ice water for 10 minutes. After adding 180 μL of 20 mg / mL p-DMAB and dispensing 180 μL to a 96-well plate, the mixture was reacted at 40 ° C. for 20 minutes, and the absorbance at 540 nm was measured with a microplate reader. The inhibition rate was calculated using the following formula with purified water as a control.

Calculation of hyaluronidase activity inhibitory effect:
The hyaluronidase activity inhibitory effect is calculated using the following formula.
Hyaluronidase inhibitory effect (%) = 1- (C-D) / (A-B) × 100
A: Absorbance when purified water is added, B: Absorbance when purified water is added and no enzyme is added
C: Absorbance when sample is added, D: Absorbance when sample is added and enzyme is not added
(5) Fibroblast activation effect test Human dermal fibroblasts were seeded in a 96-well microplate at a concentration of 7 × 10 ⁴ cells / mL at 100 μL / well. As a seeding medium, an α-MEM medium containing 5% fetal bovine serum (FBS) was used and cultured in a CO ₂ incubator (5% CO ₂ , 37 ° C.) for 24 hours. After 24 hours, each sample was replaced with α-MEM medium containing 5% FBS, and cultured for 72 hours. After removing the supernatant, MTT (3- (4,5-dimethylthiazol-2-yl) -2,5-diphenyltetrazolium bromide) was replaced with DMEM medium supplemented with 1.0 mg / mL and cultured for 4 hours. Thereafter, the produced formazan was extracted with acidic isopropanol, and the absorbance at wavelengths of 570 nm and 630 nm was measured with a plate reader. From the difference between the two measured values (absorbance at 570 nm−absorbance at 630 nm), the cell activation effect was evaluated using the following formula with the activation rate being 100% when each extraction solvent was added as a control.

Formula: Cell activation rate (%) = A / A0 × 100
A = Difference in absorbance due to sample addition
A0 = Difference in absorbance when purified water is added
(6) Maillard reaction inhibition effect test Add 100 μL of 0.5 M ribose 40 μL, 50 mg / mL lysozyme prepared with 800 μL of 0.1 M phosphate buffer (pH 7.4) and 800 μL of 0.1 M phosphate buffer (pH 7.4) to the sample tube. 60 μL of an evaluation sample whose concentration was adjusted with purified water was added. The solution was sterilized by filtration using a 0.45 μm filter in a clean bench, and reacted for 7 days in a 40 ° C. incubator. After the reaction, electrophoresis was performed on SDS-PAGE, staining was performed with 0.1% Coomassie Brilliant Blue R-250, and after decolorization, the lysozyme dimer was visually determined. In the determination, a sample using 30% BG and 50% EtOH instead of the evaluation sample was subjected to negative control, and the state when aminoguanidine sulfate was added was determined as a positive control.

(7) Involucrin production promotion test Human keratinocytes were seeded in a 96-well microplate at a density of 5 × 10 ⁴ cells / cm ² . The seeding medium was cultured in a CO ₂ incubator (5% CO ₂ , 37 ° C.) for 24 hours using a DMEM medium containing 10% fetal bovine serum (FBS). After 24 hours, each sample was replaced with DMEM medium containing 10% FBS, and cultured for 72 hours. After fixing with 4% paraformaldehyde-containing PBS, permeabilization was performed, and involucrin was immunofluorescently stained by antigen-antibody reaction. The fluorescence intensity at 12 points was measured for each well using a fluorescence plate reader. The involucrin production rate was calculated from the following formula, assuming that the control with no sample action was 100%. In addition, cell nuclei were stained with Hoechst, and the number of viable cells was also compared. The following formula was also used to calculate the viable cell ratio.

Formula: Involucrin production rate, viable cell rate (%) = (B1-B0) / (A1-A0B1-B0) × 100
A1 = fluorescence intensity with no sample added
A0 = fluorescence intensity with no sample added and no primary antibody
B1 = fluorescence intensity of sample addition
B0 = fluorescence intensity with no added primary antibody after sample addition Test result:
(1) Elastase activity inhibitory effect test (derived from pancreas)
The extract (stock solution) obtained by the method of Example 1 was made 100% and diluted with purified water for use in the test. The following table shows the activity inhibitory effect of pancreatic elastase when the sample was allowed to act at 2.5% and the 50% inhibitory concentration of the sample having a high inhibition rate.

(2) Elastase activity inhibitory effect test (derived from fibroblasts)
The following table shows the activity inhibitory effect of fibroblast-derived elastase when the sample was allowed to act at 5% and the 50% inhibitory concentration of the sample having a high inhibition rate.

  In the table, 30% BG is a 30% (w / w) mixed solvent of purified water and butylene glycol, 50% EtOH is a 50% (w / w) mixed solvent of purified water and ethanol, and 95% EtOH is purified water and ethanol. It refers to the mixed solvent. Further, 30% BG (deodorization) and 50% EtOH (deodorization) mean that the extract was deodorized with activated carbon.

  A high elastase activity inhibitory effect was observed in the extracts of cherry bark, fence, and pine needles. In particular, with respect to cherry bark and pine needles, an inhibitory effect was observed on both pancreatic elastase and fibroblast-derived elastase. In addition, for pine needles, it became clear that the effect was not significantly reduced by the deodorizing treatment.

(3) DPPH radical scavenging effect test The following table shows the DPPH radical scavenging effect when the sample was operated at 0.4% and the 50% scavenging concentration of the sample having a high scavenging effect.

  A high DPPH radical scavenging effect was observed in the extract of cherry bark and pine needles, and a DPPH radical scavenging effect was also observed in the extract of Kobaku. In addition, for pine needles, it became clear that the effect was not significantly reduced by the deodorizing treatment.

(4) Hyaluronidase activity inhibitory effect test The following table shows the hyaluronidase activity inhibitory effect when the sample was allowed to act at 2.5% and the 50% inhibitory concentration of the sample with a high inhibition rate.

A high hyaluronidase activity inhibitory effect was observed in the extract of cherry bark and pine needles. In addition, for pine needles, it became clear that the effect was not significantly reduced by the deodorizing treatment.
(5) Fibroblast activation effect test The following table shows the cell activation effect when the sample was allowed to act at 1% and 2%.

  A remarkable cell activation effect was observed in the 50% EtOH extract of cherry bark and the 95% EtOH extract of Kakitsu. It was also found that the cell activation effect was increased by increasing the concentration of almost all the crude drug extracts.

(6) Maillard reaction inhibition effect test Fig. 1 shows the result of migration of Maillard reaction product when the sample was allowed to act at 4.0%, and migration of Maillard reaction product when the sample was allowed to act at 2.0%. The results are shown in FIG. The strength of the upper band reflects the formation of lysozyme dimer, and the thinner the band, the more the formation of lysozyme dimer, that is, the Maillard reaction is suppressed. The aminoguanidine sulfate lane is the positive control.

  The table below shows the correspondence between lane numbers and samples.

A high Maillard reaction inhibitory effect was observed in the extract of cherry bark, Kobakku, and Matsuba. In addition, for pine needles, it became clear that the effect was not significantly reduced by the deodorizing treatment.
(7) Involucrin production promoting effect The following table shows the involucrin production rate and viable cell rate when the sample was allowed to act at 3%, 1%, and 0.5%.

  A high involucrin-producing effect was observed in all of the four types of herbal extracts, such as cherry bark, Kobaku, Kaki-dori and Matsuba. In particular, the pine needle extract has a remarkable effect, and it has been clarified that the effect is not significantly reduced by the deodorizing treatment.

[Example 3: Production of cosmetics]
According to the above evaluation results, examples of cosmetic preparations are shown below, but the present invention is not limited thereto.

Manufacture of milky lotion (refreshing type) containing matsuba extract and kakidoshi extract The water phase A and oil phase B in the table below are each heated to 80 ° C and dissolved. Add A to B, emulsify with an emulsifier, and cool to 35 ° C. The emulsion was subjected to high pressure treatment (25 ° C., 200 MPa, 5 Pass) to obtain an ORGANIC emulsion (fresh type) pretreatment.

  Next, the B phase in the table below is heated to 70 ° C. and dissolved by stirring. It cooled to 35 degreeC, the emulsion (fresh type) pretreatment was added and stirred, and ORGANIC emulsion (fresh type) was obtained.

Manufacture of cream (fresh type) containing matsuba extract and koboku extract Aqueous phase A and oil phase B are each heated to 80 ° C and dissolved. Add A to B, emulsify with an emulsifier, and cool to 35 ° C. The emulsion was subjected to high pressure treatment (25 ° C, 200 MPa, 5 Pass) to obtain a pretreatment of ORGANIC cream (fresh type).

  Next, the B phase in the table below is heated to 70 ° C. and dissolved by stirring. After cooling to 35 ° C, ORGANIC cream (fresh type) pretreatment was added and emulsified with an emulsifier to obtain ORGANIC cream (fresh type).

Manufacture of emulsion (ordinary type) containing oyster extract and Japanese pine extract Each of aqueous phase A and oil phase B in the table below is heated to 80 ° C and dissolved. Add A to B, emulsify with an emulsifier, and cool to 35 ° C. The emulsion was subjected to high pressure treatment (25 ° C., 200 MPa, 5 Pass) to obtain an ORGANIC emulsion (ordinary type) pretreatment.

  Next, the B phase in the table below is heated to 70 ° C. and dissolved by stirring. The mixture was cooled to 35 ° C., phase A was added and stirred to obtain ORGANIC emulsion (ordinary type).

Manufacture of a milky lotion (moist type) containing an extract of pine and persimmon extract Aqueous phase A and oil phase B in the table below are each heated to 80 ° C and dissolved. Add A to B, emulsify with an emulsifier, and cool to 35 ° C. The emulsion was subjected to high pressure treatment (25 ° C., 200 MPa, 5 Pass) to obtain an ORGANIC emulsion (moist type) pretreatment.

  Next, the B phase in the table below is heated to 70 ° C. and dissolved by stirring. The mixture was cooled to 35 ° C., phase A was added and stirred to obtain ORGANIC emulsion (moist type).

Manufacture of cream (moist type) containing matsuba extract and koboku extract Aqueous phase A and oil phase B in the table below are each heated to 80 ° C and dissolved. Add A to B, emulsify with an emulsifier, and cool to 35 ° C. The emulsion was subjected to high pressure treatment (25 ° C., 200 MPa, 5 Pass) to obtain a pretreatment of ORGANIC cream (moist type).

  Next, the B phase shown in the table below is heated to 70 ° C., dissolved by stirring, and the A phase is added and stirred. D phase is heated to 80 ° C., dissolved with stirring, and added to B phase / A phase and emulsified with an emulsifier. Furthermore, C phase was added and it cooled to 35 degreeC, and ORGANIC cream (moist type) was obtained.

Manufacture of beauty liquid (stain and dullness countermeasures) containing Spruce extract and Matsuba extract. Aqueous phase A and oil phase B in the table below are each heated to 80 ° C and dissolved. Add A to B, emulsify with an emulsifier, and cool to 35 ° C. The emulsion was subjected to high pressure treatment (25 ° C, 200MPa, 5Pass) to obtain a pretreatment of ORGANIC serum (anti-stain and dullness).

  Next, the B phase in the table below is heated to 70 ° C. and dissolved by stirring. The mixture was cooled to 35 ° C, phase A was added, and the mixture was stirred to obtain ORGANIC serum (anti-stain and dullness measures).

Manufacture of beauty liquid (wrinkle and sagging countermeasure) containing Spruce extract and Japanese pine extract Extract water phase A and oil phase B in the table below, respectively, by heating to 80 ° C and dissolving. Add A to B, emulsify with an emulsifier, and cool to 35 ° C. The emulsion was subjected to high-pressure treatment (25 ° C, 200MPa, 5Pass) to obtain a pretreatment of ORGANIC serum (wrinkle / sagging measures).

  Next, the B phase in the table below is heated to 70 ° C. and dissolved by stirring. Cooled to 35 ° C, added Phase A and stirred to obtain ORGANIC serum (wrinkle / sagging measures).

Manufacture of eye cream containing Matsuba extract, Kakidoshi extract, and Koboku extract. Phase B in the table below is heated to 70 ° C, dissolved with stirring, and phase A (the above-mentioned “ORGANIC cream (fresh type) pretreatment” and “ Add ORGANIC emulsion (moist type) pretreatment ") and stir. D phase is heated to 80 ° C, dissolved by stirring, and added to B phase / A phase and emulsified with an emulsifier. Furthermore, C phase was added and it cooled to 35 degreeC, and ORGANIC eye cream was obtained.

Manufacture of cleansing oil containing matsuba extract, persimmon extract, kokuboku extract, and persimmon extract Heat phase A in the table below to 80 ° C and stir to dissolve. It cooled to 40 degreeC, added the B phase, and stirred with the emulsifier, and ORGANIC cleansing oil was obtained.

[Example 4: Production of cosmetics (comparative example)]
As a comparative example, a cosmetic was prepared in which none of the extracts of cherry bark, Kobaku, Kaki-dori, or Matsuba was added.

Comparative Example Manufacture of serum (anti-stain and dullness) Water phase A and oil phase B in the table below are each heated to 80 ° C and dissolved. Add A to B, emulsify with an emulsifier, and cool to 35 ° C. The emulsion was subjected to high pressure treatment (25 ° C., 200 MPa, 5 Pass) to obtain a pre-treatment for a comparative cosmetic liquid (anti-stain / dullness).

  Next, the B phase in the table below is heated to 70 ° C. and dissolved by stirring. The mixture was cooled to 35 ° C., phase A was added, and the mixture was stirred to obtain a comparative beauty serum (stain / dullness countermeasure).

This comparative example is a comparative example for the “ORGANIC beauty essence (stain / dullness countermeasure)” containing the Spruce extract and the Matsuba extract of Example 3.
Comparative Example Manufacture of serum (wrinkle / sagging measures) Water phase A and oil phase B in the table below are each heated to 80 ° C and dissolved. Add A to B, emulsify with an emulsifier, and cool to 35 ° C. The emulsion was subjected to high-pressure treatment (25 ° C, 200MPa, 5Pass) to obtain a pre-treatment for a comparative cosmetic liquid (wrinkle / sagging measures).

  Next, the B phase in the table below is heated to 70 ° C. and dissolved by stirring. After cooling to 35 ° C., Phase A was added and stirred to obtain a comparative cosmetic liquid (wrinkle / sagging measures).

This comparative example is a comparative example for the “ORGANIC beauty essence (wrinkle / sagging countermeasure)” containing the Spruce extract and the Matsuba extract of Example 3.
[Example 5: Sensory test for cosmetics]
The sensory test of the manufactured cosmetics was conducted by 20 panelists (women in their 20s and 60s). An appropriate amount (0.2 to 0.5 ml) of the produced serum was taken in the hand, and then applied to the skin after washing the face half by half on the right and left faces. When used continuously for about one month twice a day, compared to the comparative example, inflammation, wrinkles, blemishes and dullness were improved and an anti-aging effect was seen.

Claims (15)

  An external composition for skin comprising a herbal extract obtained from one or more selected from cherry bark, Kobaku, Kaki-dori and Matsuba. The external composition for skin according to claim 1, comprising a pine needle extract, and further comprising one or more herbal extracts selected from Kakitsu, Kobaku, and cherry bark.   Maillard reaction inhibitor containing as an active ingredient any of cherry bark extract, pine needle extract, or Kokaku extract.   The external composition for skin which contains the Maillard reaction inhibitor which uses either of the extract of Claim 3 as an active ingredient independently or in combination.   An involucrin production promoter containing as an active ingredient any one of pine needle extract, cherry bark extract, Kobaku extract, and fence extract.   An external composition for skin containing an involucrin production promoter containing any one of the extracts according to claim 5 alone or in combination.   An antioxidant containing any one of a cherry bark extract, a magnolia extract, or a pine needle extract as an active ingredient.   A composition for external use containing an antioxidant alone or in combination with any one of the extracts according to claim 7 as an active ingredient.   An elastase activity inhibitor containing either a cherry bark extract or a pine needle extract as an active ingredient.   An external composition for skin containing an elastase activity inhibitor containing any one of the extracts according to claim 9 alone or in combination.   A hyaluronidase activity inhibitor containing either a cherry bark extract or a pine needle extract as an active ingredient.   A composition for external use containing a hyaluronidase activity inhibitor containing any one of the extracts according to claim 11 alone or in combination.   A fibroblast activator containing either a cherry bark extract or a fence extract as an active ingredient.   A composition for external use containing a fibroblast activator containing any one of the extracts according to claim 13 alone or in combination.   The external composition for skin according to claim 1, 4, 6, 8, 10, 12, or 14, which is a cosmetic or a medicated cosmetic.