JP2013144654A - Composition for suppressing decrease in contraction force of collagen, containing extract of flower part of prunus mume - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a decrease inhibitor for contraction force of collagen, capable of suppressing skin aging by suppressing collagen contraction by collagen crosslinking formation based on a glycation reaction in a skin tissue.SOLUTION: A composition for suppressing the decrease in contraction force of collagen contains the extract of a flower part of a Prunus mume as an active ingredient.

Description

  The present invention relates to a composition containing an extract of a ume flower, and more specifically, the present invention relates to a composition for suppressing a decrease in collagen contraction force comprising an extract of a flower of a genus Rosaceae as an active ingredient. It is about.

The skin is an organ that covers the outer surface of the body and, despite its thickness of about 2 mm, occupies about 16% of the body weight and has a protective function inside the body.
The skin has a structure in which the epidermis composed of the skin layer, stratum corneum, granule layer, basal layer and spiny layer; dermis composed of collagen, elastin, fibroblasts and matrix; and subcutaneous tissue containing subcutaneous fat laminated Have

In the epidermis, the basal cells that make up the basal layer in the lowermost layer divide to form new epidermal cells, which are gradually pushed up toward the surface of the body, forming into spinous layers, granule cells, and keratinocytes. It is known that the cells move to the stratum corneum while changing their functions sequentially, and finally the horny cells form a flat mica-like shape and peel off from the surface of the skin.
Melanocytes that produce melanin pigments are interspersed between basal cells, producing melanin pigments in response to ultraviolet rays, which are also known to absorb ultraviolet rays and protect the dermis from ultraviolet rays. ing.

In addition, the dermal tissue that occupies most of the skin is filled with protein fibers and matrix, and most of these protein fibers are occupied by fibers called collagen. It is an ingredient that supports and also plays a role in supporting elastin which gives the skin elasticity.
The matrix filled between the collagen fibers and the elastin fibers is composed of a proteoglycan in which mucopolysaccharide and protein are bound, and also includes the above-described collagen fibers and elastin fibers and fibroblasts that produce the substrate.

In Japan, which is said to have the longest life expectancy in the world, aging over the age of real people is a problem.
The aging of the skin and body is caused by excessive exposure to ultraviolet rays, cross-linking formation due to collagen oxidation reaction, etc., and glycation in the body (Maillard reaction: Non-Patent Document 1) is enhanced with age, and sugar and protein and / or It is also suggested that amino acids react and irreversible advanced glycation end products (AGEs) accumulate in the body, affecting the onset and progression of aging, diabetes aging, aging-related diseases, etc. Has been.

The dermis part is composed of proteins such as collagen and elastin, of which collagen accounts for about 70% of the dermis, and this dermis part is deeply involved in the softness of the skin.
In the dermis part, elastin exists so as to be entangled with collagen, and this collagen and elastin create a three-dimensional structure, and a jelly-like substance such as hyaluronic acid, which is a moisturizing ingredient, fills the skin, creating elasticity and firmness of the skin Yes.

The saccharification reaction is considered to consist of the following two stages.
That is, in the first stage, glycation (Maillard reaction) occurs in the collagen portion of the skin protein, and the amino group of the lysine residue or the guanidyl group of the arginine residue in the protein and the carbonyl group of the sugar are non-enzymatically. This is a pre-reaction in which a reversible Schiff base is formed by reaction, and this Schiff base becomes a stable Amadori compound by the Amadori transition.
The second stage is a late reaction in which the formed Amadori compound undergoes various reactions such as dehydration, condensation, oxidation, and transfer, and finally becomes irreversible final glycation products (AGEs) (non-reactive) Patent Document 2).

  When glycation occurs in the collagen part of the skin protein, the amino group of the lysine residue or the guanidyl group of the arginine residue in the protein and the carbonyl group of the sugar react non-enzymatically, and after passing through the Schiff base and the Amadori compound Thus, a cross-linked structure connecting the proteins is formed. When this cross-linked structure is formed, it is believed that the range of movement of the molecule is limited, the molecular structure is hardened, the original elasticity of the skin is lost, and the skin is aged.

  Furthermore, since the cross-linked product is judged to be a foreign substance due to the cross-linking of collagen and elastin, the secretion amount of degrading enzymes (collagenase and elastase) increases, so that not only the cross-linked product but also normal collagen and elastin are easily decomposed. It has been found that firmness and elasticity are further lost, the skin becomes brittle, and further, wrinkles, tarmi, kusumi, etc. are generated (Non-patent Documents 3 and 4).

For the purpose of preventing skin aging, a topical skin preparation has been proposed that normalizes the keratinization of the skin epidermis by blending liposomes encapsulating a lotus extract (Patent Document 1).
In addition, one or two kinds selected from hydrolyzed casein, L-lysine or L-arginine for the purpose of inhibiting collagen cross-linking formation which can prevent skin aging by inhibiting collagen cross-linking formation due to saccharification reaction or oxidation reaction A collagen cross-linking inhibitor containing the above has been proposed (Patent Document 2).

Japanese Patent Publication No.8-18994 JP 2001-39816 A

Kazuo Kondo, active oxygen taught by specialists, PHP Institute (2004) Yukio Shigeta et al., Glycation of Protein AGE Basic and Clinical, Medical School (1997) Daisaburo Fujiki, Mechanism of aging and life, Natsume (2001) Daisaburo Fujiki, Collagen, Kyoritsu Shuppan (1997)

  An object of the present invention is to provide a composition that suppresses skin aging by suppressing collagen contraction due to collagen cross-linking formation based on a saccharification reaction in skin tissue.

  In order to solve the above-mentioned problems, the present inventors diligently searched for a new material having a collagen contraction force reduction inhibitory action, and found that it has an excellent effect on the extract of the flower part of ume (Prunus mume). The headline and the present invention were completed.

  ADVANTAGE OF THE INVENTION According to this invention, the composition for a collagen contraction force fall suppression characterized by including the extract of a ume flower part as an active ingredient is provided.

  The composition for suppressing a decrease in collagen contraction force according to the present invention containing an extract of a ume flower part as an active ingredient has an effect of suppressing the skin aging phenomenon by effectively suppressing the contraction of collagen.

It is the figure which showed the effect with respect to the collagen area by the white plum flower methanol extract shown in Table 1 by the sample density | concentration (microgram / mL) used for the collagen gel area (%) on the vertical axis | shaft, and the horizontal axis | shaft.

The composition for suppressing a decrease in collagen contraction force of the present invention contains an extract of a ume flower part as an active ingredient.
Ume is a member of the genus Rosaceae, and both horticultural varieties and fruit varieties can be used, and the production area is not limited.

  The ume flower used as the extraction material in the present invention means the whole flower including petals, stamens, pistils and buds, and any of the flower that has flowered and the bud before flowering can be used. In addition, as the flower of the ume flower or the bud before flowering, either a dried flower or a raw flower may be used. From the viewpoint of easy acquisition and storage of the material, the ume flower bud is dried. It is particularly preferable to use a product.

  Solvents used for the extraction of ume flowers according to the present invention are water, water-containing or non-water-containing lower alcohols such as methanol, ethanol, n-propanol, i-propanol, n-butanol, sec-butanol and t-butanol. In view of extraction efficiency and cost, methanol or ethanol is preferable.

These extraction solvents are used in an amount of about 2 to 15 times (weight), preferably about 4 to 13 times (weight), more preferably about 6 to 12 times (weight) with respect to the extraction material.
The extraction temperature can be arbitrarily set between room temperature and the boiling point of the solvent. For example, the extraction material is used as an extraction solvent at a temperature of 50 ° C. to the boiling point of the extraction solvent under shaking or non-shaking or refluxing. It is appropriate to carry out by dipping.

In the case of extracting the extraction material with heating under reflux, it is appropriate to carry out for about 30 minutes to 4 hours. In addition, although it is possible to extract at a temperature lower than 50 ° C., it is preferable to extract for a longer time than the above time.
Although extraction operation may be performed only once about the same material, it is preferable from the point of extraction efficiency to repeat several times, for example about 2 to 5 times.

The extract obtained by filtering the solid from the cooled extraction mixture may be concentrated by a conventional method to obtain an extract.
Concentration is preferably performed at a low temperature and under reduced pressure, and may be performed until the extract is dried to obtain a dry extract.

The extract may be used as it is to prepare the composition of the present invention, but may also be used as a powder or lyophilized product. Solids such as these powders or freeze-dried products can be obtained by methods known in the art.
Therefore, the extract in the present invention includes both an extract and a solid product obtained by solidifying the extract.

  The extract may be subjected to a purification treatment before and after concentration. As the purification treatment, a chromatographic method, an ion exchange chromatographic method, a partition extraction with a solvent, or the like can be employed alone or in combination. For example, as a chromatographic method, a column chromatography, a thin layer chromatography, a high performance liquid chromatography, a centrifugal liquid chromatography or the like using a normal phase or reverse phase carrier or an ion exchange resin, or a combination thereof is performed. Is mentioned.

In this case, purification conditions such as a carrier and an elution solvent can be appropriately selected according to various chromatographies.
Among these, the extract can be concentrated to obtain an extract, and this extract can be distributed using water and a non-hydratable organic solvent to obtain a water-soluble fraction or an organic solvent-soluble fraction.

Examples of the non-hydratable organic solvent include ethyl acetate, n-butanol, hexane, chloroform and the like. Among them, ethyl acetate is particularly preferable from the viewpoint of distribution efficiency.
That is, the extract obtained by concentration or the concentrated residue obtained by concentration to dryness can be distributed using ethyl acetate and water to obtain a water-soluble fraction or an ethyl acetate-soluble fraction.

  The partition extraction can be performed according to a conventional method such as a stirring or shaking distribution method or a droplet counter-current distribution method usually performed in the art. For example, it is performed by adding about 0.1 to 10 times (volume) (1:10 to 10: 1) of ethyl acetate and water to an extract or the like at room temperature, with shaking or without shaking. Is appropriate. Further, the organic solvent-soluble fraction thus obtained may be subjected to the purification treatment as described above. That is, the organic solvent-soluble fraction obtained above can be further purified by reverse-phase chromatography or reverse-phase high performance chromatography alone or in combination.

  The extract of a ume flower part according to the present invention has an action of suppressing a decrease in collagen contraction force, and the composition containing the extract is used as an inhibitor of a decrease in collagen contraction force. It can be used for prevention and / or treatment of decline.

  In addition, the ume flower part extract according to the present invention can be directly used as a soft drink when extracted with water, and when extracted with methanol or ethanol, the extraction solvent is completely distilled off under reduced pressure. Thus obtained solid or lyophilized product can be used as a food additive.

  The extract of the present invention can be used as it is, diluted with an appropriate medium, or by a known method in the field of pharmaceutical production, such as a solid agent (for example, powder, granule, tablet, capsule), semi-solid. It can be formulated into various forms such as an agent or liquid.

  An appropriate carrier may be added to these preparations, and examples of such a carrier include pharmaceutically acceptable excipients, binders or disintegrants.

  Examples of excipients include sugar alcohols such as erythritol, maltitol, mannitol, sorbitol, xylitol, and lactitol, purified sucrose, sucrose, trehalose, lactose, reduced maltose water candy, powdered reduced maltose water candy, glucose, maltose, and the like. Examples include saccharides, corn starch, crystalline cellulose, powdered cellulose, calcium monohydrogen phosphate, calcium hydrogen phosphate, anhydrous calcium hydrogen phosphate, calcium lactate, precipitated calcium carbonate, light anhydrous silicic acid, hydrous silicon dioxide, silicon dioxide and the like.

  Examples of the binder include gum arabic powder, hydroxyethylcellulose, hydroxypropylcellulose (HPC), hydroxypropylmethylcellulose (HPMC), cellulose derivatives such as methylcellulose, carboxymethylcellulose, carboxymethylethylcellulose, povidone (PVP), vinylpyrrolidone copolymer Examples include coalescence (copolyvidone), polyvinyl alcohol (PVA), acrylic polymer, pullulan, dextrin, pregelatinized starch, hydroxypropyl starch, tragacanth powder, crystalline cellulose, low-substituted hydroxypropylcellulose (L-HPC), and the like. .

  Examples of the disintegrant include croscarmellose sodium, low-substituted hydroxypropyl cellulose, carmellose calcium, crospovidone, corn starch, carboxymethyl starch sodium, hydroxypropyl starch, and partially pregelatinized starch.

In the above preparation, other additives such as lubricants, fluidizers, antioxidants, preservatives, colorants, pH adjusters, sweeteners, fragrances, surfactants, plasticizers and the like are appropriately added. May be used.
Such additives include lubricants (eg, magnesium stearate, calcium stearate, talc, sucrose fatty acid ester, glycerin fatty acid ester, hydrogenated oil, macrogol 6000, etc.), fluidizing agents (eg, light anhydrous silica Acids, hydrous silicon dioxide, kaolin, etc.), antioxidants (eg, dibutylhydroxytoluene (BHT), propyl gallate, butylhydroxyanisole (BHA), tocopherol, citric acid, etc.), preservatives (paraoxybenzoates, etc.) ),

Colorant (for example, riboflavin, vitamin B ₁₂ , titanium oxide, yellow ferric oxide, ferric oxide, edible red No. 2, edible red No. 3, edible red No. 102, edible red No. 104, edible red No. 105, edible red No. 106, Edible Yellow No. 4, Edible Yellow No. 5, Edible Green No. 3, Edible Blue No. 1, Edible Blue No. 2, Copper Chlorophyll Sodium, Copper Chlorophyll, etc.), pH regulator (eg, sodium hydroxide, sodium citrate) , Hydrochloric acid, sodium bicarbonate, sodium carbonate, calcium lactate, phosphoric acid, dipotassium phosphate, sodium hydrogen phosphate, potassium dihydrogen phosphate, sodium dihydrogen phosphate)

Sweetening agents (eg, sucrose, mannitol, D-sorbitol, xylitol, aspartame, stevia, dipotassium glycyrrhizinate, acesulfame K, sucralose, etc.), flavors (eg, L-menthol, peppermint oil, eucalyptus oil, orange oil, Clove oil, turpentine oil, fennel oil, vanillin, etc.), surfactant (polyoxyethylene hydrogenated castor oil, glyceryl monostearate, sorbitan fatty acid ester (such as sorbitan monostearate, sorbitan monolaurate), polyoxyethylene polyoxypropylene, And polysorbates, sodium lauryl sulfate, macrogol, sucrose fatty acid ester, etc.) and plasticizers (triethyl citrate, polyethylene glycol, triacetin, cetanol, etc.).
In addition, a corrigent, an adsorbent, a preservative, a wetting agent, an antistatic agent and the like may be added as appropriate.

  If necessary, the solid agent is a water-soluble base (or polymer) such as hydroxypropylcellulose, hydroxypropylmethylcellulose, or povidone, a water-insoluble base (or polymer) such as ethylcellulose, an enteric base, or a gastric group. You may coat with an agent, saccharides, etc.

  Said solid agent can be manufactured by a conventional method. For example, the active ingredient and the carrier may be mixed to prepare a powder, or the active ingredient and the carrier may be mixed and granulated, and the granulated product may be sized and granulated (fine granule or granule if necessary) A tablet can be prepared by tableting a mixture (particularly a mixture of a granulated product and a carrier) containing the granulated product. Capsules can be prepared by filling capsules with the above granules.

As a carrier in the case of a liquid agent, an aqueous medium (purified water, ethanol-containing purified water, etc.), alcohols (ethanol, glycerin, etc.), a water-soluble polymer and the like are used.
As a carrier in the case of a semi-solid preparation, an oily base (lipids such as vegetable oil, petrolatum, liquid paraffin, etc.), a hydrophilic base (emulsion base) and the like are used.

  The above liquid preparation can be prepared by dissolving or dispersing an active ingredient in a carrier, filtering or sterilizing if necessary, and filling a predetermined container. A semi-solid preparation can also be prepared by a conventional method, for example, by mixing an active ingredient and a carrier, sterilizing if necessary, filling a predetermined container, or applying to a substrate.

  The amount of the extract used as an active ingredient varies depending on the form, age of the user, skin condition, etc., but when used as a solid or semi-solid, 1 to 3 times a day for adults. It is about 0.003g-0.3g per time.

  In addition, the extract according to the present invention can be in the form of parenteral topical or systemic skin cosmetics, bath preparations, skin patch sheets and the like.

  Specific examples include basic cosmetics such as lotions, emulsions, creams, ointments, gels, lotions, oils, packs, mists and facial cosmetic sheets, makeup cosmetics such as foundations and blushers.

  In the composition of the present invention, in addition to the above active ingredients, other active ingredients may be optionally used in combination as long as the effects are not impaired.

  Hereinafter, the present invention will be described in more detail with reference to Examples such as Examples, Test Examples, Comparative Test Examples, and Formulation Examples, but the present invention is not limited thereto.

Example 1
Extraction method 2.0 kg of dried buds immediately before flowering of Prunus mume was extracted with 20 kg of methanol at 80 ° C. with heating under reflux (3 hours × 4 times). The extract was filtered and concentrated to dryness under reduced pressure to give the methanol extract (607.95 g, 30.40% yield). Hereinafter, this extract was used as a sample.

Test example 1
Cell matrix IA (made by Nitta Gelatin) 4.8 mL, Hanks buffer (DIBCO, Hanks' Balanced Salt Solution (HBSS) (10 ×)) 0.6 mL, liquid collagen reconstitution buffer (sodium hydroxide 50 mM) 0.6 mL of sodium bicarbonate 260 mM, HEPES 200 mM) was mixed on ice, and 0.8 mL of this mixture was dispensed into 6 wells of a 12-well plate. The plate was incubated at 37 ° C. for 2 hours to gel the collagen.

  Of each well in which gelled cell matrix IA (collagen) is contained in 5 wells of each gelled well, 500 mM D (−) ribose solution dissolved in 0.15M phosphate buffer is added. 1 mL each was overlaid. Samples of each concentration (0, 3, 10, 30 and 100 μg) were added to these 5 wells.

The remaining 1 well was overlaid with only 0.15 M phosphate buffer as a blank.
This was incubated at 37 ° C. for 5 days. After completion of the incubation, the overlaid solution was removed, the collagen gel was washed twice with phosphate buffered saline (PBS), and then 2 mL of PBS was overlaid on each well.

This was incubated at 37 ° C. for 1 day. After completion of the incubation, the overlaid PBS was removed, the collagen gel was washed twice with Dulbecco's modified Eagle medium (DMEM medium), and 1 × 10 ⁵ fibroblast cells (1 × 10 ⁵ cells suspended in 1 mL of DMEM medium) NB1RGB).
This was incubated at 37 ° C. for 2 days (24 hours after seeding, the cells were sufficiently adhered to the collagen gel, and then the collagen gel was detached from the plate).

  Next, each collagen gel was transferred to a 6-well plate in which 2 mL of PBS was dispensed in each well, and this plate was scanned with a scanner (Gp-9800F, manufactured by Epson Corporation), and the captured image was subjected to image analysis. Each collagen gel image is extracted with software (Photoshop (registered trademark), Adobe (registered trademark)), and the gel area is extracted with image analysis software (Image J, National Institute of Health, USA). It was measured.

Comparative Test Example 1
It is known to have an inhibitory effect on the formation of AGEs, and the inhibitory effect on protein cross-linking and polymerization formation has been confirmed. Aminoguanidine, whose development was stopped by former Yamanouchi Pharmaceutical Co., Ltd. in 1998, was used as a comparative sample. The collagen gel area was measured by the same test method as above.

Test results The results of measuring the collagen gel surface area in each test using the above samples are shown in Table 1 below.
In the table below, the collagen gel area using samples at various concentrations is expressed as a percentage of the blank collagen gel area (number of examples n = 5).

  FIG. 1 is a graph showing the results of the white plum blossom methanol extract shown in Table 1 above with the collagen gel area (%) on the vertical axis and the sample concentration (μg / mL) used on the horizontal axis.

  From the above results, it can be seen that the extract according to the present invention has a four-fold or more powerful inhibitory effect on collagen contraction force reduction compared to aminoguanidine.

Example 2
10 parts by weight of the extract obtained in Example 1 was mixed with 25 parts by weight of lactose and filled into a gelatin capsule to obtain a gelatin capsule containing 500 mg of the extract in one capsule.

Example 3
A gelatin capsule containing 50 mg of extract in one capsule was obtained in the same manner as in Example 2 except that 1 part by weight of ume extract was used.

に従って、常法により化粧水１ｋｇを調製した。 Example 4
Preparation of lotion The following formula:

1 kg of skin lotion was prepared according to a conventional method.

Examples 5-8
1 kg of lotion was prepared in the same manner as in Example 4 except that 0.01, 0.1, 1 and 5% by weight of ume extract were used.

に従って、常法により乳液１ｋｇを調製した。 Example 9
Preparation of emulsion The following formula:

1 kg of emulsion was prepared by a conventional method.

Examples 10-13
1 kg of milky lotion was produced in the same manner as in Example 9 except that 0.01, 0.1, 1 and 5% by weight of ume extract were used.

に従って、常法によりクリーム１ｋｇを調製した。 Example 14
Cream preparation The following formula:

1 kg of cream was prepared according to a conventional method.

Examples 15-18
1 kg of cream was produced in the same manner as in Example 14 except that 0.01, 0.1, 1 and 5% by weight of ume extract were used, respectively.

に従って、常法によりファンデーション１ｋｇを調製した。 Example 19
Preparation of foundation The following formulation:

1 kg of foundation was prepared by a conventional method.

Examples 20-23
1 kg of foundation was produced in the same manner as in Example 19 except that 0.01, 0.1, 1 and 5% by weight of ume extract were used, respectively.

  The composition for suppressing a decrease in collagen contraction force containing an extract of ume flowers according to the present invention as an active ingredient can be used as a composition for preventing or treating skin aging, or as a cosmetic.

Claims (4)

  A composition for suppressing a decrease in collagen contractility, comprising an extract of a ume flower part as an active ingredient.   The composition according to claim 1, wherein the flower portion of the ume is a ume bud or a flower that has bloomed.   The composition according to claim 1 or 2, wherein the ume flower part is a dried product of ume buds.   The composition according to any one of claims 1 to 3, wherein the extract is an extract obtained by extracting a flower part of ume with a lower alcohol or water.

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