TWI582110B - Platelet-derived growth factor-BB produces hyperactivity agents, and is associated with leaf-derived stem cell-producing accelerators, stem cell stabilizers, and dermal regenerators - Google Patents

Description

Platelet-derived growth factor-BB produces a hypertrophic agent, and a leaf cell stem cell production promoter, a stem cell stabilizer, and a dermal regenerative agent

The present invention relates to a platelet-derived growth factor-BB (PDGF-BB)-producing agent, and a leaf cell stem cell production promoter and a stem cell stabilization device comprising the PDGF-BB-producing agent. Agent, and dermal regenerative agent.

A stem cell is a cell which has both the ability to differentiate into cells which differentiate into a plurality of cells, and the two kinds of properties which are capable of generating the same self-replication ability of the same cell as the cell by cell division. The stem cell line derived from the embryo of the initial stage of production of the fertilized egg is called an embryonic stem cell (ES cell, Embryonic Stem cell). Although human ES cells are expected to be used in regenerative medicine, the formation of new human ES cells cannot be confirmed due to the ethical problem of using fertilized eggs.

In recent years, artificially pluripotent stem cells (iPS cells, induced Pluripotent Stem cells) have also attracted attention as cells having properties similar to those of ES cells. However, in the formation of iPS cells, there are many problems in terms of canceration of cells, formation efficiency, and the like. On the other hand, since the body stem cells having the ability to differentiate into a specific tissue are obtained from the body tissues of the patients themselves, such as bone marrow, there is no ethical problem such as embryonic stem cells.

It is known that epidermal stem cells are present in the epidermal base layer of the skin (Non-Patent Document 1), and it is reported that hair follicle epithelial stem cells (Non-Patent Document 2) or skin pigment stem cells are present in a region called a convex region of the hair follicle (Non-patent literature) 3). On the other hand, the leather is though In the fiber component containing collagen as a main body, there is a fibroblast formed into a slender spindle shape, but it is not clear whether or not stem cells are present in fibroblasts of the dermis. Further, although it is known that there are a plurality of skin-derived precursors (SKP) which are differentiated into a series of cells such as fat, glial, cartilage, and muscle in the dermis (Non-Patent Document 4), the dermal fiber is not clearly defined. The association of mother cells with SKP.

Leaf cell stem cells isolated from bone marrow as fibroblast precursor cells (Non-Patent Document 5) differentiate into various cells belonging to the mesenchymal system (osteocytes, muscle cells, chondrocytes, sputum cells, fat cells, etc.) Therefore, it is expected to be applied to regenerative medicine such as reshaping of bones or blood vessels and muscles. Recently, it has become clear that there is a possibility that there are many tissues having mesenchymal tissues, and mesenchymal stem cells can be isolated from fat, cord blood, placenta, and the like (Non-Patent Documents 6 to 8).

According to recent insights, mesenchymal stem cells are known to exist as extravascular cells (Pericyte, ectolog) in the blood vessels of the whole body, and play a role in vascular stabilization or tissue constant maintenance (Non-Patent Literature) 9 and 10).

In addition, when the blood vessel is damaged at or near the site of the tissue damage, the leaf cell stem cells are separated from the blood vessel as the extracellular extracellular cells (the outer cell) and are proliferated to supply the lost cells (Non-Patent Documents 11 to 14). Further, the bioactive factor is released to protect the tissue (Non-Patent Documents 15 to 19), and the repair and regeneration of the damaged tissue are performed. In addition to the effects of angiogenesis or anti-apoptosis, these secreted factors also strongly inhibit the action of immunity (Non-Patent Documents 21 and 22), and inhibit the destruction of damaged tissues via T cells or B cells ( Non-patent documents 9 and 22).

Further, it is known that mesenchymal stem cells also exhibit anti-fibrotic effects (Non-Patent Documents 23 and 24) or effects against multiple sclerosis or diabetes (Non-Patent Document 9).

On the other hand, it is gradually clear that chronic inflammation is a common pathology common to various diseases (for example, metabolic syndrome, arteriosclerotic disease, cancer, neurodegenerative disease, autoimmune disease, etc.) (Non-Patent Document 25). For example, reports of endothelial cell function due to chronic inflammation The disorder or the insulin resistance is a cause of various diseases such as diabetes or arteriosclerotic diseases (Non-Patent Document 26). Further, it is also clear that the fat tissue itself which is obese has a transition to inflammation (Non-Patent Documents 27 to 29). Since chronic inflammation is produced around the blood vessels, it is considered that there is a flaw in the interaction between the phylogenetic stem cells and the blood vessels between the extravascular cells (outer cells) in the chronic inflammation.

Based on the above findings, it can be considered that as long as the promotion or stabilization of mesenchymal stem cells can be achieved, vascular stabilization, tissue structural maintenance, repair of damaged tissues, regeneration, anti-fibrosis, multiple sclerosis or It is extremely effective for various purposes such as prevention and treatment of various diseases such as diabetes, prevention and improvement of various states based on chronic inflammation such as metabolic syndrome.

The present inventors have reported that mesenchymal stem cells are also present in the dermis, and a method for efficiently isolating mesenchymal stem cells from the dermis has been established (Japanese Patent Application No. 2009-213291). In consideration of the action of the mesenchymal stem cells, it is considered to be effective for improving or regenerating the state of the dermis by stabilizing or causing hyperstimulation between the phylloblastic stem cells in the dermis.

Further, the present inventors have confirmed in more detail the presence of mesenchymal stem cells in the dermis or subcutaneous fat, and found that platelet-derived growth factor-BB (PDGF-BB) is associated with the localization of mesenchymal stem cells, and further It is clear that the production of PDGF-BB in vascular endothelial cells contributes to the development and stabilization of mesenchymal stem cells (Japanese Patent Application No. 2010-209705).

Platelet-derived growth factor (PDGF) is a proliferation factor involved in the regulation of proliferation and proliferation of mesenchymal stem cells such as fibroblasts, smooth muscle cells, and glial cells, and is produced by various cells such as epithelial cells or endothelial cells. At least four kinds of PDGF-A, B, C and D exist in PDGF, but the A chain and the B chain form a homo- or heterodimeric structure by forming a disulfide bond, and have three isoforms (PDGF- AA, AB, BB). It is known that PDGF expresses its physiological function via PDGF receptor (PDGFR), which is a tyrosine kinase-associated type. use. The gene of PDGF-B is well known and can be cloned (Non-Patent Document 30).

It is considered that as long as the component which is effective for the production of PDGF-BB can be found, it is possible to promote the production of mesenchymal stem cells or stabilize the stem cells, and it can be effectively used for the above various uses and the like.

[Previous Technical Literature] [Non-patent literature]

Non-Patent Document 1: Watt FM, J Dermatol Sci, 28: 173-180, 2002

Non-Patent Document 2: Cotsarelis G et al., Cell, 57: 201-209, 1989

Non-Patent Document 3: Nishimura EK et al., Nature, 416: 854-860, 2002

Non-Patent Document 4: Wong CE al., J Cell Biol, 175: 1005-1015, 2006

Non-Patent Document 5: Pittenger MF et al., Science, 284: 143-147, 1999

Non-Patent Document 6: Park KW et al., Cell Metab, 8: 454-457, 2008

Non-Patent Document 7: Flynn A et al., Cytotherapy, 9:717-726, 2007

Non-Patent Document 8: lgura K et al., Cytotherapy, 6: 543-553, 2004

Non-Patent Document 9: da Silva Meirelles L et al, Stem Cells, 2008 Sep; 26 (9): 2287-2299

Non-Patent Document 10: da Silva Meirelles L et al., J Cell Sci, 2006; 119: 2204-2213

Non-Patent Document 11: Dai WD et al., Circulation, 2005; 112: 2I4-223

Non-Patent Document 12: Fazel S et al., J Thorac Cardiovasc Surg, 2005; 130: 1310-1318

Non-Patent Document 13: Noiseux N et al., Mol Ther, 2006; 14: 840-850

Non-Patent Document 14: Zhao LR et al., Exp Neurol, 2002; 174:11-20

Non-Patent Document 15: Gnecchi M et al., Nat Med, 2005; 11: 367-368

Non-Patent Document 16: Kinnaird T et al., Circ Res, 2004; 94: 678-685

Non-Patent Document 17: Kinnaird T et al., Circulation, 2004; 109:1543- 1549

Non-Patent Document 18: Tang YL et al., Ann Thorac Surg, 2005; 80: 229-237

Non-Patent Document 19: Zhang M et al., FASEB J, 2007; 21: 3197-3207

Non-Patent Document 20: Le Blanc K et al., J Intern Med, 2007; 262:509-525

Non-Patent Document 21: Uccelli A et al., Trends Immunol, 2007; 28:219-226

Non-Patent Document 22: Caplan Al et al., J Cell Biochem, 2006; 98: 1076-1084

Non-Patent Document 23: Fang BJ et al., Transplantation, 2004; 78: 83-88

Non-Patent Document 24: Ortiz LA et al., Proc Natl Acad Sci USA, 2003; 100: 8407-841

Non-Patent Document 25: Xiaochuan Jiahong, Experimental Medicine, 28: 1680-1687, 2010

Non-Patent Document 26: Medzhitov R, Nature, 454: 428-35, 2008

Non-Patent Document 27: Hotamisligil GS, Nature, 444 (7121): 860-7, 2006

Non-Patent Document 28: Wellen KE et al., J Clin Invest, 115 (5): 1111-9, 2005

Non-Patent Document 29: Li Bo Xiao Xiang, Experimental Medicine, 28:1717-1723, 2010

Non-Patent Document 30: Dalla-Favera R et al., Nature, 292: 31-35, 1981

The present invention has been made in view of the above circumstances, and it is an object of the invention to provide an agent effective for the production of PDGF-BB, and to provide an agent effective for promoting the production of mesenchymal stem cells and/or stabilization thereof.

The inventors of the present invention conducted repeated studies on various materials and screened out the agents for the production of PDGF-BB, and found that each plant source of cranberry, mangosteen, and astragalus, as well as inositol and inositol phosphate showed remarkable results, respectively. The PDGF-BB produces a hyperactive effect to achieve the present invention.

Accordingly, this application contains the following inventions:

[1] A platelet-derived growth factor-BB (PDGF-BB)-producing agent comprising one or two selected from the group consisting of a bilberry source, a mangosteen source, a scutellaria source, an inositol, and an inositol phosphate. The above components are used as an active ingredient.

[2] The PDGF-BB according to the above [1] produces a sputum agent which contains inositol and further contains a yeast extract.

[3] The PDGF-BB according to [1] or [2] above produces a sputum agent, wherein the inositol is an inositol phosphate derived from rice bran.

[4] A mesenchymal stem cell production promoting agent comprising the PDGF-BB producing a hyperreactive agent according to any one of the above [1] to [3].

[5] A stem cell stabilizer comprising the PDGF-BB-producing agent according to any one of the above [1] to [3].

[6] A dermal regenerating agent comprising the PDGF-BB producing a stimulating agent according to any one of the above [1] to [3].

According to the present invention, an agent effective for the production of PDGF-BB can be provided, and an agent effective for promoting or stabilizing the production of mesenchymal stem cells can be provided by using the above agent.

[Bilberry Source]

Bilberry (scientific name: Vaccinium vitis-idaea L.) is a plant of the genus Bilberry. In the present invention, as a source of bilberry, use: bilberry flower, flower spike, ear, peel, fruit, stem, leaf, branch, branch, trunk, bark, rhizome, root bark, root, seed or The person who has been pulverized by the whole grass is extracted by a solvent or the like after pulverization, or is decomposed by treatment such as enzymatic treatment or mechanical treatment after pulverization.

In the case of extracting bilberry, the extraction solvent may be used arbitrarily as long as it is a solvent for extraction, and may be used alone or in combination: water, methanol, ethanol, propylene glycol, 1,3-butylene glycol, or C. An alcohol such as a triol or an organic solvent such as chloroform, dichloroethane, carbon tetrachloride, acetone or ethyl acetate.

The extraction can be carried out at room temperature or under heating (for example, using a heated solvent such as warm water or hot water).

The extract obtained by the extraction with the above solvent may be used as it is, or the concentrated extract may be used, or may be used, or the extract may be obtained by an adsorption method such as an ion exchange resin to remove impurities, or a porous polymer may be used. The column (for example, Amberlite XAD-2) is adsorbed and then precipitated and concentrated in methanol or ethanol to obtain. Further, an extract obtained by extraction by a known distribution method (for example, a distribution method using water/ethyl acetate or the like) may be used.

When the bilberry is decomposed, "single cell treatment" which selectively decomposes the intercellular substance and maintains the cell morphology can be performed. For example, the method described in Japanese Patent Laid-Open Publication No. 2002-193734 can be referred to. Specifically, it refers to a treatment in which, by enzymatic decomposition or mechanical decomposition, the cell is selectively decomposed or destroyed, and the cell is intervened, and the cell is decomposed into cell units while maintaining cell morphology. Forming a porridge, liquid composition or freeze-dried product. In order to carry out the above selective decomposition, an enzyme treatment can be used. For example, it can be obtained by isolation, isolation, or purification from an enzyme raw material such as the genus Rizopus or the genus Aspergillus by a generally known method, or a commercially available product can also be used. When the monocellular enzymes are allowed to function in the bilberry, It is preferred to function under optimum conditions such as the optimum temperature of the enzyme to be used and the optimum pH, and the minimum amount required for use. Here, the optimum temperature of the single cell enzyme is 30 to 45 ° C, and the optimum pH is 4 to 6. Further, the portion which becomes a residue after the enzymatic reaction can be removed, for example, by using a sieve of about 20 mesh to obtain a bilberry source which can be contained in the composition for external use.

Further, in addition to the suspension of the above-described single cellified plant, a single cell slurry obtained by dehydrating the suspension by a method such as centrifugation or a dried product thereof may be used in the present invention.

To date, bilberry has been used as a cosmetic agent for bactericidal, antibacterial, and dandruff prevention (Japanese Patent Laid-Open No. 2002-193734) (Japanese Patent Laid-Open No. 61-238719). ), an inhibitor of elastase activity (Japanese Patent Laid-Open No. 2002-363088), an active enzyme eliminator (Japanese Patent Laid-Open No. 2002-363027), a melanin production inhibitor (Japanese Patent Laid-Open No. 2002-363057), and lipid peroxide production inhibition Agent (Japanese Patent Laid-Open No. 2002-363089), inhibitor of collagenase activity (Japanese Patent Laid-Open No. 2003-12531), inhibitor of hyaluronidase activity (Japanese Patent Laid-Open No. 2003-73287), inhibitor of metalloproteinase expression (Japan) Patent Laid-Open No. 2002-193738), an anti-glycation agent (Japanese Patent Laid-Open No. 2004-505007), and an anti-skin agent for skin disorders such as skin hypertrophy and skin sclerosis (Japanese Patent Laid-Open No. 2005-306850), but the more The case where the orange source has a PDGF-BB production promoting action, a mesenchymal stem cell production promoting action, a stem cell stabilizing action, and a dermis stabilization effect has not been known so far, and these effects are not known by the present inventors. Find.

[Mangosteen source]

Mangosteen (scientific name: Garcinia mangostana) is a plant of the genus Euphorbia.

In the present invention, as a source of mangosteen, the extract of the mangosteen fruit, peel, fruit, stem, leaf, branch, branch, trunk, bark, rhizome, root bark, root, seed or whole grass can be used. Or those which are extracted by a solvent or the like after pulverization.

In the case of extracting mangosteen, the extracting solvent can be used arbitrarily as long as it is a solvent for extraction, and can be obtained, for example, by extracting with the following solvent and distilling off the solvent: methanol, ethanol, propanol, and isopropyl A lower alcohol such as an alcohol, butanol or isobutanol or an aqueous lower alcohol, a polyhydric alcohol such as propylene glycol or 1,3-butanediol, or an organic solvent such as an aqueous polyol, acetone or ethyl acetate.

The extraction can be carried out at room temperature or under heating (for example, using a heated solvent such as warm water or hot water).

Further, an enzyme may be added to the solvent to carry out an extraction treatment. By adding an enzyme, the cell tissue of the fruit can be disintegrated, thereby further improving the extraction efficiency. As the enzyme, a cell tissue disintegrating enzyme is preferably used. Examples of such an enzyme include pectinase, cellulase, hemicellulase, α-amylase, and phytase. These enzymes may be used alone or in combination of two or more.

The extract obtained by using the above extraction solvent can be used as it is, or the extraction solvent can be distilled off and dried as needed.

So far, Mangosteen has been used as a UV absorber (Japanese Patent Laid-Open No. 9-87155), MMPs (matrix metalloproteinases, matrix metalloproteinases) inhibitors (Japanese Patent Laid-Open No. 2003-252745), and fibroblast activators (Japanese Patent Special In the case of 2006-249051), it is known that the production of xanthones derivatives and the action of collagen are produced (Japanese Patent Laid-Open No. 2009-84169), but the source of mangosteen has a promoting effect on PDGF-BB, The situation in which the stem cell stem cell production promoting action, the stem cell stabilizing action, and the dermis stabilization effect have not been known so far, and these effects were first discovered by the present inventors and the like.

[黄芩源物]

Astragalus (scientific name: Scutellaria baicalensis Georgi, Labiatae) is a plant of the genus Astragalus. In the present invention, as a source of scutellaria, the ear, peel, fruit, stem, leaf, branch, branch, trunk, bark, rhizome, root bark, root, and species of the scutellaria can be used. The person who has been pulverized after the whole or the whole grass is pulverized, or is extracted by a solvent or the like after pulverization.

In the case of extracting scutellaria, the extraction solvent may be used arbitrarily as long as it is usually used for plant extraction or the like, and examples thereof include water, lower alcohol such as methanol, ethanol, isopropanol or n-butanol, propylene glycol, and the like. A polyol such as 1,3-butanediol, or a hydrate of the alcohols, a hydrocarbon solvent such as n-hexane or toluene. These may be used alone or in combination of any two or more. Among them, a lower alcohol such as methanol or ethanol is preferably used.

The extraction can be carried out at room temperature or under heating (for example, using a heated solvent such as warm water or hot water).

Further, an enzyme may be added to the solvent to carry out an extraction treatment. By adding an enzyme, the cell tissue of the fruit can be disintegrated, thereby further improving the extraction efficiency. As the enzyme, a cell tissue disintegrating enzyme is preferably used. Examples of such an enzyme include pectinase, cellulase, hemicellulase, α-amylase, and phytase. These enzymes may be used alone or in combination of two or more.

The extract obtained by using the above extraction solvent can be used as it is, or the extraction solvent can be distilled off and dried as needed.

To date, Astragalus has been used as a dermal papilla activator (Japanese Patent Laid-Open No. 11-240823), a hyaluronic acid production energy increasing agent (Japanese Patent Laid-Open No. 10-95735), and an immunosuppressive agent for preventing ultraviolet skin immune function decline (Japan) Patent No. 11-71295), an anti-inflammatory agent (Japanese Patent Laid-Open No. 2006-8536), and an antioxidant (Japanese Patent Laid-Open No. Hei 5-238925), but the source of Astragalus membranaceus has a PDGF-BB production promoting effect, and a mesenchymal The case where stem cell production promoting action, stem cell stabilization, and dermal stabilizing action have not been known so far, and these effects were first discovered by the present inventors and the like.

[inositol, inositol phosphate]

It is known that inositol is called vitamin B, and is obtained by biosynthesis from glucose even in an organism. Inositol is also included in plants such as grains, alfalfa, beans, and fruits. Inositol is usually present in the form of this hydroxyphosphorylation (inositol phosphate). Also in the present invention This inositol phosphate can be used. Various stereoisomers of inositol are included in the inositol used in the present invention. As a stereoisomer of inositol, there are cis-inositol (1,2,3,4,5,6/0-inositol), epi-inositol (1,2,3,4,5/6- Inositol), allo-inositol (1,2,3,4/5,6-inositol), myo-inositol (1,2,3,5/4,6-inositol), muco-inositol (1,2,4,5/3,6-inositol), neo-inositol (1,2,3/4,5,6-inositol), chiro-inositol (1,2,4/3 , 5,6-inositol), and scyllo-inositol (1,3,5/2,4,6-inositol). Examples of the inositol phosphate include a compound obtained by phosphorylating one or two or more of the six hydroxyl groups of inositol. Among them, a compound obtained by phosphorylating all of the six hydroxyl groups of inositol, that is, phytic acid, is preferred. The inositol or inositol phosphate used in the present invention is preferably derived from rice bran. Further, any salt of inositol phosphate can also be used in the present invention.

According to an embodiment of the present invention, inositol and inositol phosphate can be used by dissolving a monomer powder in a medium and appropriately diluting it.

Inositol and inositol phosphate can also increase PDGF-BB production by combining with other substances. As the other substance, a yeast extract can be mentioned, and for example, Biodyne (registered trademark) EMPP (manufactured by Arch Personal Care Products L.P.) can be used. In the case of using the extract, the extract can be added to the medium so as to be about 0.1% by concentration conversion from the dried residual component of the extract.

To date, inositol and inositol phosphate have been used as a cosmetic composition for aging skin containing inositol and/or for stress-bearing skin (Japanese Patent Laid-Open No. 204-501069), containing inositol phosphate (especially phytic acid and A cosmetic composition for reducing or preventing signs of fatty edema (Japanese Patent Laid-Open No. Hei 8-253406), and a composition for skin moisturizing and/or skin protection and/or skin anti-aging (Japanese Patent Special) Open the case of 2010-150258). In addition, as a vitamin B, there is a skin external preparation for cell activation containing vitamins B2 and B6 (Japanese Patent Laid-Open No. Hei 9-241146), and an antioxidant containing vitamin B (Japanese Patent Laid-Open No. Hei 7-277939 and Japanese Patent No. Open 205306831, etc.). However, vitamin B or inositol or inositol phosphate has a PDGF-BB production-promoting effect, mesenchymal stem cell production promoting action, stem cell stabilization, and dermal stabilization. It is not known that these effects were first discovered by the inventors and the like.

[PDGF-BB produces a hypertrophic agent, a mesenchymal stem cell production promoter, a stem cell stabilizer, and a dermal regenerative agent]

The PDGF-BB producing agent of the present invention contains one or more selected from the group consisting of a bilberry source, a mangosteen source, a scutellaria source, an inositol, and an inositol phosphate as an active ingredient. Further, the leaf cell stem cell production promoting agent, the stem cell stabilizer, and the dermal regenerating agent according to the present invention comprise the PDGF-BB producing hypertrophy agent of the present invention containing the above active ingredient. The PDGF-BB of the present invention produces a stimulating agent, a mesenchymal stem cell production promoting agent, a stem cell stabilizer, and a dermal regenerating agent (hereinafter referred to as "the agent of the present invention"), which may contain the above-mentioned active ingredient alone. Any one of them may be used in any combination and in a ratio of two or more.

As described above, according to the findings of the present inventors, PDGF-BB is related to the localization of mesenchymal stem cells, and the production of mesenchymal stem cells can be achieved by hyperactive PDGF-BB in vascular endothelial cells. And stabilization (Japanese Patent Patent No. 2010-209705). That is, the PDGF-BB producing stimulating agent of the present invention can be extremely effectively used for the purpose of hyperproliferation and stabilization of mesenchymal stem cells.

Further, as described above, it is known that the hyperplasia and stabilization of mesenchymal stem cells are stable to blood vessels, maintenance of tissue constants, repair of damaged tissues, regeneration (especially regeneration of dermis), anti-fibrosis, and multiple occurrences. The prevention and treatment of various diseases such as sclerosing disease and diabetes are extremely effective in the prevention and improvement of various states based on chronic inflammation such as metabolic syndrome. Therefore, the leaf system stem cell production promoting agent and stem cell stabilizer of the present invention using the PDGF-BB of the present invention to produce a sputum agent can be extremely effectively used for such applications.

The agent of the present invention may be a composition obtained by combining the above-mentioned active ingredient with one or more other components such as an excipient, a carrier, and/or a diluent. The composition or form of the composition is arbitrary, and may be appropriately selected depending on conditions such as the active ingredient or the use. The The composition can be produced according to its dosage form by a suitable combination with an excipient, a carrier, and/or a diluent, and the like, and by a conventional method.

The agent of the present invention can be incorporated into various foods and drinks, feeds (pet foods, etc.) to be ingested by humans and animals. Further, it may be formulated into a cosmetic or the like for use in humans and animals, or as a pharmaceutical preparation for humans and animals.

Specifically, when the agent of the present invention is formulated in a food or beverage, a feed, or the like, the amount of the plant body or the extract thereof (dry mass) may be appropriately selected depending on the type, purpose, form, usage, and the like. Decide. For example, it can be formulated in such a manner that the amount of the plant or its extract per day is 0.5 mg to 1 g (dry residual component) and the cranberry extract is 0.5 mg to 3 g. In particular, in the case of use as a food or drink for health care, in order to give full play to the specific effects of the active ingredient of the present invention, it is preferred that the adult geranium extract is 10 mg to 500 mg (dry residual ingredient) per day, and the cranberry extract is obtained. Ingestion is carried out in the form of 10 mg to 1.5 g (dry residual component).

The form of the food or drink or the feed may be in any form, and may be, for example, formed into a pellet, a pellet, a slurry, a gel, a solid or a liquid. In the above-mentioned form, various known substances, such as a binder, a disintegrating agent, a thickening agent, a dispersing agent, a resorbing agent, a flavoring agent, a buffering agent, and a surfactant, which are contained in foods and drinks, etc., can be suitably contained. Excipients such as dissolution aids, preservatives, emulsifiers, isotonic agents, stabilizers or pH adjusters.

In the case where the agent of the present invention is used for a cosmetic, the amount (dry mass) of the plant body or the extract thereof can be appropriately determined depending on the kind, purpose, form, usage, and the like of the above. For example, the hollyhock extract and the cranberry extract may be separately formulated in a total amount of the cosmetic material in an amount of 0.00001% to 50% (in terms of dry mass), and preferably 0.0001% to 5% (in terms of dry mass).

In addition to the above-mentioned components, it may be appropriately formulated into a skin external preparation such as a cosmetic or a pharmaceutical, as needed, within a range that does not impair the effects of the present invention. For example, antioxidants, oils, ultraviolet defense agents, surfactants, tackifiers, alcohols, powder components, colored materials, aqueous components, water, various skin nutrients, and the like.

Further, it may be appropriately formulated: a metal ion chelating agent such as disodium ethylenediaminetetraacetate, trisodium ethylenediaminetetraacetate, sodium citrate, sodium polyphosphate, sodium metaphosphate, gluconic acid, methylparaben, and nibble. Preservatives such as periethyl ethyl ester and paraben, caffeine, citric acid, verapamil, tranexamic acid and its derivatives, licorice extract, licorice, papaya fruit hot water extract, various Natural medicine, acetic acid vitamin E, glycyrrhizic acid and its derivatives or its salts, vitamin C, magnesium ascorbyl phosphate, ascorbyl glucoside, arbutin, kojic acid and other whitening agents, glucose, fructose, mannose, sucrose, seaweed Sugars such as sugar.

Further, the external preparation for skin can be widely used for cosmetics, quasi-drugs and the like used for the outer skin, and is particularly preferably used in cosmetics. The dosage form may be any one suitable for application to the skin, and a solution system, a solubilizing system, an emulsification system, a powder dispersion system, a water-oil two-layer system, a water-oil-powder three-layer system, an ointment, a lotion, and a coagulating solution may be used. Any dosage form such as glue or aerosol.

When the agent of the present invention is used as a cosmetic, it is preferably used as a lotion, lotion, foundation, lipstick, lipstick, facial cleanser, massage cream, peeling mask, hand cream, hand powder, shower gel, It can be used in the form of body lotion, body cream, bathing lotion, etc., and can be appropriately formulated at this time, and components which are usually added and formulated when forming such forms, such as a moisturizer, a fragrance, a solubilizing agent, a stabilizer, and an ultraviolet absorber. , UV scattering agents and other ingredients.

When the agent of the present invention is used as an external preparation, the amount of the plant body or the extract thereof (dry mass) can be appropriately determined depending on the type, purpose, form, usage, and the like. For example, it is preferably 0.00001% to 50% (calculated as dry mass) in the total amount of the cosmetic, and is preferably 0.0001% to 5% (converted to dry mass) in other foods and drinks.

In the case where the agent of the present invention is used as a quasi-drug, the preparation can be preferably used by oral or parenteral (intravenous administration, intraperitoneal administration, etc.). The dosage form is also arbitrary, examples If it can be suitably prepared by a known method, a solid preparation for oral administration such as a tablet, a granule, a powder or a capsule, or an oral liquid preparation such as an internal liquid preparation or a syrup preparation, or a non-oral liquid such as an injection preparation Any form such as preparation. In these pharmaceutical preparations, a commonly used binder, disintegrant, viscosity enhancer, dispersant, resorption enhancer, flavoring agent, buffering agent, surfactant, dissolution aid, and preservative can also be suitably used. Excipients such as emulsifiers, isotonic agents, stabilizers or pH adjusters.

When it is an external preparation, it can be widely used for various ointments and the like, but it is preferably used as an emulsion, a suspension, an emulsion, a liquid, an ointment, a patch, and the like. Furthermore, the form in which the agent of the present invention can be used is not limited to the form and form.

For the expression of the PDGF-BB gene in the leaf cell stem cells when the agent of the present invention is administered, for example, the amount of expression can be determined by measuring the amount of PDGF-BB and evaluated. Preferably, the assay is carried out by using an antibody specific for PDGF-BB and by a method known in the art, for example, immunostaining using fluorescent substances, pigments, enzymes, etc., western blotting, immunoassay methods such as ELISA. (Enzyme-linked immunosorbent assay), RIA (Radio Immunoassay) and various methods. Further, for example, the amount of mRNA encoding PDGF-B can be measured by extracting total RNA (Ribonucleic Acid, ribonucleic acid) in mesenchymal stem cells, and the amount of expression can be determined and evaluated. The extraction of mRNA and the determination of the amount thereof are also well known in the art. For example, the quantification of RNA can be performed by a quantitative polymerase chain reaction (PCR) method, such as a real-time polymerase chain reaction (RT-PCR, Reverse Transcription-Polymerase). Chain Reaction). Selection of primers suitable for RT-PCR can be carried out by methods well known to those skilled in the art.

[Examples]

The invention will now be described in more detail by way of examples. Furthermore, the present invention is not limited to this.

[sample of evaluation]

The following was used as a sample for evaluation of the PDGF-BB production.

‧ Inositol:

After the commercially available inositol powder (manufactured by Wako Pure Chemical Industries, Ltd.: inositol) was dissolved in PBS (Phosphate Buffered Saline, phosphate buffer), it was used in an amount of 10 ppm with respect to the following medium.

‧Phytic acid:

Commercially available phytic acid (manufactured by Nacalai Tesque, Inc.: 50% aqueous solution) was used in an amount of 10 ppm with respect to the following medium.

‧ Astragalus extract:

An extract obtained by extracting 70% by volume of ethanol (a mixture of water and ethanol in a volume ratio of 3:7) to remove the root of the scutellaria outer skin was used. The extract was dried and stored, and dissolved in 70% by volume of ethanol before use, and then used in an amount of 15 ppm based on the following medium (in terms of dry weight of the extract).

‧Cranberry CRS (cell release system):

A suspension obtained by adding 1,3-BG to a cell suspension obtained by treating bilberry leaves with a cytolysis enzyme (Macerozyme A) at 40% by volume. The suspension was stored as a liquid, and was used in an amount of 30 ppm with respect to the following medium (in terms of dry weight of the extract).

‧ Mangosteen Bark Extract:

An extract obtained by extracting the bark of mangosteen with 70% by volume of 1,3-BG (a mixture of water and 1,3-BG in a volume ratio of 3:7) was used. The extract was dried and stored, and dissolved in 70% by volume of 1,3-BG before use, and then used in an amount of 10 ppm based on the following medium (in terms of dry weight of the extract).

[Evaluation of hyperactivity of PDGF-BB in vascular endothelial cells]

HUVEC (Human Umbilical Vein Endothelial Cells) were subcultured in EGM-2 medium (Sanguang pure drug), and the passage 4th generation cells were suspended in VEGF-free-free ( Physico-dermal growth factor-A, vascular endothelial growth factor-A) in Humedia-EG2 medium (Kurabo), sown in a coating of 20-well multi-plate (Asahi Glass) coated with collagen at 5% CO _{In the} presence of ₂ , culture was carried out at 37 ° C for 3 to 5 days until the cells reached collection. The above various samples were added so as to have the above-mentioned concentrations, or they were exchanged in Humedia-EG2 medium (Kurabo) to which a solvent for dissolving various samples to be evaluated was added, and further cultured for 2 days. The cultured cells were subjected to mRNA extraction and purification using an RNA extraction reagent MagNA Pure LC mRNA HS kit (Roche) and an automatic nucleic acid extraction apparatus MagNA Pure LC 1.0 Instruments (Roche) according to the instructions provided. For each sample, mRNA of the same capacity was placed in a mold, and PDGF-B was carried out using the primer pair of the following SEQ ID NOs: 1 and 2, the reaction reagent QuantiFast SYBR Green RT-PCR Kit (Qiagen), and the reaction device LightGycler (Roche). One step of the gene quantifies immediate (RT)-PCR. The composition conditions are based on Qiagen's operating instructions. Further, the conditions of the RT-PCR were set to 50 ° C for 20 minutes in the RT reaction, 95 ° C for 15 minutes in the initial modification, and 94 ° C for 15 seconds in the modification, and were set in the annealing. 60 ° C, 20 seconds, set to 72 ° C, 30 seconds in the extension. Furthermore, G3PDH was used as an internal standard (primer pair of SEQ ID NOs: 3 and 4), and used to correct the amount of mRNA in the control group.

PDGF-B:

Forward introduction: 5'-CCTGGCATGCAAGTGTGA-3' (serial number 1)

Reverse primer: 5'-CCAATGGTCACCCGATTT-3' (serial number 2)

G3PDH:

Forward introduction: 5'-GCACCGTCAAGGCTGAGAAC-3' (serial number 3)

Reverse primer: 5'-ATGGTGGTGAAGACGCCAGT-3' (serial number 4)

[Evaluation results]

The ratio of the expression amount of the PDGF-BB mRNA obtained by each of the above samples to the amount of the expression obtained by dissolving the solvent of each sample to be evaluated according to the above evaluation order Shown in the table below. Based on the results below, it was found that these components have an activity of making PDGF-BB exhibit hyperactivity.

<110> Shiseido Co., Ltd.

<120> Platelet-derived growth factor-BB produces a hypertrophic agent and a leaf cell stem cell Producing accelerators, stem cell stabilizers, and dermal regenerants

<130> SSD-Z669-PCT

<140> 100123401

<141> 2011-07-01

<160> 4

<170> PatentIn version 3.5

<210> 1

<211> 18

<212> DNA

<213> Artificial sequence

<220>

<223> PDGF-B forward primer

<400> 1

<210> 2

<211> 18

<212> DNA

<213> Artificial sequence

<220>

<223> PDGF-B reverse primer

<400> 2

<210> 3

<211> 20

<212> DNA

<213> Artificial sequence

<220>

<223> G3PDH forward primer

<400> 3

<210> 4

<211> 20

<212> DNA

<213> Artificial sequence

<220>

<223> G3PDH reverse primer

<400> 4

Claims (3)

A use of one or two selected from the group consisting of inositol and inositol phosphate for the production of a platelet-derived growth factor-BB (PDGF-BB)-producing agent. The use of claim 1, wherein the PDGF-BB produces a sputum agent and further comprises a yeast extract. The use of claim 1 or 2, wherein the inositol or inositol phosphate is from rice bran.