JP2018123130A - Cosmetic composition and mitochondrial transfer promoting agent - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide cosmetic compositions used as novel cosmetics, drugs, foods, and the like which are effective in the prevention and/or improvement of symptoms and conditions associated with dysfunction and quality deterioration of intracellular mitochondria closely involved in aging, environmental factors, stress, and the like.SOLUTION: The present invention is a cosmetic composition containing a mitochondrial transfer promoting agent. The mitochondria transfer promoting agent comprises at least one water soluble active ingredient selected from the group consisting of Phellodendron amurense extract, Coptis root extract, Prunus domestica extract, Curcuma domestica Valeton extract, Melia azadirachta extract, Hamamelis virginiana extract, Capsicum annuum extract, an Althaea officinalis extract, Calendula officinalis extract, Aesculus hippocastanum extract, Zingiber officinale extract, Rosa centifolia extract, Poria Sclerotium extract, Eucalyptus extract, Citrus junos extract, Coix lacryma-joli extract, Lavandula extract, Malus pumila extract, Citrus limon extract, Scutellaria baicalensis extract, Aloe extract, puerariae radix extract, Asiasarum root extract, Rehmannia glutinosa extract, Perilla frutescens var. crispa extract, Prunus extract, Matricaria chamomilla extract, Gambir extract, Arnica montana extract, Sophora angustifolia extract, Ruscus aculeatus extract, Luffa aegyptiaca extract, Centaurea cyanus extract, An Aspalathus linearis extract, Prunus persica extract, Angelica keiskei extract, Urtica dioica extract, Foeniculum vulgare extract, Malva sylvestris var. mauritiana extract, Ononis spinosa extract, Olea europaea L. extract, Prunus armeniaca extract, Satsuma orange extract, Tussilago farfara extract, Prunus mume extract, Gynostemma pentaphyllum extract, Chlorophyta extract, Rhodophyta extract, Phaeophyceae extract, Lentinula edodes extract, Cordyceps sinensis extract, Litchi chinensis extract, water-soluble collagen, and royal jelly extract.SELECTED DRAWING: None

Description

  The present invention relates to a cosmetic composition and a mitochondrial transfer promoter.

  Mitochondria are intracellular organelles that produce the energy necessary for various intracellular activities. It is known that this mitochondrion is impaired in function and quality due to environmental factors such as aging, ultraviolet rays, stress, and genetic background. In addition, it is also known that intracellular mitochondrial function declines in various diseases such as inflammatory diseases and mitochondrial diseases (see Non-Patent Document 1). When such mitochondrial dysfunction and deterioration of quality are caused, energy production in vivo is weakened, and it is also pointed out that active oxygen and lipid peroxidation products may be generated in cells. Thus, researches for preventing aging by a mitochondrial dysfunction inhibitor using a specific compound have been carried out (see Patent Document 1).

JP 2007-55923 A

Nature, 2011 Jan 13; 469 (7329), p. 221-5

  Under such circumstances, new cosmetics, drugs, and foods that are effective in preventing and improving symptoms and conditions associated with impaired cellular mitochondrial dysfunction and quality that are closely related to aging, environmental factors, stress, etc. Etc. are required. Then, this invention makes it a subject to provide the composition effective in prevention and improvement of the said symptom and state.

  As a result of various studies by the present inventors in order to solve the above problems, it has been found that water-soluble components such as specific plant extracts, algae extracts, and fungal extracts have an effect of promoting mitochondrial transfer between cells. . In particular, these water-soluble components exhibit an excellent mitochondrial transfer promoting effect among specific cells involved in the structure of skin tissue. The composition containing such a specific water-soluble component is prevention / improvement of symptoms or conditions caused by the presence of cells in which mitochondrial dysfunction or deterioration is caused by aging, environmental factors, stress, etc., It can be suitably used for rejuvenation such as maintaining a healthy state and improving skin firmness.

That is, the gist of the present invention is as described below.
[1] A cosmetic composition containing a mitochondrial transfer promoter.
[2] Mitochondrial transfer promoters include yellowfin extract, auren extract, prune extract, turmeric extract, Melia aziracta extract, Hamelis extract, capsicum extract, altea extract, red pepper extract, horse chestnut extract, ginger extract, centifolia rose extract , Bukuryu extract, Eucalyptus extract, Yuzu extract, pearl extract, Lavender extract, Apple extract, Lemon extract, Aloe extract, Aloe extract, Cascon extract, Saishin extract, Giant extract, Perilla extract, Sakura extract, Chamomile extract, Acacia yak extract, Arnica extract, Clara extract, butcher bloom extract, loofah extract, cornflower extract, rooibos extract, peach extract, ashitaba extract, nettle , Fennel extract, mallow extract, onionis extract, olive extract, apricot extract, Citrus unshiu extract, Japanese dandelion extract, plum extract, green alga extract, red algae extract, red algae extract, brown algae extract, shiitake extract, fuyumina tsutake mushroom extract, litchi extract, The cosmetic composition according to [1], comprising at least one water-soluble active ingredient (A) selected from the group consisting of water-soluble collagen and royal jelly extract.
[3] The water-soluble active ingredient (A) is yellowfin bark extract, laurel root extract, pruned degradation product, turmeric rhizome extract, Melia azadilacta leaf extract, Hamelis leaf extract, red pepper fruit extract, Altea root extract, Tokinsenka flower Extract, horse chestnut seed extract, ginger rhizome extract, centifolia rose flower extract, budweed extract, eucalyptus leaf extract, yuzu fruit extract, pearl seed extract, lavender flower extract, apple fruit extract, lemon fruit extract, argon extract, aloe extract, Cascon extract, Saishin extract, cherry leaf extract, chamomile flower extract, asenya extract, arnica flower extract, clara root extract, butcher bloom root extract, loofah extract, cornflower flower extract, rooibos extract, peach leaf extract, ashitaba / Stem extract, nettle leaf extract, fennel fruit extract, mallow flower extract, onionis extract, olive leaf extract, apricot seed extract, ginger root extract, perilla leaf extract, Satsuma mandarin peel extract, dandelion flower extract, ume fruit extract, amacha eel extract The cosmetic composition according to [2], which is at least one ingredient selected from the group consisting of chlorella extract, cypress extract, wakame extract, shiitake extract, fuyumushinatsu salmon extract, litchi pattern extract, and water-soluble collagen object.
[4] Water-soluble active ingredient (A) is yellowfin bark extract, laurel root extract, prune degradation product, turmeric rhizome extract, Melia azajirakuta leaf extract, Hamelis leaf extract, capsicum fruit extract, Altea root extract, Tokinsenka flower The cosmetic composition according to [3], which is at least one plant extract selected from the group consisting of an extract and a horse chestnut seed extract.
[5] The cosmetic composition according to any one of [1] to [4], which is used to promote mitochondrial transfer.
[6] The cosmetic composition according to any one of [1] to [5], which is used to promote mitochondrial transfer in skin tissue.
[7] Used to promote mitochondrial transfer between one or two cells selected from the group consisting of dermal fibroblasts, epidermal keratinocytes, adipose-derived mesenchymal stem cells, epidermal stem cells and dermal stem cells The cosmetic composition according to any one of [1] to [6].
[8] The cosmetic composition according to any one of [1] to [7], which is used as an external composition for skin.
[9] The cosmetic composition according to any one of [1] to [7], which is used as an oral composition.
[10] Yellowfin extract, Auren extract, Prunes extract, Turmeric extract, Melia aziracta extract, Hamelis extract, Pepper extract, Altea extract, Toki-senka extract, Horse chestnut extract, Ginger extract, Centifolia rose extract, Buddle extract, Eucalyptus extract , Yuzu extract, pearl barley extract, lavender extract, apple extract, lemon extract, dragon gourd extract, aloe extract, cuckoo extract, saicin extract, geranium extract, perilla extract, cherry extract, chamomile extract, asenya extract, arnica extract, clara extract, butcher bloom extract , Loofah extract, cornflower extract, rooibos extract, peach extract, ashitaba extract, nettle extract, fennel extract, mallow , Onionis extract, olive extract, apricot extract, Citrus unshiu extract, dandelion extract, ume extract, green tea extract, green algae extract, red algae extract, brown algae extract, shiitake extract, fuyushinatsutake mushroom extract, litchi extract, water-soluble collagen, and royal jelly A mitochondrial transfer promoter for a cosmetic composition comprising at least one water-soluble active ingredient (A) selected from the group consisting of extracts.
[11] The cosmetic composition according to any one of [1] to [9], further comprising a component that imparts migration of fat-derived stem cells.
[12] The cosmetic composition according to any one of [1] to [9], further comprising a collagen production functional component.
[13] Used to promote collagen production, recovery of skin elasticity, regeneration of elasticity, promotion of cell regeneration, recovery / improvement of skin aging symptoms, [1] to [9], [11], [12] The cosmetic composition according to any one of [12].

  The cosmetic composition of the present invention effectively promotes mitochondrial transfer between cells by containing a mitochondrial transfer promoter, and from normal cells to normal cells from which mitochondrial dysfunction or quality deterioration has occurred. It can replenish mitochondria and can help aerobic respiration of those cells. Therefore, rejuvenation (skin aging) such as prevention / improvement of symptoms or conditions caused by the presence of cells with mitochondrial dysfunction or quality deterioration in the body, maintenance of health condition, improvement of skin elasticity (recovery) (Symptom recovery / improvement) and the like. Further, it is suitably used for promoting collagen production, regeneration of elasticity, and cell regeneration.

It is a figure which shows the change of the gene expression (SOD2) in a human fibroblast by mitochondrial transfer. It is a figure which shows the change of the gene expression (PPARGC1) in a human fibroblast by mitochondrial transfer. It is a figure which shows the effect with respect to collagen production of the substance which has a mitochondria transfer promotion effect (human fibroblast).

  Hereinafter, the cosmetic composition and the mitochondrial transfer promoter of the present invention will be described in detail.

<Beauty composition>
The cosmetic composition of the present invention contains a mitochondrial transfer promoter. “Mitochondrial transfer” refers to a phenomenon in which mitochondria move from one cell to another. Here, the type of cells to which mitochondria migrate may be between the same type of cells or between different types of cells. This transition may be performed, for example, through a tunnel called a tunneling nanotube that connects cells, but the transition path is not particularly limited, and may be a path through exocytosis and endocytosis. Good. In addition to mitochondria itself, mitochondria itself includes mitochondrial genes (DNA, RNA), various proteins, and complexes of these with other organs such as other proteins and endoplasmic reticulum. May be. The “mitochondrial transfer promoter” refers to an agent that has an effect of promoting mitochondrial transfer occurring between cells. Here, the effect of promoting mitochondrial transfer includes any meaning of increasing the supply ability of cells that supply mitochondria and increasing the ability to receive mitochondria. The cosmetic composition of the present invention refers to a composition used for the purpose of making the appearance beautiful (for example, beauty such as beautiful skin). The cosmetic composition of the present invention may contain other components in addition to the essential component mitochondrial transfer promoter as long as the effects of the present invention are not impaired.

[Mitochondrial transfer promoter]
The mitochondrial transfer promoter of the present invention contains the following water-soluble active ingredient (A). The water-soluble active ingredient (A) can effectively promote mitochondrial transfer between cells, and can replenish normal mitochondria from healthy cells to cells in which mitochondrial dysfunction or quality deterioration has occurred, Can help aerobic respiration of those cells. Therefore, it is suitable for rejuvenation such as prevention / amelioration of symptoms or conditions caused by the presence of cells with mitochondrial dysfunction or quality degradation in vivo, maintenance of health condition, improvement of skin firmness, etc. Can be used. Therefore, the mitochondrial transfer promoter of the present invention is suitably used for a cosmetic composition.

Examples of the water-soluble active ingredient (A) include the following ingredients:
Yellowfin, Auren, Prunes (also known as Prunus), Turmeric, Melia azadiracta (also referred to as Neem), Hamelis, Pepper, Altea, Dendrobinus, Horse Chestnut, Ginger, Centifolia rose, Bakuryo, Eucalyptus, Yuzu, Barley, Lavender, Apple, Lemon, Ogon, Aloe, Cascon, Saishin, Giant, Perilla, Sakura, Chamomile, Acacia, Arnica, Clara, Butcher Bloom, Loofah, Cornflower, Rooibos, Peach, Soybean, Ashitaba, Nettle, Fennel, Zeniaoi, Plant extracts such as onionis, olives, apricots, citrus mandarin, dandelion, ume, and candy
Extracts of algae (including seaweeds and freshwater algae) such as green algae (for example, chlorella), red algae, brown algae (for example, hijiki, seaweed, etc.);
Fungal extracts such as shiitake mushrooms, fuyumushinatsusatake, litchi (also referred to as garlic mushroom);
Various extracts derived from water-soluble collagen and royal jelly extract.
The mitochondrial transfer promoter of the present invention may contain one of these water-soluble active ingredients (A) alone or in combination of two or more.

More preferably, as the water-soluble active ingredient (A), yellowfin bark extract, auren root extract, pruned degradation product, turmeric rhizome extract, Melia aziracta leaf extract (also referred to as neem leaf extract), Hamelis leaf extract, chili pepper Fruit extract, Altea root extract, Cinnamon flower extract, Horse chestnut seed extract, Ginger rhizome extract, Centifolia rose flower extract, Bulboon extract, Eucalyptus leaf extract, Yuzu fruit extract, pearl seed extract, lavender flower extract, apple fruit extract, Lemon fruit extract, Ogon extract, Aloe extract, Cascon extract, Saishin extract, Giant extract, Perilla extract, Cherry leaf extract, Chamomile flower extract, Acacia extract, Arnica flower extract, Clara root extract, Butcher bloom root extract, F Mae extract, cornflower flower extract, rooibos extract, peach leaf extract, ashitaba leaf / stem extract, nettle leaf extract, fennel fruit extract, mallow flower extract, onionis extract, olive leaf extract, apricot seed extract, ginger root extract, perilla leaf extract, Plant extracts such as Citrus unshiu peel extract, dandelion flower extract, ume fruit extract, candy extract, etc .;
Seaweed or freshwater algae algae extract such as green algae extract (chlorella extract), red algae extract, brown algae extract (hijiki extract, seaweed extract);
Fungal extracts such as shiitake extract, fuyumushinatsukutake extract, litchi pattern extract;
Examples include water-soluble collagen.

  The mitochondrial transfer promoter of the present invention preferably contains the above plant extract from the viewpoint of the mitochondrial transfer promoting effect among the water-soluble active ingredients (A). Among the above plant extracts, yellowfin bark extract, oren root extract, pruned degradation product, turmeric rhizome extract, Melia aziracta leaf extract, Hamelis leaf extract, capsicum fruit extract, altea root extract, cinnamon flower extract, horse chestnut seed extract Are more preferable, and yellowfin bark extract, auren root extract, and a prune degradation product are more preferable.

  In the present invention, the extract is not particularly limited as long as the component is extracted from a plant or the like in a solvent. Although it will not be limited if it is a component which can be extracted from a plant etc. as said component, Usually, the component which melt | dissolves in a solvent is meant. Moreover, even if it does not melt | dissolve in a solvent, as long as it is a component which is a liquid independently, those can also be included in an extract. Moreover, in this invention, an extract and an extract are used synonymously.

  In the present invention, the extraction method of the extract is not particularly limited, and a known method can be appropriately used depending on the type of plant or the like. Specifically, the solvent for extraction includes water; organic solvents such as ethanol, butylene glycol (for example, 1,3-butylene glycol), propylene glycol, methanol, glycerin, and the like. Considering that the extract is used as an active ingredient of the composition of the present invention, and from the viewpoint that a higher effect of the present invention can be expected, it is preferable to use water, ethanol, butylene glycol or the like as the extraction solvent. . Moreover, it is also possible to extract without using a solvent, for example, it can obtain by heating and pressing a plant. Moreover, the extract used for this invention may be processed by enzymatic decomposition, fermentation, hydrolysis, etc. Furthermore, a commercial item can also be used as said water-soluble active ingredient (A).

  The content of the mitochondrial transfer promoter in the cosmetic composition of the present invention is not particularly limited as long as the effect of the present invention can be obtained. In terms of the water-soluble active ingredient (A), 0.0001% by weight or more is preferable, 0.0005% by weight or more is more preferable, and 0.001% by weight or more is more preferable. Moreover, 25 weight% or less is preferable, 20 weight% or less is more preferable, 15 weight% or less is further more preferable, and 10 weight% or less is especially preferable. If it is the said range, sufficient said mitochondrial transfer promoter (water-soluble active ingredient (A)) sufficient to obtain the mitochondrial transfer promotion effect between cells will be contained. In addition, the content of the mitochondrial transfer promoter in the cosmetic composition of the present invention is preferably 0.0001 to 25% by weight, more preferably 0.0005 to 20% by weight in terms of the water-soluble active ingredient (A). 0.001 to 15% by weight is more preferable, and 0.001 to 10% by weight is particularly preferable.

  The cosmetic composition of the present invention has an effect of promoting mitochondrial transfer occurring between cells by containing a mitochondrial transfer promoter containing the water-soluble active ingredient (A). Here, the effect of promoting mitochondrial transfer includes any meaning of increasing the supply ability of cells that supply mitochondria and increasing the ability to receive mitochondria. The cells to which the mitochondrial transfer agent and cosmetic composition of the present invention can be effective are not particularly limited, but cells that have decreased mitochondrial function due to aging, stress, inflammation, etc., and cause mitochondrial transfer to those cells It is a highly capable cell. Specifically, but not limited to, skin (dermal) fibroblasts, skin epithelial cells, epidermal keratinocytes, epidermal stem cells, dermal stem cells, pigment cells, adipocytes, hair matrix cells, hair papilla cells, vascular endothelial cells, Neurons, brown adipocytes, Langerhans cells, Merkel cells, immune cells (macrophages, dendritic cells, etc.), myocytes, corneal epithelial cells, conjunctival epithelial cells, goblet cells, embryonic cells / embryonic stem (ES) cells / iPS Examples include stem cells such as cells, germ stem cells, ova, sperm, hematopoietic stem cells, corneal epithelial stem cells, conjunctival epithelial stem cells, corneal stem cells, conjunctival stem cells, and mesenchymal stem cells (for example, adipose-derived mesenchymal stem cells).

  The mitochondrial transfer promoter and cosmetic composition of the present invention exhibit an excellent effect of promoting mitochondrial transfer, particularly between cells of skin tissue. Here, the skin tissue is a concept including a subcutaneous tissue. It is well known that mitochondria are involved in intracellular energy production, and it is known that aging of the tissue is promoted if the function or quality of mitochondria deteriorates due to aging, environmental factors, stress, etc. It has been. Therefore, according to the mitochondrial transfer promoter and cosmetic composition of the present invention, since a high mitochondrial transfer promoting effect is exhibited between cells of skin tissue, weakened skin fibroblasts and epidermal keratinocytes are activated, Their original cell functions are restored, for example, the ability to produce extracellular matrix (ECM) components such as collagen and the ability to proliferate cells, the source of elasticity is regenerated, and the elasticity of the skin is restored. I can expect. In fact, as shown in the Examples, in cells (receptor cells) that have undergone mitochondrial transfer, the expression level of antioxidant enzyme (SOD2) increases, and the expression of a gene (PPARGC1) that is an indicator of cell activation is expressed. Is also enhanced. Furthermore, from healthy cells and cells whose mitochondrial transfer ability (ability to transfer intracellular mitochondria to other cells) is enhanced by the mitochondrial transfer promoter and cosmetic composition of the present invention, By supplying new mitochondria to the epidermal keratinocytes, the mitochondria in the cells rejuvenate, acquire resistance to oxidative stress, improve skin tissue aging symptoms such as wrinkles, sagging and dullness of the skin, Effective for rejuvenation. Furthermore, the mitochondrial transfer promoter and cosmetic composition of the present invention can be expected to exert the same effect on stem cells such as epidermal stem cells and dermal stem cells present in the skin tissue. Can rejuvenate healthy skin. The mitochondrial transfer promoter and cosmetic composition of the present invention are used for promoting collagen production, recovery of skin elasticity, regeneration of the source of elasticity, promotion of cell regeneration, and recovery / improvement of skin aging symptoms.

  The mitochondrial transfer promoter and cosmetic composition of the present invention are suitably used as an external composition for skin because a high mitochondrial transfer promoting effect is exerted particularly on skin tissue. In addition, since it is expected to have an effect even after being taken into the body, it is also suitably used as an oral composition for supplements and foods. In addition to the mitochondrial transfer promoter containing the water-soluble active ingredient (A) effective for promoting mitochondrial transfer, the cosmetic composition of the present invention may contain other components depending on the application.

  In addition to the mitochondrial transfer promoter containing the water-soluble active ingredient (A), the cosmetic composition of the present invention can be prepared by mixing various components described below according to a conventional method.

[Skin external composition]
The cosmetic composition of the present invention is used as an external composition for skin such as cosmetics, cleaning agents, pharmaceuticals, quasi-drugs, and bath preparations. Specifically, basic cosmetics such as skin lotion, milky lotion, gel, cream, cosmetic liquid, pack, mask, hand cream, body lotion, body cream, etc .; sunscreen cosmetics; makeup cosmetics; Makeup removers, body shampoos, shampoos, rinses, treatments, etc .; scalp compositions such as hair restorers and hair growth agents; oral compositions such as toothpastes; wound ointments, acne external preparations, poultices, etc. Examples include pharmaceuticals and quasi drugs. When the cosmetic composition of the present invention is used as an external composition for skin, the other components other than the mitochondrial transfer promoter are, for example, bases / carriers, additives (interfaces) as long as the effects of the present invention are not impaired. Active agents, thickeners, preservatives, pH adjusters, chelating agents, stabilizers, irritation reducers, preservatives, colorants, dispersants, fragrances, pearlescent agents, and the like.

  Although it does not specifically limit as a base or a carrier, For example, paraffin, liquid paraffin, squalane, white wax, gelled hydrocarbon (Plastibase etc.), ozokerite, ceresin, petrolatum, hard fat, microcrystalline wax, α-olefin oligomer And hydrocarbons such as light liquid paraffin; fatty acids such as lauric acid, myristic acid, palmitic acid, stearic acid, behenic acid, and isostearic acid; glyceryl tri-2-ethylhexanoate (trioctanoin), and tri ( Tri-fatty acid glycerides such as glyceryl (caprylic acid / capric acid); higher alcohols such as cetanol, stearyl alcohol, and behenyl alcohol; methyl polysiloxane, highly polymerized methyl polysiloxane, dimethylsiloxane methyl (Polyoxyethylene) siloxane / methyl (polyoxypropylene) siloxane copolymer, dimethylsiloxane / methyl (polyoxyethylene) siloxane copolymer, dimethylsiloxane / methyl (polyoxypropylene) siloxane copolymer, polyoxyethylene / Methyl polysiloxane copolymer, poly (oxyethylene / oxypropylene) / methyl polysiloxane copolymer, dimethyl siloxane / methyl cetyloxy siloxane copolymer, dimethyl siloxane / methyl stearoxy siloxane copolymer, alkyl acrylate copolymer Combined methylpolysiloxane ester, crosslinked methylpolysiloxane, crosslinked methylphenylpolysiloxane, crosslinked polyether-modified silicone, crosslinked alkylpolyether-modified silicone, crosslinked alcohol Kill-modified silicone, decamethylcyclopentasiloxane, ethyltrisiloxane, methyltrimethicone, methylsiloxane network polymer, polyoxyethylene / methylpolysiloxane copolymer, methylhydrogenpolysiloxane, triethoxysilylethylpolydimethylsiloxyethylhexyl dimethicone And silicone oils such as dimethylpolysiloxane; ethylene glycol monoacetate, ethylene glycol diacetate, triethylene glycol diacetate, hexylene glycol diacetate, and 2-methyl-2-propene-1,1- Glycol acetates such as diol diacetate; triethylene glycol divalerate, 2,2,4-trimethyl-1,3-pentanediol monoisobutyrate, and 2,2 Glycol esters such as 4-trimethyl-1,3-pentanediol diisobutyrate; ethylene glycol diacrylate, diethylene glycol diacrylate, propylene glycol monoacrylate, 2,2-dimethyl-trimethylene glycol diacrylate, and Glycol acrylates such as 1,3-butylene glycol diacrylate; glycol dinitrates such as ethylene glycol dinitrate, diethylene glycol dinitrate, triethylene glycol dinitrate, and propylene glycol dinitrate; 2,2 ′-[1 , 4-phenylenedioxy] diethanol; cellulose derivatives such as ethylcellulose, hydroxypropylcellulose, and hydroxypropylmethylcellulose; poly Nylpyrrolidone; carrageenan; polyvinyl butyrate; polyethylene glycol; dioxane; butylene glycol adipic acid polyester; isopropyl myristate, octyldodecyl myristate, isopropyl palmitate, cetyl palmitate, isononyl isononanoate, and pentaerythrisol tetra-2-ethylhexanoate Esters such as elite; polysaccharides such as dextrin and maltodextrin; lower alcohols such as ethanol and isopropanol; ethylene glycol monomethyl ether, ethylene glycol monoethyl ether, ethylene glycol monopropyl ether, diethylene glycol monomethyl ether, diethylene glycol Monoethyl ether, diethylene glycol monopropyl ether And glycol bases such as ter, diethylene glycol monobutyl ether, propylene glycol monoethyl ether, propylene glycol monopropyl ether, dipropylene glycol monoethyl ether, and dipropylene glycol monopropyl ether; and aqueous bases such as water.

  Examples of the surfactant include sorbitan monoisostearate, sorbitan monolaurate, sorbitan monopalmitate, sorbitan monostearate, diglycerol sorbitan penta-2-ethylhexylate, and diglycerol sorbitan tetra-2-ethylhexylate. Sorbitan fatty acid esters such as: glyceryl monostearate and glyceryl fatty acids such as glyceryl monostearate malate; polyglyceryl fatty acids such as polyglyceryl monostearate, polyglyceryl monoisostearate, and polyglyceryl diisostearate; monostearin Propylene glycol fatty acid esters such as propylene glycol acid; polyoxyethylene hydrogenated castor oil 40 (HCO-40), polyoxyethylene Hydrogenated castor oil derivatives such as hydrogenated castor oil 50 (HCO-50), polyoxyethylene hydrogenated castor oil 60 (HCO-60), and polyoxyethylene hydrogenated castor oil 80; polyoxyethylene (20) sorbitan monolaurate Polyoxys such as (polysorbate 20), polyoxyethylene (20) sorbitan monostearate (polysorbate 60), polyoxyethylene (20) sorbitan monooleate (polysorbate 80), and polyoxyethylene (20) sorbitan isostearate Ethylene sorbitan fatty acid esters; polyoxyethylene mono coconut oil fatty acid glyceryl; glycerin alkyl ether; alkyl glucoside; polyoxyalkylene alkyl ether such as polyoxyethylene cetyl ether; Amines such as rilamine and oleylamine; silicone surfactants such as polyoxyethylene-methylpolysiloxane copolymer, lauryl PEG-9 polydimethylsiloxyethyl dimethicone, and PEG-9 polydimethylsiloxyethyl dimethicone; Can be mentioned.

  Examples of the thickener include guar gum, locust bean gum, carrageenan, xanthan gum, dextran, methylcellulose, ethylcellulose, carboxymethylcellulose, hydroxymethylcellulose, hydroxyethylcellulose, hydroxypropylcellulose, hydroxypropylmethylcellulose, sodium alginate, propylene glycol alginate, polyvinyl Alcohol, polyvinyl pyrrolidone, polyvinyl methyl ether, carboxy vinyl polymer, alkyl methacrylate copolymer, sodium polyacrylate, polyethylene glycol, bentonite, dextrin fatty acid ester, pectin, (hydroxyethyl acrylate / acryloyldimethyltaurine Na) copolymer The Chill distearyl ammonium hectorite, (ammonium acryloyldimethyltaurate / vinylpyrrolidone) copolymer, polyethylene glycol distearate, triisostearate, ethylene glycol, and triisostearate, polyoxyethylene (20) include methyl glucoside and the like.

  Examples of preservatives or preservatives include benzoic acid, sodium benzoate, dehydroacetic acid, sodium dehydroacetate, isobutyl paraoxybenzoate, isopropyl paraoxybenzoate, butyl paraoxybenzoate, ethyl paraoxybenzoate, propyl paraoxybenzoate, Examples include benzyl paraoxybenzoate, methyl paraoxybenzoate, and phenoxyethanol.

  Examples of pH adjusters include inorganic acids (hydrochloric acid, sulfuric acid, phosphoric acid, polyphosphoric acid, boric acid, etc.), organic acids (lactic acid, acetic acid, citric acid, sodium citrate, tartaric acid, malic acid, succinic acid, succinic acid, etc. Acid sodium, oxalic acid, gluconic acid, fumaric acid, propionic acid, acetic acid, aspartic acid, epsilon-aminocaproic acid, glutamic acid, and aminoethylsulfonic acid), gluconolactone, ammonium acetate, inorganic base (sodium hydrogen carbonate, carbonic acid) Sodium, potassium hydroxide, sodium hydroxide, calcium hydroxide, and magnesium hydroxide), and organic bases (monoethanolamine, triethanolamine, diisopropanolamine, triisopropanolamine, lysine, and the like).

  Examples of chelating agents include ethylenediaminetetraacetic acid (edetic acid), ethylenediaminetetraacetic acid salt (sodium salt (sodium edetate: Japanese Pharmacopoeia, EDTA-2Na, etc.), potassium salt, etc.), phytic acid, gluconic acid, polyphosphorus An acid, metaphosphoric acid, etc. are mentioned.

  Examples of the stabilizer include sodium polyacrylate, dibutylhydroxytoluene, and butylhydroxyanisole.

  Examples of the stimulus reducing agent include licorice extract, sodium alginate, and 2-methacryloyloxyethyl phosphorylcholine.

  Examples of the colorant include inorganic pigments and natural pigments.

  Examples of the pearl luster imparting agent include ethylene glycol distearate, ethylene glycol monostearate, and triethylene glycol distearate.

  In the cosmetic composition as an external composition for skin, for example, an antioxidant component, an anti-aging component, an anti-inflammatory component, a whitening component, a keratin softening component, a cell activation component, and vitamins, as long as the effects of the present invention are not hindered. , Blood circulation promoting component, moisturizing component, component having prevention and / or repair action of DNA damage, UV absorbing component, UV scattering component, cleaning component, antibacterial component, astringent component, fat-derived stem cell migration imparting component (adipose-derived stem cell And other active ingredients that can be added to cosmetics, such as a collagen-producing functional ingredient. Other active ingredients can be used singly or in combination of two or more.

  Antioxidant components include, for example, components derived from plants (eg, grape, ginseng, or comfrey) (excluding water-soluble active ingredients (A)); proanthocyanidins, tocopherols and derivatives thereof, ascorbic acid and derivatives thereof , Hesperidin, glucosyl hesperidin, ergothioneine, sodium bisulfite, erythorbic acid and its salts, flavonoids, glutathione, glutathione peroxidase, glutathione-S-transferase, catalase, superoxide dismutase, thioredoxin, taurine, thiotaurine, and hypotaurine .

  Antiaging components include, for example, hydrolyzed soy protein, retinoid (retinol and its derivatives, retinoic acid, retinal, etc.), pangamic acid, kinetin, ursolic acid, turmeric extract, sphingosine derivative, silicon, silicic acid, N-methyl -L-serine, mevalonolactone and the like.

  Anti-inflammatory components include, for example, components derived from plants (eg, Comfrey) (excluding water-soluble active ingredients (A)); allantoin, calamine, glycyrrhizic acid or derivatives thereof, glycyrrhetinic acid or derivatives thereof, zinc oxide, Examples include guaiazulene, pyridoxine hydrochloride, menthol, camphor, turpentine oil, indomethacin, and salicylic acid or derivatives thereof.

  Examples of the whitening component include arbutin, hydroquinone, kojic acid, ellagic acid, phytic acid, lucinol, chamomile ET, ascorbic acid or a derivative thereof, vitamin E or a derivative thereof, pantothenic acid or a derivative thereof, tranexamic acid, and a whitening action. Plant components (for example, plant extracts or essential oils) other than the water-soluble active ingredient (A) they have are listed.

  Examples of the keratin soft component include lanolin, urea, phytic acid, lactic acid, lactate, glycolic acid, salicylic acid, malic acid, and citric acid.

  Examples of the cell activation component include components derived from plants (for example, bilberry) (excluding the water-soluble active component (A)); amino acids such as γ-aminobutyric acid and ε-aminoproic acid; retinol and its derivatives. Vitamins such as thiamine, riboflavin, pyridoxine hydrochloride, and pantothenic acids; α-hydroxy acids such as glycolic acid and lactic acid; tannins; flavonoids; saponins;

  Vitamins include, for example, retinol derivatives such as retinol, retinol acetate and retinol palmitate, retinal, retinoic acid, methyl retinoic acid, ethyl retinoic acid, retinol retinoic acid, d-δ-tocopheryl retinoate, α-tocotic acid. Vitamin A such as ferryl retinoate and β-tocopheryl retinoate; provitamin A such as β-carotene, α-carotene, γ-carotene, δ-carotene, lycopene, zeaxanthin, cryptoxanthine, and echinone; δ Vitamin Es such as tocopherol, α-tocopherol, β-tocopherol, dl-α-tocopherol succinate, dl-α-tocopherol calcium succinate, δ-tocopherol and tocopherol nicotinate; riboflavin, flavinmo Vitamin B2s such as nucleotide, flavin adenine dinucleotide, riboflavin butyrate, riboflavin tetrabutyrate, sodium riboflavin 5′-phosphate, and riboflavin tetranicotinate; methyl nicotinate, nicotinic acid, nicotinamide, and the like Nicotinic acids; ascorbyl stearate, L-ascorbyl dipalmitate, ascorbyl tetraisopalmitate (ascorbyl tetra-2-hexyldecanoate), ascorbic acid, sodium ascorbate, dehydroascorbic acid, sodium ascorbate phosphate, ascorbic acid phosphate Vitamin C such as ester magnesium and ascorbic acid glucoside; methyl hesperidin, ergocalciferol, and cholecalcif Vitamin D such as erol; Vitamin K such as phylloquinone and farnoquinone; dibenzoyl thiamine, dibenzoyl thiamine hydrochloride, thiamine hydrochloride, thiamine cetyl hydrochloride, thiamine thiocyanate, thiamine lauryl hydrochloride, thiamine nitrate, thiamine Monophosphate, thiamine lysine salt, thiamine triphosphate, thiamine monophosphate phosphate, thiamine monophosphate, thiamine diphosphate, thiamine diphosphate hydrochloride, thiamine triphosphate, and thiamine triphosphate monophosphate Vitamin B1 such as salt; Vitamin B6 such as pyridoxine hydrochloride, pyridoxine acetate, pyridoxal hydrochloride, 5'-pyridoxal phosphate, and pyridoxamine hydrochloride, cyanocobalamin, hydroxocobalamin And folic acid such as folic acid and pteroylglutamic acid; pantothenic acid, calcium pantothenate, pantothenyl alcohol (panthenol), D-pantethein, D-panthetin, coenzyme A, and Pantothenic acids such as pantothenyl ethyl ether; biotins such as biotin and biocytin; and vitamin-like agents such as carnitine, ferulic acid, α-lipoic acid, orotic acid, and γ-oryzanol.

  Examples of the blood circulation promoting component include plants (for example, ginseng, ginkgo, enamel, Dutch oak, carrot, burdock, rice, hawthorn, hawthorn, gerbil, nematode, assembly, thyme, clove, tonin, garlic, butcher bloom, Ingredients (excluding water-soluble active ingredient (A)) derived from grapes, buttons, maronier, melissa, yuzu, yokoinin, rosemary, rosehip, chimpi, touki, spruce, peach, apricot, walnut, or corn; Acid tocopherol; glucosyl hesperidin; and hesperidin.

  Examples of moisturizing components include components derived from plants (eg, Tigaya) (excluding the water-soluble active ingredient (A)); alanine, serine, leucine, isoleucine, threonine, glycine, proline, hydroxyproline, glucosamine, and theanine Amino acids such as and derivatives thereof; proteins such as collagen, gelatin, and elastin, peptides, or hydrolysates thereof; glycerin, 1,3-butylene glycol, propylene glycol, polyethylene glycol, dipropylene glycol, and diglycerin Sugar alcohols such as sorbitol; phospholipids such as lecithin and hydrogenated lecithin; hyaluronic acid, sodium hyaluronate, acetyl hyaluronic acid, sodium acetyl hyaluronate, heparin, Mucopolysaccharides such as chondroitin; NMF-derived components such as lactic acid, sodium pyrrolidonecarboxylate, and urea; polyglutamic acid; polymers having a phospholipid polar group such as MPC polymer (for example, LIPIDURE (registered trademark)); Examples thereof include polyoxypropylene methyl glucoside; trimethyl glycine (betaine); hydroxyethyl urea; acrylic acid / acrylamide / dimethyl diallylammonium chloride copolymer; and sorbitol.

  Ingredients having the action of preventing and / or repairing DNA damage include, for example, ingredients derived from animals (eg, Artemia); ingredients derived from plants (eg, cat's claw) (excluding water-soluble active ingredients (A)) ); And nucleic acid components such as DNA, DNA salts, RNA, and RNA salts.

  Examples of the UV absorbing component include 2-methoxyhexyl paramethoxycinnamate, 2- [4- (diethylamino) -2-hydroxybenzoyl] benzoic acid hexyl ester, 2,4,6-tris [4- (2-ethylhexyl). Oxycarbonyl) anilino] -1,3,5-triazine, 2-ethylhexyl dimethoxybenzylideneoxoimidazolidinepropionate, and 2,4-bis-[{4- (2-ethylhexyloxy) -2-hydroxy} -phenyl] -6- (4-methoxyphenyl) -1,3,5-triazine and the like.

  Examples of the ultraviolet scattering component include inorganic compounds such as zinc oxide, titanium oxide, iron oxide, cerium oxide, zirconium oxide, titanium silicate, zinc silicate, anhydrous silicic acid, cerium silicate, and hydrous silicic acid. . Also, those inorganic compounds coated with an inorganic powder such as hydrous silicate, aluminum hydroxide, mica, or talc, and those inorganic compounds as resin powders such as polyamide, polyethylene, polyester, polystyrene, or nylon Examples include composites and those obtained by treating those inorganic compounds with silicon oil or fatty acid aluminum salts.

  Examples of the cleaning component include polyoxyalkylene alkyl (or alkenyl) ether sulfate, alkyl (or alkenyl) sulfate, higher fatty acid salt, ether carboxylate, amide ether carboxylate, alkyl phosphate ester salt, N- Anionic surfactants such as acyl amino acid salts, polyoxyalkylene fatty acid amide ether sulfates, acylated isethionates, and acylated taurates; amine oxides, glycerin fatty acid esters, sorbitan fatty acid esters, alkyl saccharides, polyoxyalkylene alkyl ethers, Nonionic surfactants such as fatty acid alkanolamides and polyoxyalkylene hydrogenated castor oil; mono- or di-long chains optionally having alkylene oxide attached thereto and having straight-chain or branched long-chain alkyl groups Alkyl cationic surfactants such as quaternary ammonium salts; carboxymethyl betaine, sulfobetaine, imidazolinium betaine, and amphoteric surfactants such as amide betaine.

  Antibacterial components include, for example, chlorhexidine, salicylic acid, benzalkonium chloride, acrinol, ethanol, benzethonium chloride, cresol, gluconic acid and its derivatives, popidone iodine, potassium iodide, iodine, isopropylmethylphenol, triclocarban, triclosan, photosensitizer No. 101, Photosensitive Element 201, Paraben, Phenoxyethanol, 1,2-pentanediol, and alkyldiaminoglycine hydrochloride.

  Examples of astringent components include metal salts such as alum, chlorohydroxyaluminum, aluminum chloride, allantoin aluminum salt, zinc sulfate, and potassium aluminum sulfate; and organic acids such as tannic acid, citric acid, lactic acid, and succinic acid be able to.

  Examples of components that impart migration of adipose-derived stem cells and functional components for collagen production include monosaccharides, disaccharides, oligosaccharides, polysaccharides, mucopolysaccharides, collagens, oligopeptides (dipeptides, tripeptides, tetrapeptides) And their derivatives), hydrophilic biological substances such as polypeptides, dipropylene glycol, 1,2-butylene glycol, 1,3-butylene glycol, propylene glycol, 1,3-propanediol, 1,2-pentanediol And polyol compounds having two or more hydroxyl groups in an alkyl group having 3 to 6 carbon atoms such as 1,2-hexanediol, ethylene glycol and diethylene glycol.

  When the cosmetic composition of the present invention is a composition for external use on the skin, the content of the mitochondrial transfer promoter in terms of the water-soluble active ingredient (A) is 0.0001% by weight based on the whole composition for external use on the skin. The above is preferable, 0.0005% by weight or more is more preferable, and 0.001% by weight or more is more preferable. Moreover, 10 weight% or less is preferable, 5 weight% or less is more preferable, 3 weight% or less is further more preferable, and 1 weight% or less is especially preferable. If it is the said range, sufficient said water-soluble active ingredient (A) that the mitochondrial transfer promotion effect is acquired in skin tissue will be contained. Moreover, 0.0001-10 weight% is preferable, as for content of the water-soluble active ingredient (A) conversion of the said mitochondrial transfer promoter in the external composition for skin, 0.0005-5 weight% is more preferable, 0.001 -3% by weight is more preferable, and 0.001-1% by weight is particularly preferable.

  What is necessary is just to apply | coat the skin external composition of this invention by the normal usage method and use frequency of a skin external composition. For example, the external composition for skin of the present invention is applied to a skin site where a symptom or condition (for example, reduced elasticity, wrinkle, sagging, dullness, etc.) due to mitochondrial dysfunction or quality deterioration is concerned. An appropriate amount (for example, about 0.05 to 5 g) can be applied several times a day (about 1 to 3 times per day) by means of application or the like so as to penetrate the skin.

  The use target of the cosmetic composition as the composition for external use of the skin of the present invention is not particularly limited, but symptoms or conditions caused by mitochondrial dysfunction or deterioration, such as skin tension or reduced elasticity, wrinkles, tarmi, Suitable subjects include humans who have a decrease in gloss or skin texture, humans who have the above symptoms due to a decrease in collagen production, humans who are concerned about skin aging symptoms, and the elderly. It can also be applied to young adults, including children and adolescents, and healthy adults, and it may be affected by mitochondrial dysfunction and deterioration due to aging, ultraviolet rays, sunburn, or excessive washing. Symptoms and the like caused by a decrease, wrinkle, tarmi, gloss reduction, skin texture decrease, collagen production decrease, etc. can be prevented, or progression can be suppressed. The cosmetic composition as the external composition for skin of the present invention is used for promoting collagen production, recovery of skin elasticity, regeneration of the source of elasticity, promotion of cell regeneration, and recovery / improvement of skin aging symptoms.

[Oral composition]
The cosmetic composition of the present invention is a functional label food, a health function food (including a specific health food (including a disease risk reduction label, a standard specification type), a conditional specific health food, or a nutritional function food), health Oral compositions such as foods and nutritional supplements (including balanced diets, supplements, etc.) can be used.

  The dosage form is not particularly limited, but the above-mentioned mitochondrial transfer promoter (containing the water-soluble active ingredient (A)), and an excipient or additive commonly used in foods are blended with this, In the form of pills, granules, powders, powders, capsules (including soft capsules), wettable powders, emulsions, liquids, jellies, gels, extracts, or elixirs, using known methods It can also be formulated. Of these, preparations such as tablets, pills, powders, granules, capsules, liquids, jellies, gels, and drops are preferred because they are easy to take and carry. Excipients or additives include fats and oils, stabilizers, emulsifiers, dispersants, suspending agents, fragrances, thickeners, sweeteners, colorants, fragrances, preservatives, antioxidants, and organic acids. Can be mentioned.

  Examples of the fats and oils include natural vegetable oils such as peanut oil, cocoa butter, rice germ oil, perilla oil, linseed oil, etc., hardened oils thereof, and glycerides of fatty acids (including medium chain fatty acids) (glycerides, diglycerides, triglycerides, etc.) , And beeswax.

  Examples of the emulsifier, dispersant, suspending agent, or stabilizer include polyhydric alcohols such as polyethylene glycol, propylene glycol, glycerin, and sorbitol; glycerin fatty acid esters; and sodium carboxymethyl cellulose.

  Examples of the thickener include locust bean gum, guar gum, xanthan gum, tamarind gum, pectin, and carrageenan.

  Examples of the sweetener include sucrose, fructose, maltose, erythritol, trehalose, maltitol, licorice extract, stevia processed sweetener, aspartame, acesulfame potassium, sorbitol, xylitol and the like.

Examples of the preservative include benzoic acid or a salt thereof, sorbic acid or a salt thereof, butyl paraoxybenzoate, isobutyl paraoxybenzoate, isopropyl paraoxybenzoate, and ethyl paraoxybenzoate.

  Examples of the antioxidant include ascorbic acid, sodium ascorbate, ascorbic acid stearate, sodium ascorbate stearate, ascorbyl palmitate, bisulfite, sodium hyposulfite, sulfur dioxide, disodium EDTA calcium, erythorbine Examples include acid, sodium erythorbate, and tocopherol.

  Examples of the organic acid include citric acid, succinic acid, tartaric acid, aspartic acid, lactic acid, malic acid, malonic acid, fumaric acid, and maleic acid.

  An excipient | filler and a food additive can be used individually by 1 type or in combination of 2 or more types.

  In addition, a cosmetic composition as an oral composition can be prepared by adding the mitochondrial transfer promoter (containing the water-soluble active ingredient (A)) to general foods. Examples of foods include milk, milk drinks, lactic acid bacteria drinks, soft drinks with fruit juice, carbonated drinks, vegetable juice drinks, tea drinks, fruit juice drinks, ion drinks, sports drinks, functional drinks, vitamin supplement drinks, nutrition supplement balance drinks , Beverages such as jelly beverages, alcoholic beverages, and soups; puddings such as custard pudding, milk pudding, and pudding with fruit juice, jellys, dressings, and gel-like confectionery such as creams, chewing gum, and balloons Gum-like gums (plate gum, sugar-coated grain gum, etc.), chocolates, soft candy (including caramel, nougat, gummy candy, marshmallow, etc.), caramels such as toffee, biscuits (hard biscuits, soft Biscuits, soft cookies, crackers, and semi-biscuit biscuits) and cakes Confectionery and the like; ice cream, ice milk, lacto, sherbet, and frozen desserts such as frozen desserts; bread; noodles; and pet food, and the like.

  In the cosmetic composition as an oral composition, a further active ingredient may be blended within a range not impeding the effects of the present invention. For example, among the antioxidant components, anti-aging components, anti-inflammatory components, whitening components, vitamins, blood circulation promoting components and the like exemplified as components that can be blended in the external composition for skin, any component suitable for oral use is blended can do. These active ingredients can be used individually by 1 type or in combination of 2 or more types.

  When the cosmetic composition of the present invention is an oral composition, the content of the mitochondrial transfer promoter is not particularly limited as long as the effect of the present invention is obtained with respect to the whole oral composition. Although not intended, in terms of the water-soluble active ingredient (A), 0.0001% by weight or more is preferable, 0.001% by weight or more is more preferable, 0.005% by weight or more is further preferable, 0.01% by weight The above is particularly preferable. Moreover, 25 weight% or less is preferable, 20 weight% or less is more preferable, 15 weight% or less is further more preferable, and 10 weight% or less is especially preferable. If it is the said range, sufficient said water-soluble active ingredient (A) that the mitochondrial transfer promotion effect between cells will be acquired will be contained. Moreover, 0.0001-25 weight% is preferable, as for content of the said water-soluble active ingredient (A) conversion of the said mitochondrial transfer promoter in an oral composition, 0.001-20 weight% is more preferable, 0.005 -15 wt% is more preferred, and 0.01-10 wt% is particularly preferred.

  For oral compositions, in addition to the humans listed as preferred application targets of the composition for external application to the skin, humans with reduced ability to produce collagen in vivo due to mitochondrial dysfunction or quality deterioration, for example, corneas such as corneal ulcers Patients with disorders; rheumatism; joint disorders such as arthritis such as osteoarthritis and osteoarthritis; or patients with inflammatory diseases and the like are also suitable subjects. In addition, healthy individuals such as children, young people including adolescents and healthy adults can be targeted. Moreover, it is possible to prevent or suppress the progression of skin tension or elasticity, wrinkles, tarmi, gloss, or skin texture due to aging, ultraviolet rays, sunburn, or excessive washing. . The cosmetic composition of the present invention as an oral composition can be used for promotion of collagen production, recovery of skin elasticity, regeneration of the source of elasticity, promotion of cell regeneration, recovery and improvement of skin aging symptoms. .

[Evaluation method of mitochondrial transfer promoting effect]
Examples of the method for evaluating the mitochondrial transfer promoting effect of the mitochondrial transfer promoting agent or cosmetic composition of the present invention include the methods described below. That is, when multiple cells are co-cultured, the percentage of cells receiving mitochondrial transfer (%) in the entire recipient cell is confirmed, and the mitochondrial transfer rate (%) is calculated. Depending on the degree, a method for evaluating the effect of promoting mitochondrial transfer can be mentioned.

  Specifically, a step of measuring the mitochondrial transfer rate in cells treated with a sample to be examined for the presence or absence of the effect of promoting mitochondrial transfer (hereinafter also referred to as “step 1”), and the obtained mitochondrial transfer rate is used as a reference mitochondria. It includes at least a step for comparison with the transfer rate (control) (hereinafter also referred to as “step 2”).

  Below, each process is demonstrated in detail.

[Step 1: Measuring mitochondrial transfer rate]
Cells that supply the mitochondria are stained with a mitochondrial staining reagent, and cells that receive the mitochondrion are stained with a cytoplasmic staining reagent. Staining of cells with these reagents is performed, for example, by incubating at room temperature to 37 ° C. for a predetermined time. Examples of the mitochondrial staining reagent include CytoPainter Cell Tracking Staining Kit-Green Fluorescence (manufactured by Abcam), MitoTracker Mitochondition-Selective Green Green (manufactured by Molecular Probes, etc.). Examples of the cytoplasm staining reagent include CellTrace (trademark). Far Red Cell Proliferation Kit (manufactured by Invitrogen), CellTrace Violet Cell Proliferation Kit, for flow cytometry (manufactured by Lifetech.), CellTrace Red Cell Profiter Kit . After staining, the cells are washed with HBSS (+) or HBSS (−), etc., detached with an enzyme such as trypsin, EDTA, etc., washed with HBSS (+) or HBSS (−), etc. Cells are seeded in the same plate (96 well plate etc.). Both cells are co-cultured, a sample to be evaluated is added, and the cells are cultured for about 4 to 24 hours. Thereafter, a nuclear staining reagent is added and incubated at room temperature to 37 ° C. for a predetermined time, and an image is taken with an imaging system such as ImageXpress. As the nuclear staining reagent, for example, DAPI, DRAQ5 (manufactured by biostatus), Hoechst 33342 (manufactured by DOJINDO) or the like can be used. Analysis can be performed using a cell count program or the like to calculate the mitochondrial transfer rate.

  Further, only one of the mitochondrial supply side or reception side cells is cultured for 4 to 24 hours in a medium to which the sample to be evaluated is added, and the cells are washed with HBSS (+) or HBSS (-), and then trypsin or the like. Cells are detached with EDTA or other enzyme. The cells on both the supply side and the reception side are mixed and seeded in one plate (well) and co-cultured for 4 to 24 hours. Thereafter, a nuclear staining reagent is added in the same manner as described above, and incubated at room temperature to 37 ° C. for a predetermined time (for example, about 15 minutes), and an image is taken with a confocal laser image system such as ImageXpress. Analysis can be performed using a cell count program or the like, and the mitochondrial transfer rate can be calculated. By treating only one of the cells on the mitochondrial supply side or the reception side with the sample, it is possible to confirm which cell acts on the cell to exert the effect of promoting mitochondrial transfer.

Analysis with the above cell count program can be performed as follows.
(1) Observe with a filter that recognizes nuclear staining such as Cy5 filter or DAPI filter, and recognize and measure the nucleus (= measurement of cell number).
(2) Observe with a filter that recognizes recipient cell staining, such as Cy5 filter or DAPI filter, and recognize and measure cytoplasm that is a cell that receives mitochondria (receptor side) (Red stained or Violet stained cells = Acceptor).
(3) Observation with a filter that recognizes mitochondrial staining, such as a GFP filter or a FITC filter, to recognize and measure cells (supply side) having mitochondria (Green stained or MitoTracker stained cells = supply side).
(4) The mitochondrial transfer rate (%) can be calculated by the following formula.
Further, the cytotoxicity of the drug can be evaluated by measuring the number of cells in (1).
Formula: (number of cells in (2) and (3) / number of cells in (2)) × 100

[Step 2: Step of comparing with reference mitochondrial transfer rate (%)]
In this step, the mitochondrial transfer rate (%) of the cells obtained in step 1 is compared with the mitochondrial transfer rate (%) calculated in the control experiment without addition of the sample. In many cases, when the mitochondrial transfer rate of the obtained cells is higher than the control mitochondrial transfer rate determined as a reference, the sample is judged to have an effect of promoting mitochondrial transfer. Conversely, if the mitochondrial transfer rate of the cells obtained in step 1 is lower than the control mitochondrial transfer rate, the sample is judged to have a mitochondrial transfer inhibitory effect.

  Hereinafter, the present invention will be described in more detail based on examples, but the present invention is not limited to these examples.

Example 1 NHDF (Human Skin Fibroblast) x NHDF (Human Skin Fibroblast)
The effect of each extract on mitochondrial transfer between NHDFs (human skin fibroblasts) was examined by the following method.

(Evaluation test of mitochondrial transfer ability)
Cryopreserved NHDF (human skin fibroblasts) was put to sleep in a T75 flask, cultured for 1 day, and then washed 3 times with HBSS (+). Mitochondrial supply side cells were added with mitochondrial staining reagent for 30 minutes, and recipient side cells were added with cytoplasmic staining reagent for 1 hour, stained in a 37 ° C. incubator, and then DMEM (10% FBS, 1% AB). ) Replaced with medium. The next day, after washing 3 times with HBSS (+), the cells were detached with trypsin / EDTA, seeded on a 96-well plate at 2,000 cells / well on both the supply side and the reception side, and co-cultured in a 37 ° C. incubator overnight. At that time, each extract was added at a final concentration of 1/1000. On the next day, the nuclear staining reagent was diluted in PBS (−) and replaced with the medium in the well. After staining at room temperature for 15 minutes, an image was taken with ImageXpress (16 fields / well). Each image was analyzed with a cell count program and the mitochondrial transfer rate was calculated. Table 1 shows the results for the extract that had an effect of increasing the mitochondrial transfer rate compared to the control (100%) to which each extract was not added.

Analysis using the cell count program was performed as follows.
(1) Observation with a filter that recognizes nuclear staining, such as a DAPI filter, to recognize and measure the nucleus (= measurement of the number of cells).
(2) Observe with a filter that recognizes recipient cell staining, such as a Cy5 filter, and recognize and measure the cytoplasm that is the cell (receptor side) that receives mitochondria (Red-stained cells = recipient side).
(3) Observe with a filter that recognizes mitochondrial staining, such as a FITC filter, to recognize and measure cells with mitochondria (supply side) (Green-stained cells = supply side).
(4) The mitochondrial transfer rate (%) can be calculated by the following formula.
Further, the cytotoxicity of the drug can be evaluated by measuring the number of cells in (1).
Formula: (number of cells in (2) and (3) / number of cells in (2)) × 100

(Examination of pretreatment of mitochondrial donor cell or recipient cell)
Several types of extracts having a particularly high reproducibility in the mitochondrial transfer rate were selected in the above test, and further evaluation was made as to whether these extracts act on the cells on the supply side or on the reception side.
Cryopreserved NHDF (human skin fibroblasts) was asleep, seeded on 6-well plates (3 × 10 ⁴ cells / well), and cultured for 4 days. After washing 3 times with HBSS (+), the mitochondrial staining reagent was added to the cells on the mitochondrial supply side for 30 minutes, the cytoplasmic staining reagent was added to the cells on the receiving side and stained in a 37 ° C. incubator, The medium was replaced with DMEM (10% FBS, 1% AB) medium. Next, turmeric rhizome extract, yellowfin bark extract, and Melia azadilacta leaf extract were added at a final concentration of 1/1000 to cells on the supply side or reception side, and cultured overnight in a 37 ° C. incubator. In addition, the same test was conducted for dipotassium glycyrrhizinate, which is the main component of licorice extract, and berberine sulfate, which is the main component of yellowfin bark extract. The next day, after washing three times with HBSS (+), the cells are detached with trypsin / EDTA, seeded on a 96-well plate so that both the supply side and the reception side become 2,000 cells / well, and untreated cells (2,000 cells / well) ). The cells were cultured overnight in a 37 ° C. incubator. The next day, the nuclear staining reagent was diluted in PBS (−) and replaced with the medium in the well. After staining at room temperature for 15 minutes, an image was taken with ImageXpress (16 fields / well). Analysis with a cell count program and mitochondrial transfer rates were calculated. The count program is the same as above. Table 2 shows the results of the pretreatment of the extract that had the effect of increasing the mitochondrial transfer rate compared to the control (100%) to which each extract was not added.

The cells, medium, drug, and staining reagent used in the above experiment are shown below.
Cells: NHDF human dermal fibroblasts: Human Dermal Fibroblast (KURABO KF-4009)
Medium: DMEM (Gibco 11995065) + 10% FBS + 1% AB * During cell culture and co-culture * AB: Antibiotic-Antimicotic (Gibco)
Drug: Trypsin / EDTA (manufactured by KURABO) HK-3120
Mitochondrial staining reagent: Mito Tracker (registered trademark) Green FM (M7514 manufactured by Molecular Probes) 1 / 7,000 dilution in HBSS (+) (final 70 nM)
Cytoplasmic staining reagent: CellTrace ™ Far Red Cell Proliferation Kit (Invitrogen ™ C34564) 1/1000 diluted in HBSS (+) (final 5 uM)
Nuclear staining reagent: Hoechst 33342 (manufactured by DOJINDO 346-07951) 1/1000 dilution

  It has been found that the extracts shown in Table 1 have an effect of increasing the mitochondrial transfer rate between human skin fibroblasts. Among them, turmeric rhizome extract, yellowfin bark extract, Hamelis leaf extract, and Melia azadiracta leaf extract were found to remarkably increase the mitochondrial transfer rate between human fibroblasts with high reproducibility. Furthermore, as shown in Table 2, it was found that the turmeric rhizome extract and the melia azadiracta leaf extract showed excellent effects particularly on the skin fibroblasts on the side of supplying mitochondria. Dipotassium glycyrrhizinate also showed a similar effect. On the other hand, it was found that yellowfin bark extract has an excellent effect on dermal fibroblasts on the side receiving mitochondria. Berberine sulfate also showed similar effects. From this result, it can be expected that the mitochondrial transfer ability can be more effectively enhanced by using a combination of extracts having a high tendency to act on the supply side and extracts having a high tendency to act on the receiving side.

Example 2 NHEK (human epidermal keratinocytes) x NHEK (human epidermal keratinocytes)
The effect of each extract on mitochondrial transfer between NHEK (human epidermal keratinocytes) was examined by the following method.

(Evaluation test of mitochondrial transfer ability)
Cryopreserved NHEK (human epidermal keratinocytes) was asleep in a T75 flask and cultured for 4 days. After washing three times with HBSS (−), the mitochondrial staining reagent was added to the cells on the mitochondrial supply side for 30 minutes, the cytoplasmic staining reagent was added to the cells on the receiving side and stained in a 37 ° C. incubator, Replaced with Humdia KG2 medium. The next day, after washing three times with HBSS (−), the cells were detached with trypsin / EDTA, seeded on a 96-well plate at 2,000 cells / well on both the supply side and the reception side, and co-cultured. At that time, the extract was added at a final concentration of 1/1000 and cultured overnight in a 37 ° C. incubator. The next day, the nuclear staining reagent was diluted in PBS (−) and replaced with the medium in the well. After staining at room temperature for 15 minutes, an image was taken with ImageXpress (16 fields / well). Each image was analyzed with a cell count program and the mitochondrial transfer rate was calculated. The cell count program is the same as described above. Table 3 shows the results of the extract having an effect of increasing the mitochondrial transfer rate compared to the control (100%) to which each extract was not added.

(Examination of pretreatment of mitochondrial donor cell or recipient cell)
Several types of extracts having a particularly high reproducibility in the mitochondrial transfer rate were selected in the above test, and further evaluation was made as to whether these extracts act on the cells on the supply side or on the reception side.
Cryopreserved NHEK (human epidermal keratinocytes) was asleep, seeded on a 12-well plate (2 × 10 ⁴ cells / well), and cultured for 5 days. After washing three times with HBSS (−), the mitochondrial staining reagent was added to the cells on the mitochondrial supply side for 30 minutes, the cytoplasmic staining reagent was added to the cells on the receiving side and stained in a 37 ° C. incubator, Replaced with Humdia KG2 medium. Next, Altea root extract and yellowfin bark extract were added at a final concentration of 1/1000 to the cells on the supply side or the reception side, and the cells were cultured overnight in a 37 ° C. incubator. A similar test was conducted for berberine sulfate, which is the main component of yellowfin bark extract, and curcumin, which is the main component of turmeric rhizome extract. The next day, after washing 3 times with HBSS (−), the cells are detached with trypsin / EDTA, seeded on a 96-well plate so that the supply side and the reception side are 2,000 cells / well, and untreated cells (2,000 cells / well). ). The cells were cultured overnight in a 37 ° C. incubator. The next day, the nuclear staining reagent was diluted in PBS (−) and replaced with the medium in the well. After staining at room temperature for 15 minutes, an image was taken with ImageXpress (16 fields / well). Analysis with a cell count program and mitochondrial transfer rates were calculated. The count program is the same as above. Table 4 shows the results of the pretreatment of the extract that had the effect of increasing the mitochondrial transfer rate compared to the control (100%) to which each extract was not added.

The cells, medium, drug, and staining reagent used in the above experiment are shown below.
Cells: NHEK (Human epidermal keratinocytes) KK-4009 (manufactured by KURABO)
Medium: Humdia-KG2 medium KK-2150S (manufactured by KURABO) During cell culture and co-culture
Drug: Trypsin / EDTA (manufactured by KURABO) HK-3120
Mitochondrial staining reagent: Mito Tracker® Green FM (molecular probes M7514) 1 / 7,000 diluted in HBSS (−) (final 70 nM)
Cytoplasmic staining reagent: CellTrace ™ Far Red Cell Proliferation Kit (Invitrogen ™ C34564) 1/1000 diluted in HBSS (−) (final 5 uM)
Nuclear staining reagent: Hoechst 33342 (manufactured by DOJINDO 346-07951) 1/1000 dilution

  The extracts shown in Table 3 were found to have an effect of increasing the mitochondrial transfer rate between human epidermal keratinocytes. Among them, it has been found that Altea root extract, yellowfin bark extract, auren root extract, red pepper fruit extract, and horse chestnut seed extract are reproducible and remarkably increased between human epidermis keratinocytes. Furthermore, as shown in Table 4, it was found that the Altea root extract and yellowfin bark extract showed an excellent effect especially on the epidermal keratinocytes on the side receiving mitochondria. Berberine sulfate and curcumin also showed similar effects.

<< Example 3 >> AD-MSC (adipose-derived mesenchymal stem cells) x NHDF (human skin fibroblasts)
The effect of each extract on mitochondrial transfer between AD-MSC (adipose-derived mesenchymal stem cells) and NHDF (human skin fibroblasts) was examined by the following method.

(Evaluation test of mitochondrial transfer ability)
Cryopreserved AD-MSC (adipose-derived mesenchymal stem cells) and NHDF (human skin fibroblasts) were put to sleep in a T75 flask and cultured for 5 days. After washing 3 times with HBSS (+), the mitochondrial staining reagent was added to the cells on the mitochondrial supply side for 30 minutes, the cytoplasmic staining reagent was added to the cells on the receiving side and stained in a 37 ° C. incubator, AD-MSC was replaced with Stem life medium, and NHDF was replaced with DMEM (10% FBS, 1% AB) medium. The next day, after washing 3 times with HBSS (+), the cells were detached with trypsin / EDTA, seeded on a 96-well plate at 2,000 cells / well on both the supply side and the reception side, and co-cultured. At that time, the extract was added at a final concentration of 1/1000 and cultured overnight in a 37 ° C. incubator. The next day, the nuclear staining reagent was diluted in PBS (−) and replaced with the medium in the well. After staining at room temperature for 15 minutes, an image was taken with ImageXpress (16 fields / well). Each image was analyzed with a cell count program and the mitochondrial transfer rate was calculated. The cell count program is the same as described above.

(Examination of pretreatment of mitochondrial donor cell or recipient cell)
Several types of extracts having a particularly high reproducibility in the mitochondrial transfer rate were selected in the above test, and further evaluation was made as to whether these extracts act on the cells on the supply side or on the reception side.
Cryopreserved AD-MSC (adipose-derived mesenchymal stem cells) is seeded on a 12-well plate (3.6 × 10 ⁴ cells / well), NHDF (human skin fibroblasts) is asleep, and seeded on a 12-well plate (8. 5 × 10 ⁴ cells / well) and cultured for 4 days. After washing 3 times with HBSS (+), the mitochondrial staining reagent was added to the cells on the mitochondrial supply side for 30 minutes, the cytoplasmic staining reagent was added to the cells on the receiving side and stained in a 37 ° C. incubator, AD-MSC was replaced with Stem life medium, and NHDF was replaced with DMEM (10% FBS, 1% AB) medium. Subsequently, yellowfin bark extract and prune degradation product were added to the cells to be the supply side or the reception side at a final concentration of 1/1000, and cultured overnight in a 37 ° C. incubator. The same test was conducted for curcumin, which is the main component of turmeric rhizome extract. The next day, after washing three times with HBSS (+), the cells are detached with trypsin / EDTA, seeded on a 96-well plate so that both the supply side and the reception side become 2,000 cells / well, and untreated cells (2,000 cells / well) ). The cells were cultured overnight in a 37 ° C. incubator. The next day, the nuclear staining reagent was diluted in PBS (−) and replaced with the medium in the well. After staining at room temperature for 15 minutes, an image was taken with ImageXpress (16 fields / well). Analysis with a cell count program and mitochondrial transfer rates were calculated. The count program is the same as above.

The cells, medium, drug, and staining reagent used in the above experiment are shown below.
Cell: AD-MSC (Adipose-derived mesenchymal stem cell) Adipose derived stem cell (Life Line KW-4109)
NHDF human dermal fibroblasts: Human Dermal Fibroblast (KFRABO KF-4009)
Culture medium:
(1) Stem life MSC Comp Kit (Life Line LL-0034) (AD culture)
(2) DMEM (Gibco 11995065) + 10% FBS + 1% AB ^* (when NHDF cells are cultured or co-cultured)
^* AB: Antibiotic-Antimycotic (Gibco)
Drug: Trypsin / EDTA (manufactured by KURABO) HK-3120
Mitochondrial staining reagent: Mito Tracker® Green FM (molecular probes M7514) 1 / 7,000 diluted in HBSS (+) (final 70 nM)
Cytoplasmic staining reagent: CellTrace ™ Far Red Cell Proliferation Kit (Invitrogen ™ C34564) 1/1000 diluted in HBSS (+) (final 5 uM)
Nuclear staining reagent: Hoechst 33342 (manufactured by DOJINDO 346-07951) 1/1000 dilution

  The extracts shown in Table 5 were found to have the effect of increasing the mitochondrial transfer rate between fat-derived mesenchymal stem cells and dermal fibroblasts. Among them, turmeric rhizome extract, yellowfin bark extract, auren root extract, eucalyptus flower extract, and prunes degradation products have a reproducible and reproducible mitochondrial transfer rate between adipose-derived mesenchymal stem cells and human skin fibroblasts. I found it to rise. Furthermore, as shown in Table 6, it was found that the prune degradation product showed an excellent effect particularly on the adipose-derived mesenchymal stem cells on the side of supplying mitochondria. On the other hand, it was found that yellowfin bark extract has an excellent effect on dermal fibroblasts on the side receiving mitochondria. Curcumin also showed a similar effect. Therefore, it can be expected that the mitochondrial transfer ability can be enhanced more effectively by using a combination of extracts having a high tendency to act on the supply side and extracts having a high tendency to act on the receiving side.

Example 4 NHDF (human skin fibroblasts) x NHEK (human epidermal keratinocytes)
The effect of each extract on mitochondrial transfer between NHDF (human skin fibroblasts) and NHEK (human epidermal keratinocytes) was examined by the following method.

(Evaluation test of mitochondrial transfer ability)
Cryopreserved NHDF (human skin fibroblasts) and NHEK (human epidermal keratinocytes) were asleep in a T75 flask and cultured for 5 days. NHDF was washed with HBSS (+), NHEK was washed with HBSS (−) three times, mitochondrial staining reagent was added to cells on the mitochondrial supply side, and cytoplasmic staining reagent was added to cells on the receiving side for 30 minutes. After staining in a 37 ° C. incubator for a time, NHDF was replaced with DMEM (10% FBS, 1% AB) medium, and NHEK was replaced with Humedia KG2 medium. The next day, NHDF was washed with HBSS (+), NHEK was washed three times with HBSS (−), the cells were detached with trypsin / EDTA, and seeded on a 96-well plate so that the supply side and the receiving side were 2,000 cells / well. Co-cultured. At that time, the extract was added at a final concentration of 1/1000 and cultured overnight in a 37 ° C. incubator. The next day, the nuclear staining reagent was diluted in PBS (−) and replaced with the medium in the well. After staining at room temperature for 15 minutes, an image was taken with ImageXpress (16 fields / well). Each image was analyzed with a cell count program and the mitochondrial transfer rate was calculated. The cell count program is the same as described above.

(Examination of pretreatment of mitochondrial donor cell or recipient cell)
Several types of extracts having a particularly high reproducibility in the mitochondrial transfer rate were selected in the above test, and further evaluation was made as to whether these extracts act on the cells on the supply side or on the reception side.
Cryopreservation NHDF were seeded at seeding (human dermal fibroblasts) in 6well plates ^(5x10 4 cells / well) into T25 flasks 1.5x10 ⁵ cells / well), separately NHEK (human epidermal keratinocytes) After falling asleep, seeded on a 6-well plate ( ⁴ × 10 ⁴ cells / well), seeded in a T25 flask at 2 × 10 ⁵ cells / well), and cultured for 4 days. NHDF was washed with HBSS (+), NHEK was washed with HBSS (−) three times, mitochondrial staining reagent was added to cells on the mitochondrial supply side, and cytoplasmic staining reagent was added to cells on the receiving side for 30 minutes. After staining in a 37 ° C. incubator for a time, NHDF was replaced with DMEM (10% FBS, 1% AB) medium, and NHEK was replaced with Humedia KG2 medium. Next, yellowfin bark extract was added at a final concentration of 1/1000 to cells serving as a supply side or a reception side, and cultured overnight in a 37 ° C. incubator. The same test was conducted for curcumin, which is the main component of turmeric rhizome extract. The next day, NHDF was washed with HBSS (+), NHEK was washed three times with HBSS (−), the cells were detached with trypsin / EDTA, and seeded on a 96-well plate so that the supply side and the receiving side were 2,000 cells / well. Co-cultured with untreated cells (2,000 cells / well). The cells were cultured overnight in a 37 ° C. incubator. The next day, the nuclear staining reagent was diluted in PBS (−) and replaced with the medium in the well. After staining at room temperature for 15 minutes, an image was taken with ImageXpress (16 fields / well). Analysis with a cell count program and mitochondrial transfer rates were calculated. The count program is the same as above.

The cells, medium, drug, and staining reagent used in the above experiment are shown below.
Cells: NHDF human dermal fibroblasts: Human Dermal Fibroblast (KURABO KF-4009)
NHEK (human epidermal keratinocytes) KK-4209 (manufactured by KURABO)
Culture medium:
(1) Humdia-KG2 medium KK-2150S (manufactured by KURABO) (when NHEK cells are cultured)
(2) DMEM (Gibco 11995065) + 10% FBS + 1% AB ^* (when NHDF cells are cultured)
^* AB: Antibiotic-Antimycotic (Gibco)
(3) Medium in which the above two types of medium are mixed at 1: 1 (when co-cultured)
Drug: Trypsin / EDTA (manufactured by KURABO) HK-3120
Mitochondrial staining reagent: Mito Tracker® Green FM (molecular probes M7514) 1 / 7,000 diluted in HBSS (+) (final 70 nM)
Cytoplasmic staining reagent: CellTrace ™ Far Red Cell Proliferation Kit (Invitrogen ™ C34564) 1/1000 diluted in HBSS (−) (final 5 uM)
Nuclear staining reagent: Hoechst 33342 (manufactured by DOJINDO 346-07951) 1/1000 dilution

  The extracts shown in Table 7 were found to have a high mitochondrial transfer promoting effect between dermal fibroblasts and epidermal keratinocytes. Among them, yellowfin bark extract and auren root extract were found to remarkably increase the mitochondrial transfer rate between human skin fibroblasts and human epidermal keratinocytes with high reproducibility. Furthermore, as shown in Table 8, it was found that yellowfin bark extract has an excellent effect on keratinocytes on the side that receives mitochondria. Curcumin also showed a similar effect.

<Examination of the effect of mitochondrial transfer on cells>
Changes in the expression of genes (SOD2 and PPARGC1) were examined for the purpose of confirming what kind of functional change occurred in the cells that received mitochondrial transfer (receiving cells).

  The mitochondrial supply side is AD-MSC (human adipose-derived mesenchymal stem cell), the reception side is NHDF (human skin fibroblast), and the cells stored at −80 ° C. are sluken into the flask at the recommended seeding density. The cells were cultured in a 37 ° C., 5% CO 2 incubator for a day. On the 4th day, only NHDF was washed twice with PBS (−), cytoplasmic staining reagent was added, and staining was performed in an incubator for 1 hour. After staining, the plate was washed twice with PBS (−), replaced with a normal medium, and simultaneously added with rotenone / DMSO (final 1 uM) and cultured overnight in an incubator. At this time, NHDF stained with cytoplasm without adding rotenone was also prepared as a control comparative sample. On the 5th day, both AD-MSC and NHDF (with and without rotenone) were washed twice with PBS (−), trypsin was added to detach the cells from the culture flask, and NHDF was added 1: 1 to AD-MSC. Mixed at a ratio and co-cultured. At this time, single culture NHDF (with or without rotenone) was also prepared as a comparative sample with co-culture. On day 6, cells were detached in the same manner as described above, washed twice with 1% BSA / PBS (−), suspended in 1% BSA / PBS, and passed through a cell strainer. Thereafter, using a cell sorter (SH800Z manufactured by SONY), the receiving NHDFs were sorted using the fluorescence of the cytoplasm staining reagent as an index. Sorted cells were collected with a 15 mL falcon tube. After sorting, a total of 4 types of single culture NHDF (with and without rotenone) and co-culture NHDF (with and without rotenone) were washed twice with PBS (-), RNA was extracted using an RNA extraction kit, and cDNA CDNA was synthesized using a synthesis kit. Using this as a sample, the expression of SOD2 and PPARGC1 was confirmed by real-time PCR. GAPDH was used as an endogenous control for gene expression. The results are shown in FIG. 1 (SOD2) and FIG. 2 (PPARGC1).

  The cells, media, drugs, and equipment used in the above experiment are as follows.

cell:
AD-MSC (adipose-derived mesenchymal stem cell HMSC-Ad): Adipose derived stem cell (Life Line FC-0034);
NHDF human dermal fibroblasts: Human Dermal Fibroblast (KFRABO KF-4009)

Culture medium:
During AD culture: Stem life MSC Comp Kit (Life Line LL-0034)
During NHDF culture and co-culture: DMEM (Gibco) + 10% FBS + 1% AB *
* AB: Antibiotic-Antimycotic (Gibco)

reagent:
Cytoplasmic staining reagent: CellTrace Violet Cell Proliferation Kit, for flow cytometry (Invitrogen) C34557 1/1000 diluted in HBSS (+) (final 5 uM)
Rotenon: (SIGMA) R8875-1G
Trypsin / EDTA: (manufactured by KURABO) HK-3120
BSA: (made by SIGMA) Bovine Serum Albumin A2153-50G
RNA extraction kit: (QIAGEN) RNeasy Mini Kit (250) 74106
cDNA synthesis kit: (manufactured by TOYOBO) RiverTra Ace (registered trademark) qPCR Master Mix with gDNA Remover FSQ-301
Real-time PCR reagent: (Applied biosystems) TaqMan (registered trademark) Fast Advanced Master Mix 4444557
TaqMan probe: (Applied biosystems) SOD2, PPARGC1, GAPDH

machine:
CELL SORTER: (made by SONY) SH800Z
Thermal Cycler: (Applied Biosystems) 2720
Real-time PCR System: Quant Studio (Applied biosystems)

  SOD2 is an antioxidant enzyme localized in mitochondria and functions in mitochondria, and thus may contribute to recovery of mitochondrial dysfunction. As a result of the gene expression this time, as shown in FIG. 1, the SOD2 gene expression was higher in the co-culture group than in the single culture group. In addition, in the damaged cells treated with rotenone, the expression of SOD2 was significantly increased by co-culture. From the above, it was confirmed that mitochondrial transfer occurs in damaged cells, leading to functional recovery. In addition, as shown in FIG. 2, PPARGC1, which is an index for increasing the amount of mitochondria, showed the same tendency as SOD2. That is, since the expression of PPARGC1 increased, it was confirmed that cell activation was caused by mitochondrial transfer.

<< Examination of the effects of prune degradation products and aloe extract on collagen production ability of human fibroblasts >>
As a result of the above test, it was found that the prune degradation product and the aloe extract have an effect of promoting mitochondrial transfer. In order to examine the effects of these substances on collagen production, the following tests were conducted. That is, cryopreserved human fibroblasts (manufactured by Kurabo Industries Inc.) were put to sleep in a recommended seeding density cell number flask and cultured in a 37 ° C., 5% CO ₂ incubator. On the 4th day of culture, the cells were washed twice with PBS (−), trypsin was added to detach the cells from the culture flask, and DMEM / 10% FBS / 1% AB medium (Gibco) was used on a 48-well plate. And 3 × 10 ⁴ cells / well. After 6 hours, the medium was replaced with DMEM / 0.1% FBS / 1% AB. The next day, prune degradation product (final 0.002%), aloe extract (final 0.002%), complex A (tripeptide-1-copper final 0.1 mg / ml, collagen final 1 mg / ml, caprooil tetrapeptide -3 (final 0.075 mg / ml) and ascorbic acid (final 1 mg / ml) were added to the cells, and the cell supernatant was recovered after 52 hours.
Protein amount measurement (collagen amount measurement) was measured according to the protocol using R & D systems Human Pro-Collagen I alpha 1 Duo Set ELISA by ELISA method. As the sample, the recovered supernatant was used at 150-fold dilution. The results are shown in FIG.

The cells, media, and reagents used in this study are as follows.
Cell: NHDF human dermal fibroblast: Human Dermal Fibroblast (KURABO KF-4009)
Medium: DMEM (Gibco) + 10% FBS + 1% AB *
* AB: Antibiotic-Antimycotic (Gibco)
Reagent: Trypsin / EDTA (manufactured by KURABO) HK-3120
Commercially available cosmetic raw materials were used for the prune degradation products and aloe extract.

  As shown in FIG. 1, complex A (contains tripeptide-1-copper, collagen, caprooil tetrapeptide-3, which are known to have an effect of imparting migration of adipose-derived stem cells and a function of producing collagen. ) Collagen production promoting effect in human fibroblasts was observed by itself, but when the prune degradation product and aloe extract having the mitochondrial transfer promoting effect were added, the effect was remarkably enhanced.

  In the same manner as above, dipropylene glycol, 1,2-butylene glycol, 1,3-butylene glycol, propylene glycol, 1,3-propanediol, 1,2-pentanediol, 1,2-hexanediol, ethylene Polyol compounds having 2 or more hydroxyl groups in C3-6 alkyl groups such as glycol and diethylene glycol, monosaccharides, disaccharides, oligosaccharides, polysaccharides, mucopolysaccharides, collagens, oligopeptides, polypeptides, etc. The material of the present invention, which is a substance having an effect of imparting migration of fat-derived stem cells (an effect of causing migration or promoting migration of fat-derived stem cells), such as a hydrophilic biological material of the present invention, or a material having a known collagen production function Combined with materials having mitochondrial transfer function, collagen in cells It is possible to increase the raw amount.

  Formulation examples (formulation examples) of the cosmetic composition of the present invention are shown below. The following are all compositions for external use on the skin and can be prepared by conventional methods. The present invention is not limited to these pharmaceutical formulation examples.

  The mitochondrial transfer promoter and cosmetic composition of the present invention prevent symptoms and conditions caused by including cells in which mitochondrial dysfunction or deterioration occurs due to aging, environmental factors, stress, etc. -It can be suitably used for rejuvenation such as improvement, maintenance of health condition, and improvement of skin firmness.

Claims (10)

  A cosmetic composition containing a mitochondrial transfer promoter.   Mitochondrial transfer promoters include yellowfin extract, auren extract, prune extract, turmeric extract, Melia aziracta extract, Hamelis extract, capsicum extract, altea extract, red pepper extract, horse chestnut extract, ginger extract, centifolia rose extract, bucurium extract , Eucalyptus extract, yuzu extract, barley extract, lavender extract, apple extract, lemon extract, gourd extract, aloe extract, cuckoo extract, saicin extract, ginger extract, perilla extract, cherry extract, chamomile extract, asenyaku extract, arnica extract, clara extract, Butcher bloom extract, loofah extract, cornflower extract, rooibos extract, peach extract, ashitaba extract, nettle extract, Oyster extract, mallow extract, onionis extract, olive extract, apricot extract, Satsuma mandarin extract, Japanese dandelion extract, plum extract, green tea extract, green algae extract, red algae extract, brown algae extract, shiitake extract, fuyumina tsutake mushroom extract, litchi extract, water-soluble The cosmetic composition according to claim 1, comprising at least one water-soluble active ingredient (A) selected from the group consisting of soluble collagen and royal jelly extract.   Water-soluble active ingredient (A) is yellowfin bark extract, auren root extract, prune degradation product, turmeric rhizome extract, Melia aziracta leaf extract, Hamelis leaf extract, capsicum fruit extract, Altea root extract, Tokinsenka flower extract, Atlantic Tochinoki seed extract, ginger rhizome extract, centifolia rose flower extract, budweed extract, eucalyptus leaf extract, yuzu fruit extract, pearl seed extract, lavender flower extract, apple fruit extract, lemon fruit extract, argon extract, aloe extract, cuckoo extract, Saisin extract, Sakura leaf extract, Chamomile flower extract, Acacia catechu extract, Arnica flower extract, Clara root extract, Butcher bloom root extract, Loofah extract, Cornflower flower extract, Rooibos extract, Peach leaf extract, Ashitaba leaf / stem Kiss, nettle leaf extract, fennel fruit extract, mallow flower extract, onionis extract, olive leaf extract, apricot seed extract, ginger root extract, perilla leaf extract, citrus peel extract, dandelion flower extract, ume fruit extract, amacha eel extract, chlorella The cosmetic composition according to claim 2, wherein the cosmetic composition is at least one component selected from the group consisting of an extract, a hinoki extract, a seaweed extract, a shiitake extract, a cypress extract, a lychee pattern extract, and a water-soluble collagen.   Water-soluble active ingredient (A) is yellowfin bark extract, auren root extract, prune degradation product, turmeric rhizome extract, Melia aziracta leaf extract, Hamelis leaf extract, capsicum fruit extract, Altea root extract, Tokinsenka flower extract, and The cosmetic composition according to claim 3, which is at least one plant extract selected from the group consisting of horse chestnut seed extract.   The cosmetic composition according to any one of claims 1 to 4, which is used for promoting mitochondrial transfer.   The cosmetic composition according to any one of claims 1 to 5, which is used to promote mitochondrial transfer in skin tissue.   It is used to promote mitochondrial transfer between one or two cells selected from the group consisting of dermal fibroblasts, epidermal keratinocytes, adipose-derived mesenchymal stem cells, epidermal stem cells and dermal stem cells. The cosmetic composition according to any one of 1 to 6.   The cosmetic composition according to any one of claims 1 to 7, which is used as an external composition for skin.   The cosmetic composition according to any one of claims 1 to 7, which is used as an oral composition.   Yellowfin extract, Auren extract, Prune extract, Turmeric extract, Melia azadilacta extract, Hamelis extract, Pepper extract, Altea extract, Toki-senka extract, Horse chestnut extract, Ginger extract, Centifolia rose extract, Buddle extract, Eucalyptus extract, Yuzu extract, Pearl barley extract, lavender extract, apple extract, lemon extract, argon extract, aloe extract, cuckoo extract, saicin extract, dianthus extract, perilla extract, cherry extract, chamomile extract, asenya extract, arnica extract, clara extract, butcher bloom extract, loofah extract , Cornflower extract, rooibos extract, peach extract, ashitaba extract, nettle extract, fennel extract, mallow extract, o From varnish extract, olive extract, apricot extract, Citrus unshiu extract, dandelion extract, ume extract, green algae extract, green algae extract, red algae extract, brown algae extract, shiitake extract, agaric extract, litchi extract, water-soluble collagen, and royal jelly extract A mitochondrial transfer promoter for a cosmetic composition comprising at least one water-soluble active ingredient (A) selected from the group consisting of