JP2008239549A - Extracellular matrix production improver - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide an extracellular matrix production improver, having an excellent extracellular matrix production promotion effect on cells, especially collagen or hyaluronic acid production promotion effect and satisfactorily standing a long-term use, especially an agent for improving collagen production and an agent for improving hyaluronic acid production. <P>SOLUTION: The extracellular matrix production improver comprises a polyamine as an active ingredient. A method for using the extracellular matrix production improver as a cosmetic, a quasi-medicine (skin care preparation, bath medicine, hair grower etc.), a food and beverage or a medicine comprising it as an active ingredient is provided. <P>COPYRIGHT: (C)2009,JPO&INPIT

Description

  The present invention relates to substances and methods for improving extracellular matrix production of various cells.

  Various diseases are deeply related to a decrease in the division rate and cell function of all dividing cells. For example, the skin is dermis and the epidermis is composed of epidermal cells, fibroblasts, and extracellular matrices such as elastin, collagen, and hyaluronic acid that support these extracellular skin structures. In young skin, skin properties such as flexibility are ensured by maintaining the homeostasis of these skin tissue interactions, and the skin is glossy, tightened, transparent, and maintained moist. . However, ultraviolet rays, dryness, stress, etc. cause a decrease in the function of the extracellular matrix and fibroblasts in particular, and as a result, the skin properties such as skin flexibility decline, and the skin loses its gloss, firmness, and transparency, Symptoms such as roughening, wrinkles and dullness occur.

  Examples of substances having an extracellular matrix production promoting action include substances that promote collagen production, such as licorice leaf extract (Patent Document 1), lotus germ extract (Patent Document 2), hydrolyzed potato protein ( Patent document 3), persimmon leaf extract or hawthorn fruit extract (patent document 4), substances having an action of promoting hyaluronic acid production include rice bran solvent extract (patent document 5), Iridae crocus saffron Extracts (Patent Document 6), cucurbitaceae plant seed extracts (Patent Document 7), and the like are known, and some of these are used as quasi-drugs and cosmetics as collagen production promoters or hyaluronic acid production promoters. However, it cannot be said that the effect is sufficient, and an extracellular matrix production promoter that exhibits a satisfactory effect has not been obtained.

Polyamine is a generic term for aliphatic hydrocarbons having two or more primary amino groups, and is a natural product that exists universally in the living body, and more than 20 types of polyamines have been found. Typical polyamines include putrescine, spermidine and spermine. The main physiological functions of polyamines are as follows: (1) Stabilization and structural changes of nucleic acids by interaction with nucleic acids (2) Promotion of various nucleic acid synthesis systems (3) Activation of protein synthesis systems (4) Cell membrane Stabilization and enhancement of membrane permeability of substances (5) Elimination of active oxygen (6) Promotion of cell growth is known, but it promotes extracellular matrix production and collagen or hyaluronic acid production in cells Was not known at all.
JP 2000-191498 A JP 2002-29980 A Japanese Patent Laid-Open No. 2005-263689 JP 2006-160629 A JP 2005-272433 A JP-A-2005-206510 JP 2006-273815 A

  An object of the present invention is to provide an extracellular matrix production improver that has an excellent extracellular matrix production improving effect on cells and has safety sufficient to withstand long-term use. It is to provide a method for use in quasi-drugs (external preparations for skin, bath preparations, hair restorers, etc.), foods and drinks, and pharmaceuticals.

  As a result of diligent efforts to achieve the above object, the present inventors have found that polyamine is effective as an extracellular matrix production improver for cells, and have completed the present invention. That is, the present invention has the following configuration.

1. An agent for improving extracellular matrix production, comprising polyamine as an active ingredient.
2. An extracellular matrix production improver, wherein the extracellular matrix production improver is a collagen production improver and / or a hyaluronic acid production improver.
3. 1. The extracellular matrix production improver according to 1 or 2, wherein the polyamine is at least one compound selected from the group consisting of aliphatic hydrocarbons having two or more primary amino groups.
4). Polyamine is 1,3-diaminopropane, putrescine, cadaverine, cardine, spermidine, homospermidine, aminopropyl cadaverine, theremin, spermine, thermospermine, canabalmin, aminopentylnorspermidine, N, N-bis (aminopropyl) cadaverine, The extracellular matrix production according to any one of 1 to 3 above, which is at least one compound selected from the group consisting of homospermine, cardopentamine, homocardopentamine, cardohexamine and homocardohexamine. Improver.
5. The extracellular matrix production improver according to any one of 1 to 4 above, wherein the polyamine is at least one compound selected from the group consisting of putrescine, spermidine and spermine.
6). Cosmetics containing the extracellular matrix production enhancer according to any one of 1 to 5 as an active ingredient.
7). A quasi-drug containing the extracellular matrix production enhancer according to any one of 1 to 5 as an active ingredient.
8). Food / beverage products containing the extracellular matrix production improver in any one of said 1-5 as an active ingredient.
9. A pharmaceutical comprising the extracellular matrix production enhancer according to any one of 1 to 5 as an active ingredient.
10. A method for improving extracellular matrix production, comprising a step of bringing a polyamine into contact with an animal.
11. A method for improving collagen production, comprising a step of bringing a polyamine into contact with an animal.
12 11. The method for improving collagen production as described in 11 above, wherein the polyamine is at least one compound selected from the group consisting of aliphatic hydrocarbons having two or more primary amino groups.
13. Polyamine is 1,3-diaminopropane, putrescine, cadaverine, cardine, spermidine, homospermidine, aminopropyl cadaverine, theremin, spermine, thermospermine, canabalmin, aminopentylnorspermidine, N, N-bis (aminopropyl) cadaverine, 11. The method for improving collagen production according to 11 or 12 above, which is at least one compound selected from the group consisting of homospermine, cardopentamine, homocardopentamine, cardohexamine and homocardohexamine.
14 The method for improving collagen production according to any one of 11 to 13, wherein the polyamine is at least one compound selected from the group consisting of putrescine, spermidine and spermine.
15. A method for improving hyaluronic acid production, comprising a step of bringing a polyamine into contact with an animal.
16. 15. The method for improving hyaluronic acid production as described in 15 above, wherein the polyamine is at least one compound selected from the group consisting of aliphatic hydrocarbons having two or more primary amino groups.
17. Polyamine is 1,3-diaminopropane, putrescine, cadaverine, cardine, spermidine, homospermidine, aminopropyl cadaverine, theremin, spermine, thermospermine, canabalmin, aminopentylnorspermidine, N, N-bis (aminopropyl) cadaverine, 15. The method for improving hyaluronic acid production as described in 15 or 16 above, which is at least one compound selected from the group consisting of homospermine, cardopentamine, homocardopentamine, cardohexamine and homocardohexamine.
18. The method for improving hyaluronic acid production according to any one of 15 to 17 above, wherein the polyamine is at least one compound selected from the group consisting of putrescine, spermidine and spermine.

  According to the present invention, it is expected to have an excellent extracellular matrix production improving action with safety, and provides cosmetics and quasi-drugs (skin external preparations, bath preparations, hair growth agents, etc.), foods and drinks, and pharmaceuticals containing polyamine as an active ingredient. can do.

In the present invention, “enhancement of extracellular matrix production” means to improve the production of extracellular matrix such as elastin, collagen, hyaluronic acid, etc. that supports the extracellular skin structure. A substance that improves extracellular matrix production in this way is called an “extracellular matrix production enhancer”. "Beautiful skin" is realized by improving extracellular matrix production.
“Extracellular matrix production improvers” include “elastin production improvers”, “collagen production improvers” and “hyaluronic acid production improvers”. In particular, the “collagen production improver” refers to a substance that improves the production of collagen among the “extracellular matrix production improvers”. The “hyaluronic acid production enhancer” refers to a substance that improves the production of hyaluronic acid among the “extracellular matrix production enhancers”.

  In the present invention, “activation” refers to reducing the decrease in cell function or cell activity by actively increasing or maintaining the cell function or cell activity of the animal. For example, in skin cells, by reducing the decrease in skin cell function associated with the accumulation of structural changes in the sublamellar due to photoaging, skin wrinkles, sagging, hardening, dullness, etc. can be prevented and improved to give gloss and transparency. , Refers to maintaining a smooth, youthful and healthy skin condition. In dermal papilla or hair matrix cells, it refers to suppressing hair loss by maintaining the hair cycle by reducing the decrease in function of hair milk or hair matrix cells due to stress and hormone balance.

  In the present invention, “polyamine” is a general term for aliphatic hydrocarbons having two or more primary amino groups, and is a natural product that exists universally in the living body, and more than 20 types of polyamines have been found. For example, 1,3-diaminopropane, putrescine, cadaverine, cardine, spermidine, homospermidine, aminopropyl cadaverine, theremin, spermine, thermospermine, canabalmin, aminopentylnorspermidine, N, N-bis (aminopropyl) cadaverine, homo Examples include spermine, cardopentamine, homocardopentamine, cardohexamine, and homocardohexamine. Typical polyamines include putrescine, spermidine and spermine.

  In the present invention, “putrescine” is one of typical polyamines and is a general natural product that exists universally in living organisms, and is an aliphatic hydrocarbon compound having two primary amino groups. . “Cadaverine” is one of the typical polyamines and is a general natural product that exists universally in living organisms, and is an aliphatic hydrocarbon compound having two primary amino groups. “Spermidine” is one of the typical polyamines and is a general natural product ubiquitously present in living organisms, and is an aliphatic hydrocarbon compound having three primary amino groups. “Spermine” is one of the typical polyamines and is a general natural product universally present in living organisms, and is an aliphatic hydrocarbon compound having four primary amino groups.

  The term “contact with animals” as used in the present invention refers to all dosage forms (ampoules, capsules, powders, granules, pills, tablets, solids, liquids) of polyamines for humans and livestock. , Gel, foam, emulsion, cream, ointment, sheet, mousse, etc.) for external application, external treatment, oral administration, and the like. For example, polyamines can be blended in cosmetics, quasi drugs, foods and drinks, pharmaceuticals, feeds, etc. and brought into contact with animals.

  The main physiological functions of polyamines are: (1) stabilization and structural changes of nucleic acids by interaction with nucleic acids, (2) promotion of various nucleic acid synthesis systems, (3) activation of protein synthesis systems, ( 4) Stabilization of cell membranes, enhancement of membrane permeability of substances, (5) elimination of active oxygen, etc. are known.

  Polyamines can be obtained from fish larvae (Japanese Patent No. 3518778), animal sperm, milk material (Japanese Patent Laid-Open No. 2001-8663), yeast (Japanese Patent Laid-Open No. 2000-245493), and the like. It can also be obtained from chemical synthesis (JP-A-7-277917), chemical synthesis by enzymatic reaction or microbial metabolism (Metabolism, Vol. 9, No. 11, 1010-1017, 1972), commercial products, etc.

  The polyamine may be used as an extracellular matrix production improver as it is, but it is preferably used by blending it into cosmetics / quasi-drugs (external preparations for skin, bath preparations, hair growth agents, etc.), foods and pharmaceuticals. The concentration at which the polyamine is blended (M: mol / liter) is determined by the degree of absorption, the degree of action, the product form, the frequency of use, etc., and is not particularly limited, but is usually 0.00001 to 100 mM, preferably It is 0.00005-75 mM, More preferably, it is 0.0001-50 mM.

  The extracellular matrix production improver of the present invention is used for cosmetics, quasi-drugs, foods and drinks, pharmaceuticals, etc. within the range that does not impair the effects of the present invention as required in addition to the essential components of polyamine. Components and additives can be used in combination.

  For example, as fats and oils, avocado oil, almond oil, fennel oil, sesame oil, olive oil, orange oil, orange rafa oil, sesame oil, cocoa butter, chamomile oil, carrot oil, cucumber oil, beef fat, beef tallow fatty acid, cucumber nut oil, Safflower oil, soybean oil, camellia oil, corn oil, rapeseed oil, persic oil, castor oil, cottonseed oil, peanut oil, turtle oil, mink oil, egg yolk oil, cacao butter, palm oil, palm kernel oil, molasses, palm oil Beef tallow, pork tallow, hydrogenated oil, hydrogenated castor oil, and the like.

  Examples of waxes include beeswax, carnauba wax, whale wax, lanolin, liquid lanolin, reduced lanolin, hard lanolin, candelilla wax, montan wax, shellac wax, and the like.

  Examples of the mineral oil include liquid paraffin, petrolatum, paraffin, ozokelide, ceresin, microcristan wax, polyethylene powder, squalene, squalane and pristane.

  Examples of fatty acids include lauric acid, myristic acid, palmitic acid, stearic acid, behenic acid, oleic acid, 12-hydroxystearic acid, undecylenic acid, tall oil, lanolin fatty acid and other natural fatty acids, isononanoic acid, caproic acid, 2- Synthetic fatty acids such as ethyl butanoic acid, isopentanoic acid, 2-methylpentanoic acid, 2-ethylhexanoic acid and isopentanoic acid are exemplified.

  Alcohols include natural alcohols such as ethanol, isopyropanol, lauryl alcohol, cetanol, stearyl alcohol, oleyl alcohol, lanolin alcohol, cholesterol, phytosterol, synthetic alcohols such as 2-hexyldecanol, isostearyl alcohol, 2-octyldodecanol, oxidation Ethylene, ethylene glycol, diethylene glycol, triethylene glycol, ethylene glycol monoethyl ether, ethylene glycol monobutyl ether, diethylene glycol monomethyl ether, diethylene glycol monoethyl ether, polyethylene glycol, propylene oxide, propylene glycol, polypropylene glycol, 1,3-butylene glycol, Glycerin, batyl Alcohol, pentaerythritol, sorbitol, mannitol, glucose, polyhydric alcohols such as sucrose and the like.

  Esters include isopropyl myristate, isopropyl palmitate, butyl stearate, hexyl laurate, myristyl myristate, oleyl oleate, decyl oleate, octyldodecyl myristate, hexyl decyl dimethyloctanoate, cetyl lactate, myristyl lactate, Examples include diethyl phthalate, dibutyl phthalate, lanolin acetate, ethylene glycol monostearate, propylene glycol monostearate, propylene glycol dioleate, and the like.

  Examples of the metal soap include aluminum stearate, magnesium stearate, zinc stearate, calcium stearate, zinc palmitate, magnesium myristate, zinc laurate, and zinc undecylate.

  Gum and water-soluble polymer compounds include gum arabic, benzoin gum, danmar gum, guaiac oil, Irish moss, Karaya gum, tragacanth gum, carob gum, quinseed, agar, casein, dextrin, gelatin, pectin, starch, carrageenan, carboxyalkyl Chitin, chitosan, hydroxyalkylchitin, low molecular chitosan, chitosan salt, sulfated chitin, phosphorylated chitin, alginic acid and its salt, hyaluronic acid and its salt, chondroitin sulfate, heparin, ethylcellulose, methylcellulose, carboxymethylcellulose, carboxyethylcellulose, carboxy Sodium ethylcellulose, hydroxyethylcellulose, hydroxypropylcellulose, nitrocellulose, crystalline cellulose, polyvinyl chloride Alcohol, polyvinyl methyl ether, polyvinyl pyrrolidone, polyvinyl methacrylate, polyacrylic acid salts, polyalkylene oxide or crosslinked polymer thereof such as polyethylene oxide or polypropylene oxide, carboxyvinyl polymers, such as polyethyleneimine.

  Surfactants include anionic surfactant (carboxylate, sulfonate, sulfate ester salt, phosphate ester salt), cationic surfactant (amine salt, quaternary ammonium salt), amphoteric surfactant (carboxylic acid) Type amphoteric surfactant, sulfate ester type amphoteric surfactant, sulfonic acid type amphoteric surfactant, phosphate ester type amphoteric surfactant), nonionic surfactant (ether type nonionic surfactant, ether ester type non-type) Ionic surfactants, ester-type nonionic surfactants, block polymer-type nonionic surfactants, nitrogen-containing nonionic surfactants), other surfactants (natural surfactants, derivatives of protein hydrolysates, Examples thereof include polymer surfactants, surfactants containing titanium and silicon, and fluorocarbon surfactants.

  As vitamins, retinol, retinal (vitamin A1), dehydroretinal (vitamin A2), carotene, lycopene (provitamin A), vitamin B group, thiamine hydrochloride, thiamine sulfate (vitamin B1), Riboflavin (vitamin B2), pyridoxine (vitamin B6), cyanocobalamin (vitamin B12), folic acid, nicotinic acids, pantothenic acids, biotins, choline, inositols, vitamin C group, ascorbic acid and its derivatives, vitamin D group , Ergocalciferol (vitamin D2), cholecalciferol (vitamin D3), dihydrotaxosterol, vitamin E group, tocopherol and its derivatives, ubiquinones, vitamin K group, phytonadione (vitamin K1), menaquinone (bi Tamine K2), menadione (vitamin K3), menadiol (vitamin K4) and the like.

  As amino acids, valine, leucine, isoleucine, threonine, methionine, phenylalanine, tryptophan, lysine, glycine, alanine, asparagine, glutamine, serine, cysteine, cystine, tyrosine, proline, hydroxyproline, aspartic acid, glutamic acid, hydroxylysine, arginine Ornithine, histidine and the like, and sulfates, phosphates, nitrates, citrates, and amino acid derivatives such as pyrrolidone carboxylic acid.

  Whitening agents include ascorbic acid or derivatives thereof, sulfur, placental hydrolysate, ellagic acid or derivatives thereof, kojic acid or derivatives thereof, glucosamine or derivatives thereof, arbutin or derivatives thereof, hydroxycinnamic acid or derivatives thereof, glutathione, arnica Extract, Ogon extract, Sakuhakuhi extract, Psycho extract, Bowfu extract, Manntake mycelium culture or its extract, Linden extract, Peach leaf extract, Ages extract, Kujin extract, Gyuyu extract, Toki extract, Yokuinin extract, Oyster leaf extract, Daio extract , Button pipi extract, clam extract, horse chestnut extract, hypericum extract, oil-soluble licorice extract and the like.

  As humectants, hyaluronic acid, polyglutamic acid, serine, glycine, threonine, alanine, collagen, hydrolyzed collagen, hydronectin, fibronectin, keratin, elastin, royal jelly, chondroitin sulfate heparin, glycerophospholipid, glyceroglycolipid, sphingophospholipid , Glycosphingolipid, linoleic acid or esters thereof, eicosapentaenoic acid or esters thereof, pectin, bifidobacteria fermentation product, lactic acid fermentation product, yeast extract, litchi mycelium culture or extract thereof, wheat germ oil, avocado Oil, rice germ oil, jojoba oil, soybean phospholipid, γ-oryzanol, bellows oyster extract, yokuinin extract, siamese extract, typhoid extract, diatomaceous earth extract, beetle aloe extract, burdock extract, mannen wax extract Arnica extract, such as wheat bran, and the like.

  As hair restorer, pentadecanoic acid glyceride, coleus extract, gentian extract, pine coconut extract, royal jelly extract, kumazasa extract, t-flavanone, 6-benzylaminopurine, assembly extract, carpronium chloride, minoxidil, finasteride, adenosine, nicotinamide, Examples include mulberry root extract, diau extract and 5-aminolevulinic acid.

  As an extract or extract of animal or plant, herbal medicine, Asenyaku (Asen Yaku), Ashitaba, Acerola, Altea, Arnica, Avocado, Achacha (Amacha), Aloe, Aloe Vera, Nettle, Ginkgo (Ginkgo leaf, Ginkgo), Fennel ( Ayaka), Turmeric (Usukin), Usubasaicin (Spicy), Japanese apricot (Plum), Vulture, Uwaurushi, Neubara (Natural Fruit), Hikikoshi (Elongation Grass), Ogi (Twilight), Koganebana (Ougon), Yamazakura (Cherry Blossom) ), Yellowfin, yellow orchid, ginseng, carrot, helicopter, licorice, Dutch pepper, orange, yellow-bellied (dianthus), crabs (summer hay), tsurudukudami (what heads), Enju ), Mugwort (guy leaves), scallops (莪 朮), kudu (katsune), valerian (yoshikusa) ), Chamomile, yellow-crowned cucumber (Gurone), Chinese mugwort (licorice), licorice (licorice), Japanese dandelion (Japanese winter flower, Japanese winter leaf), raspberry, kiwi fruit, Japanese cypress (flowerflower), chrysanthemum (Chrysanthemum flower) (Fruit), citrus fruit (fruit), tachibana (tachibana bark), cucumber, udo or shishiudo (clam alive, solitary), apricot (apricot jelly), wolfberry (earthbone bark, coconut, coconut leaf), Clara (bitter) ), Camphor, cinnamon, grapefruit fruit, Nikkei (Kaikin), Kaigai (Kei-Gai), Ebisu gussa (Keiko), Maruba morning glory or morning glory (ken cow), safflower (red flower), gobishi (penta), comfrey, copaiba, gardenia (Yamako), Gentiana, Honoki (Koh-pak), Hinata Taikozuchi (cow knee), Goshuyu (Kurean), Burdock, Korean ginger (Gomi) Child), Rice, Rice bran, Wheat, Mishima Saiko (Saiko), Saffron, Savonso, Hawthorn (Yamazako), Salamander (Samurai), Salvia, Sanchinin ginseng (Thirty-seven ginseng), Shiitake mushroom (Shibai), Giou ( Ground yellow), shikunshi (messenger), purple (purple), perilla (purple leaves, shiso), oysters (pepper), peonies (glaze), psyllium (carrots, carrots), ginger (ginger), ginger (ginger)菖蒲), hornworm (Sadako), Shimotsuke, white birch, honeysuckle (gold and silver flowers, Shinobu), Ivy, Achillea millefolium, elderberry, red bean, elderberry (bone graft), mallow, senkyu (river cucumber), Assembly (this medicine), mulberry (mulberry white skin, mulberry leaf), jujube (large coral), soybean, taranki, chikutsujinjin (bamboo ginseng), Hanasuge (knowledge) ), Warmoko (land), Dokudami (10 medicines), Fuyumushinatsusatake (Cycium Cordyceps), red pepper, physalis (Torone), Tachikakuso, Ryokucha (green tea), kocha (tea), clove (clove), Eunshu mikan (Chenhide), camellia, camellia, capsicum (banban), Touki (toll), red snapper, daidai (orange peel), walnut mushroom (gem), corn (south fur), eucommia (Tochu, Tochu), tomato , Nanten (Southern fruit), garlic (large sun), barley (malt), hakusen (white birch), janohi (barley winter), parsley, batata, mint (light load), hamamelis, rose, loquat leaves (bamboo leaves) ), Matsuhodo, Grape or its leaves, Loofah, Bodaiju, Button (peony skin), Hops, Maikai (My buds), Pine leaves, Marronnier , Mannenrou, Mukuroji, Melissa, Merirot, Bokeh (Kiso), Palms, Peach (Momojin, Momoha), Giant oysters (Ishiboshi), Areca (Mellow wax), Mejijiki (Masuhagi), Cornflower, Yukinoshita (Tiger ear grass), Prunus (Plum), Yachabushi (Yakuin), pearl barley (Yokuinin), mugwort, lavender, apple fruit, garlic mushroom (ganoderma), lemon fruit, forsythia (rendoku), forsythia, genokosho (old lady), ashridokoro (roth) Root), chicken crest, cattle / human placenta extract, pig / cow stomach, duodenum, or intestinal extract or its degradation product, water-soluble collagen, water-soluble collagen derivative, collagen hydrolyzate, elastin, elastin water Degradation product, water-soluble elastin derivative, silk protein, silk protein degradation product, bovine blood Such as protein decomposition products and the like.

  Examples of microorganism culture metabolites include yeast extract, zinc-containing yeast extract, germanium-containing yeast extract, selenium-containing yeast extract, magnesium-containing yeast extract, rice fermentation extract, Euglena extract, and lactic acid fermented milk powder.

  Examples of the α-hydroxy acid include glycolic acid, citric acid, malic acid, tartaric acid, and lactic acid.

  Inorganic pigments include silicic anhydride, magnesium silicate, talc, kaolin, bentonite, mica, mica titanium, bismuth oxychloride, zirconium oxide, magnesium oxide, zinc oxide, titanium oxide, calcium carbonate, magnesium carbonate, yellow iron oxide, Examples include Bengala, black iron oxide, Gunjo, chromium oxide, chromium hydroxide, carbon black, and calamine.

  Examples of ultraviolet absorbers include p-aminobenzoic acid derivatives, salicylic acid derivatives, anthranilic acid derivatives, coumarin derivatives, amino acid compounds, benzotriazole derivatives, tetrazole derivatives, imidazoline derivatives, pyrimidine derivatives, dioxane derivatives, camphor derivatives, furan derivatives, Examples include pyrone derivatives, nucleic acid derivatives, allantoin derivatives, nicotinic acid derivatives, vitamin B6 derivatives, oxybenzone, benzophenone, guaiazulene, shikonin, baicalin, baicalein, and berberine.

  Astringents include lactic acid, tartaric acid, succinic acid, citric acid, allantoin, zinc chloride, zinc sulfate, zinc oxide, calamine, zinc p-phenolsulfonate, potassium aluminum sulfate, resorcin, ferric chloride, tannic acid, etc. Can be mentioned.

  Antioxidants include ascorbic acid and its salts, stearic acid ester, tocopherol and its ester derivatives, nordihydrogua cetelenic acid, butylhydroxytoluene (BHT), butylhydroxyanisole (BHA), parahydroxyanisole, propyl gallate , Sesamol, sesamorin, gossypol, etc.

  Anti-inflammatory agents include ictamol, indomethacin, kaolin, salicylic acid, sodium salicylate, methyl salicylate, acetylsalicylic acid, diphenhydramine hydrochloride, d or dl-camphor, hydrocortisone, guaiazulene, camazulene, chlorpheniramine maleate, glycyrrhizic acid and its salts, Examples thereof include glycyrrhetinic acid and a salt thereof.

  Examples of the disinfectant / disinfectant include acrinol, sulfur, benzalkonium chloride, benzethonium chloride, methyl rosaniline chloride, cresol, calcium gluconate, chlorhexidine gluconate, sulfamine, mercurochrome, lactoferrin or a hydrolyzate thereof.

  For hair, selenium disulfide, alkylisoquinolinium bromide, zinc pyrithione, biphenamine, thianthol, castari tincture, ginger tincture, pepper tincture, quinine hydrochloride, strong aqueous ammonia, potassium bromate, sodium bromate, thiol Examples include glycolic acid.

  Perfumes include natural animal flavors such as musk, civet, castorium, ambergris, anise essential oil, angelica essential oil, Iran essential oil, Iris essential oil, fennel essential oil, orange essential oil, cananga essential oil, caraway essential oil, cardamom essential oil, guayakwood essential oil, cumin Essential oil, black letter essential oil, cinnamon essential oil, cinnamon essential oil, geranium essential oil, copaiba balsam essential oil, coriandel essential oil, perilla essential oil, cedarwood essential oil, citronella essential oil, jasmine essential oil, gingergrass essential oil, cedar essential oil, spearmint essential oil, western peppermint essential oil, large Perfume essential oil, tuberose essential oil, clove essential oil, orange flower essential oil, winter green essential oil, trout balsam essential oil, buttery essential oil, rose essential oil, palmarosa essential oil, persimmon essential oil, hiba essential oil, sandalwood essential oil, petit gren essential oil, bay essential oil, vetiva essential oil, bergamot Oil, Peru balsam essential oil, Boadrose essential oil, melamine essential oil, mandarin essential oil, eucalyptus essential oil, lime essential oil, lavender essential oil, linaloe essential oil, lemongrass essential oil, lemon essential oil, rosemary essential oil, Japanese mint essence essential oil, etc. Examples include synthetic fragrances.

  As dyes and colorants, red cabbage dye, red rice dye, red dye, anato dye, squid dye, turmeric dye, enju dye, krill dye, amber dye, caramel, gold, silver, gardenia dye, corn dye, onion dye , Tamarind pigment, spirulina pigment, buckwheat whole plant pigment, cherry pigment, laver pigment, hibiscus pigment, grape juice pigment, marigold pigment, purple potato pigment, purple yam pigment, lac pigment, rutin and the like.

  Examples of the sweetener include sugar, sweet tea, fructose, arabinose, galactose, xylose, mannose, maltose, honey, glucose, miraculin, monelin and the like.

  Examples of the nutrient fortifier include shell calcined calcium, cyanocolabamine, yeast, wheat germ, soybean germ, egg yolk powder, hemicellulose, heme iron and the like.

  In addition, hormones, sequestering agents, pH adjusters, chelating agents, antiseptic / antibacterial agents, refreshing agents, stabilizers, emulsifiers, animal / plant proteins and their degradation products, animal / plant polysaccharides and their Degradation product, animal / plant glycoprotein and its degradation product, blood flow promoter, anti-inflammatory agent / antiallergic agent, cell activator, keratolytic agent, wound treatment agent, foam enhancer, thickener, oral preparation, Deodorizing / deodorizing agents, bittering agents, seasonings, enzymes and the like.

  The dosage form of the present invention is arbitrary, ampoules, capsules, powders, granules, pills, tablets, solids, liquids, gels, bubbles, emulsions, creams, ointments, sheets It can be used in combination with quasi-drugs such as mousse, cosmetics for skin and hair, bath preparations, foods and drinks, and pharmaceuticals.

  Specific examples of cosmetics and quasi-drugs include basic cosmetics such as internal and external pharmaceutical preparations, lotions, emulsions, creams, ointments, lotions, oils, packs, facial cleansers, skin cleansers, shampoos, etc. , Rinse, hair treatment, hair cream, pomade, hair spray, hairdressing, permanent, hair art, hair coloring, hair growth, hair restoration, etc., foundation, white powder, funny, lipstick, blush, eye shadow, eye Makeup cosmetics such as liners, mascara, eyebrows, eyelashes, finishing cosmetics such as nail polishes, perfumes, bath preparations, other toothpastes, mouth fresheners, gargles, liquid odors and deodorants, hygiene Goods, sanitary cotton, wet tissue and the like.

  Examples of foods and beverages include soft drinks, carbonated drinks, nutrition drinks, fruit drinks, lactic acid drinks, ice cream, ice sherbet, shaved ice and other frozen desserts, buckwheat, udon, harsame, gyoza skin, shumai skin, chinese food Noodles such as noodles, instant noodles, candy, candy, gum, chocolate, tablet confectionery, snack confectionery, biscuits, jelly, jam, cream, baked confectionery, bakery confectionery, crab, salmon, clams, tuna, sardines, shrimp, Seafood such as skipjack, mackerel, whale, oyster, saury, squid, red scallop, scallop, abalone, sea urchin, salmon roe, tocobushi, fishery products such as kamaboko, ham, sausage, milk, processed milk, fermented milk, etc. Fats and oils for products, salad oil, tempura oil, margarine, mayonnaise, shortening, whipped cream, dressing, etc. Processed foods, sauces, sauces and other seasonings, curry, stew, oyakodon, rice bowls, miscellaneous cooking, Chinese rice bowls, bowls, tempura, eel rice bowls, hayashi rice, oden, mabodorf, beef bowl, meat sauce, egg soup, omelet rice, dumplings Retort pouch foods such as shumai, hamburger and meatballs, various forms of health and nutritional supplements, health functional foods, tablets, capsules, drinks, troches and the like.

  Although the extracellular matrix production improver of the present invention is suitably applied to humans, it can also be applied to animals other than humans as long as each effect can be expected.

  EXAMPLES The present invention will be described more specifically with reference to the following examples. However, these are merely examples and do not limit the scope of the present invention.

(Polyamine analysis method)
Animal, plant, microorganism, extract and extract, extracellular matrix production improver characterized by containing polyamine, cosmetics characterized by containing polyamine as an active ingredient, polyamine as an active ingredient The quasi-pharmaceutical products characterized by the above, the food and drink products characterized by containing polyamine as an active ingredient, the polyamine content contained in pharmaceutical products characterized by containing polyamine as an active ingredient by the following method You can investigate. Polyamines include free polyamines, compound polyamines, and conjugated polyamines, which can be analyzed with different extraction methods (Plant Cell Physiol., 43 (2), 196-206, 2002, J. Nutr. Biochem. 4, 66-70, 1993, Biosci. Biotech. Biochem., 61 (9), 1582-1584, 1997). As a specific example, a method for analyzing plant seed free polyamine will be described in detail. About 0.1 to 1.0 g of soybean seeds were diluted with an internal standard solution (1,6-hexaneamine or 1,7-diaminoheptane, internal standard amount = 7.5 or 12 nmol) and a 5% aqueous perchloric acid solution (sample raw 5-20 mL per 1.0 g body weight) and thoroughly triturate and extract at room temperature using a Polytron mixer. The grinding solution is centrifuged at 4 ° C., 35,000 × g for 20 minutes to collect the supernatant, and this solution is used as a free polyamine solution. Add 100-400 μL of free polyamine solution (polyamine composition, purified polyamine composition), 200 μL of saturated sodium carbonate aqueous solution, 200 μL of dansyl chloride / acetone solution (10 mg / mL) to a microtube with a screw cap and mix gently. To do. After tightly closing the stopper of the tube, it is covered with aluminum foil and heated in a water bath at 60 ° C. for 1 hour for dansylation. After allowing the tube to cool, 200 μL of an aqueous proline solution (100 mg / mL) is added and mixed. Cover with aluminum foil and reheat in water bath for 30 minutes. After cooling, nitrogen gas is blown to remove acetone, and then 600 μL of toluene is added and mixed vigorously. After leaving the tube to separate into two phases, the upper toluene layer is fractionated into 300 μL microtubes. Nitrogen gas is blown onto the collected toluene to completely remove the toluene. 200 μL of methanol is added to the tube to dissolve the dansylated free polyamine. The amount of free polyamines of putrescine, spermidine, and spermine is analyzed by an internal standard method using high-performance liquid chromatography connected to a fluorescence detector (excitation wavelength: 365 nm, emission wavelength: 510 nm). As the HPLC column, μ Bondapak C18 (manufactured by Waters: 0273324, 3.9 × 300 mm, particle diameter 10 μm) is used. The polyamine content in the sample is calculated by obtaining the peak areas of each polyamine and the internal standard from the standard solution and the HPLC chart of the sample, respectively.

(Example 1) Evaluation using normal human skin fibroblasts Evaluation was performed according to the following procedure. Normal human skin fibroblasts were seeded in a 48-well microplate so that the number of normal human skin fibroblasts was 2.0 × 10 ⁴ per well. The seeding medium used was Dulbecco's modified Eagle medium (DMEM) supplemented with 10% fetal bovine serum. After culturing at 37 ° C. in a carbon dioxide concentration of 5 vol% for 24 hours, the culture medium was replaced with a test medium to which a polyamine group sample having an arbitrary concentration was added, and further cultured for 48 hours. Subsequently, the medium containing 3- (4,5-dimethyl-2-thiazolyl) -2,5-diphenyltetrazolium bromide (MTT) was replaced with a medium containing 100 μg / mL and cultured for 3 hours. Formazan generated by the opening of the tetrazolium ring was removed. Extraction was performed with 2-propanol, and absorbance at 550 nm was measured with a microplate reader. At the same time, the absorbance at 650 nm was measured as turbidity, and the cell activation effect was evaluated by the difference between the two measured values. The evaluation results are shown in FIG. 1 as relative values with the cell activation effect in the control (water addition) as 100.

  As is clear from FIG. 1, it was confirmed that the medium supplemented with spermine, spermidine, and putrescine had a high cell activation / anti-aging effect on normal human skin fibroblasts. In particular, when 0.0001 mM, 0.001 mM, 0.01 mM, 0.1 mM putrescine, 0.0001 mM, 0.001 mM, 0.01 mM spermidine, 0.001 mM, 0.01 mM spermine are added, Compared to the blank (water), a significant cell activation effect of 20% or more was observed in all cases. From the above, it is clear that these polyamines have an excellent cell activating effect, and by applying them to the skin, they exhibit extremely excellent effects, such as skin wrinkles, sagging caused by ultraviolet exposure, Dullness and roughening can be effectively improved.

(Example 2) Evaluation using human hair papilla cells-Human hair papilla cell culture method Human hair papilla cells (THPC-001) total kit (HDPC total kit: THPCK-001, manufacturer: Cell Applications Inc. USA) , Import sales company: Toyobo Co., Ltd.), and cultured human hair papilla cells by a conventional method. Dispense PCGM medium for suspending thawed cells into 10 mL and 15 mL centrifuge tubes, and incubate with ice. That is, a vial containing frozen human hair dermal papilla cells (THPC-001) is rapidly thawed in a 37 ° C. constant temperature bath. About 1 mL of PCGM medium is gradually dropped into this vial to dilute DMSO, and the entire amount is transferred to a centrifuge tube containing PCGM medium and suspended. The suspended cells are centrifuged at 4 ° C. and 1000 rpm for 5 minutes in a cooling low centrifuge. Carefully aspirate the precipitated cells and resuspend in 1 mL PCGM medium. The whole amount is planted in a T-75 flask coated with a collagen solution, and placed in an incubator kept at 37 ° C. under a carbon dioxide concentration of 5 vol% under static humidity for stationary culture. After 1 day, the medium is changed. Thereafter, the culture medium is changed every other day and subcultured. The PCGM medium components were 250 mL of PCGM basal medium containing 1% FBS, 2.5 mL of bovine pituitary extract (BPE) 100-fold diluted solution, 2.5 mL of fetal calf serum (FCS) 100-fold diluted solution, 1.25 mL of a 200-fold diluted solution of insulin, transferrin, triiodothyronine solution (ITT) and 1.25 mL of a 200-fold diluted solution of siloprotein solution (Cyp) were used.

-Evaluation of dermal papilla cell activation effect Evaluation was performed according to the following procedure. Human hair papilla cells were seeded in a 48-well microplate at 2.0 × 10 ⁴ cells per well. The seeding medium used was Dulbecco's modified Eagle medium (DMEM) supplemented with 1 wt% fetal bovine serum. After culturing for 24 hours, the culture medium was replaced with a test medium to which a polyamine group sample having an arbitrary concentration was added, and further cultured for 48 hours. Subsequently, the medium containing 100 μg / mL of 3- (4,5-dimethyl-2-thiazolyl) -2,5-diphenyltetrazolium bromide (MTT) was replaced and cultured for 3 hours. Formazan produced by the opening of the tetrazolium ring was removed. Extraction was performed with 2-propanol, and absorbance at 550 nm was measured with a microplate reader. At the same time, the absorbance at 650 nm was measured as turbidity, and the cell activation effect was evaluated by the difference between the two measured values. The evaluation results are shown in FIG. 2 as relative values with the cell activation effect in the blank with no sample taken as 100.

  As is clear from FIG. 2, it was confirmed that the medium supplemented with spermidine, putrescine, and spermine has a high cell activation effect on human hair papilla cells. In particular, when 0.001 mM, 0.01 mM, 0.1 mM putrescine, 0.01 mM, and 0.1 mM spermidine were added, 20% or more significant cells compared to the blank (water). Activation and anti-aging effects were observed. From the above, it is clear that these polyamines have an excellent cell activating effect, and by applying them to the skin, they exhibit extremely excellent effects, such as skin wrinkles, sagging caused by ultraviolet exposure, Dullness and roughening can be effectively improved. Furthermore, since an excellent cell activation effect was confirmed in human hair papilla cells, polyamine can effectively improve stress formation, hormone balance, hair formation inhibition caused by UV exposure, and the like.

(Example 3) Evaluation of collagen production ability by polyamine Evaluation was performed according to the following procedure. Normal human dermal fibroblasts were seeded in a 48-well microplate so that the number of normal human skin fibroblasts was 1.0 × 10 ⁵ per well. The seeding medium used was Dulbecco's modified Eagle medium (DMEM) supplemented with 1% fetal bovine serum. After culturing at 37 ° C. in a carbon dioxide concentration of 5 vol% for 24 hours, after washing twice with PBS (−), the medium was replaced with a serum-free medium to which a polyamine group sample of an arbitrary concentration was added, and the conditions were further maintained for 2 days. And cultured. From the culture supernatant, type I procollagen C-terminal peptide (Procollagen type I carboxyterminal propeptide: PIP) produced by human fibroblasts was measured with Procollagen type I C-peptide (PIP) EIA Kit (TaKaRa). Collagen production acceleration rate was determined by measuring standard products using the above ELISA kit, creating a calibration curve from the results, and determining the amount of collagen produced when the sample was added and the amount of collagen produced when no sample was added. Evaluation was performed by calculating the collagen production amount at the time of addition as 100%.

  As is clear from FIG. 3, it was recognized that the collagen production activity (collagen production improving action) of human skin was significantly increased in the medium to which putrescine, spermidine and spermine were added. The concentration is indicated by polyamine concentration. In all the polyamine concentration groups, a significant collagen production improvement effect of 20% or more was recognized compared to the blank (water). By applying polyamine to the skin, fibroblast decay is reduced, collagen production is improved, and the amount of collagen can be maintained. As a result, it is possible to effectively improve wrinkles, sagging, dullness, roughness and the like caused by aging, exposure to ultraviolet rays and the like.

(Example 4) Evaluation of hyaluronic acid production ability by polyamine Polyamine was putrescine, spermidine and spermine, respectively. The evaluation was performed according to the following procedure. Normal human dermal fibroblasts were seeded in a 48-well microplate so that the number of normal human skin fibroblasts was 1.0 × 10 ⁵ per well. The seeding medium used was Dulbecco's modified Eagle medium (DMEM) supplemented with 1% fetal bovine serum. After culturing at 37 ° C. in a carbon dioxide concentration of 5 vol% for 24 hours, after washing twice with PBS (−), the medium was replaced with a serum-free medium to which a sample having an arbitrary polyamine concentration was added. The cells were cultured for 6 days under the same conditions. Hyaluronic acid in the culture supernatant was measured. The cells on the plate were lysed with a cell lysate containing 0.5% Triton-X100, and after recovery, the amount of protein was measured (Bio Rad, protein assay kit), which was used as an indicator of the number of cells. For the measurement of hyaluronic acid, hyaluronic acid produced by normal human skin fibroblasts was measured by a sandwich method using hyaluronic acid binding protein (HABP) using a hyaluronic acid plate “Chugai” (Fujirebio). When the amount of hyaluronic acid of a sample to which no polyamine was added (control) was taken as 100, the amount of hyaluronic acid of each polyamine was taken as the production rate.

  As is clear from FIGS. 4, 5, and 6, it was recognized that the hyaluronic acid production rate of human skin was significantly increased in the medium added with each polyamine. In all the polyamine concentration groups, a significant improvement in hyaluronic acid production was observed as compared with the control. By applying polyamine to the skin, the deterioration of fibroblasts can be reduced, the production of hyaluronic acid can be improved, and the amount of hyaluronic acid can be maintained. As a result, it is possible to effectively improve skin wrinkles, sagging, dullness, roughness caused by exposure to ultraviolet rays and the like.

(Example 5) Manufacture of essence The essence of the composition shown below was manufactured by the conventional method. As a control, a serum containing no polyamine was also produced by a conventional method.
(Composition) (wt%)
Sorbit 4.0
Dipropylene glycol 6.0
Polyethylene glycol 1500 5.0
POE (20) oleyl alcohol ether 0.5
Sucrose fatty acid ester 0.2
Methylcellulose 0.2
Spermidine 0.01
Total amount of purified water is 100

(Example 6) Manufacture of emulsion The emulsion of the composition shown below was manufactured by the conventional method. As a control, an emulsion containing no polyamine was also produced by a conventional method.
(Composition) (wt%)
Glyceryl ether 1.5
Sucrose fatty acid ester 1.5
Sorbitan monostearate 1.0
Squalane 7.5
Dipropylene glycol 5.0
Spermidine 0.01
Total amount of purified water is 100

(Example 7) Production of cream A cream having the following composition was produced by a conventional method. As a control, a cream containing no polyamine was also produced by a conventional method.
(Composition) (wt%)
Propylene glycol 6.0
Dibutyl phthalate 19.0
Stearic acid 5.0
Glycerol monostearate 5.0
Sorbitan monostearate 12.0
Polyethylene sorbitan monostearate 38.0
Sodium edetate 0.03
Spermidine 0.01
Total amount of purified water is 100

(Example 8) Sensory evaluation Sensory evaluation was performed using Examples 5-7. In addition, the comparative example which does not contain a polyamine was also evaluated simultaneously. For sensory evaluation, 20 panelists aged 40 to 60 years who are worried about wrinkles, sagging, dullness, etc. are treated as a group, and the examples and comparative examples are used twice a day for 3 months continuously. A questionnaire survey was conducted on the skin condition after a month.

  Table 1 shows the number of evaluators in each item as a result of sensory evaluation. More than 90% of the panel in the example compared to the comparative example not containing polyamine replied that the skin's skin / gloss was recovered and wrinkles were reduced. It was shown that. Further, similar results such as a recovery effect of skin symptoms were obtained even when the spermidine compounding concentration was 0.1% by weight.

(Example 9) Manufacture of essence The essence of the composition shown below was manufactured by the conventional method. As a control, a serum containing no polyamine was also produced by a conventional method.
(Composition) (wt%)
Sorbit 4.0
Dipropylene glycol 6.0
Polyethylene glycol 1500 5.0
POE (20) oleyl alcohol ether 0.5
Sucrose fatty acid ester 0.2
Methylcellulose 0.2
Putrescine 0.05
Total amount of purified water is 100

(Example 10) Manufacture of emulsion The emulsion of the composition shown below was manufactured by the conventional method. As a control, an emulsion containing no polyamine was also produced by a conventional method.
(Composition) (wt%)
Glyceryl ether 1.5
Sucrose fatty acid ester 1.5
Sorbitan monostearate 1.0
Squalane 7.5
Dipropylene glycol 5.0
Putrescine 0.05
Total amount of purified water is 100

(Example 11) Production of cream A cream having the composition shown below was produced by a conventional method. As a control, a cream containing no polyamine was also produced by a conventional method.
(Composition) (wt%)
Propylene glycol 6.0
Dibutyl phthalate 19.0
Stearic acid 5.0
Glycerol monostearate 5.0
Sorbitan monostearate 12.0
Polyethylene sorbitan monostearate 38.0
Sodium edetate 0.03
Putrescine 0.05
Total amount of purified water is 100

(Example 12) Sensory evaluation Sensory evaluation was performed using Examples 9-11. In addition, the comparative example which does not contain a polyamine was also evaluated simultaneously. For sensory evaluation, a group of 20 to 40-year-old panel worried about wrinkles and other symptoms was taken as a group, and the examples and comparative examples were used twice a day for 3 months, and the skin after 3 months. We conducted a questionnaire survey on the condition.

  As a result of sensory evaluation, the number of evaluators in each item is shown in Table 2. More than 90% of the panel in the example compared to the comparative example not containing polyamine replied that the skin's skin / gloss was recovered and wrinkles were reduced. It was shown that. Furthermore, similar results such as the effect of recovering skin symptoms were obtained even when the putrescine concentration was 0.5% by weight.

(Example 13) Manufacture of essence The essence of the composition shown below was manufactured by the conventional method. As a control, a serum containing no polyamine was also produced by a conventional method.
(Composition) (wt%)
Sorbit 4.0
Dipropylene glycol 6.0
Polyethylene glycol 1500 5.0
POE (20) oleyl alcohol ether 0.5
Sucrose fatty acid ester 0.2
Methylcellulose 0.2
Spermine 0.01
Total amount of purified water is 100

(Example 14) Manufacture of emulsion The emulsion of the composition shown below was manufactured by the conventional method. As a control, an emulsion containing no polyamine was also produced by a conventional method.
(Composition) (wt%)
Glyceryl ether 1.5
Sucrose fatty acid ester 1.5
Sorbitan monostearate 1.0
Squalane 7.5
Dipropylene glycol 5.0
Spermine 0.01
Total amount of purified water is 100

(Example 15) Production of cream A cream having the composition shown below was produced by a conventional method. As a control, a cream containing no polyamine was also produced by a conventional method.
(Composition) (wt%)
Propylene glycol 6.0
Dibutyl phthalate 19.0
Stearic acid 5.0
Glycerol monostearate 5.0
Sorbitan monostearate 12.0
Polyethylene sorbitan monostearate 38.0
Sodium edetate 0.03
Spermine 0.01
Total amount of purified water is 100

(Example 16) Sensory evaluation Sensory evaluation was performed using Examples 13-15. In addition, the comparative example which does not contain a polyamine was also evaluated simultaneously. For sensory evaluation, 20 panelists aged 40 to 60 years who are worried about wrinkles, sagging, dullness, etc. are treated as a group, and the examples and comparative examples are used twice a day for 3 months continuously. A questionnaire survey was conducted on the skin condition after a month.

  Table 3 shows the number of evaluators in each item as a result of the sensory evaluation. More than 90% of the panel in the example compared to the comparative example not containing polyamine replied that the skin's skin / gloss was recovered and wrinkles were reduced. It was shown that. Further, similar results such as a recovery effect of skin symptoms were obtained even when the spermine compounding concentration was 0.1% by weight.

(Example 17) Evaluation using normal human skin fibroblasts Evaluation was performed according to the following procedure. Normal human skin fibroblasts were seeded in a 48-well microplate so that the number of normal human skin fibroblasts was 2.0 × 10 ⁴ per well. The seeding medium used was Dulbecco's modified Eagle medium (DMEM) supplemented with 10% fetal bovine serum. After culturing at 37 ° C. in a carbon dioxide concentration of 5 vol% for 24 hours, the cells were cultured for 2 hours in DMEM containing 10 μg / ml mitomycin C (antibiotic, manufactured by Nacalai Tesque). Since mitomycin C has an action of inhibiting DNA replication, cell proliferation is suppressed by the treatment. Thereafter, after washing with PBS, the medium was replaced with a test medium to which a polyamine group sample having an arbitrary concentration was added, and further cultured for 48 hours. Subsequently, WST-8 [2- (2-methoxy-4-nitrophenyl) -3- (4-nitrophenyl) -5- (2,4-disulfophenyl) -2H-tetrazolium, monosodium salt, manufactured by Nacalai Tesque, Inc.] 100 μg / The medium was replaced with a medium containing mL and cultured for 3 hours. Formazan produced by the opening of the tetrazolium ring was extracted with 2-propanol, and the absorbance at 450 nm was measured with a microplate reader. At the same time, the absorbance at 600 nm was measured as turbidity, and the cell activation effect was evaluated by the difference between the two measured values. The evaluation results are shown in FIG. 7 as relative values with the cell activation effect in the control (water addition) as 100.

  As is clear from FIG. 7, in the medium to which putrescine, spermidine, and spermine were added despite the suppression of cell growth with mitomycin C, the cell activation activity was significantly enhanced against normal human skin fibroblasts. It was found to have. Cell growth activity is known as a physiological function of polyamines, but cell activation activity was shown by adding various polyamines despite the suppression of cell growth by mitomycin C having cell growth arresting activity. From these results, it was found for the first time that various polyamines have cell activation / cell activation action without cell proliferation (increase in cell number) or cell regeneration. From the above, it is clear that these polyamines have an excellent cell activation (cell activation) action, and when applied to the skin, they exhibit extremely excellent effects, such as aging and exposure to ultraviolet rays. It is possible to effectively improve skin wrinkles and sagging caused by the like.

(Example 18) Evaluation of collagen production ability by polyamine Evaluation was performed according to the following procedure. Normal human dermal fibroblasts were seeded in a 48-well microplate so that the number of normal human skin fibroblasts was 1.0 × 10 ⁵ per well. The seeding medium used was Dulbecco's modified Eagle medium (DMEM) supplemented with 1% fetal bovine serum. After culturing at 37 ° C. in a carbon dioxide concentration of 5 vol% for 24 hours, after washing twice with PBS (−), the medium was replaced with a serum-free medium to which a polyamine group sample of an arbitrary concentration was added, and further 3 days, 6 days Culture was performed under the same conditions. From the culture supernatant, type I procollagen C-terminal peptide (Procollagen type I carboxyterminal propeptide: PIP) produced by human fibroblasts was measured with Procollagen type I C-peptide (PIP) EIA Kit (TaKaRa). The cells on the plate were lysed with a cell lysate containing 0.5% Triton-X100, and after recovery, the amount of protein was measured (Bio Rad, protein assay kit), which was used as an indicator of the number of cells. Collagen production acceleration rate is measured with the above-mentioned ELISA kit for the standard product, and a calibration curve is created from the results. From the calibration curve, the amount of collagen produced when the sample is added per protein (number of cells) and the collagen when no sample is added The production amount was determined, and the collagen production amount when no sample was added was calculated as 100% for evaluation.

  As is clear from FIGS. 8 and 9, it was recognized that the collagen production rate of human skin was significantly increased per cell in the medium supplemented with putrescine, spermidine and spermine. The concentration is indicated by polyamine concentration. Compared with the blank (water) in all polyamine concentration groups, any significant collagen production improving action was recognized. Cell proliferation is known as a physiological function of polyamines, but various polyamines showed an effect of improving collagen production per cell using the amount of protein as an indicator. It was discovered for the first time from this result that it has an effect of improving collagen production (extracellular matrix production) without being accompanied. In other words, it has been clarified that various polyamines have an effect of increasing the amount of collagen produced per cell. From the above, by applying these polyamines to the skin, the decline of fibroblasts can be reduced, collagen production can be improved, and the amount of collagen can be maintained. As a result, it is possible to effectively improve skin wrinkles, sagging, dullness, etc. caused by exposure to ultraviolet rays.

(Example 19) Manufacture of essence The essence of the composition shown below was manufactured by the conventional method. As a control, a serum containing no polyamine was also produced by a conventional method.
(Composition) (wt%)
Sorbit 4.0
Dipropylene glycol 6.0
Polyethylene glycol 1500 5.0
POE (20) oleyl alcohol ether 0.5
Sucrose fatty acid ester 0.2
Methylcellulose 0.2
Spermidine 0.01
Putrescine 0.01
Spermine 0.005
Total amount of purified water is 100

(Example 20) Manufacture of emulsion The emulsion of the composition shown below was manufactured by the conventional method. As a control, an emulsion containing no polyamine was also produced by a conventional method.
(Composition) (wt%)
Glyceryl ether 1.5
Sucrose fatty acid ester 1.5
Sorbitan monostearate 1.0
Squalane 7.5
Dipropylene glycol 5.0
Spermidine 0.01
Putrescine 0.01
Spermine 0.005
Total amount of purified water is 100

(Example 21) Production of cream A cream having the following composition was produced by a conventional method. As a control, a cream containing no polyamine was also produced by a conventional method.
(Composition) (wt%)
Propylene glycol 6.0
Dibutyl phthalate 19.0
Stearic acid 5.0
Glycerol monostearate 5.0
Sorbitan monostearate 12.0
Polyethylene sorbitan monostearate 38.0
Sodium edetate 0.03
Spermidine 0.01
Putrescine 0.01
Spermine 0.005
Total amount of purified water is 100

(Example 22) Sensory evaluation Sensory evaluation was performed using Examples 19-21. In addition, the comparative example which does not contain a polyamine was also evaluated simultaneously. For sensory evaluation, 20 panelists aged 40 to 60 years who are worried about wrinkles, sagging, dullness, etc. are treated as a group, and the examples and comparative examples are used twice a day for 3 months continuously. A questionnaire survey was conducted on the skin condition after a month.

  Table 4 shows the number of evaluators in each item as a result of the sensory evaluation. More than 90% of the panels that mixed and blended each polyamine than the comparative example that did not contain polyamine replied that skin elasticity, gloss was restored, and wrinkle reduction was observed. It was shown that there was a symptom recovery effect. Furthermore, it has been confirmed for the first time that the effects of improving the recovery of skin symptoms are better when the polyamines are mixed and mixed than when the polyamines are mixed alone.

  According to the present invention, it is expected to have an excellent extracellular matrix production improving action with safety, and provides cosmetics and quasi-drugs (skin external preparations, bath preparations, hair growth agents, etc.), foods and drinks, and pharmaceuticals containing polyamine as an active ingredient. This is expected to make a significant contribution to industry.

Activation activity by polyamine using human skin fibroblasts Activation activity by polyamine using human hair papilla cells Collagen production improving activity by polyamine (2 days culture) Hyaluronic acid production enhancing activity by polyamine (1 day culture) Hyaluronic acid production improving activity by polyamine (3 days culture) Hyaluronic acid production improving activity by polyamine (6 days culture) Activation activity by polyamine using human skin fibroblasts Collagen production improving activity by polyamine (3 days culture) Collagen production improving activity by polyamine (6 days culture)

Claims (18)

  An agent for improving extracellular matrix production, comprising polyamine as an active ingredient.   An extracellular matrix production improver, wherein the extracellular matrix production improver is a collagen production improver and / or a hyaluronic acid production improver.   The improvement in extracellular matrix production according to claim 1 or 2, wherein the polyamine is at least one compound selected from the group consisting of aliphatic hydrocarbons having two or more primary amino groups. Agent.   Polyamine is 1,3-diaminopropane, putrescine, cadaverine, cardine, spermidine, homospermidine, aminopropyl cadaverine, theremin, spermine, thermospermine, canabalmin, aminopentylnorspermidine, N, N-bis (aminopropyl) cadaverine, The cell according to any one of claims 1 to 3, wherein the cell is at least one compound selected from the group consisting of homospermine, cardopentamine, homocardopentamine, cardohexamine and homocardohexamine. Outer matrix production improver.   The extracellular matrix production enhancer according to any one of claims 1 to 4, wherein the polyamine is at least one compound selected from the group consisting of putrescine, spermidine and spermine.   Cosmetics containing the extracellular matrix production improver according to any one of claims 1 to 5 as an active ingredient.   A quasi-drug containing the extracellular matrix production enhancer according to any one of claims 1 to 5 as an active ingredient.   Food / beverage products containing the extracellular matrix production improver in any one of Claims 1-5 as an active ingredient.   A pharmaceutical comprising the extracellular matrix production improver according to any one of claims 1 to 5 as an active ingredient.   A method for improving extracellular matrix production, comprising a step of bringing a polyamine into contact with an animal.   A method for improving collagen production, comprising a step of bringing a polyamine into contact with an animal.   The method for improving collagen production according to claim 11, wherein the polyamine is at least one compound selected from the group consisting of aliphatic hydrocarbons having two or more primary amino groups.   Polyamine is 1,3-diaminopropane, putrescine, cadaverine, cardine, spermidine, homospermidine, aminopropyl cadaverine, theremin, spermine, thermospermine, canabalmin, aminopentylnorspermidine, N, N-bis (aminopropyl) cadaverine, The method for improving collagen production according to claim 11 or 12, which is at least one compound selected from the group consisting of homospermine, cardopentamine, homocardopentamine, cardohexamine and homocardohexamine. .   The method for improving collagen production according to any one of claims 11 to 13, wherein the polyamine is at least one compound selected from the group consisting of putrescine, spermidine and spermine.   A method for improving hyaluronic acid production, comprising a step of bringing a polyamine into contact with an animal.   The method for improving hyaluronic acid production according to claim 15, wherein the polyamine is at least one compound selected from the group consisting of aliphatic hydrocarbons having two or more primary amino groups.   Polyamine is 1,3-diaminopropane, putrescine, cadaverine, cardine, spermidine, homospermidine, aminopropyl cadaverine, theremin, spermine, thermospermine, canabalmin, aminopentylnorspermidine, N, N-bis (aminopropyl) cadaverine, The hyaluronic acid production improvement according to claim 15 or 16, which is at least one compound selected from the group consisting of homospermine, cardopentamine, homocardopentamine, cardohexamine and homocardohexamine. Method.   The method for improving hyaluronic acid production according to any one of claims 15 to 17, wherein the polyamine is at least one compound selected from the group consisting of putrescine, spermidine and spermine.