JP6050573B2 - LTBP-4 production promoter - Google Patents

Description

  The present invention relates to an LTBP-4 (Latent TGFβ-binding protein-4) production promoter containing an algal extract or a plant extract.

  The LTBP (Latent TGFβ-binding protein) family is a protein belonging to the same superfamily as the fibrin protein that is involved in blood coagulation. By binding inactive latent TGFβ, TGFβ Is stored in the matrix in an inactive state, so that TGFβ can be mobilized from there when needed.

  Among such proteins, LTBP-4 is known to be associated with several diseases and functions in vivo. That is, in addition to the relationship between LTBP-4 expression and cancer progression (Non-Patent Document 1), it has been reported that LTBP-4 promotes the action of elastin deposition during elastic fiber formation in and outside the body. (Patent Document 1). Furthermore, there are reports that pulmonary emphysema is observed in mutant LTBP-4 (LTBP4-/-) mice and the characteristics of late-onset emphysema that develops in relation to human COPD are acquired (Non-patent Document 2 and Patent Document 2). ).

JP 2011-26236 A International Publication No. WO03 / 015505

Bertucci F., et al. Oncogene. 23: 1377-91, 2004 Sterner-Kock A., et al. Genes Dev. 16: 2264-2273, 2002

  An object of the present invention is to provide a production promoter for LTBP-4, which is a useful protein having such physiological activity.

  As a result of intensive studies to solve the above problems, the present inventors have found that an algae extract or a plant extract has an ability to promote LTBP-4 production and have completed the present invention.

  That is, the present invention relates to an LTBP-4 production promoter containing an algal extract or plant extract.

  The LTBP-4 production promoter may contain an extract of red algae, green algae, or brown algae as an active ingredient.

  The above red algae are Acrocaetium, Chinolemo, Oxenaceae, Cyprididae, Erythropertis, Oenothera, Kakrate, Coralidae, Dulce, Proboscisus, Rhodogorgon, Benimidae, Kaginori, Kagykenori It may be at least one selected from the group consisting of algae belonging to the order of Coleoptera, Cerambycidae, Cerambycidae, Pleurotus, Scorpaenidae, Itabyosa, and Cryptera.

  The green algae are: Mongolia, Yellow-winged, Sayamidro, Ketophora, Ketopertis, Chlorella, Microtamnion, Trevoxia, Kawanori, Mamiera, Monomastics, Dryomotics, Sumires Selected from the group consisting of algae belonging to the order of: Chlorodendron, Doricomastics, Nephrocellemis, Himidoridae, Aosa, Shiogusa, Ipomoea, Midorige, Iwauta, Milla, Anemone, and Casano There may be at least one.

  The brown algae is at least one selected from the group consisting of algae belonging to the order of Isogawala, Ajigusa, Nagamatsumo, Cyprus, Muchimo, Keyamamo, Urushigusa, Kombu, Ishige, and Hibamata obtain.

  The plant extract is Lamiaceae, Leguminosae, Crestedaceae, Pteraceae, Tadeidae, Grapeaceae, Asteraceae, Gentianaceae, Muraceae, White Jellyfish, Lotus, Aoi, Citrus, Ginger, Rubiaceae It may be at least one selected from the group consisting of plants belonging to the above.

  The present invention also relates to a composition for promoting LTBP-4 production, which contains any of the above LTBP-4 production promoters.

  The composition for promoting LTBP-4 production can be a cosmetic.

  The LTBP-4 production promoting composition may be a food.

  The present invention provides an LTBP-4 production promoter containing an algal or plant extract as an active ingredient.

It is the graph showing the effect of the LTBP-4 production promoter (corresponding to Table 1) containing different kinds of algae or plant extracts compared to the control (left). It is the graph showing the effect of the LTBP-4 production promoter (corresponding to Table 2) containing extracts of different types of algae or plants in comparison with the control (left). It is the graph which represented the effect of the LTBP-4 production promoter (corresponding to Table 3) containing the extract of different kinds of algae or plants in comparison with the control (left).

  LTBP-4 used in the present invention refers to a polypeptide derived from the nucleotide sequence set forth in SEQ ID NO: 1 (GenBank accession number AF051344) or an ortholog thereof, or mutants thereof (including SNP and haplotype). The ortholog of LTBP-4 is not particularly limited, and may be derived from, for example, any animal, preferably a mammal. Examples of mammals include cattle, sheep, pigs, goats, monkeys, rabbits, rats, hamsters, guinea pigs and mice. LTBP-4 is a secreted protein and the signal sequence can be removed by processing. In the method of the present invention, either LTBP-4 from which the signal sequence has been removed or LTBP-4 from which the signal sequence has not been removed can be used, but those from which the signal sequence has been removed are preferred. Further, the LTBP-4 mutant is not particularly limited. For example, the LTBP-4 mutant hybridizes with a DNA having a sequence complementary to the base sequence shown in SEQ ID NO: 1 under stringent conditions and is a DNA encoding LTBP-4. Examples of the polypeptide derived from the polypeptide encoded by the base sequence shown in SEQ ID NO: 1 include a sequence in which one to several amino acids are deleted, substituted, or added.

  In addition to the relationship between its expression and cancer progression, LTBP-4 has been reported to promote the action of elastin deposition during elastic fiber formation in and outside the body. Furthermore, the possibility of treatment of emphysema, and treatment of late-onset emphysema that develops in relation to human COPD has been suggested.

  Elastic fibers are extracellular fibers that take on the “elastic” property in tissues that return to their original state when stretched. Elasticity is found in many tissues of the body and is an essential property for the function of tissues rich in stretchable tissues such as lungs, arteries and skin. Elastic fibers are thought to be formed by depositing and crosslinking elastin around microfibrils. Elastin is a highly hydrophobic protein consisting of about 750 amino acids, and elastin deposition is known to involve proteins such as LTBP-4 that promote its action.

  Since elastic fibers are deteriorated or broken by aging or by external factors such as protease secreted from inflammatory cells, there are various states or diseases in which regeneration of elastic fibers is desired. Conditions or diseases for which elastic fiber regeneration is desired include emphysema, vascular injury, skin laxity, wounds, elastic fiber degradation (eg, caused by aging or UV rays, skin sagging), rough skin, arteriosclerosis, aorta Aneurysm, age-related macular degeneration, perineal hernia, anal hernia, basilar aneurysm, gastrointestinal motility disorder, pressure ulcer, etc.

  The present invention relates to a substance that promotes the production of LTBP-4 that is also involved in the formation of such elastic fibers, and such an LTBP-4 production promoting action is found in algae or plant extracts, An LTBP-4 production promoter comprising an algae or plant extract as an active ingredient is provided.

  The algal extract referred to in the present invention mainly refers to an extract extracted from brown algae, red algae or green algae by a usual method. Here, the red algae is, for example, the order of the genus Oleoptera (Oxenaceae, Funolino bogei), Ezozoenidae (Anemone quercivorus, Pleurotus serrata, Fusanori, Hirasanori, Nisehusanori, Ragara, Benimo zouk, Ezozoen) ) , Kainori, Chikinori, Suginori, Kotojitsu no Mata, Oobatsu No Mata, Tsuno Mata, Ibottsu No Mata, Akabaginanso, Atomugi Nori, Hirohanokakrate, Kintoki, Fushi Totoki, Centipede, Nikumukade, Utsuromukade, Hitomitsu, Katanari, Nagakintoki, Tambanori, Fudarak, Hiramkade, Hirakintoki, Sumidakade, Obakintoki, Hijirimen, Tsurutsuru, Matsunori, Masuhito , Coconut rose, key swordfish, seidai rose, tachiibara, hirohanotosakamodoki, hosobanotosakamodoki, usuginu, ikanoashi, etsukiiwakawa, kijinoo, saimi, okitsunori, harigane, mizusuna ), Pleurotus (Shiramo, Mizoogonori, Beniogonori, Fusiclenori, Birch Steller's sea bream, scorpionate (Villus serrata, Cyprinus, Caenopus, Taoyagisou, Fushitsunagi, Kosushifushitsunagi, Fukutsunagi, Nisebara Nori, Masagoshibari), Locust (Futagigis, Egonori, Togegisu, Hagisugi, Hagizugi Saeda, Benihiba, Nagabugugusa, Langeria, Yaleus Banori, Susius Banori, Kagios Banori, Tachius Beni, Hasujiginu, Togenori, Tsukubiwazo, Benzomo, Yuna, Yanaginori, Makuri, Sozozosozode , Fujimatsumo, Drosophila chinensis, Isomurasaki, Kozanemo), Acrocaetium, Chi It refers to algae belonging to the order of Norimo, Erythropertis, Euphorbiaceae, Dulce, Rhodogorgon, Kaginori, Kawamoku, Itabusa.

  Green algae are: Hibidoro (Siwaransomodoki, Ranodomodoki), Aosasa (Nagaaosa, Button Aosa, Ribbon Aosa, Bowuaonori, Usubaaoori, Anaaaosa, Amiaosa, Yabregusa) Cairomomo, Giant Pterosa, Caterpillar, Assamirisiogusa, Chasiogusa), Ipomoea (Siriomidro), Midorigea (Aomogusa, Magamata, Duckweed), Tamavalonia, Tamagobaronia, Ovalonia, Iwauta, Chileoptera Itogenomayuhaki, Mayuhakimo, Hime butterfly), mil eyes (Nanban High Mill, Higemil, Nezashimir, Sakibutomi, Nagamil, Mill, Motelle Mill, Hiramil, Tamil, Kobushimiru, Cromil), Anemone (Katahanohanemo, Ohananemo, Nankaihanemo), Casanori (Mizutama, Fudenoho, Ryukyugasa, Casanori, Isosugina), Ohigemawara eyes, Yokomado , Ketopertis, Chlorella, Microtamnion, Trevoxia, Kawanori, Mamiera, Monomastics, Dreamomastic, Sumiremo, Umikadamamo, Chlorodendron, Dreamomastic, Nephrothermis, etc. The algae to which it belongs.

  Brown algae are: Isogawala (Matsumo, Kinirohammon), Amygusa (Ezoyahaz, Yahazugusa, Herayahaz, Shiwayahaz, Ajigusa, Itoamiji, Fukurimamiji, Phaeosagi, Sanadagusa, Umichiwasa, Osamu Osa (Chromo, Futomoku, Nebarimo, Shiwanokawa), Crayfish eyes (Watamo, Hosookuwatamamo, Fukuronori, Kagomenori, Iwahige, Havanori, Habanori, Kayamo-doki, Kayanori), Whip-eye (Polypoda), Shigusa-Ru (Ishigusa-guru) Tobacco rush), Kombu eyes (Wakame, Tsurumo, Sujime, Kajime, Kurome, Arame, Macombu, Hosome Kombu), Shigego (Ishige, Iroro), Hibamata (Uganomok, Yaban) Say click, Joromoku, Sunabikimoku, Fushisujimoku, Sargassum, hijiki, Oba saw-Mok, Isomoku, akamoku, Nokogirimoku, Tamahahakimoku, Narasamo, Hiranejimoku, Yatsumatamoku, Mametawara, Oobamoku, Umitoranoo, Azumanejimoku, Yoremokumodoki, Endoumoku, Rappamoku), and the like.

  In the present invention, the effect of promoting the production of LTBP-4 was found in algae or plant extracts. Extracts from algae or plants may be used singly or in combination of two or more. Such an algae or plant is not limited, and an extract is a water-soluble component, an acid-soluble component, an alkali-soluble component from an algae or a plant, as long as the production promoting action of LTBP-4 is not impaired. An organic solvent soluble component.

  In the present invention, the component having the action of promoting the production of LTBP-4 is not particularly limited as long as it can be extracted from algae and plants. For example, brown algae may contain fucoidan, red algae may contain carrageenan and / or silicon, and green algae may contain chlorophyll α and / or β.

  The LTBP-4 production promoting action in the present invention is also found in plant extracts. The “plant extract” in the present specification includes, in addition to components extracted from plants, plant extract fermented liquids obtained by coexisting components extracted from plants and, for example, lactic acid bacteria. Plants include, for example, mallow, rhododendron, echidaceae, cruciferous, iridaceae, gramineae, nettleaceae, cranaceae, cichlidae, araliaceae, urchinaceae, cucurbitaceae, plantain, hypericaceae, asteraceae , Asteraceae, buttercup, camphoraceae, buckthorn, mulberry, poppy, scorpionaceae, scorpionaceae, araceae, primrose, perilla, shiraneae, ginger, ginger, hydrangea, hydrangea Family, violet family, celery family, spring mackerel family, aceae family, boxwood family, azalea family, camellia family, dodami family, cypress family, eggplant family, dianthus family, lotus family, rose family, bumblebee family, convolvulaceae family, cypress family, Grapeaceae, beech, diatomaceae, button, matabidae, pine, legume, man Examples include plants belonging to the family Cactaceae, Citrusaceae, Citricaceae, Sphagnumaceae, Purpleaceae, Barberry, Yamanoidae, Saxifragaceae, Lilyaceae, Orchidaceae, Gentianaceae, Lempoxaceae, Nodusaceae.

  Such plant extracts are not particularly limited, but include osconus urgonum extract, genus saffronaceae tabebuia impetiginosa, leguminous licorice root extract, buckthorn jujube fruit extract, matabidae kiwi seed extract, laceaceae Eye Leaf, Compositae Arch Chalk Leaf Extract, Tabebuia Impetiginosa Bark Extract, Gentian Gentiana Root Extract, Muraceae Shikon Extract, Murasaki Murasaki Root Extract, White Jellyfish Extract or Polysaccharide-Containing White Jellyfish Extract , Aceraceae Baobab leaf extract or hydrolyzed Baobab leaf extract, Citrus spruce or bitter orange peel extract, Citrus spruce extract / bitter orange peel extract, Ginger department moon peach leaf extract, and Acaenaceae asenyaku Extract 1 type or 2 types or more selected from the group which consists of. In addition, when the said plant extract is what can be obtained commercially, the said commercially available plant extract can also be used simply.

  The contents of the present invention will be described in detail below.

  As the algae or plant extract, a crude product of an extract from a natural product may be used as it is, or a product obtained by separating and purifying a crude product may be used.

  The method for obtaining the algae or plant extract is not particularly limited, and a normal extraction method is employed. The crude extract is a cutting machine, slicer, etc., for example, whole algae or whole plants or specific parts thereof (roots, stems, stems, bark, buds, leaves, flowers, fruits, seeds, etc.). What is shredded by a cutter / peeler or the like can be obtained by extracting with water, hot water, or a solvent obtained by combining organic solvents alone or in combination. Examples of the organic solvent include alcohols such as methanol, ethanol, 1-propanol, 2-propanol, 1-butanol and 2-butanol, esters such as methyl acetate and ethyl acetate, ketones such as acetone, formaldehyde, dimethyl sulfoxide and the like. Can be mentioned.

  In addition, a solvent at room temperature or room temperature can be used as the solvent, but a heating solvent or a thermal solvent (a solvent heated near the boiling point) can also be used.

  Furthermore, extraction can be performed under acid and alkali conditions. Examples of the acid include inorganic acids such as hydrochloric acid, sulfuric acid, nitric acid, and phosphoric acid, and organic acids such as acetic acid, citric acid, oxalic acid, succinic acid, formic acid, and propionic acid. As the alkali, an alkali inorganic salt is preferably used, and examples thereof include potassium hydroxide and sodium hydroxide.

  It is also possible to use a freeze-drying powdering method, a high pressure method or an ultrahigh pressure method. In the high-pressure method, the cells in the algae or plant are pressurized and crushed by high-pressure heat treatment at 120 ° C. for 4 hours.

  An example of the extraction conditions for the crude extract is given. When the extraction solvent is an organic solvent, the extraction temperature is not particularly limited, but preferably 1 to 100 ° C, more preferably 20 to 100 ° C, with stirring for 1 to 24 hours, preferably 1 hour to 10 hours. Do. Further, when the extraction solvent is water, the temperature at the time of extraction is not particularly limited as long as it is equal to or lower than the boiling point of the solvent. However, since an extraction method using pressurized hot water may be used, about 0 to 200 ° C. is preferable. About 20 to 130 ° C. is more preferable. Also, the reaction time is not particularly limited, but it may be set to several seconds to several days, more preferably 30 minutes to 6 hours.

  The reaction temperature and reaction time may be appropriately selected depending on the type and concentration of the solvent.

  If it is extracted in this way, the component which has LTBP-4 production promotion ability can fully be extracted.

  The obtained extract is centrifuged to obtain the supernatant as an extract. Such an extract contains a water-soluble, acidic-soluble, alkali-soluble, or organic solvent-soluble polysaccharide fraction.

  As a method for separating and purifying an algae or plant extract from a crude product of the extract, for example, a method of adding a quaternary ammonium salt such as cetylpyridinium chloride to precipitate the algae or plant extract and washing with an alcohol solution There is also a method using chromatography using an anion exchange resin such as DEAE cellulose.

  The algae or plant extract solution thus extracted and purified can be used as it is, or can be diluted with a diluting solution or concentrated to be used as an extract. Or it can be prepared in the form of a paste and used.

  The LTBP-4 production promoter comprising the algae or plant extract of the present invention as an active ingredient can be used for the treatment of diseases caused by a decrease in the expression level of LTBP-4. For example, there are possibilities for treatment of cancer, emphysema, and late-onset emphysema that develops in relation to human COPD. It is also useful for the treatment of diseases related to the formation of elastic fibers that are thought to involve LTBP-4 production. A disease associated with elastic fiber formation is a condition or disease in which elastic fiber regeneration is desired, such as pulmonary emphysema, vascular injury, skin laxity, wounds, elastic fiber degradation (eg, caused by aging or ultraviolet radiation). ), Rough skin, arteriosclerosis, aortic aneurysm, age-related macular degeneration, perineal hernia, anal hernia, basilar aneurysm, gastrointestinal motility disorder, pressure ulcer, etc. Furthermore, it is useful for cosmetic purposes such as prevention and improvement of skin sagging and wrinkles. The LTBP-4 production promoter of the present invention is also useful as a cell culture reagent having the ability to regenerate elastic fibers. Furthermore, it is useful for producing artificial tissues such as artificial skin, skin models, and artificial blood vessels. Furthermore, LTBP-4 itself, which has not been fully elucidated, can be applied to research, and is considered useful in a very wide range.

  The LTBP-4 production promoter of the present invention can be provided in the form of any preparation or composition that can be widely used as a pharmaceutical, quasi-drug, food, cosmetic, etc., but preferably as a functional food or cosmetic. Available formulation or composition.

  Regardless of whether the composition containing the LTBP-4 production promoter of the present invention takes any form of pharmaceuticals, quasi-drugs, foods, and cosmetics, the algae or plant extract content is converted to dry weight. For example, it may be about 0.000001 to 99% by weight, more preferably about 0.00001 to 30% by weight, still more preferably about 0.0001 to 10% by weight, particularly preferably 0.001 to 5% by weight. It is. If it is the said range, sufficient LTBP-4 production promotion effect is seen by use of a pharmaceutical, a quasi-drug, a foodstuff, and cosmetics.

  When the present invention is used for pharmaceuticals or health foods, the dose of algae or plant extract can be 0.0001 to 5000 mg / kg per day for adults in dry weight. If it is the said range, the improvement or prevention of the disease or condition which requires LTBP-4 production is attained.

  Skin topical form

  The LTBP-4 production promoter comprising the algae or plant extract (crude extract or purified product) of the present invention as an active ingredient can take a form known as an external preparation form of cosmetics or quasi drugs. . As such forms, for example, liquids, suspensions, emulsions, creams, ointments, gels, liniments, lotions, aerosols, powders, poultices, nonwoven fabrics and the like were impregnated with chemicals. Examples thereof include a sheet agent and a stick agent such as a lipstick. Among these, the liquid, suspension, emulsion, cream, ointment, gel, lotion, and gel are preferably used. By setting it as this form, the elastic fiber formation effect can fully be exhibited. For skin external preparations, in addition to algal or plant extracts, known additives that are added to skin external preparations (cosmetics, quasi-drugs, pharmaceuticals) within a range that does not impair the effects of the present invention, for example, , Surfactants, oils, alcohols, humectants, thickeners, preservatives (preservatives), antioxidants, chelating agents, pH adjusters, stabilizers, dispersants, fragrances, colorants, pigments, pearls Gloss imparting agent, blood circulation promoting component, moisturizing component, UV absorbing component, UV scattering component, cleaning component, antibacterial component, astringent component, vitamins, amino acids, anti-aging component, anti-inflammatory component, whitening component, keratin softening component, cell An activation component, water, etc. can be mix | blended. An additive can be used individually by 1 type or in combination of 2 or more types.

  In addition, the specific use of the cosmetic composition is not particularly limited, and basic cosmetics such as lotion, milky lotion, cream, cosmetic liquid, sunscreen cosmetics, packs, hand creams, body lotions, body creams; Cosmetics for cleaning such as face wash, makeup remover, body shampoo, shampoo, rinse; makeup cosmetics such as foundation, makeup base, lip balm, lipstick, teak color;

<Base or carrier>
Bases or carriers include liquid paraffin, squalane, gelled hydrocarbons (such as plastibase), ozokerite, α-olefin oligomers, hydrocarbons such as light liquid paraffin; methyl polysiloxane, cross-linked methyl polysiloxane, highly polymerized methyl Polysiloxane, Cyclic silicone, Alkyl-modified silicone, Cross-linked alkyl-modified silicone, Amino-modified silicone, Polyether-modified silicone, Polyglycerin-modified silicone, Cross-linked polyether-modified silicone, Cross-linked alkyl polyether-modified silicone, Silicone / alkyl chain copolymer Modified polyether-modified silicone, silicone / alkyl chain co-modified polyglycerin-modified silicone, polyether-modified branched silicone, polyglycerin-modified branched silicone, acrylic silicone, Silicone oils such as phenyl-modified silicones and silicone resins; cellulose derivatives such as ethyl cellulose, hydroxypropyl cellulose, hydroxypropyl methylcellulose; polyvinylpyrrolidone; carrageenan; polyvinyl butyrate; polyethylene glycol; dioxane; butylene glycol adipate polyester; isopropyl myristate , Esters such as octyldodecyl myristate, isopropyl palmitate, cetyl palmitate, isononyl isononanoate, pentaerythritol tetra-2-ethylhexanoate; polysaccharides such as dextrin and maltodextrin; lower levels such as ethanol and isopropanol Alcohol; ethylene glycol monomethyl ether, ethylene glycol monoethyl ether , Ethylene glycol monopropyl ether, diethylene glycol monomethyl ether, diethylene glycol monoethyl ether, diethylene glycol monopropyl ether, diethylene glycol monobutyl ether, propylene glycol monoethyl ether, propylene glycol monopropyl ether, dipropylene glycol monoethyl ether, dipropylene glycol monopropyl Examples include glycol ethers such as ethers; polyhydric alcohols such as polyethylene glycol, propylene glycol, 1,3-butylene glycol, glycerin, and isoprene glycol; and aqueous bases such as water.

  A base or a support | carrier can be used individually by 1 type or in combination of 2 or more types.

<Additives>
Examples of the antioxidant that can be contained in the composition for external use of the present invention include dibutylhydroxytoluene, butylhydroxyanisole, sorbic acid, sodium sulfite, ascorbic acid, erythorbic acid, L-cysteine hydrochloride and the like.

  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; propylene glycol fatty acid esters such as propylene glycol monostearate; polyoxyethylene hydrogenated castor oil 40 (HCO-40), polyoxyethylene hydrogenated castor oil 50 (HCO-50), polyoxyethylene hardened Hardened castor oil derivatives such as castor oil 60 (HCO-60), polyoxyethylene hydrogenated castor oil 80; polyoxyethylene (20) sorbitan (polysorbate 20) monolaurate, monostearate Polyoxyethylene sorbitan fatty acid esters such as polyoxyethylene (20) sorbitan (polysorbate 60), polyoxyethylene monooleate (20) sorbitan (polysorbate 80), polyoxyethylene (20) sorbitan isostearate; Glyceryl alkyl ether; alkyl glucoside; polyoxyalkylene alkyl ether such as polyoxyethylene cetyl ether; amines such as stearylamine and oleylamine; polyoxyethylene / methylpolysiloxane copolymer; And silicone surfactants such as lauryl PEG-9 polydimethylsiloxyethyl dimethicone and PEG-9 polydimethylsiloxyethyl dimethicone.

  Examples of the thickener include guar gum, locust bean gum, carrageenan, xanthan gum, carboxymethylcellulose, hydroxymethylcellulose, hydroxyethylcellulose, hydroxypropylcellulose, hydroxypropylmethylcellulose, polyvinyl alcohol, polyvinylpyrrolidone, carboxyvinyl polymer, alkyl methacrylate methacrylate. Copolymers, polyethylene glycol, bentonite, (hydroxyethyl acrylate / acryloyldimethyltaurine Na) copolymer, (acryloyldimethyltaurine ammonium / vinylpyrrolidone) copolymer, and the like.

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

Examples of pH adjusters include inorganic acids (hydrochloric acid, sulfuric acid, etc.), organic acids (lactic acid, sodium lactate, citric acid, sodium citrate, succinic acid, sodium succinate, etc.), inorganic bases (potassium hydroxide, sodium hydroxide, etc.) ) And organic bases (such as triethanolamine, diisopropanolamine, and triisopropanolamine).
Examples of the chelating agent include EDTA / disodium salt, EDTA / calcium disodium salt, and the like.

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

  An additive can be used individually by 1 type or in combination of 2 or more types.

<Other active ingredients>
The composition for external use of this invention can contain another active ingredient in the range which does not impair the effect of this invention. Specific examples of active ingredients include moisturizing ingredients, anti-inflammatory ingredients, antibacterial ingredients, vitamins, peptides or derivatives thereof, amino acids or derivatives thereof, cell activation ingredients, anti-aging ingredients, blood circulation promoting ingredients, and the like.

  Internal form

  The algae or plant extract, which is an active ingredient of the present invention, is mixed with a pharmaceutically acceptable carrier, and is a solid preparation such as a tablet, capsule, granule or powder; or a liquid preparation such as a syrup or injection. Can also be administered orally or parenterally. These preparations may contain conventional additives. As the pharmaceutically acceptable carrier, various organic or inorganic carrier substances commonly used as pharmaceutical materials are used. Excipients, lubricants, binders, disintegrants in solid preparations; solvents, dissolution aids in liquid preparations , Suspending agents, isotonic agents, buffers, soothing agents and the like. Further, if necessary, preparation additives such as preservatives, antioxidants, colorants, sweeteners and the like can be used.

  Preferable examples of the excipient include sugar alcohols such as D-sorbitol, mannitol and xylitol, sugars such as glucose, sucrose, lactose and fructose, crystalline cellulose, carmellose sodium, croscarmellose sodium, calcium hydrogen phosphate , Wheat starch, rice starch, corn starch, potato starch, dextrin, β-cyclodextrin, light silicic acid anhydride, titanium oxide, magnesium aluminate metasilicate, talc, kaolin and the like.

  Preferable examples of the binder include cellulose derivatives such as methyl cellulose, ethyl cellulose, hydroxypropyl cellulose, hydroxypropyl methyl cellulose, polyvinyl pyrrolidone, polyvinyl alcohol, acrylic polymer, gelatin, gum arabic, pullulan, pregelatinized starch, agar , Tragacanth, sodium alginate, propylene glycol alginate, and the like.

  Preferable examples of the disintegrant include starch, low-substituted hydroxypropylcellulose, carboxymethylcellulose calcium, croscarmellose sodium, hydroxypropyl starch, and partially pregelatinized starch. Preferable examples of the solvent include water for injection, alcohol, propylene glycol, macrogol, sesame oil, corn oil and the like.

  Preferable examples of the lubricant include stearic acid, magnesium stearate, calcium stearate, polyoxyl stearate, cetanol, talc, hydrogenated oil, sucrose fatty acid ester, dimethylpolysiloxane, beeswax, and white beeswax.

  Preferable examples of the solubilizer include polyethylene glycol, propylene glycol, D-mannitol, benzyl benzoate, ethanol, trisaminomethane, cholesterol, triethanolamine, sodium carbonate, sodium citrate and the like. Suitable examples of the suspending agent include surfactants such as stearyltriethanolamine, sodium lauryl sulfate, laurylaminopropionic acid, lecithin, benzalkonium chloride, benzethonium chloride, glyceryl monostearate; for example, polyvinyl alcohol, Examples thereof include hydrophilic polymers such as polyvinylpyrrolidone, sodium carboxymethylcellulose, methylcellulose, hydroxymethylcellulose, hydroxyethylcellulose, and hydroxypropylcellulose.

  Preferable examples of the isotonic agent include sodium chloride, glycerin, D-mannitol and the like. Preferable examples of the buffer include buffer solutions such as phosphate, acetate, carbonate, citrate and the like. Preferable examples of the soothing agent include benzyl alcohol. Preferable examples of the preservative include paraoxybenzoates, chlorobutanol, benzyl alcohol, phenethyl alcohol, dehydroacetic acid, sorbic acid and the like. Preferable examples of the antioxidant include sulfite and ascorbic acid.

  The method for producing the liquid preparation or solid preparation of the present invention is not particularly limited as long as it includes a step of producing a liquid preparation or solid preparation by mixing an algae or plant extract with a preparation raw material. The method can be used.

Example of solid preparation:
For example, a method of kneading a formulation composition containing an algae or plant extract and a formulation raw material, and pulverizing and sizing the extruded granulated product by passing through a screen, kneading the formulation composition After stirring and granulating by adding water and molding with a vertical granulator, pulverizing and sieving using a comil, and after compressing the formulation composition with a roller compactor, pulverizing and sieving with a roll granulator, A method of fluidized bed drying after stirring granulation is exemplified.

  In addition, for example, in the case of producing by direct compression, after mixing an algae or plant extract and a pharmaceutical composition containing a pharmaceutical raw material, it may be directly put into a tableting machine for tableting.

  Food

  The LTBP-4 production promoter containing the algal or plant extract of the present invention can be provided by being contained in a food or functional food. Examples of such foods or functional foods include cooked rice; various kinds of noodles including soba, udon, harusame, Chinese noodles, instant noodles, cup noodles; soft drinks, carbonated drinks, nutrition drinks, fruit drinks, lactic acid drinks, sports drinks Beverages such as Carrero, stew, various soups; Ice cream, ice sorbet, shaved ice and other frozen desserts; Strawberries, cookies, candy, gum, chocolate, tablet confectionery, snacks, biscuits, jellies, jams, creams, and other baked goods Sweets such as kamaboko, hampen, processed food such as ham, sausage; dairy products such as processed milk and fermented milk; fats and oils such as salad oil, tempura oil, margarine, mayonnaise, shortening, whipped cream, dressing Processed foods; seasonings such as sauces, dressings, miso, soy sauce, sauces; soups Stew, salad, delicatessen, sprinkle, pickles; health and nutrition supplement other various forms is illustrated.

  Furthermore, supplements (scattering, granules, soft capsules, hard capsules, tablets, chewable tablets, quick-disintegrating tablets, syrups, liquids, etc.) containing the LTBP-4 production promoter of the present invention may be prepared.

  Moreover, the LTBP-4 production promoter of this invention can also be contained with respect to animal foods, such as a pet.

  Additives are added to the food as needed. Examples of such additives include glucose, fructose, sucrose, maltose, sorbitol, stevioside, rubusoside, corn syrup, lactose, mannitol, dextrin, citric acid, sodium citrate, tartaric acid, malic acid, succinic acid, lactic acid L-ascorbic acid, dl-α-tocopherol, sodium erythorbate, glycerol, propylene glycol, glycerol fatty acid ester, polyglycerol fatty acid ester, sucrose fatty acid ester, sorbitan fatty acid ester, gum arabic, carrageenan, casein, gelatin, pectin, Examples include agar, vitamin C, vitamin B, vitamin E, nicotinic acid amide, calcium pantothenate, amino acids, calcium salts, surfactants, pigments, fragrances, and preservatives.

  It is also possible to prepare LTBP-4 in vitro using the LTBP-4 production promoter containing the algae or plant extract of the present invention, and to use it for the study of LTBP-4 itself and elastic fibers. The LTBP-4 production promoter of the present invention can be used to produce LTBP-4 in vitro and to produce artificial tissues such as artificial elastic fiber culture reagents, artificial skin, skin models, and artificial blood vessels. When used for such in vitro research and application, it is also preferable to use the algal or plant extract as it is.

  In order to produce LTBP-4 in vitro, a cell containing LTBP-4 can be prepared in vitro by preparing a composition containing about 1% to 70% by dry weight as extracted from an algal or plant extract. Coexist. Such a composition can be used as it is as an LTBP-4 production promoter of the present invention. A specific method for producing LTBP-4 includes culturing cells capable of producing LTBP-4 in contact with the LTBP-4 production promoter of the present invention. Cultured cells typically secrete LTBP-4 extracellularly. Alternatively, LTBP-4 production can be promoted in vivo. The cell to be used is a cell derived from any animal, preferably a mammal. Such cells are cells derived from tissues where LTBP-4 is present, such as skin, blood vessels, lungs, uterus, heart, kidney, pancreas, testis, ovary, small intestine, colon, cartilage, fibroblasts, smooth muscle Examples include cells such as cells, epithelial cells, and endothelial cells. Preferably, dermal fibroblasts or vascular smooth muscle cells are used. For in vitro LTBP-4 production, cells obtained by culturing undifferentiated cells such as primary cultured cells derived from living bodies, established cell lines, and stem cells can also be used.

  Cells that can produce LTBP-4 are originally cells that do not have LTBP-4 production ability or cannot express sufficient amounts, and produce LTBP-4 by gene transfer. Cells that have been manipulated in this way are also included. By introducing a vector capable of expressing the LTBP-4 gene into a cell, a cell having acquired LTBP-4 production ability or a cell having improved LTBP-4 production ability can be produced.

  The elastic fiber formation promoter of the present invention may further contain a DANCE (developmental arteries and neural crest epidermal growth factor (EGF) -like) expression enhancing factor (DANCE inducing factor). DANCE expression enhancing factor can be used, for example, DANCE expression enhancement such as Nanten quail extract, winged bean extract, phytic acid and physiologically acceptable salts thereof, and L-hydroxyproline and physiologically acceptable salts thereof Factor: DANCE expression vector, DANCE inducing factor expression vector and the like.

  Next, the present invention will be specifically described with reference to test examples and examples, but the present invention is not limited to the following examples.

[Examples 1 to 23, Comparative Examples 1 to 6] (various algae and plant extracts)
It used for the evaluation test of LTBP-4 production promotion as it was using the extract of various commercially available algae and plants shown in Tables 1-3.

[Test Example 1] (Evaluation test of LTBP-4 production promotion)

Test example 1:
The test was carried out using the direct adsorption method (direct ELISA method).
First, 3500 human fibroblasts were seeded per well in a 96-well plate, and subculture medium (DMEM (purchased from Dulbecco's Modified Eagle Medium, Invitrogen), 10% fetal bovine serum (purchased from FBS, JRH Bioscience) 2 mM glutamine, 100 units / mL penicillin, 100 g / mL streptomycin) at 37 ° C. and 5% CO ₂ for 48 hours. Furthermore, a serum-free medium, that is, DMEM / F12, 2 mM glutamine, 100 units / mL penicillin, 100 g / mL streptomycin was used, and in this serum-free medium, Examples 1 to 23 and Comparative Examples 1 to 6 were used. The obtained extract (described in Tables 1 to 3) was cultured at 37 ° C. and 5% CO ₂ for 3 days with or without addition of 0.001% by weight of the extract. The cultured fibroblast supernatant (or cell lysate) was added to 96 wells and allowed to stand at 4 ° C. overnight to form a stationary phase. Subsequently, the wells were washed with PBS, blocked after adding Block Ace (purchased from DS Pharma Biomedical) 4 times with PBS to the wells, and then anti-LTBP-4 antibody solution (from Santa Cruz). Purchased SC-30360) was added to each well and allowed to react for 1 hour at room temperature. Further, the wells were washed with PBS, a secondary antibody solution labeled with HRP (ZyMax81-1620 purchased from Zymed Laboratories. Inc.) was added and allowed to react at room temperature for 1 hour, and then the chromogenic substrate was reacted for 10 minutes. I let you. The color intensity after the reaction was stopped was measured using an absorptiometer (OD450 nm), the numbers expressed in comparison with the control are shown in Tables 1 to 3, and further graphed are shown in FIGS.

藻類または植物の抽出物は、優れたLTBP-4産生促進作用を有していることが示された。特に、紅藻類または緑藻類から得られる抽出物、あるいは、植物のうちでも、茶花乾燥エキス、バオバブなどを始め、非常に優れた効果を有しているものが見出された。

Algal or plant extracts have been shown to have an excellent LTBP-4 production promoting effect. In particular, an extract obtained from red algae or green algae, or among plants, has been found to have a very excellent effect, such as dried tea flower extract and baobab.

[Composition Preparation Example 1]
Cosmetic composition The unit of the numerical value in the Example of the following cosmetic compositions is "weight%".

[Composition Preparation Example 2]
Oral Composition In the formulation examples of the following food compositions, the weight of the algae or plant extract is the dry weight.
In the whole formulation of the food composition, the algae or plant extract is preferably an ethanol extract.

  This drink is taken once every three meals a day.

  Three tablets of this tablet are taken every three meals a day.

Claims (10)

  The LTBP-4 production promoter which contains the extract of Geridium curcilaineum, Kaguibaranori, or asparagopsis almata as an active ingredient.   The composition for LTBP-4 production promotion containing the LTBP-4 production promoter of Claim 1.   The composition for promoting the production of LTBP-4 according to claim 2, wherein the extract is contained in an amount of 0.000001 to 99% by weight in terms of dry weight with respect to the total amount.   The composition for promoting the production of LTBP-4 according to claim 2 or 3, which is a cosmetic.   The composition for promoting LTBP-4 production according to claim 2 or 3, which is a food. A topical composition comprising an extract of Chlorella, Gentiana, Igis, Geridium Curatiginaum, Pervetianacariculata, Kaguibaranori, Imosemiru, Asparagopsis armata, Condor crisps, and moon peach leaf and DANCE expression enhancing factor (however, , Excluding extracts of Condor crisps and topical compositions containing phytic acid) . The topical application according to claim 6, wherein the DANCE expression enhancing factor includes a nankazura extract, winged bean extract, phytic acid or a physiologically acceptable salt thereof, or L-hydroxyproline or a physiologically acceptable salt thereof. Composition (excluding an extract of Condor crisps and an external composition containing phytic acid) .   The composition for external use according to claim 6 or 7, wherein the composition for external use is for promoting production of LTBP-4. Chlorella, gentian, Igisu, Bae behenate thia canary Kurata, Kagiibaranori, Imosemiru, co command pulse Crisp scan, or monthly artificial elastic fiber formation promoting agent containing as an active ingredient an extract of peach leaf.   Promote LTBP-4 production in vitro using extracts of Chlorella, Gentian, Igis, Geridium curtyraineum, Pervetia canariculata, Kaguibaranori, Imosemiru, Asparagopsis armata, Condor crisps, or moon peach leaves How to make.

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