JP4800870B2 - Matrix metalloproteinase inhibitor - Google Patents

Description

  The present invention relates to a matrix metalloprotease inhibitor. More specifically, it has an inhibitory effect on matrix metalloproteinases (MMPs) activity that greatly affects skin wrinkles, sagging, and elasticity, and it inhibits matrix metalloproteinases that maintain skin tension and suppress aging. And an anti-aging cosmetic containing the matrix metalloproteinase inhibitor. Matrix metaprotease inhibitors are used particularly in fields such as quasi drugs and cosmetics.

  Skin aging is divided into physiological aging due to aging and photoaging that is emphasized by ultraviolet rays. It is said that photoaging is proportional to the intensity and time of exposure to ultraviolet rays, and up to about 80% of the spots and wrinkles that are cosmetically concerned are said to be physiological aging and heterogeneous photoaging. The structure of the skin consists of an epidermal layer and a dermal layer, with a basement membrane between them. The basement membrane is mainly composed of type IV collagen and laminin, and the structure is held by anchoring fibrils composed of type VII collagen. The dermis layer is composed of a macromolecular network called an extracellular matrix, and is mainly composed of intricately intertwined types I, III, V, VI, collagen, elastin, proteoglycan, fibronectin and the like. From previous studies, the amount of collagen in photoaged skin has decreased significantly compared to physiologically aged skin, but there was no significant difference in the amount of collagen produced in cultured fibroblasts from sun-exposed and coated skin. Rather, it has been reported that the proliferation activity of cultured fibroblasts in photo-aged skin is higher than that of physiologically-aged skin, and the amount of collagen production is also increased. Therefore, the decrease in collagen during photoaging is considered to be due to the fact that the degradation of collagen exceeds the synthesis, and suppressing collagen degradation is considered to be effective in suppressing wrinkle formation (Non-patent Document 1). In addition, histological studies have reported that exposure to ultraviolet rays damages the basement membrane (rupture and multiplexing) and dermal matrix components such as collagen degenerate and decrease, and matrix metalloprotease is a factor that causes this change. Has been pointed out (Non-Patent Documents 2 and 3). Matrix metalloproteases have collagenase (MMP-1; degradation of type I and type III collagen), gelatinase (MMP-2, MMP-9; type IV, type V collagen, laminin, elastin due to differences in structure and substrate specificity. Degradation), stromelysin (MMP-3, MMP-10; proteoglycan, type IV collagen, laminin degradation), membrane-bound matrix metalloprotease (MMP-14, MMP-15, MMP-16, MMP-24) And other molecular species are known. In recent years, it has been suggested that epidermal gelatinase induces wrinkle formation and that basement membrane injury is involved in the process (Non-patent Document 3).

  Usually, the activity of MMPs including gelatinase is strictly regulated by the production and activation of proenzymes and the action of TIMPs (TIMP-1, TIMP-2, TIMP-3, TIMP-4), which are endogenous inhibitors of MMPs. It is known to be adjusted. As functions of TIMPs, in addition to forming a complex with active MMP 1: 1 and inhibiting its activity, TIMP-1 is progelatinase B and TIMP-2 is stromal procollagenase or progelatinase A It is known to form a complex with the proenzyme and inhibit the self-activation of these proenzymes. In addition, it has been reported that the activation of MMPs involves membrane-bound MMPs present on the cell surface (Non-patent Document 4).

  Skin aging due to aging and sun exposure increases MMPs activity, which cannot be controlled by TIMPs present in the body, and promotes degradation of the extracellular matrix. In studies using keratinocytes, MMP-9 secretion is induced by UV-B irradiation but not by UV-A (Non-patent Document 5). In studies using cultured fibroblasts, UV- It has been reported that the production of collagenase and stromelysin is enhanced by B irradiation (Non-patent Document 6). Chronic UV radiation has also been reported to induce gelatinase (MMP-2, MMP-9) production in the skin, causing basement membrane damage at the epidermal dermal junction in line with wrinkle formation ( Non-patent documents 2, 3).

  Therefore, in skin aging due to aging and sun exposure, it is considered that MMPs activity exceeds TIMPs activity, and MMPs activity and TIMPs are suppressed by inhibiting MMPs production, inhibiting MMPs activation, promoting TIMPs production or enhancing TIMPs activity. Returning the balance of activity to normal is considered important for preventing skin aging. Further, inhibition of activation of MMPs such as gelatinase by inhibition of membrane-bound MMPs is considered to be effective in normalizing the balance between MMPs activity and TIMPs activity. By the above method, a drug that directly or indirectly inhibits MMPs activity is considered to be useful as an effective anti-aging drug.

Studies have been made so far to find drugs having an inhibitory action on MMPs. For example, flavones, anthocyanidins, catechins, and tannin compounds have been found to have activity (Patent Documents 1 to 5). However, for example, the only catechins that exhibit gelatinase inhibitory activity by reversibly binding to MMP-2 are epicatechin gallate (ECg) and epigalloylcatechin gallate (EGCg), and catechin, epicatechin, epi No inhibitory activity has been found in galloylcatechin, and it has been pointed out that the presence of the galloyl group at position 3 is important for the inhibition of gelatinase activity by ECg and EGCg (Non-patent Document 7). On the other hand, MMP-2 and MMP-9 inhibitory activity has been found in caffeic acid and prenyl ester of caffeic acid (Non-patent Document 8), but MMPs inhibitory activity has not been known so far in polymers of caffeic acid. In addition, since there was no galloyl group in the structure of the caffeic acid polymer, it could not be expected that the caffeic acid polymer exhibited strong MMPs inhibitory activity.
Danichiro, Ichiro, Fragrance Journal, 11-17 (1998) Inomata S. et al., J Inv Dermatol, 120, 1-7 (2003) Jun Amano, Biotherapy, 19, 404-410 (2005) Lewalle JM. Et al., J Cell Physiol, 165, 475-483 (1995) Onoue S. et al., J Dermatol Sci, 33, 105-111 (2003) Brenneisen P. et al., Photochem Photobiol, 64, 877-885 (1996) Cheng XW. Et al., Arch Biochem Biophys, 415, 126-132 (2003) Chung TW. Et al., FASEB J, 18, 1670-1681 (2004) JP-A-8-104628 JP 2000-080035 A JP 2000-226329 A JP 2000-344672 A Japanese Patent Laying-Open No. 2005-075805

  An object of the present invention is to provide an excellent matrix metalloprotease inhibitor and an anti-aging cosmetic comprising the matrix metalloprotease inhibitor as an active ingredient.

The gist of the present invention is as follows.
(1) A matrix metalloproteinase inhibitor containing, as an active ingredient, a cultured cell extract of Murasaki genus plant or a processed product thereof.
(2) A matrix metalloprotease inhibitor containing a caffeic acid polymer as an active ingredient.
(3) In the above (2), the caffeic acid polymer is one or more selected from rosmarinic acid or a salt thereof, lysospermic acid or a salt thereof, lysospermic acid B or a salt thereof, and epilabdocin or a salt thereof. The matrix metalloprotease inhibitor described.
(4) The matrix metalloproteinase inhibitor according to any one of (1) to (3), wherein the matrix metalloprotease inhibitor is a gelatinase inhibitor, a membrane-bound matrix metalloprotease inhibitor, or a TIMPs activity enhancer.
(5) A skin basement membrane degradation inhibitor containing the matrix metalloproteinase inhibitor according to any one of (1) to (4) as an active ingredient.

(6) A laminin degradation inhibitor containing the matrix metalloprotease inhibitor according to any one of (1) to (4) as an active ingredient.
(7) An elastin degradation inhibitor comprising the matrix metalloprotease inhibitor according to any one of (1) to (4) as an active ingredient.
(8) A matrix metalloprotease inhibitor composition containing the following components (A) and (B).
(A) One or more medicinal components selected from the group consisting of a cell extract of a plant belonging to the genus Murasaki, or a processed product thereof, or a caffeic acid polymer (B) a whitening agent, an antioxidant, an anti-inflammatory agent, an antibacterial agent One or more medicinal components selected from the group consisting of agents, cell activators, humectants, and peripheral vascular blood flow promoters (9) The matrix metalloprotease according to any one of (1) to (8) above An anti-aging cosmetic comprising at least one selected from an inhibitor, a matrix metalloproteinase inhibitor composition, a skin basement membrane degradation inhibitor, a laminin degradation inhibitor, and an elastin degradation inhibitor.

  ADVANTAGE OF THE INVENTION According to this invention, the anti-aging cosmetics which use the outstanding matrix metalloprotease inhibitor and this matrix metalloprotease inhibitor as an active ingredient can be provided.

The present invention will be described in detail below.
The method for preparing the extract of the cell extract of the genus Murasaki used in the present invention is not particularly limited, but the cultured cell is established from a part of the plant body such as a leaf, root, or stem of the genus Lithospermum. This cell or culture solution is obtained by drying and then extracting. Examples of the plants of the genus Lithospermum that serve as raw materials for establishing cultured cells include purple (Lithospermum erythrorhizon et. Lithospermum zollingeri A.DC.) and the like. Among them, purple (Lithospermum erythrorhizon et. Zucc.) Is preferable. In addition, the production area of these genus plants is not particularly limited. The roots of purple are called purple roots, and it is known in Kampo that the extract has anti-inflammatory, anti-edema and anti-tumor effects.

  Although it is not particularly limited as a method for culturing cultured cells of the genus Murasaki and the established cultured cells, as an example of preferred culture, callus is aseptically obtained from a part of the tissue of the genus Murasaki, for example, cotyledons, Furthermore, a strain in which the content of p-O-β-D-glucosylbenzoic acid (PHBOG) is maintained at a high level is obtained, and the cells are subcultured every 14 days to proliferate the cells. Examples thereof include a method of obtaining a cell line having a high content of caffeic acid polymer, which is cultured in a caffeic acid polymer production induction medium, for example, a liquid medium of M-9 and does not exhibit a purple color derived from naphthoquinone.

  In preparing an extract from Murasaki plant culture cells, examples of the extraction solvent include water, lower monohydric alcohols (such as methanol, ethanol, 1-propanol, 2-propanol, 1-butanol, and 2-butanol), and liquid polyvalents. Alcohol (glycerin, propylene glycol, 1,3-butylene glycol, etc.), lower alkyl ester (ethyl acetate, etc.), hydrocarbon (benzene, hexane, pentane, etc.), ketones (acetone, methyl ethyl ketone, etc.), ethers (diethyl ether) , Tetrahydrofuran, dipropyl ether, etc.), acetonitrile and the like, and one kind or two or more kinds can be used.

  As an example of a preferable extraction method, ethanol or water having a water content of 0 to 100 vol% is used, and the mixture is extracted at room temperature or heated for 1 to 5 days, followed by filtration, and the obtained filtrate is further added for one week. A method in which the mixture is allowed to stand for aging and then filtered again.

  The extract thus obtained can be used as it is as an active ingredient such as the matrix metalloprotease inhibitor of the present invention. In addition, the extracted extract is treated with various purification means such as ion exchange chromatography, gel filtration chromatography, dialysis, etc., and used as a processed product with further enhanced activity by obtaining an active fraction using matrix metalloprotease inhibitory activity as an index. May be.

  The caffeic acid polymer used in the present invention is a compound containing a structure in which two or more molecules of caffeic acid are polymerized or a salt thereof, and preferred examples thereof include, but are not particularly limited to, for example, rosmarinic acid, lysospermine Examples thereof include acid, lysospermic acid B, epilabdocin, lysospermic acid magnesium salt, lysospermic acid B magnesium salt, lysospermic acid ammonium / potassium salt, and lysospermic acid B calcium / magnesium salt.

  The ratio of the component (A) and the component (B) in the matrix metalloprotease inhibitor composition of the present invention is not particularly limited and can be selected within a wide range, but the component (A ) May be 0.001 to 10% by weight, preferably 0.01 to 5% by weight, and component (B) may be 0.01 to 10% by weight, preferably 0.1 to 5% by weight. .

  The matrix metalloprotease inhibitor of the present invention only needs to inhibit the enzyme activity of the matrix metalloprotease. For example, the gelatinase inhibitor is a compound or composition that selectively exhibits inhibitory activity against gelatinase, and is a membrane-bound type. The matrix metalloprotease inhibitor may be any compound or composition exhibiting an activity that inhibits the enzyme activity of the membrane-bound matrix metalloprotease, and its shape is not particularly limited. The TIMPs activity enhancer is a compound or composition having a function of enhancing the “inhibition of MMP activity by complex formation with MMP”, which is a function of TIMPs, and its shape is not particularly limited.

  In addition to the matrix metalloprotease inhibitor of the present invention, the skin basement membrane degradation inhibitor, laminin degradation inhibitor, elastin degradation inhibitor and cosmetics containing the matrix metalloproteinase inhibitor of the present invention as an active ingredient are usually used for pharmaceuticals and cosmetics. For example, powder components, liquid fats and oils, solid fats and oils, waxes, hydrocarbon oils, higher fatty acids, ester oils, silicone oils, anionic surfactants, cationic surfactants, amphoteric surfactants, nonionic interfaces Activators, water-soluble polymers, thickeners, UV absorbers, sequestering agents, alcohols, sugars, amino acids, organic amines, polymer emulsions, pH adjusters, vitamins, antioxidants, antioxidant aids, A moisturizer, water, whitening agent, anti-inflammatory agent, antibacterial agent, cell activator, peripheral vascular blood flow promoter, etc. are appropriately blended as necessary, Jo, emulsified, solid, powder, paste, film-like, aerosol-like cosmetics, can be prepared by generally known methods depending on the dosage form intended. Specific ingredients that can be formulated are listed below, and the matrix metalloprotease inhibitor of the present invention and any one or more of the following components are blended, and the skin basement membrane degradation inhibitor of the present invention, laminin Degradation inhibitors, elastin degradation inhibitors and cosmetics can be prepared.

  Examples of the powder component include inorganic powders (for example, talc, kaolin, mica, sericite (sericite), muscovite, phlogopite, synthetic mica, saucite, biotite, vermiculite, magnesium carbonate, calcium carbonate, silicic acid. Aluminum, barium silicate, calcium silicate, magnesium silicate, strontium silicate, metal tungstate, magnesium, silica, zeolite, barium sulfate, calcined calcium sulfate (baked gypsum), calcium phosphate, fluorine apatite, hydroxyapatite, ceramic powder Organic powder (eg, polyamide resin powder (nylon powder), polyethylene powder, polymethyl methacrylate powder, polystyrene powder, copolymer resin powder of styrene and acrylic acid, benzoguanamine resin powder, poly-4 Ethylene powder, cellulose powder, metal soap (eg, zinc myristate, calcium palmitate, aluminum stearate); inorganic white pigment (eg, titanium dioxide, zinc oxide, etc.); inorganic red pigment (eg, iron oxide) (Bengara), iron titanate, etc.); inorganic brown pigments (eg, γ-iron oxide, etc.); inorganic yellow pigments (eg, yellow iron oxide, ocher, etc.); inorganic black pigments (eg, black iron oxide, Low-order titanium oxide, etc.); inorganic purple pigments (eg, manganese violet, cobalt violet, etc.); inorganic green pigments (eg, chromium oxide, chromium hydroxide, cobalt titanate, etc.); inorganic blue pigments (eg, ultramarine blue) Pearl pigments (eg, titanium oxide coated mica, titanium oxide coated bismuth oxychloride, titanium oxide coated) Lac, colored titanium oxide coated mica, bismuth oxychloride, fish scale foil, etc.); metal powder pigments (eg, aluminum powder, copper powder, etc.); organic pigments such as zirconium, barium, or aluminum lake (eg, red 201, red 202) No., Red No. 204, Red No. 205, Red No. 220, Red No. 226, Red No. 228, Red No. 405, Orange No. 203, Orange No. 204, Yellow No. 205, Yellow No. 401, Blue No. 404, Red No. 3, Red 104, Red 106, Red 227, Red 230, Red 401, Red 505, Orange 205, Yellow 4, Yellow 5, Yellow 202, Yellow 203, Green 3 and Blue 1 Etc.); natural pigments (for example, chlorophyll, β-carotene, etc.) and the like.

  Examples of liquid oils include avocado oil, camellia oil, turtle oil, macadamia nut oil, corn oil, mink oil, olive oil, rapeseed oil, egg yolk oil, sesame oil, persic oil, wheat germ oil, southern castor oil, castor oil, linseed oil , Safflower oil, cottonseed oil, eno oil, soybean oil, peanut oil, tea seed oil, kaya oil, rice bran oil, cinnagiri oil, Japanese kiri oil, jojoba oil, germ oil, triglycerin and the like.

  Examples of the solid fat include cacao butter, coconut oil, hardened coconut oil, palm oil, palm kernel oil, owl kernel oil, hardened oil, owl, and hardened castor oil.

  As the wax, for example, beeswax, candelilla wax, cotton wax, carnauba wax, bayberry wax, ibota wax, whale wax, montan wax, nuka wax, lanolin, kapok wax, liquid lanolin, sugar cane wax, lanolin fatty acid isopropyl, lauryl hexyl, reduced lanolin, jojoba wax, Examples include hard lanolin, shellac wax, POE lanolin alcohol ether, POE lanolin alcohol acetate, POE cholesterol ether, lanolin fatty acid polyethylene glycol, POE hydrogenated lanolin alcohol ether, and the like.

  Examples of the hydrocarbon oil include liquid paraffin, ozokerite, squalane, pristane, paraffin, ceresin, squalene, petrolatum, microcrystalline wax, and the like.

  Examples of higher fatty acids include lauric acid, myristic acid, palmitic acid, stearic acid, behenic acid, oleic acid, undecylenic acid, tall oil fatty acid, isostearic acid, linoleic acid, linolenic acid, eicosapentaenoic acid (EPA), docosahexaenoic acid (DHA) etc. are mentioned.

  Examples of ester oils include isopropyl myristate, cetyl octanoate, octyldodecyl myristate, isopropyl palmitate, butyl stearate, hexyl laurate, myristyl myristate, decyl oleate, hexyl decyl dimethyloctanoate, cetyl lactate, lactic acid Myristyl, lanolin acetate, isocetyl stearate, isocetyl isostearate, cholesteryl 12-hydroxystearate, ethylene glycol di-2-ethylhexanoate, dipentaerythritol fatty acid ester, n-alkyl glycol monoisostearate, neopentyl glycol dicaprate, Diisostearyl malate, glycerin di-2-heptylundecanoate, trimethylolpropane tri-2-ethylhexanoate, triiso Trimethylolpropane teaate, pentaerythritol tetra-2-ethylhexanoate, glycerin tri-2-ethylhexanoate, glyceryl trioctanoate, glyceryl triisopalmitate, trimethylolpropane triisostearate, cetyl 2-ethylhexanoate, 2-ethylhexyl palmitate, glyceryl trimyristate, tri-2-heptylundecanoic acid glyceride, castor oil fatty acid methyl ester, oleyl oleate, acetoglyceride, 2-heptylundecyl palmitate, diisobutyl adipate, N-lauroyl-L -Glutamic acid-2-octyldodecyl ester, di-2-heptylundecyl adipate, ethyl laurate, di-2-ethylhexyl sebacate, 2-hexyl myristate Rudeshiru, palmitic acid 2-hexyl decyl, 2-hexyldecyl adipate, diisopropyl sebacate, 2-ethylhexyl succinate, and triethyl citrate.

  Examples of the silicone oil include linear polysiloxanes (for example, dimethylpolysiloxane, methylphenylpolysiloxane, diphenylpolysiloxane, etc.); cyclic polysiloxanes (for example, octamethylcyclotetrasiloxane, decamethylcyclopentasiloxane, dodecamethylcyclohexyl). And silicone resins that form a three-dimensional network structure, various modified polysiloxanes (amino-modified polysiloxane, polyether-modified polysiloxane, alkyl-modified polysiloxane, fluorine-modified polysiloxane, etc.) It is done.

  Anionic surfactants include, for example, fatty acid soaps (eg, sodium laurate, sodium palmitate, etc.); higher alkyl sulfates (eg, sodium lauryl sulfate, potassium lauryl sulfate, etc.); alkyl ether sulfates (eg, POE-lauryl sulfate triethanolamine, POE-sodium lauryl sulfate, etc.); N-acyl sarcosine acid (eg, sodium lauroyl sarcosine, etc.); higher fatty acid amide sulfonates (eg, sodium N-myristoyl-N-methyl taurate, palm Oil fatty acid methyl taurine sodium, lauryl methyl taurine sodium, etc .; Phosphate ester salt (POE-oleyl ether sodium phosphate, POE-stearyl ether phosphate, etc.); Sulfosuccinate (for example, Sodium di-2-ethylhexyl sulfosuccinate, sodium monolauroyl monoethanolamide polyoxyethylene sodium sulfosuccinate, sodium lauryl polypropylene glycol sulfosuccinate, etc .; alkyl benzene sulfonates (eg, sodium linear dodecyl benzene sulfonate, triso linear dodecyl benzene sulfonate) Ethanolamine, linear dodecyl benzene sulfonic acid, etc.); higher fatty acid ester sulfates (for example, hydrogenated coconut oil fatty acid sodium glycerin sulfate, etc.); N-acyl glutamate (for example, monosodium N-lauroyl glutamate, di-N-stearoyl glutamate) Sodium, N-myristoyl-L-monosodium glutamate, etc.); sulfated oil (eg funnel oil); POE-A Kill ether carboxylic acid; POE-alkyl allyl ether carboxylate; α-olefin sulfonate; higher fatty acid ester sulfonate; secondary alcohol sulfate ester; higher fatty acid alkylolamide sulfate ester; lauroyl monoethanolamide succinic acid Sodium; N-palmitoyl aspartate ditriethanolamine; sodium caseinate and the like.

  Examples of the cationic surfactant include alkyltrimethylammonium salts (eg, stearyltrimethylammonium chloride, lauryltrimethylammonium chloride, etc.); alkylpyridinium salts (eg, cetylpyridinium chloride, etc.); distearyldimethylammonium dialkyldimethylammonium chloride; Poly (N, N′-dimethyl-3,5-methylenepiperidinium chloride); alkyl quaternary ammonium salt; alkyldimethylbenzylammonium salt; alkylisoquinolinium salt; dialkyl morpholinium salt; POE-alkylamine; Examples include alkylamine salts; polyamine fatty acid derivatives; amyl alcohol fatty acid derivatives; benzalkonium chloride; benzethonium chloride and the like.

  Examples of amphoteric surfactants include imidazoline-based amphoteric surfactants (eg, 2-undecyl-N, N, N- (hydroxyethylcarboxymethyl) -2-imidazoline sodium, 2-cocoyl-2-imidazolinium hydroxide). Side-1-carboxyethyloxy disodium salt, etc.); betaine surfactants (for example, 2-heptadecyl-N-carboxymethyl-N-hydroxyethylimidazolinium betaine, lauryldimethylaminoacetic acid betaine, alkylbetaine, amide betaine) , Sulfobetaine, etc.).

  Examples of the lipophilic nonionic surfactant include sorbitan fatty acid esters (for example, sorbitan monooleate, sorbitan monoisostearate, sorbitan monolaurate, sorbitan monopalmitate, sorbitan monostearate, sorbitan sesquioleate, Sorbitan trioleate, diglycerol sorbitan penta-2-ethylhexylate, diglycerol sorbitan tetra-2-ethylhexyl); glycerin polyglycerin fatty acids (for example, mono-cotton oil fatty acid glycerin, mono-erucic acid glycerin, sesquioleate glycerin, monostearin) Glycerin acid, α, α′-oleic acid pyroglutamate glycerin, monostearate glycerin malate, etc.); propylene glycol fatty acid esters (for example, And propylene glycol monostearate); hardened castor oil derivatives; glycerin alkyl ethers and the like.

  Examples of the hydrophilic nonionic surfactant include POE-sorbitan fatty acid esters (for example, POE-sorbitan monooleate, POE-sorbitan monostearate, POE-sorbitan monooleate, POE-sorbitan tetraoleate, etc.); POE sorbite fatty acid esters (eg, POE-sorbite monolaurate, POE-sorbite monooleate, POE-sorbite pentaoleate, POE-sorbite monostearate, etc.); POE-glycerin fatty acid esters (eg, POE-glycerin) Monostearate, POE-glycerol monoisostearate, POE-monooleate such as POE-glycerol triisostearate, etc.); POE-fatty acid esters (eg, POE-distearate, PO Monodiolates, ethylene glycol distearate, etc.); POE-alkyl ethers (eg POE-lauryl ether, POE-oleyl ether, POE-stearyl ether, POE-behenyl ether, POE-2-octyldecyl ether, POE-cholestanol) Ethers, etc.); Pluronic types (for example, Pluronic etc.); POE • POP-alkyl ethers (for example, POE • POP-cetyl ether, POE • POP-2-decyltetradecyl ether, POE • POP-monobutyl ether, POE) POP-hydrogenated lanolin, POE / POP-glycerin ether, etc.); Tetra POE / tetra-POP-ethylenediamine condensates (eg, Tetronic, etc.); POE-castor oil hardened castor oil derivative ( For example, POE-castor oil, POE-hardened castor oil, POE-hardened castor oil monoisostearate, POE-hardened castor oil triisostearate, POE-hardened castor oil monopyroglutamic acid monoisostearic acid diester, POE-hardened castor Oil maleic acid, etc.); POE-beeswax lanolin derivatives (eg POE-sorbite beeswax etc.); alkanolamides (eg coconut oil fatty acid diethanolamide, lauric acid monoethanolamide, fatty acid isopropanolamide etc.); POE-propylene glycol fatty acid Examples include esters; POE-alkylamines; POE-fatty acid amides; sucrose fatty acid esters; alkyl ethoxydimethylamine oxides;

  Examples of natural water-soluble polymers include plant-based polymers (for example, gum arabic, gum tragacanth, galactan, guar gum, carob gum, caraya gum, carrageenan, pectin, agar, quince seed (malmello), alge colloid (guckweed extract), starch (Rice, corn, potato, wheat), glycyrrhizic acid); microbial polymer (eg, xanthan gum, dextran, succinoglucan, pullulan, etc.); animal polymer (eg, collagen, casein, albumin, gelatin, etc.), etc. Is mentioned.

  Semi-synthetic water-soluble polymers include, for example, starch polymers (eg, carboxymethyl starch, methylhydroxypropyl starch, etc.); cellulose polymers (methylcellulose, ethylcellulose, methylhydroxypropylcellulose, hydroxyethylcellulose, sodium cellulose sulfate) Hydroxypropylcellulose, carboxymethylcellulose, sodium carboxymethylcellulose, crystalline cellulose, cellulose powder and the like); alginic acid polymers (for example, sodium alginate, propylene glycol alginate, etc.) and the like.

  Synthetic water-soluble polymers include, for example, vinyl polymers (eg, polyvinyl alcohol, polyvinyl methyl ether, polyvinyl pyrrolidone, carboxyvinyl polymer); polyoxyethylene polymers (eg, polyethylene glycol 20,000, 40). , 000, 60,000 polyoxyethylene polyoxypropylene copolymer, etc.); acrylic polymers (for example, sodium polyacrylate, polyethyl acrylate, polyacrylamide, etc.); polyethyleneimine; cationic polymers, and the like.

  Examples of the thickener include gum arabic, carrageenan, caraya gum, gum tragacanth, carob gum, quince seed (malmello), casein, dextrin, gelatin, sodium pectate, sodium alginate, methylcellulose, ethylcellulose, CMC, hydroxyethylcellulose, hydroxypropyl Cellulose, PVA, PVM, PVP, sodium polyacrylate, carboxyvinyl polymer, locust bean gum, guar gum, tamarind gum, cellulose dialkyldimethylammonium sulfate, xanthan gum, magnesium aluminum silicate, bentonite, hectorite, AlMg silicate (Beegum), Examples thereof include laponite and silicic anhydride.

Examples of the ultraviolet absorber include the following compounds.
(1) Benzoic acid ultraviolet absorbers, for example, paraaminobenzoic acid (hereinafter abbreviated as PABA), PABA monoglycerin ester, N, N-dipropoxy PABA ethyl ester, N, N-diethoxy PABA ethyl ester, N, N-dimethyl PABA ethyl ester, N, N-dimethyl PABA butyl ester, N, N-dimethyl PABA ethyl ester and the like.
(2) Anthranilic acid-based ultraviolet absorbers such as homomenthyl-N-acetylanthranilate.
(3) Salicylic acid ultraviolet absorbers such as amyl salicylate, menthyl salicylate, homomenthyl salicylate, octyl salicylate, phenyl salicylate, benzyl salicylate, p-isopropanol phenyl salicylate, and the like.

(4) Cinnamic acid-based ultraviolet absorbers For example, octyl cinnamate, ethyl-4-isopropyl cinnamate, methyl-2,5-diisopropyl cinnamate, ethyl-2,4-diisopropyl cinnamate, methyl-2,4- Diisopropyl cinnamate, propyl-p-methoxycinnamate, isopropyl-p-methoxycinnamate, isoamyl-p-methoxycinnamate, octyl-p-methoxycinnamate (2-ethylhexyl-p-methoxycinnamate), 2-ethoxy Ethyl-p-methoxycinnamate, cyclohexyl-p-methoxycinnamate, ethyl-α-cyano-β-phenylcinnamate, 2-ethylhexyl-α-cyano-β-phenylcinnamate, glyceryl mono-2-ethylhexanoyl -Diparamet Such as Shishin'nameto.
(5) Triazine-based ultraviolet absorber, for example, bisresorcinyl triazine.
More specifically, bis {[4- (2-ethylhexyloxy) -2-hydroxy] phenyl} -6- (4-methoxyphenyl) 1,3,5-triazine, 2,4,6-tris { 4- (2-ethylhexyloxycarbonyl) anilino} 1,3,5-triazine and the like.
(6) Other ultraviolet absorbers For example, 3- (4′-methylbenzylidene) -d, l-camphor, 3-benzylidene-d, l-camphor, 2-phenyl-5-methylbenzoxazole, 2-hydroxy -4-methoxyphenylbenzotriazole, 2- (2'-hydroxy-5'-t-octylphenyl) benzotriazole, 2- (2-hydroxy-5-methylphenyl) benzotriazole, dibenzalazine, dianisoylmethane, 4- Pyridazine derivatives such as methoxy-4′-t-butyldibenzoylmethane, 5- (3,3-dimethyl-2-norbornylidene) -3-pentan-2-one, dimorpholinopyridazinone and the like.

  Examples of the sequestering agent include 1-hydroxyethane-1,1-diphosphonic acid, 1-hydroxyethane-1,1-diphosphonic acid tetrasodium salt, disodium edetate, trisodium edetate, and tetrasodium edetate. Sodium citrate, sodium polyphosphate, sodium metaphosphate, gluconic acid, phosphoric acid, citric acid, ascorbic acid, succinic acid, edetic acid, trisodium ethylenediaminehydroxyethyl triacetate and the like.

  Examples of the lower alcohol include ethanol, propanol, isopropanol, isobutyl alcohol, t-butyl alcohol and the like.

  Examples of the polyhydric alcohol include dihydric alcohols (for example, ethylene glycol, propylene glycol, trimethylene glycol, 1,2-butylene glycol, 1,3-butylene glycol, tetramethylene glycol, 2,3-butylene glycol, penta Methylene glycol, 2-butene-1,4-diol, hexylene glycol, octylene glycol, etc.); trihydric alcohols (eg, glycerin, trimethylolpropane, 1,2,6-hexanetriol, etc.); For example, pentaerythritol etc .; pentavalent alcohol (eg xylitol etc.); hexavalent alcohol (eg sorbitol, mannitol etc.); polyhydric alcohol polymer (eg diethylene glycol, dipropylene glycol, triethyl) Glycol, polypropylene glycol, tetraethylene glycol, diglycerin, polyethylene glycol, triglycerin, tetraglycerin, polyglycerin, etc.); divalent alcohol ethers (for example, ethylene glycol monomethyl ether, ethylene glycol monoethyl ether, ethylene glycol monobutyl ether) , Ethylene glycol monophenyl ether, ethylene glycol monohexyl ether, ethylene glycol mono 2-methylhexyl ether, ethylene glycol isoamyl ether, ethylene glycol benzyl ether, ethylene glycol isopropyl ether, ethylene glycol dimethyl ether, ethylene glycol diethyl ether, ethylene glycol dibutyl ether , Propi Glycol monomethyl ether, propylene glycol monoethyl ether, propylene glycol monobutyl ether, propylene glycol isopropyl ether, etc.); polyhydric alcohol polymer alkyl ethers (for example, diethylene glycol monomethyl ether, diethylene glycol monoethyl ether, diethylene glycol monobutyl ether, diethylene glycol dimethyl ether, Diethylene glycol diethyl ether, diethylene glycol butyl ether, diethylene glycol methyl ethyl ether, triethylene glycol monomethyl ether, triethylene glycol monoethyl ether, dipropylene glycol methyl ether, dipropylene glycol ethyl ether, dipropylene glycol buty Dihydric alcohol ether ester (for example, ethylene glycol monomethyl ether acetate, ethylene glycol monoethyl ether acetate, ethylene glycol monobutyl ether acetate, ethylene glycol monophenyl ether acetate, diethylene glycol monoethyl ether acetate, diethylene glycol monobutyl ether acetate, propylene) Glycol monomethyl ether acetate, propylene glycol monoethyl ether acetate, propylene glycol monopropyl ether acetate, propylene glycol monophenyl ether acetate, etc.); glycerin monoalkyl ether (eg, chimyl alcohol, ceralkyl alcohol, batyl alcohol, etc.); sugar alcohol (Eg, sorbitol, maltitol, maltotriose, mannitol, sucrose, erythritol, glucose, fructose, amylolytic sugar, maltose, xylitol, amylolytic sugar reducing alcohol, etc.); glycolide; tetrahydrofurfuryl alcohol; POE -Tetrahydrofurfuryl alcohol; POP-butyl ether; POP / POE-butyl ether; Tripolyoxypropylene glycerin ether; POP-glycerin ether; POP-glycerin ether phosphate; POP / POE-pentane erythritol ether, polyglycerin and the like.

  Examples of higher alcohols include linear alcohols (eg, lauryl alcohol, cetyl alcohol, stearyl alcohol, behenyl alcohol, myristyl alcohol, oleyl alcohol, cetostearyl alcohol); branched chain alcohols (eg, monostearyl glycerin ether (batyl alcohol) ), 2-decyltetradecinol, lanolin alcohol, cholesterol, phytosterol, hexyldodecanol, isostearyl alcohol, octyldodecanol and the like.

  Examples of monosaccharides include tricarbon sugars (eg, D-glyceryl aldehyde, dihydroxyacetone, etc.); tetracarbon sugars (eg, D-erythrose, D-erythrulose, D-treose, erythritol, etc.); pentose sugars (eg, L-arabinose, D-xylose, L-lyxose, D-arabinose, D-ribose, D-ribulose, D-xylulose, L-xylulose, etc .; hexose (eg, D-glucose, D-talose, D) -Bucicose, D-galactose, D-fructose, L-galactose, L-mannose, D-tagatose, etc.); pentose sugar (eg, aldoheptose, heproose, etc.); For example, 2-deoxy-D-ribose, 6-deoxy-L-galactose, 6-deoxy-L Mannose, etc.]; amino sugars (eg, D-glucosamine, D-galactosamine, sialic acid, aminouronic acid, muramic acid, etc.); uronic acids (eg, D-glucuronic acid, D-mannuronic acid, L-guluronic acid, D- Galacturonic acid, L-iduronic acid, etc.).

  Examples of the oligosaccharide include sucrose, umbelliferose, lactose, planteose, isolicnose, α, α-trehalose, raffinose, lycnose, umbilicin, stachyose verbus course, and the like.

  Examples of the polysaccharide include cellulose, quince seed, chondroitin sulfate, starch, galactan, dermatan sulfate, glycogen, gum arabic, heparan sulfate, hyaluronic acid, tragacanth gum, keratan sulfate, chondroitin, xanthan gum, mucoitin sulfate, guar gum, dextran, kerato sulfate. , Locust bean gum, succinoglucan, caronic acid and the like.

  Examples of amino acids include neutral amino acids (eg, threonine, cysteine, etc.); basic amino acids (eg, hydroxylysine, etc.) and the like. Examples of the amino acid derivative include acyl sarcosine sodium (lauroyl sarcosine sodium), acyl glutamate, acyl β-alanine sodium, glutathione, and pyrrolidone carboxylic acid.

  Examples of the organic amine include monoethanolamine, diethanolamine, triethanolamine, morpholine, triisopropanolamine, 2-amino-2-methyl-1,3-propanediol, 2-amino-2-methyl-1-propanol, and the like. Is mentioned.

  Examples of the polymer emulsion include an acrylic resin emulsion, a polyethyl acrylate emulsion, an acrylic resin liquid, a polyacryl alkyl ester emulsion, a polyvinyl acetate resin emulsion, and a natural rubber latex.

  Examples of the pH adjuster include buffers such as lactic acid-sodium lactate, citric acid-sodium citrate, and succinic acid-sodium succinate.

  Examples of vitamins include vitamin A, B1, B2, B6, C, E and derivatives thereof, pantothenic acid and derivatives thereof, biotin and the like.

  Antioxidants include, for example, vitamin E and its derivatives (dl-α (β, γ) -tocopherol, dl-α-tocopherol acetate, nicotinic acid-dl-α-tocopherol, linoleic acid-dl-α-tocopherol, Tocopherols such as dl-α-tocopherol succinate and derivatives thereof, ubiquinones, etc., vitamin A and derivatives thereof (retinol palmitate, retinol acetate such as retinol and derivatives thereof, retinal such as dehydroretinal and derivatives thereof), carotenoids (Carotene, lycopene, astaxanthin, etc.), vitamin B and its derivatives (thiamine hydrochloride, thiamine sulfate, riboflavin, riboflavin acetate, pyridoxine hydrochloride, pyridoxine dioctanoate, flavin adenine dinucleotide, cyanocobalamin, leaf Nicotinic acid amide, nicotinic acid such as benzyl nicotinate, choline, etc.), vitamin C and its derivatives (L-ascorbic acid alkyl ester such as L-ascorbyl dipalmitate and L-ascorbyl tetraisopalmitate, L- Ascorbic acid phosphate, L-ascorbic acid sulfate, etc.), vitamin D and its derivatives (ergocalciferol, cholecalciferol, dihydroxystannal etc.), rutin and its derivatives, thiotaurine, taurine, hydroquinone and its derivatives, histidine Catechin and its derivatives, grabrizine, glabrene, liquiritin, isoliquiritin and licorice extract containing these, glutathione and its derivatives, dibutylhydroxytoluene, butylhydroxyanisole, Gallic acid and its derivatives, cholesterol and its derivatives, superoxide dismutase, mannitol, cucumber extract, quette extract, gentian (gentian) extract, gentian extract, hawthorn extract, peony extract, ginkgo biloba extract, koganebana ( (Ougon) extract, carrot extract, maika (maika, hermanus) extract, sunpens (kawara ketsumei) extract, tormentilla extract, parsley extract, grape extract, button (button pi) extract, mokka (bokeh) extract , Melissa extract, Yashajitsu (Yasha) extract, Yukinoshita extract, Rosemary (mannenrou) extract, lettuce extract, tea extract (Oolong tea, black tea, green tea, etc.), microbial fermentation metabolites, seaweed extract, spirit Turf extract, eggshell membrane extract, placenta Distillate include swingle extract or the like. (In parentheses, plant aliases and herbal medicine names are listed.)

  Examples of the antioxidant assistant include phosphoric acid, citric acid, ascorbic acid, maleic acid, malonic acid, succinic acid, fumaric acid, kephalin, hexametaphosphate, phytic acid, and ethylenediaminetetraacetic acid.

  Examples of humectants include polyethylene glycol, propylene glycol, glycerin, 1,3-butylene glycol, xylitol, sorbitol, maltitol, chondroitin sulfate, hyaluronic acid, mucoitin sulfate, caronic acid, atelocollagen, cholesteryl-12-hydroxystearate, and lactic acid. Examples thereof include sodium, bile salt, dl-pyrrolidone carboxylate, short-chain soluble collagen, diglycerin (EO) PO adduct, Izayoi rose extract, yarrow extract, and mellote extract.

  As a whitening agent, vitamin C and its derivatives (L-ascorbic acid alkyl esters such as L-ascorbyl dipalmitate, L-ascorbyl tetraisopalmitate, L-ascorbic acid phosphate, L-ascorbic acid sulfate, etc.) , Placenta extract, glabrizine, glabrene, liquiritin, isoliquiritin and licorice extract, yokuinin (barley) extract, koganebana (ogon) extract, seaweed extract (comb, macombu, wakame, hijiki, hibamata, sujime Brown algae, such as Tuna, Tuna, Tsuruarame, Chigaiso, Honda Rawa, Giant Kelp; Tengusa, Tsunomatsu, Sugiori, Usubanori, Asakusanori, Matsunori, Tosakamatsu, Funori, Ogonori, Kamensou, Igis, Egori, etc. Algae: Chlorella, Aonori, Donariella, Chlorococcus, Anaaaosa, Kawanoori, Marimo, Shiogusa, Casanori, Futojuzumo, Tamajuzumo, Aegusa, Aomidoro and other green algae; Spirulina and other cyanobacteria, etc.), biclen extract, grape extract, grape extract , Wheat extract, tomato extract, carotenoids (carotene, lycopene, astaxanthin, etc.), agarose, oligosaccharides, neoagarobiose, hydroquinone and its derivatives, cysteine and its derivatives, asparagus extract, ibukitorano extract, Neurose extract, Ezocogi extract, Pea extract, Chamomile extract, Caquette extract, Orange extract, Raspberry extract, Kiwi extract, Clara extract, Coffee extract, Gogo Oil, Sesame oil, Sesame extract, Rice extract, Rice extract, Saisin extract, Hawthorn extract, Sunpens extract, Peonies extract, Shirayuri extract, Mulberry extract, Toki extract , Beech extract, beech sprout extract, black currant extract, hop extract, maika (maikai, hamanasu) extract, mokka (bokeh) extract, yukinoshita extract, tea extract (Oolong tea, black tea, green tea, etc. ), Ganoderma extract, microbial fermentation metabolite, soybean extract, molasses extract, Rakan fruit extract and the like. (In parentheses, plant aliases and herbal medicine names are listed.)

  Anti-inflammatory agents include glycyrrhizic acid and its derivatives, glycyrrhetinic acid and its derivatives, vitamin B and its derivatives (thiamine hydrochloride, thiamine sulfate, riboflavin, riboflavin acetate, pyridoxine hydrochloride, pyridoxine dioctanoate, flavin adenine dinucleotide , Nicotinic acid such as cyanocobalamin, folic acid, nicotinic acid amide, benzyl nicotinate, choline, tranexamic acid, thiotaurine, hypotaurine, aloe extract, ashitaba extract, altea extract, arnica extract, sulfur and its derivatives, nettle Extract, Ginseng extract, Turmeric extract, Yellowfin extract, Hypericum extract, Chamomile extract, Comfrey extract, Honeysuckle extract, Cucumber extract Song extract, Salvia (sage) extract, Walnut extract (Juyu) extract, Perilla extract, Birch extract, Elderberry extract, Grape extract, Mulberry extract, Eucalyptus extract, Artemisia extract, Astragalus extract Examples include extracts, chondroitin sulfate and derivatives thereof, zinc oxide, etc. (In parentheses, plant aliases, herbal medicine names, etc. are described.)

  Examples of the antibacterial agent include methyl paraben, ethyl paraben, butyl paraben, phenoxyethanol and the like.

  Examples of cell activators include carotenoids (such as carotene, lycopene, astaxanthin), vitamin A and derivatives thereof (retinol such as retinol palmitate and retinol acetate; and retinals such as dehydroretinal and derivatives thereof), vitamin C and its derivatives. Derivatives (L-ascorbic acid alkyl esters such as L-ascorbyl dipalmitate and L-ascorbyl tetraisopalmitate, L-ascorbic acid phosphate, L-ascorbic acid sulfate, etc.), vitamin B and its derivatives (thiamine hydrochloride) Salt, thiamine sulfate, riboflavin, riboflavin acetate, pyridoxine hydrochloride, pyridoxine dioctanoate, flavin adenine dinucleotide, cyanocobalamin, folic acid, nicotinamide and benzyl nicotinate (Tinic acids, choline, etc.), ribonucleic acid and salts thereof, deoxyribonucleic acid and salts thereof, α- and γ-linolenic acid, xanthine and derivatives thereof (caffeine, etc.), amino acids and derivatives thereof (serine, glutamic acid, theanine, hydroxy) Proline, pyrrolidone carboxylic acid, etc.), docosahexaenoic acid and its derivatives, eicosapentaenoic acid and its derivatives, citric acid, lactic acid, malic acid, succinic acid, almond extract, asparagus extract, apricot extract, ginkgo biloba extract , Kihada extract, barley extract, kiwi extract, cucumber extract, shiitake extract, cedar extract, assembly extract, soybean extract, jujube extract, clover extract , Pepper extract, red pepper extract, tomato extract, Garlic extract, carrot extract, hinokitiol, beech extract, grape seed oil, beech extract, beech bud extract, peach extract, eucalyptus extract, lily extract, lettuce extract, lemon extract, rosemary (Mannenrou) extract, malt root extract, animal-derived extract (mollusk and other mollusc extracts, shell extract, shellfish extract, fish extract, chicken crown extract, royal jelly, silk protein and its degradation product, placenta Extract, serum deproteinized extract, lactoferrin or degradation product thereof), yeast extract, microbial fermentation metabolite (derived from lactic acid bacteria, bifidobacteria, etc.), ganoderma extract and the like. (In parentheses, plant aliases and herbal medicine names are listed.)

  Examples of peripheral vascular blood flow promoters include vitamin E and its derivatives, assembly extract, garlic extract, ginger extract, red pepper extract, rosemary extract and the like.

  The matrix metalloprotease inhibitor of the present invention and one or more components described above can be combined to provide a multifunctional cosmetic.

  When the matrix metalloproteinase inhibitor of the present invention is blended in cosmetics or quasi drugs, the blended amount of the cultured cell extract of Murasaki spp. Or its processed product is solid relative to the total amount of cosmetics or quasi drugs. In terms of product, it is usually 0.001 to 10% by weight, preferably 0.01 to 5% by weight. The blending amount of the caffeic acid polymer is usually 0. 0% relative to the total amount of the cosmetic or quasi-drug. It is 0001 to 5% by weight, preferably 0.001 to 2% by weight.

  Hereinafter, although a reference example, an example, and a comparative example are given and the present invention is explained in detail, the present invention is not limited to these examples.

[Reference Example 1] Preparation of Murasaki cultured cell extract from Lithospermum erythrorhizon-derived cultured cells Murasaki's sterilized sterilized with 2% antiformin solution or 70% ethanol solution in advance on Linsmeier Skoog (LS) agar solid medium. A cotyledon tissue piece was placed and statically cultured in a dark place at 25 ° C. to obtain purple callus. By repeating the passage, the M-18TOM strain in which the content of p-O-β-D-glucosylbenzoic acid (PHBOG) was maintained high was obtained from a number of flasks. Next, 1 g (fresh weight) of callus of M-18TOM strain was transferred to a liquid culture medium of LS (containing 1 μM indoleacetic acid and 10 μM kinetin as plant hormones and 30 g / l sucrose as a carbon source) in a 20 ml Erlenmeyer flask. Rotating culture (amplitude 25 mm, 100 rpm) on a shaker and subcultured every 14 days to increase the growth rate of callus. In a subculture process by liquid culture, when cultivated experimentally in a liquid medium of M-9, there are a flask containing cultured cells that produce shikonin and a flask containing cultured cells that are not produced. The 18TOM strain and the cultured cells that were not produced were designated as the WM18 strain. Furthermore, while subcultured the M-18TOM strain, a cultured cell line that did not produce shikonin was isolated, and the newly obtained strain that did not produce shikonin was named TomK2.

  The TomK2 strain cultured cells thus obtained were transferred to a culture tank containing 150 L of an M-9 liquid medium and cultured with aeration and stirring at 25 ° C. for 21 days. After completion of the culture, the cultured cells and the culture solution were separated by filtration, and the obtained cultured cells were dried by ventilation at 60 ° C. To 100 g of dried Lithospermum erythrorhizon cultured cells (TomK2 strain), add 1 L of 80 vol% ethanol solution, extract at room temperature for 3 days, filter, and evaporate the solvent under reduced pressure to obtain a Murasaki cultured cell extract. Obtained.

Reference Example 2 Obtaining Caffeic Acid Polymer Murasaki cultured cell extract was subjected to low pressure column chromatography (column: φ45 × 500 mm, packing material: Wakosil 25C18 (15-30 μm), solvent: 30% aqueous methanol solution to 100% methanol. Linear gradient), fractionate the peak corresponding to each caffeic acid polymer compound (rosmarinic acid, lysospermic acid, lysopermic acid B, epilabdocin), and distill off the solvent of each fraction under reduced pressure. The desired caffeic acid polymer was obtained.

[Example 1]
9-10 weeks old, 200-220 g Wistar male rats at 25 ° C., 55 ± 5% constant temperature and humidity, light / dark 12 hours / 12 hours, solid feed (CE-2, Claire Japan) And drinking tap water freely. The lung tissue was excised from the rat, homogenized at 4 ° C. with 5 volumes of Tris-HCl buffer (50 mM, pH 7.5), and centrifuged (9000 g, 20 minutes, 4 ° C.) after supernatant. The fraction was used as a matrix metalloproteinase (MMP) enzyme source. The protein concentration was adjusted to 1.0 mg / ml using Coomassie Brilliant Blue R250, and then incubated with 0.5 mM APMA solution (50 mM Tris-HCl, 50 mM sodium chloride, pH 7.5) for 1 hour at 37 ° C. Was activated (activated MMP enzyme solution). To the activated MMP enzyme solution, lysospermic acid having a final concentration of 0.25 to 100 μM dissolved in 50 mM Tris-HCl buffer (pH 7.5) was added as a test substance, and further incubated at 37 ° C. for 1 hour. Gelatinase inhibitory activity was measured by gelatin gel zymography using as a substrate. Gelatin zymography was performed after separation of MMP groups by non-reducing SDS-PAGE electrophoresis. That is, a test solution (including an activated MMP enzyme solution) incubated with lysospermic acid was dissolved in Laemmli sample buffer excluding 2-mercaptoethanol, and 7.5% containing 1.0 mg / ml gelatin. The gel was applied to an acrylamide / bisacrylamide (29: 1) separation gel and subjected to SDS-PAGE at 200 V for 40 minutes in a Mini-Protein III apparatus (Bio-Rad). Myosin (200 kDa), β-galactosidase (116 kDa), bovine serum albumin (66 kDa) were used as molecular weight markers. After electrophoresis, the gel was washed with 0.25% Triton X-100 for 15 minutes at room temperature, twice for 10 minutes with water, and incubated with buffer (50 mM Tris-HCl (pH 7.6), 20 mM sodium chloride, 5 mM calcium chloride, 0.2%). Incubation was carried out at 37 ° C. for 18 hours in 02% Brij-58). Gelatin digestion traces observed after incubation were visualized by staining with rapid stain Coomassie Brilliant Blue kit (Nacalai Tesque). In the zymography method, the clear band observed after gel staining is consistent with the gelatinolytic activity of MMP. These transparent bands were identified as 66 kDa active MMP-2, 72 kDa latent MMP-2, 87 kDa active MMP-9, and 92 kDa latent MMP-9, respectively. The digestion trace by gelatin zymography is taken into a PC with a scanner (EPSON ES-2200), quantified by a quantitative image analysis method (L Process V2.0 and Image Gauge V4.0, FUJI PHOTOFILM Co Ltd.), and Sigma Stat statistical Statistical processing was performed using software ver.2.03 (SPSS Inc., IL, USA). Each measured value was displayed as an average value, and a significant difference was examined by a Kluskal-Wallis ANOVA rank test (p <0.05). FIG. 1 shows the results of inhibition of gelatin digestion of lysospermic acid against active MMP-2. In addition, Table 1 shows IC ₅₀ values obtained from FIG.

[Comparative Example 1]
The same procedure as described in Example 1 was performed except that the test substance was caffeic acid. The results are shown in Table 1.

[Example 2]
The same procedure as described in Example 1 was performed except that the test substance was rosmarinic acid, lysospermic acid B or epilabdocin. The results are shown in Table 1.

[Comparative Example 2]
The test was carried out in the same manner as in Example 1 except that the test substance was flavones nobiletin, heptamethoxyflavone, baicalein or baicalin. The results are shown in Table 1.

[Comparative Example 3]
The test substance was carried out in the same manner as described in Example 1 except that flavonols ougonin, kaempferol, quercetin or rutin were used. The results are shown in Table 1.

[Comparative Example 4]
The test was carried out in the same manner as in Example 1 except that the test substance was naringenin, naringin, hesperetin or hesperidin, which are flavanones. The results are shown in Table 1.

As is apparent from the results of FIG. 1 and Table 1, the caffeic acid polymer is MMP in comparison with caffeic acid (Comparative Example 1), flavones, flavonols and flavanones which have been reported to inhibit MMPs so far. -2 gelatin digestion activity was inhibited at a very low concentration.
[Reference Example 3] Preparation of purple root extract
30 g of Lithospermum erythrorhizon root (purple root, Tochimoto Tenkaido) was repeatedly refluxed and cooled with 100 mL of water for 1 hour three times. After filtration with a cotton plug, the water was distilled off under reduced pressure to obtain 7.15 g of a purple root extract.

[Example 3]
The same procedure as described in Example 1 was performed except that the test substance was a Murasaki cultured cell extract with a final concentration of 26.7 μg / ml to 1.67 mg / ml. The results of IC ₅₀ values are shown in Table 2.

[Comparative Example 5]
The same procedure as described in Example 3 was performed except that the test substance was a purple root extract. The results are shown in Table 2.

[Example 4]
The same procedure as described in Example 3 was performed except that the activity was that of inhibiting gelatin digestion against active MMP-9. The results are shown in Table 3.

[Comparative Example 6]
The same procedure as described in Example 4 was performed except that the test substance was a purple root extract. The results are shown in Table 3.

  As is clear from the results of Tables 2 and 3, the extract of Murasaki cultured cells showed a high inhibitory activity on MMP-2 and MMP-9 activities.

[Example 5]
As an MMP enzyme source, rat lung tissue was homogenized with 5 volumes of Tris-HCl buffer (50 mM, pH 7.5, 4 ° C.) to obtain a supernatant fraction (9000 × g, 4 ° C., 20 minutes). . This was incubated with a 0.5 mM APMA solution (50 mM Tris-HCl, 50 mM sodium chloride, pH 7.5) for 1 hour at 37 ° C. to activate the MMP group. The effect on the activated MMP group was as follows: lysospermic acid having a final concentration of 1 to 100 μM was added, followed by further incubation at 37 ° C. for 1 hour, 1.0 mg / mL gelatin, 160 ng / mL MMP-2 (TRIPLE POINT BIOLOGICS, Inc. .) Was applied to a 12.5% acrylamide / bisacrylamide (29: 1) separation gel, and SDS-PAGE was performed at 200 V for 40 minutes using a Mini-Protein III apparatus (Bio-Rad) under non-reducing conditions. . Myosin (200 kDa), β-galactosidase (116 kDa), bovine serum albumin (66 kDa) were used as molecular weight markers. After electrophoresis, the gel was washed with 2.5% Triton X-100 for 30 minutes at room temperature, twice for 10 minutes with water, and incubated in incubation buffer (50 mM Tris-HCl (pH 7.6), 20 mM sodium chloride, 5 mM calcium chloride). Incubated at 37 ° C. for 18 hours. Protein bands observed after incubation were visualized by staining with rapid stain Coomassie Brilliant Blue kit (Nacalai Tesque). In the reverse zymography method, gelatin contained in the gel is completely decomposed by MMP-2 contained at the same time. Therefore, the remaining protein is stained and confirmed as a band when MMP-2 activity is inhibited. A band with a molecular weight of 21 kDa was identified as TIMP-2 from these stained bands. This band was taken into a PC with a scanner (EPSON ES-2200) and quantified by a quantitative image analysis method (L Process V2.0 and Image Gauge V4.0, FUJI PHOTOFILM Co, Ltd.). The results are shown in FIG.

[Comparative Example 7]
The same procedure as described in Example 5 was performed except that the MMP group was not activated by APMA and lysospermic acid was not added. The results are shown in FIG.

[Comparative Example 8]
The same procedure as described in Example 5 was performed except that lysospermic acid was not added. The results are shown in FIG.
As apparent from FIG. 2, lysospermic acid was found to have an activity enhancing activity of TIMP-2.

  Hereinafter, formulation examples of various dosage forms and anti-aging cosmetics containing matrix metalloproteinase inhibitors according to the present invention will be described as production examples.

[Production Example 1] Lotion (Ingredient) (%)
(1) Glycerin 10.0
(2) 1,3-butylene glycol 6.0
(3) Murasaki cultured cell extract * 1 1.0
(4) Citric acid 0.1
(5) Sodium citrate 0.3
(6) Purified water remaining amount (7) polyoxyethylene (60EO) hydrogenated castor oil 0.5
(8) Ethanol 8.0
(9) Preservative Appropriate amount (10) Fragrance Appropriate amount * 1 What was produced in Reference Example 1 (Production method)
A. Components (1) to (6) are mixed and dissolved.
B. Components (7) to (10) are mixed and dissolved.
C. A and B were mixed and uniformed to obtain a skin lotion.

[Production Example 2] Emulsion (component) (%)
(1) Sorbitan monostearate 0.3
(2) Polyoxyethylene sorbitan monooleate (20.EO) 0.1
(3) Lipophilic glyceryl monostearate 0.2
(4) Stearic acid 0.5
(5) Cetanol 0.5
(6) Squalane 3.0
(7) Liquid paraffin 4.0
(8) Glyceryl tri-2-ethylhexanoate 2.0
(9) Methylpolysiloxane 1.0
(10) Hydrogenated soybean phospholipid 0.1
(11) Nicotinic acid dl-α-tocopherol 0.05
(12) Preservative appropriate amount (13) Carboxyvinyl polymer aqueous solution (10%) 1.0
(14) Sodium hydroxide 0.05
(15) Glycerin 5.0
(16) 1,3-butylene glycol 7.0
(17) Purified water remaining amount (18) Ethanol 5.0
(19) Murasaki cultured cell extract 0.1
(20) L-oxyproline 0.2
(21) Hemp cellulose powder 3.0
(22) Perfume proper amount (production method)
A. Ingredients (13) to (17) are heated and mixed and maintained at 70 ° C.
B. Ingredients (1) to (12) are heated and mixed and maintained at 70 ° C.
C. Add B to A, mix and uniformly emulsify.
D. After cooling C, (18) to (22) were added and mixed uniformly to obtain an emulsion.

[Production Example 3] Ointment (component) (%)
(1) Stearic acid 18.0
(2) Cetanol 4.0
(3) dl-α-tocopherol 0.2
(4) Preservative appropriate amount (5) Triethanolamine 2.0
(6) Glycerin 5.0
(7) Purified water remaining amount (8) Murasaki cultured cell extract 1.0
(9) Dipotassium glycyrrhizinate 0.5
(10) Yeast extract 0.5
(Manufacturing method)
A. A part of components (5), (6) and (7) is heat mixed and kept at 75 ° C.
B. Ingredients (1) to (4) are heated and mixed and maintained at 75 ° C.
C. Gradually add A to B.
D. (8)-(10) which melt | dissolved with the remainder of (7) was added, cooling C, and ointment was obtained.

[Production Example 4] Pack (component) (%)
(1) Polyvinyl alcohol 15.0
(2) Silicic anhydride 0.5
(3) Polyethylene glycol 0.5
(4) Polyoxypropylene methyl glucoside 5.0
(5) Glycerin 5.0
(6) Purified water remaining amount (7) Ethanol 20.0
(8) Preservative appropriate amount (9) Murasaki cultured cell extract 0.3
(10) Perfume appropriate amount (production method)
A. Components (1) to (6) are mixed, heated to 70 ° C., and stirred.
B. Mix components (7) and (8).
C. The B was added to the previous A, mixed, and then cooled to uniformly disperse (9) and (10) to obtain a pack.

[Production Example 5] Sunscreen emulsion (component) (%)
(1) Polyoxyalkylene-modified organopolysiloxane 1.0
(2) Dimethylpolysiloxane 5.0
(3) Octamethylcyclotetrasiloxane 20.0
(4) Isotridecyl isononanoate 5.0
(5) Paramethoxycinnamic acid-2-ethylhexyl 5.0
(6) Preservative appropriate amount (7) perfume appropriate amount (8) fine particle titanium oxide 10.0
(9) Particle zinc oxide 10.0
(10) Zirconium oxide 5.0
(11) Polystyrene powder 3.0
(12) Trimethylsiloxysilicic acid 0.5
(13) Dipropylene glycol 3.0
(14) Ethanol 10.0
(15) Purified water remaining amount (16) Salt 0.2
(17) Murasaki cultured cell extract 2.0
(18) Seaweed extract 3.0
(Manufacturing method)
A. Components (1) to (12) are mixed and dispersed.
B. Components (13) to (16) are mixed and dispersed.
C. Add B to A and emulsify uniformly.
D. Components (17) and (18) were added to C to obtain a sunscreen emulsion.

It is a figure which shows the gelatin digestion inhibition result of the lysospermic acid with respect to active MMP-2. It is a figure which shows the activity enhancement effect | action of TIMP-2 by a lysospermic acid. In FIG. 2, ● represents the result of Example 5, □ represents the result of Comparative Example 7, and ○ represents the result of Comparative Example 8.

Claims (6)

A matrix metalloprotease inhibitor containing, as an active ingredient, one or more caffeic acid polymers selected from lysospermic acid or a salt thereof, lysospermic acid B or a salt thereof, and epilabdocin or a salt thereof . The matrix metalloprotease inhibitor according to claim 1 , wherein the matrix metalloprotease inhibitor is a gelatinase inhibitor, a membrane-bound matrix metalloprotease inhibitor or a TIMPs activity enhancer. A skin basement membrane degradation inhibitor comprising the matrix metalloprotease inhibitor according to claim 1 or 2 as an active ingredient. A laminin degradation inhibitor comprising the matrix metalloprotease inhibitor according to claim 1 or 2 as an active ingredient. An elastin degradation inhibitor comprising the matrix metalloprotease inhibitor according to claim 1 or 2 as an active ingredient. A matrix metalloprotease inhibitor composition comprising the following components (A) and (B):
(A) Risosuperumin acid or a salt thereof, Risosuperumin acids B or salts thereof, and Epirabudoshin or one or more coffee acid polymer selected from a salt thereof (B) whitening agents, antioxidants, anti-inflammatory agents , One or more medicinal components selected from the group consisting of antibacterial agents, cell activators, moisturizers and peripheral blood flow promoters

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