JP2007246436A - Mmp-2 and/or mmp-9 inhibitor - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a new MMP-2 (matrix metalloprotease)-2 and/or MMP-9 inhibitor, to provide a disruption suppression/regeneration promoter of soft tissue adhesion, to provide a disruption suppression/regeneration promoter of connective tissual adhesion, to provide a disruption suppression/regeneration promoter of epithelial adhesion, to provide a deepening suppression/repair promoter of a periodontal pocket and to provide a tooth mobility inhibitor. <P>SOLUTION: The MMP-2 and/or MMP-9 inhibitor, disruption suppression/regeneration promoter of the soft tissue adhesion, disruption suppression/regeneration promoter of the connective tissual adhesion, disruption suppression/regeneration promoter of the epithelial adhesion, disruption suppression/regeneration promoter of the deepening suppression/repair promoter of the periodontal pocket and tooth mobility inhibitor comprising one or more kinds selected from histidine and a salt thereof as an active ingredient are provided. <P>COPYRIGHT: (C)2007,JPO&INPIT

Description

  The present invention relates to a novel MMP-2 and / or 9 inhibitor, a soft tissue adhesion destruction inhibitor / regeneration accelerator, a connective tissue adhesion destruction inhibitor / regeneration accelerator, an epithelial adhesion destruction inhibitor / regeneration accelerator, The present invention relates to a periodontal pocket deepening inhibitor / restoration promoter and a tooth movement inhibitor.

  In general, a living body has a basic skeleton formed by hard tissues such as hard bones and teeth, and soft tissues such as tendons, tendons, synovium, and periodontal ligaments that wrap and connect them to help exercise. Yes. The hard and soft tissues are connected by an adhesive structure, helping to function both tissues as a single composite rather than separate. On the other hand, it is known that matrix metalloproteases are strongly involved in alteration and reconstruction of tissues and inflammation in these attachment structures. And as a disease accompanied by those inflammations, rheumatism, arthritis, periodontal disease, etc. are mentioned. Particularly in the oral cavity, there are epithelial adhesion coming from the gingiva and connective tissue adhesion via the periodontal ligament as the attachment structure between teeth and soft tissue. These attachment structures are destroyed by diseases such as periodontal disease, and when destruction progresses, they appear as deepening of the periodontal pockets, and further progresses to tooth swaying and dropping off.

  Matrix metalloproteases (hereinafter abbreviated as MMPs) are a general term for extracellular matrix degrading enzymes characterized by having zinc (II) ions in the active site. The metabolism of extracellular matrix is mainly regulated by the balance between MMPs and tissue-derived metalloprotease inhibitors (TIMP) specific for MMPs. It is known that structural abnormalities of extracellular matrix components based on abnormal expression of MMPs and disruption of synthesis / degradation metabolic balance are associated with periodontal diseases such as periodontitis.

  As MMPs, 10 or more kinds of enzyme molecular species such as collagenase (MMP-1 and 8), stromelysin (MMP-3), gelatinase (MMP-2 and 9) are known (Non-patent Document 1). Produced by many types of cells.

  Among them, the gelatinase group (MMP-2, MMP-9) not only has gelatinolytic activity, but can also degrade type IV collagen, fibronectin, vitronectin and the like (Non-patent Document 2). It is known that it is produced by periodontal ligament cells.

  Fibronectin and vitronectin are cell adhesion proteins that constitute the intercellular matrix, and are involved in the promotion of fibroblast qualification, cell migration and maintenance such as tissue repair, and also in the periodontal ligament It has been known.

On the other hand, it has been conventionally known that histidine or a salt thereof is effective for the prevention and treatment of periodontal diseases, and oral compositions containing these are known. For example, Patent Document 1, an oral composition containing vitamin K ₂ and / or histidine hydrochloride is disclosed. Patent Document 2 includes, as an active ingredient, a combination of (I) one or more selected from lactoferrin, a lactoferrin degradation product, and a lactoferrin-related peptide and (II) a component having 1 to 3 constituent amino acids. The composition for oral cavity to contain is disclosed and histidine is mentioned as 1 type of a structural amino acid.

  The composition for oral cavity described in Patent Document 1 is expected to prevent or treat periodontal diseases based on the capillary strengthening action of histidine hydrochloride. In the example of Patent Document 1, a blood vessel strengthening action is obtained when an ointment containing 0.0001 to 10% by weight of histidine hydrochloride is applied to the rat abdomen, while the content of histidine hydrochloride is 10% by weight. It is disclosed that side effects such as mucosal detachment action occur when the amount exceeds.

Moreover, the composition for oral cavity described in Patent Document 2 is based on the bacterial endotoxin activity inhibitory effect of lactoferrin and the bacterial adhesion inhibitory effect of constituent amino acids (including histidine). It is expected to have a therapeutic effect.
JP-A 63-179823 JP 2001-89339 A Yoshiwara, Shinna: Inflammation and immunity, 2, 177-185, 1994 Tsuru, Funatsu: Biochemical Experimental Method 30, Proteolytic Enzyme I, Table I-34 J Periodontol 1995,66: 579-593 Saffarian S. et al., Science 2004, Oct 1, Vol.306, p.108-111

  In normal periodontal tissue, a shallow groove surrounded by the gingival crevicular epithelium and the enamel surface with the gingival junction epithelium crest end as the bottom is called a gingival crevice. On the other hand, in the periodontal tissues affected by periodontal diseases, the gingival sulcus is generally pathologically deepened and periodontal pockets are formed.

  Periodontal pocket formation and deepening is caused by the loss of periodontal tissue attachment to the tooth surface. Epithelial attachment and connective tissue attachment are known as modes of attachment of periodontal tissue to the tooth surface.

  When the epithelial adhesion is observed with an electron microscope, cells on the outermost layer of the gingival junction epithelium adhere to the basement membrane with hemidesmosome, and they directly adhere to the tooth surface.

  The epithelial attachment constitutes the bottom of the gingival sulcus or the gingival pocket, and when the epithelial attachment is destroyed, the pocket deepens, leading to connective tissue attachment and destruction of periodontal tissues such as periodontal ligament and bone. It is believed that the disruption of epithelial attachment can trigger the so-called subsequent severe tissue destruction.

  On the other hand, connective tissue adhesion is a mode of adhesion of periodontal tissue to cementum via collagen fibers and involves the implantation of collagen fibers in cementum. Epithelial attachment is weak attachment by hemidesmosomes, while connective tissue attachment is strong attachment by collagen fibers.

  In many cases, destruction of connective tissue adhesion is accompanied by destruction of periodontal ligament tissue and resorption of alveolar bone, and periodontal tissue is severely destroyed. If the periodontal tissue is severely destroyed, tooth swaying may occur or, in some cases, the tooth may fall off. Also, as will be described later, once connective tissue adhesion is destroyed, it is difficult to regenerate it.

  Therefore, suppression / prevention of destruction of epithelial adhesion and connective tissue adhesion is extremely important in the formation / suppression of periodontal pockets.

  The ultimate goal of periodontal disease treatment is to restore and regenerate the adhesions lost due to the destruction of periodontal tissues and the deepening of pockets. The loss / recovery of adhesion can be confirmed by, for example, a change in the attachment level (distance from the cement-enamel boundary to the pocket bottom).

  If the connective tissue attachment is destroyed, it is very difficult to regenerate the lost connective tissue attachment with common treatment. Even when there is an improvement in attachment levels, in many cases the improvement is obtained by long epithelial attachment by epithelial downgrowth and not by regeneration of connective tissue attachment. In addition, as described above, epithelial adhesion is weaker than connective tissue adhesion and cannot be said to be sufficient as a healing form.

  As a method for overcoming such a problem, a tissue guided regeneration method (GTR method) is known. The GTR method is a method in which periodontal ligament-derived cells are induced on the dentin surface by applying a membrane to acquire connective tissue adhesion with new cementum. However, it is difficult to say that the GTR method is a treatment routinely used in current dental clinics.

  Thus, in the healing of periodontal diseases, the suppression of adhesion and especially the destruction of connective tissue adhesion and the promotion of regeneration are very important issues, but there are limited ways to approach them directly. Is the current situation.

  Therefore, the present invention provides a novel MMP-2 and / or 9 inhibitor, soft tissue adhesion destruction inhibitor / regeneration accelerator, connective tissue adhesion destruction inhibitor / regeneration accelerator, epithelial adhesion destruction inhibitor / regeneration accelerator An object of the present invention is to provide an agent, a periodontal pocket deepening inhibitor / restoration promoter, and a tooth movement inhibitor.

  The inventors of the present invention have made extensive studies to achieve the above object, and as a result, histidine and salts thereof act as selective inhibitors for MMP-2 and / or MMP-9, and compositions containing these Has been found to be effective in inhibiting destruction / regeneration of soft tissue adhesion, connective tissue adhesion and epithelial adhesion, inhibiting deepening / restoration of periodontal pockets, and suppressing tooth sway, and completed the present invention. .

That is, the present invention includes the following aspects.
Item 1. An inhibitor of matrix metalloprotease (MMP) -2 and / or 9, comprising one or more selected from histidine and a salt thereof as an active ingredient.
Item 2. Item 2. The inhibitor according to Item 1, wherein the active ingredient histidine and a salt thereof are soft tissue adhesion destruction suppressing / regenerating accelerators.
Item 3. Item 3. The inhibitor according to Item 2, wherein the active ingredient histidine and a salt thereof are connective tissue adhesion destruction suppressing / regenerating accelerators.
Item 4. Item 4. The inhibitor according to Item 3, wherein the active ingredient histidine and a salt thereof are agents for suppressing or regenerating connective tissue adhesion in oral tissues.
Item 5. Item 3. The inhibitor according to Item 2, wherein the active ingredient histidine and a salt thereof are agents for suppressing or regenerating epithelial adhesion.
Item 6. Item 6. The inhibitor according to Item 5, wherein the active ingredient histidine and a salt thereof are agents that suppress or regenerate epithelial adhesion in oral tissues.
Item 7. Item 7. The inhibitor according to Items 1 to 6, wherein the active ingredient histidine and a salt thereof are agents for suppressing and restoring periodontal pocket deepening.
Item 8. Item 7. The inhibitor according to Items 1 to 6, wherein the active ingredient, histidine, and a salt thereof are tooth movement inhibitors.

  Hereinafter, the present invention will be described in more detail.

Inhibitor of MMP-2 and / or 9 of the present invention The inhibitor of MMP-2 and / or 9 of the present invention contains one or more selected from histidine and a salt thereof as an active ingredient, and MMP-2 and / or 9 Alternatively, the activity possessed by 9 (hereinafter sometimes referred to as MMP-2, 9) can be inhibited. Examples of the activity of MMP-2, 9 include type IV collagenase activity, fibronectin degrading activity, vitronectin degrading activity, laminin degrading activity, and the like.

  Further, the MMP-2 and / or 9 inhibitor of the present invention selectively inhibits the activity of MMP-2 and 9 and is the type I and III that occupies most of the collagen constituting the periodontal tissue. It does not inhibit the activities of MMP-1 and 8 that degrade type I collagen. MMPs, together with TIMP, play a role in regulating the metabolism of extracellular matrix, and are also involved in remodeling of wound sites and tissues during tissue regeneration. In particular, MMP-1 has been reported to be involved in remodeling of type I collagen, which is the most abundant in tissues, and to be a key factor for connective tissue remodeling (Non-patent Document 4). The inhibitor of MMP-2 and / or 9 of the present invention can selectively inhibit the activities of MMP-2 and 9 without suppressing these normal remodeling.

Soft Tissue Adhesion Fracture Suppression / Regeneration Accelerator The soft tissue adhesion rupture suppression / regeneration accelerator of the present invention contains at least one selected from histidine and its salts as an active ingredient and inhibits the activities of MMP-2 and 9 Thus, it is possible to suppress breakage of soft tissue adhesion and promote regeneration.

  Here, the soft tissue adhesion is an adhesion between the tooth surface and the soft tissue, specifically, an epithelial adhesion that is an adhesion between the tooth surface and the epithelium, and a bond that is an adhesion between the tooth surface and the connective tissue. Examples include tissue adhesion.

Connective tissue adhesion destruction inhibitor / regeneration accelerator The connective tissue adhesion destruction inhibitor / regeneration accelerator of the present invention contains at least one selected from histidine and a salt thereof as an active ingredient. By inhibiting the activity, particularly fibronectin and vitronectin degrading activity, it is possible to suppress the destruction of connective tissue adhesion and promote regeneration.

  Fibronectin and vitronectin are cell adhesion proteins that constitute the intercellular matrix, and are involved in the promotion of fibroblast qualification, cell migration and maintenance such as tissue repair, and also in the periodontal ligament It has been known.

  The attachment of the periodontal ligament tissue to the tooth surface and the attachment of the gingival connective tissue to the tooth surface are similar. In the present invention, the term “connective tissue attachment” includes both.

  Analysis by immunostaining revealed that fibronectin is distributed along the junction of periodontal ligament and cementum in periodontal ligament tissue, which indicates that fibronectin is a periodontal surface of periodontal collagen fibers. It is considered to play an important role in adhesion to the skin (Non-Patent Document 3).

  In the early stage of periodontal tissue repair, serous fibronectin and vitronectin are thought to play a very important role in fibroblast migration and initiation of proliferation. The induced fibroblasts are then actively involved in the formation of fibrous connective tissue whose main components are type I collagen, fibronectin and vitronectin (Non-patent Document 3).

  Furthermore, it has been reported that the newly formed periodontal ligament exhibits strong immunostaining for fibronectin and vitronectin (Non-patent Document 3).

  From these facts, it is considered that fibronectin and vitronectin play an important role in inhibiting the destruction of connective tissue adhesion in periodontal tissue and promoting regeneration.

  Therefore, the connective tissue adhesion destruction inhibitor / regeneration promoter of the present invention is effective in inhibiting connective tissue adhesion destruction / regeneration by inhibiting the fibronectin and vitronectin degradation activities of MMP-2 and 9 .

  The connective tissue adhesion destruction inhibiting / regeneration promoter of the present invention is preferably used for inhibiting connective tissue adhesion destruction / regeneration promotion in oral tissues.

Epithelial adhesion destruction inhibitor / regeneration promoter The epithelial adhesion destruction inhibitor of the present invention contains at least one selected from histidine and a salt thereof as an active ingredient, and has collagenase activity (IV It is possible to suppress the destruction of epithelial adhesion by inhibiting (type collagen degradation activity).

  The epithelial attachment (adhesion between the junctional epithelium and the tooth surface) at the bottom of the gingival sulcus and periodontal pocket is an attachment through the basement membrane and hemidesmosome, and the main component of the basement membrane is type IV collagen Are known.

  Therefore, the epithelial adhesion destruction inhibitor / regeneration promoter of the present invention is effective in inhibiting epithelial adhesion destruction / regeneration by inhibiting the collagenase activity (type IV collagen degradation activity) of MMP-2,9. It is.

  It is preferable to use the epithelial adhesion destruction inhibiting / regenerating promoter of the present invention for inhibiting epithelial adhesion destruction / regeneration in oral tissues.

Periodontal pocket deepening inhibitor / restoration promoter The periodontal pocket deepening inhibitor / restoration promoter of the present invention contains at least one selected from histidine and salts thereof as an active ingredient, and its MMP-2, 9 inhibitor Based on the effect as a soft tissue adhesion destruction inhibitor / regeneration accelerator and a connective tissue adhesion destruction inhibitor / regeneration accelerator, it is effective for inhibiting periodontal pocket deepening and restoration.

Tooth sway suppressant The tooth sway suppressor of the present invention contains at least one selected from histidine and a salt thereof as an active ingredient, its MMP-2, 9 inhibitor, a soft tissue adhesion destruction inhibitor / regeneration accelerator, binding Based on the effects of inhibiting tissue adhesion destruction / regeneration, and periodontal pocket deepening / repair promoter, it is effective in suppressing tooth movement.

  MMP-2 and / or 9 inhibitor of the present invention, soft tissue adhesion destruction inhibitor / regeneration accelerator, connective tissue adhesion destruction inhibitor / regeneration accelerator, epithelial adhesion destruction inhibitor / regeneration accelerator, periodontal pocket Deepening inhibitor / restoration promoter and tooth movement inhibitor (hereinafter, these may be collectively referred to as “inhibitor of MMP-2 and / or 9 of the present invention”) are selected from histidine and its salts One or more of these are used as active ingredients.

  The histidine and salts thereof include these optical isomers D and L.

  Examples of histidine salts, particularly L-histidine salts, include monohydrochloride, monohydrochloride monohydrate, dihydrochloride, inorganic salts such as phosphoric acid, pyrrolidone carboxylate, and gluconate hydroxycarboxylate. Examples thereof include carboxylates, inosinates, metal salts, metal complexes and the like, and hydrochlorides are preferable.

  The MMP-2 and / or MMP-9 inhibitor of the present invention is blended with various known components as necessary, and is a dispersion paste external preparation, liquid external preparation, liquid external preparation, mouthwash, mouthwash, It can be provided as an external preparation composition such as an ointment, pasta, gel, coating agent, varnish, patch, etc., membrane for tissue guided regeneration (GTR: Guided Tissue Regeneration), bone guided regeneration (GBR: Guided) Bone Regeneration) can be provided by applying, blending, kneading and impregnating medical devices with structures such as membranes, bone filling materials, and regenerative scaffolds, and powders, fine granules, granules, powders, tablets, Oral compositions such as capsules, liquids, syrups, dry syrups, inhalants, buccal disintegrants, lozenges, troches, drops, etc. can be provided, and oral compositions are particularly preferred. These external preparation compositions, medical devices, and internal use compositions can be produced by ordinary production methods. These external preparation compositions, medical devices, and internal use compositions are preferably used for the oral cavity.

  One or more compounding amounts selected from histidine and a salt thereof in the MMP-2 and / or MMP-9 inhibitor of the present invention are usually used with respect to the total amount of the external preparation composition when used in the form of the external preparation composition. It is 0.001-10 mass%, and 0.01-5 mass% is especially preferable. When the blending amount is less than 0.001% by mass, a sufficient effect cannot be obtained, and when it exceeds 10% by mass, it is disadvantageous in terms of preparation or cost. In addition, when the MMP-2 and / or MMP-9 inhibitor of the present invention is used in the form of an oral composition, it usually varies depending on the preparation, but usually one or more kinds selected from histidine and a salt thereof per day for an adult. The blending amount can be appropriately determined so that the intake amount is 10 to 4500 mg, preferably 50 to 3000 mg, and more preferably 100 to 1500 mg.

  In addition to the above-mentioned external preparation composition, medical device and internal use composition, in addition to the components of the present invention, a conventionally known bactericidal agent is blended so that various symptoms of periodontal disease or disease prevention and treatment effects can be obtained. It is expected to improve. Specific examples include quaternary ammonium salts, bisbiguanides, phenols, and non-cationic fungicides. Examples of quaternary ammonium salts include cetylpyridinium chloride, benzethonium chloride, distearyldimethylammonium chloride, stearyldimethylbenzylammonium chloride, stearyltrimethylammonium chloride, cetyltrimethylammonium chloride, lauryltrimethylammonium chloride, laurylpyridinium chloride, In particular, cetylpyridinium chloride is preferred. Bisbiguanide fungicides include bisbiguanide hexanes, bisbiguanide propyl ethers, bisbiguanide xylenes, bisbiguanide decanes, bisbiguanidodecanes, and chemically acceptable salts thereof, in particular bisbiguanide hexane. Preferred are chlorhexidine salts such as chlorhexidine gluconate and chlorhexidine hydrochloride. Examples of the phenolic fungicide include isopropylmethylphenol and hinokitiol. Examples of (halogenated) diphenyl ether fungicides include triclosan. Other fungicides include hexetidine, metronidazole, and the like. Among these fungicides, in particular, when used in combination with cetylpyridinium chloride or / and hydrochloric acid or chlorhexidine gluconate, a synergistic effect of promoting adhesion by removing periodontal pathogens that cause significant local adhesion regeneration is obtained. It is done. The blending amount of these bactericides is usually 0.001 to 0.5% by mass, particularly 0.01 to 0.1% by mass, based on the total amount of the composition.

  In the present invention, the above-mentioned external preparation composition, medical device and internal use composition include other components other than the above-mentioned essential components, for example, a surfactant, an abrasive, a wetting agent, a monohydric alcohol, depending on the form of use. Binders, fragrances, pH adjusters, preservatives, pigments, excipients, film forming agents, solubilizers, lubricants, sweeteners, corrigents, flavoring agents, etc., within a range that does not impair the effects of the present invention Can be blended as appropriate.

  For example, anionic, nonionic, cationic, and amphoteric surfactants can be used alone or in combination of two or more as surfactants. Examples of nonionic surfactants include sucrose fatty acid esters and maltose fatty acids. Sugar fatty acid esters such as esters, sugar alcohol fatty acid esters such as maltitol fatty acid esters, sorbitan fatty acid esters such as sorbitan monolaurate, polyoxyethylene sorbitan fatty acid esters such as polyoxyethylene sorbitan monolaurate and polyoxyethylene sorbitan monostearate , Fatty acid alkanolamides such as lauric acid diethanolamide, polyoxyethylene alkyl ethers such as polyoxyethylene stearyl ether, polyoxyethylene oleyl ether, monooleic acid Polyethylene glycol fatty acid esters such as reethylene glycol and polyethylene glycol monolaurate, polyglycerin fatty acid esters, polyoxyethylene glycerin fatty acid esters, polyoxyethylene fatty acid esters, alkyl glucosides, polyoxyethylene hydrogenated castor oil, glycerin fatty acid esters, polyoxyethylene A propylene block copolymer etc. are mentioned. Examples of anionic surfactants include sodium alkyl sulfates such as sodium lauryl sulfate and sodium myristyl sulfate, sodium N-acyl sarcosates such as sodium N-lauroyl sarcosinate, sodium N-myristoyl sarcosinate, and sodium N-palmitoyl glutamate. N-acyl glutamate, polyoxyethylene alkyl sulfosuccinate such as polyoxyethylene alkyl (C12 to C14) sulfosuccinate disodium, and the like. Examples of amphoteric surfactants include amino acid type, alkylbetaine type, alkylamidobetaine type, sulfobetaine type, imidazoline type and the like, preferably 2-alkyl-N-carboxymethyl-N-hydroxyethylimidazolinium betaine or And coconut oil fatty acid amidopropyl betaine. These compounding quantities are 0.05-50 mass% normally with respect to the composition whole quantity, and 0.1-5 mass% is especially preferable.

  As an abrasive, calcium carbonate, calcium phosphate, dicalcium phosphate, calcium pyrophosphate, insoluble sodium metaphosphate, titanium oxide, amorphous silica, crystalline silica, aluminosilicate, aluminum oxide, aluminum hydroxide, resin, etc. alone or in combination The above can be combined and blended. These compounding quantities are 10-60 mass% normally with respect to the composition whole quantity.

  As the wetting agent, for example, a polyhydric alcohol such as glycerin, sorbitol, polyethylene glycol, propylene glycol, ethylene glycol, hexylene glycol, 1,3-butylene glycol, polypropylene glycol, xylitol, maltitol, lactitol, etc., alone or 2 It can mix | blend combining a seed | species or more, The compounding quantity is 3-20 mass% normally with respect to the whole composition.

  Examples of the monohydric alcohol include ethanol, propyl alcohol, and isopropyl alcohol, and ethanol is particularly preferable. These monohydric alcohols can be blended singly or in combination of two or more, and the blending amount is usually 0 to 15% by mass with respect to the whole composition.

  Examples of the binder include cellulose derivatives such as carrageenan and carboxymethylcellulose, alkali metal alginates and derivatives thereof such as sodium alginate, gums such as xanthan gum, tragacanth gum and gum arabic, polyvinyl alcohol, and sodium polyacrylate. Examples of the binder include inorganic binders such as silica gel, aluminum silica gel, and bee gum. The amount of these binders is usually 0.01 to 5% by mass with respect to the total amount of the composition.

  Examples of flavoring agents include anethole, menthol, peppermint oil, spearmint oil, lemon oil, orange oil, sage oil, rosemary oil, cinnamon oil, perilla oil, winter green oil, clove oil, eucalyptus oil, pimento oil, Carvone, cinnamic aldehyde, cineol, menthone, limonene, methyl salicylate, and the like can be blended alone or in combination of two or more in a range not impairing the effects of the present invention. The compounding quantity is 0.0001-1.0 mass% normally with respect to the composition whole quantity.

  Examples of the sweetening agent include palatinit, saccharin sodium, acesulfame potassium, stevioside, neohesperidyl dihydrochalcone, glycyrrhizin, perilartine, thaumatin, asparatylphenylalanyl methyl ester, ρ-methoxycinnamic aldehyde, and the like. Can be blended alone or in combination of two or more. The compounding quantity is 0.01-1 mass% normally with respect to the whole composition, Preferably it is 0.05-0.5 mass%.

  Examples of the pH adjuster include citric acid, phosphoric acid, malic acid, pyrophosphoric acid, lactic acid, tartaric acid, pantothenic acid, glycerophosphoric acid, acetic acid, nitric acid, or a chemically possible salt or sodium hydroxide thereof. These can be blended alone or in combination of two or more so that the pH of the composition is in the range of 5-9. The compounding quantity is 0.01-2 mass% normally with respect to the whole composition.

  Excipients include calcium carbonate, silicic acid anhydride, magnesium carbonate, sucrose, lactose, starch, glucose, crystalline cellulose, mannitol, sorbit, xylitol, erythritol, palatinit, palatinose, maltitol, trehalose, lactitol, reduced Examples include starch sugar, reduced isomaltoligosaccharide, coupling sugar, gum base, gum arabic, gelatin, cetylmethylcellulose, light anhydrous silicic acid, magnesium aluminate, calcium aluminate metasilicate, sodium bicarbonate, calcium phosphate and the like. As the solubilizer, glycerin fatty acid ester, sucrose fatty acid ester, sodium lauryl sulfate, alcohol, esters, polyethylene glycol derivatives, sorbitan fatty acid esters, sulfated fatty alcohols and the like can be blended.

  As the lubricant, magnesium stearate, talc, hardened oil, or the like can be blended.

  Examples of film forming agents include shellac resin, rosin, sandalac, alkyd resin, copalite, Sumatra benzoic resin and other natural resins, polyvinyl alcohol, polyvinylpyrrolidone, polyurethane, polystyrene, polymethyl methacrylate, polyvinyl acetate, polyvinyl chloride, poly Nonionic polymers such as acrylonitrile, vinyl pyrrolidone / vinyl acetate polymer, vinyl pyrrolidone / dimethylaminoethyl methacrylate copolymer, carboxy vinyl polymer, methyl vinyl ether / maleic acid ester copolymer, acrylic acid ester / methacrylic acid ester copolymer, vinyl acetate / croton Anionic polymer such as acid copolymer, dimethylallyl ammonium chloride polymer, hydroxyethylcellulose / dimethylallylamine Such as ammonium chloride copolymers, cationic polymers such as vinylpyrrolidone quaternized dialkylaminoalkyl acrylate polymers, higher alkylbetaines, monochloroacetic acid amphoteric products of dialkylaminoethyl methacrylate polymers, octylacrylamide / butylaminoethyl methacrylate / acrylic acid ester copolymers, etc. Amphoteric polymers, fluororesins such as fluoroalkyl phosphates, silicon derivatives such as dimethylsiloxane, and the like can be blended.

  When using organic solvents in preparations such as coating agents and varnishes, remove volatile organic solvents such as ethyl acetate, dichloromethane, chloroform, alcohol, dioxane, siloxane, hexane, cyclohexane, acetone, ether, petroleum ether, and propylene carbonate. Can be blended.

  The above external preparation composition, medical device and internal use composition include vitamin E such as dl-α-tocopherol acetate, tocopherol succinate, or tocopherol nicotinate, ascorbic acids, dextranase, amylase, protease, mutanase, Enzymes such as lysozyme, lytic enzyme (lytechenzyme), antiplasmin agents such as tranexamic acid and epsilon aminocaproic acid, aluminum chlorohydroxyl allantoin, dihydrocholesterol, glycyrrhizin salts, glycyrrhetinic acid, glycerophosphate, chlorophyll, sodium chloride, caropeptide, water-soluble Inorganic phosphate compounds, fluorides such as sodium fluoride, potassium fluoride, ammonium fluoride, calcium fluoride, copper fluoride, zinc fluoride, lithium fluoride, fluoride Cesium, zirconium fluoride, tin fluoride, hydrofluoric acid, sodium monofluorophosphate, potassium monofluorophosphate, sodium titanium fluoride, potassium titanium fluoride, hexylamine hydrofluoride, laurylamine hydrofluoride, glycine Hydrofluoride, alanine hydrofluoride, fluoride such as fluorosilane, diammonium fluoride, grape seed extract, ginkgo biloba extract, oil-soluble licorice extract, mulberry bark extract, tea extract, blueberry extract, rosemary extract, birch Extract, plant extract such as koji extract, comfrey extract, bones such as PDGF (platelet-derived growth factor), IGF (insulin-like growth factor), BMP (bone formation promoting factor), FGF (fibroblast growth factor), osteopontin as well as/ Or biological tissue growth factors, enamel proteins such as amelogenin and enamelin, biological substances such as albumin, globulin, chondroitin sulfate, fibronectin, fibrinogen, and elastin, tetracyclines such as minocycline and doxycycline, and macrolides such as clarithromycin and azithromycin Antibacterial substances such as quinolones such as levofloxacin, ketolides such as tethromycin, non-steroids such as flurbiprofen, steroids such as dexamethasone, natural products such as azulene, bis Bone resorption inhibitors such as phosphonates, calcium phosphate compounds such as calcium phosphate, α tricalcium phosphate, β tricalcium phosphate, tetracalcium phosphate, hydroxyapatite and bioactive gala These can be blended alone or in combination of two or more. Among these compounds, use in combination with cysteine, cystine, glycyrrhizic acids, glycyrrhetinic acid, dl-α-tocopherol acetate, tocopherol succinate, tocopherol nicotinate, etc., ascorbic acids, lysozyme chloride, FGF, etc. As a result, synergistic destruction prevention effect and regeneration promotion effect of tissue adhesion can be obtained.

  Furthermore, natural polymers such as gelatin, collagen, konjac mannan, pullulan, chitosan, starch, dextran, albumin, alginic acid and salts thereof, polyethylene glycol, carboxyvinyl polymer, methyl vinyl ether / anhydrous are used to improve oral mucosal retention. Synthetic polymers such as maleic acid copolymer, polyvinyl pyrrolidone, lecithin such as soybean lecithin, egg yolk lecithin, semi-polymer such as polylactic acid, polyglycolic acid, cyclodextrin, polyacryl dextran, hydroxypropylmethylcellulose, hydroxypropylcellulose, carboxymethylcellulose They can be blended alone or in combination of two or more from synthetic polymers or semi-synthetic compounds.

  The MMP-2 and / or MMP-9 inhibitor of the present invention can be produced by mixing these components and according to a usual method.

  The present invention will be described more specifically with reference to test examples and examples. However, the present invention is not limited to the following test examples and examples. Further, unless otherwise specified, “%” is “mass%”.

Test Example 1 : Measurement of inhibitory activity against matrix metalloprotease (MMP) -2, 9 (gelatinase) Test compound: histidine Preparation of reaction solution Test compound as substrate buffer (5 mM CaCl ₂ , 50 mM Tris buffer containing 1 μM ZnCl ₂ The solution was added to the solution, pH 7.4) so that the final concentrations were 10 μg / ml, 30 μg / ml, and 100 μg / ml, and the test was conducted at these three concentrations.

-Cells and culture conditions Periodontal ligament fibroblasts (PDL cells) were first cultured and subcultured in Dulbecco's modified Eagle medium (DMEM) containing 10% fetal bovine serum (FBS).

-Cell treatment PDL cells were seeded in a dish having a diameter of 10 cm, cultured until confluent in 10% FBS-added DMEM medium, and then the cell surface was washed with serum-free DMEM medium. After washing, the cells were cultured in serum-free DMEM medium for 48 hours, and the culture supernatant was transferred to a filter, and centrifuged and concentrated. This concentrated solution was used for the test.

Measurement of inhibitory effect on MMP-2,9 activity Measurement of the inhibitory effect of histidine on PDL cell-derived MMP-2,9 is known as the main method for detecting the activity of MMP-2,9. Gelatin zymography was used as a substrate. Gelatin zymography is a method in which enzyme protein is separated by slab SDS-PAGE and then reacted with a substrate that coexists in the gel. Gelatinase activity is observed as a colorless band on a blue background (gelatin gel). . Slab electrophoresis was performed according to the Laemli method, and the cell supernatant prepared above was applied to an acrylamide 10% separation gel containing gelatin having a final concentration of 1 mg / ml, and then electrophoresed.

After electrophoresis, the gel was immersed in Triton buffer (2.5% Triton X-100-containing 50 mM Tris buffer, pH 7.5%), and SDS was removed while shaking for 20 minutes. Next, it was immersed in a substrate buffer solution (5 mM CaCl ₂ , 1 μM ZnCl ₂ -containing 50 mM Tris buffer solution) containing the test compound and reacted at 37 ° C. for 20 hours. After the enzyme reaction, it was stained with 0.02% Coomassie Brilliant Blue R250 and decolorized with an acetic acid / methanol aqueous solution until the band could be observed. As a negative control, a test compound to which no test compound was added was used. Moreover, the effect was compared using the dehydroabietic acid with which the inhibitory effect with respect to collagenase activity was reported (positive object).

  The MMP activity inhibitory effect was evaluated by comparing the area of the MMP activity band when the test compound was added with the area when no test compound was added (negative control). The measurement results of enzyme inhibitory activity are shown in Table 1.

Test Example 2 : Measurement of inhibitory activity against matrix metalloprotease (MMP) -1, 8 (collagenase) The inhibitory effect on collagenase activity was based on type I collagen (Life Research Institute, Limited) labeled with fluorescein isothiocyanate. The measurement was performed according to the method of Misui et al. (Japanese Journal of inflammation, Vol. 4, p. 123, 1984). That is, 0.04 units of activated human collagenase (MMP-1), 50 μg of fluorescently labeled type I collagen solution, assay buffer (0.05 M Tris-HCl buffer solution containing NaCl, CaCl ₂ , NaN ₃ , pH 7. 5) The test compound dissolved in 50 μl and 20 μl of assay buffer was mixed, and assay buffer diluted 2-fold so that the liquid volume became 200 μl was added. After reacting at 37 ° C. for 4 hours, 200 μl of ice-cooled enzyme reaction terminator / extract (0.05 M Tris-HCl buffer, pH 9.5 / ethanol containing NaCl and o-phenanthroline) was added, and the mixture was stirred for 30 minutes at room temperature. Left alone. The supernatant was collected by centrifugation and allowed to stand at room temperature for 10 minutes, and then the fluorescence intensity (excitation wavelength: 495 nm, fluorescence wavelength: 520 nm) was measured. As a control, an assay buffer diluted 2-fold was used instead of the test compound, and an assay buffer diluted 2-fold was used instead of collagenase as a blank. Moreover, the effect was compared using dehydroabietic acid with which the inhibitory effect with respect to collagenase activity was reported. The MMP-1,8 activity inhibition rate of the test compound was determined from the following formula.

Inhibition rate (%) = (1− (C−D) / (A−B)) × 100
A: fluorescence intensity of control, B: fluorescence intensity of control blank, C: fluorescence intensity of test compound, D: fluorescence intensity of test compound blank Table 2 shows the measurement results of enzyme inhibitory activity.

  From the above results, it was found that dehydroabietic acid inhibits the activities of both MMP-1, 8, MMP-2, and 9 while histidine specifically and strongly inhibits the activities of MMP-2 and 9.

Examples According to the following formulas, preparations were prepared according to conventional methods.

Example 1 Ingredients for Dispersion External Use Compounding amount (% by mass)
Dicalcium phosphate 25.0
Glycerin 15.0
Sorbitol 18.0
Sodium carboxymethylcellulose 2.0
Sodium lauryl sulfate 1.6
Carrageenan 0.4
Saccharin sodium 0.2
Fragrance 1.5
Sodium benzoate 0.5
Histidine 0.2
Dipotassium glycyrrhizinate 0.1
Sucrose fatty acid ester 0.1
Pullulan 2.0
Purified water remaining amount 100.0

Example 2 Toothpaste compounding component Compounding amount (mass%)
Histidine 3.0
Cetylpyridinium chloride 0.1
Sodium monofluorophosphate 1.0
Calcium hydrogen phosphate 20.0
Sorbit liquid 20.0
Alkyl glucoside 2.0
Hydroxyethyl cellulose 1.5
P-Hydroxybenzoate ester 0.5
Propylene glycol 3.0
Sodium lauryl sulfate 1.5
Saccharin sodium 0.1
Peppermint oil 0.3
Methyl salicylate 0.2
Anethole 0.1
Menthol 0.2
Cineol 0.3
Chitosan 2.0
Purified water remaining amount 100.0

Example 3 Mouthwashing ingredients Compounding amount (mass%)
Ethanol 5.0
Glycerin 10.0
Citric acid 0.1
Sodium citrate 0.3
Polyoxyethylene hydrogenated castor oil 5.0
Methyl paraoxybenzoate 0.3
Fragrance 0.5
Histidine 3.0
Triclosan 0.5
Lecithin 1.0
Purified water remaining amount 100.0

Example 4 Mouthwashing ingredients Compounding amount (mass%)
Ethanol 5.5
Tranexamic acid 0.1
Glycerin 5.5
Citric acid 0.05
Sodium citrate 0.4
Polyoxyethylene hydrogenated castor oil 0.8
Methyl paraoxybenzoate 0.3
Histidine 1.5
Birch extract 0.6
Glycerin fatty acid ester 0.5
Dextran 1.0
Purified water remaining amount 100.0

Example 5 Mouthwash formulation ingredients Amount (mass%)
Ethanol 11.0
Glycerin 7.0
Citric acid 0.03
Sodium citrate 0.2
Polyoxyethylene hydrogenated castor oil 0.7
Lauric acid diethanolamide 1.0
Methyl paraoxybenzoate 0.3
Histidine 1.0
Cetylpyridinium chloride 0.45
Purified water remaining amount 100.0

Example 6 Pasta preparation compounding amount (mass%)
Liquid paraffin 11.0
Cetanol 9.0
Glycerin 20.0
Sorbitan monopalmitate 1.0
Polyoxyethylene sorbitan monostearate 4.0
Sodium lauryl sulfate 0.3
Benzethonium chloride 0.2
Tocopherol nicotinate 1.0
Cetylbenzethonium chloride 0.1
Methyl salicylate 0.15
Saccharin 0.05
Fragrance 0.3
Rosemary extract 0.3
α-Amylin 0.08
Histidine 0.5
Lecithin 2.0
Purified water remaining amount 100.0

Example 7 Tablet compounding ingredients Compounding amount (mass%)
Xylitol 69.8
Maltitol 21.0
Gum arabic 1.5
Sucrose fatty acid ester 2.5
Powder flavor 1.0
Citric acid 4.0
Histidine 0.2
Total amount 100.0

Example 8 Lozenge blending component Blending amount (mass%)
Palatinit 57.4
Xylitol 20.0
Aspartame 0.05
Histidine 0.7
Rosemary extract 0.2
Perfume 0.08
Colorant 0.8
Purified water remaining amount 100.0

Example 9 Membrane for tissue induction regeneration method It has a two-layer structure mainly composed of polylactic acid, the thickness of one sheet is 0.1 mm, the total thickness of the film is 0.4 mm, and the diameter is formed on the upper surface of the film. A 0.1 mm circular hole is drilled, the porosity is 40%, and the bottom surface has a rectangular hole with a side of 0.2 mm, and the porosity is 33%. A membrane was created. A 1.0% water-soluble gel of hydroxymethylpropylcellulose with 5% histidine was applied to the lower surface of the film to prepare a membrane for tissue induction regeneration method as an example of the present invention.

Claims (8)

An inhibitor of matrix metalloprotease (MMP) -2 and / or 9, comprising one or more selected from histidine and a salt thereof as an active ingredient. The inhibitor according to claim 1, wherein the active ingredient histidine and a salt thereof are soft tissue adhesion destruction suppressing / regenerating accelerators. The inhibitor according to claim 2, wherein the active ingredient histidine and a salt thereof are connective tissue adhesion destruction suppressing / regenerating accelerators. The inhibitor according to claim 3, wherein the active ingredient histidine and a salt thereof are agents for inhibiting destruction / regeneration of connective tissue adhesion in oral tissues. The inhibitor according to claim 2, wherein the active ingredient histidine and a salt thereof are epithelial adhesion destruction inhibitor / regeneration promoter. The inhibitor according to claim 5, wherein the active ingredient histidine and a salt thereof are agents for suppressing or regenerating epithelial adhesion in oral tissues. The inhibitor according to claim 1, wherein the active ingredient histidine and a salt thereof are periodontal pocket deepening suppression / restoration promoters. The inhibitor according to any one of claims 1 to 6, wherein the active ingredient histidine and a salt thereof are tooth movement inhibitors.