Glycation stress inhibitor

Abstract

An object of the present invention is to provide a novel glycation stress inhibitor having an activity of inhibiting glycation stress occurring in vivo.SOLUTION: The glycation stress inhibitor, the glycation reaction inhibitor or the inflammatory reaction inhibitor for inhibiting the inflammatory reaction caused by AGEs contains one or more kinds selected from the group consisting of glycyrrhizic acids, allantoins, azulenes, pyridoxine hydrochloride, hinokitiols, copper chlorophylls, lysozyme chloride and epsilon-aminocaproic acid. Each agent is useful as an external preparation, an agent for oral cavity, a gingival care agent, and a periodontal disease preventive agent.SELECTED DRAWING: None

Description

[Technical Field]

[0001] The present invention relates to an agent for suppressing glycative stress. [Background technology]

[0002] The Maillard reaction (glycation reaction) between reducing sugars such as glucose and proteins produces glycation products, a phenomenon that has long been known as the browning phenomenon of foods. This glycation reaction also occurs in living organisms, and the substances produced by the glycation reaction are called advanced glycation end products (hereinafter referred to as AGEs). The production of AGEs is an irreversible reaction, and the produced AGEs are excreted from the body through metabolism. However, when the metabolic rate slows due to aging or other factors, they tend to accumulate in various tissues in the body (Patent Document 1).

[0003] When AGEs bind to RAGE (Receptor for AGEs; AGE receptor) expressed on the cell membranes of cells such as monocytes / macrophages, vascular endothelial cells, and fibroblasts, inflammation is induced by factors such as AGE-induced oxidative stress, activation of NF-κB activity, and activation of intracellular signals such as mitogen-activated protein kinase (MAPK) and protein kinase C (PKC). This AGE-induced inflammatory response is called glycative stress or AGE-RAGE interaction and is known to cause a variety of symptoms. Glycative stress is used to describe not only the inflammatory response caused by AGEs but also the biological stresses and various effects that occur during the AGE production process. For example, glycative stress contributes to browning and dullness of the skin, and hyperglycemia can lead to cataracts, vascular disorders, and renal dysfunction (Non-Patent Document 1). Therefore, it is extremely important to prevent or suppress glycative stress.

[0004] Patent Document 1 describes that volatile oils from citrus fruits have Maillard reaction inhibitory activity and AGE-protein crosslinker cleavage activity. Patent Document 2 describes that Coptis japonica extract, Phellodendron bark extract, and Sanshichi extract have AGE production inhibitory effect and are safe materials that do not irritate the skin, and therefore can be used in various skin cosmetics, bath additives, etc. to prevent skin aging. Patent Document 3 describes that extracts of plants from the genus Pinellia inhibit the binding of AGEs and RAGE, thereby suppressing inflammation caused by AGEs. Patent Document 4 describes that a combination of quercetin and sulforaphane is effective against AGE-induced inflammation by suppressing the expression of RAGE.

[0005] Meanwhile, various anti-inflammatory ingredients have been known for some time. For example, allantoin is known to promote skin proliferation and wound healing, and to have anti-inflammatory effects, but its mechanism of action is unknown. Dipotassium glycyrrhizinate is widely used as an anti-inflammatory ingredient, but its mechanism of action is inhibition of the arachidonic acid cascade. The arachidonic acid cascade is a metabolic pathway that produces inflammatory substances such as prostaglandins and thromboxanes using arachidonic acid derived from phospholipids that constitute cell membranes. Dipotassium glycyrrhizinate exerts its anti-inflammatory effect by inhibiting the activation of enzymes such as phospholipase A and lipoxygenase, thereby suppressing the production of phospholipid-derived inflammatory substances. Pyridoxine hydrochloride is a vitamin B6 vitamer that is most abundant in vitamin B6 supplements. Pyridoxine hydrochloride is known to improve metabolism and suppress inflammation in the skin and mucous membranes when pyridoxine hydrochloride is deficient, but it does not have any inhibitory effect on inflammation caused by other factors (Non-Patent Documents 2 and 3). Hinokitiol is known to have antibacterial activity against bacteria and fungi, as well as anti-inflammatory effects. This anti-inflammatory effect is known to be due to its inhibitory effect on 11β-hydroxysteroid dehydrogenase (11β-HSD), a local cortisol-modifying enzyme (Non-Patent Document 4). Epsilon aminocaproic acid is known to have hemostatic and anti-inflammatory effects, which are known to be due to its antiplasmin effect. [Prior art documents] [Patent documents]

[0006] [Patent Document 1] Japanese Patent Application Laid-Open No. 2004-35424 [Patent Document 2] Japanese Patent Application Laid-Open No. 2016-210729 [Patent Document 3] Patent Publication No. 2021-31424 [Patent Document 4] Japanese Patent Application Publication No. 2017-145236 [Non-patent literature]

[0007] [Non-Patent Document 1] Yagi et al., "Evaluation of Glycative Stress and Anti-Glycative Effects," Oleoscience, Vol. 18, No. 2 (2018), pp. 67-73 [Non-patent document 2] Chiang EP et al.,Arthritis Research & Therapy 2005,7:R1254-R1262 [Non-patent document 3] Chiang EP et al.,Arthritis Research & Therapy 2005,7:R1404-R1411 [Non-patent document 4] Itoi-Ochi et al., Program of the 42nd Annual Meeting of the Japanese Society for Investigative Dermatology, Page 225 (2017) Summary of the Invention [Problem to be solved by the invention]

[0008] However, because the anti-inflammatory mechanism of each ingredient differs from the mechanism of onset of AGE-induced inflammation, it cannot be said that they are effective against AGE-induced inflammatory responses just because they are anti-inflammatory ingredients.The object of the present invention is to provide new glycative stress inhibitors that have activity in suppressing glycative stress that occurs in the body, particularly inhibitory effects on AGE production, glycation reactions, and AGE-induced inflammation, and in particular, inhibitors of AGE production, glycation reactions, and AGE-induced inflammation. [Means for solving the problem]

[0009] The present invention provides the following [1] to

[12] . [1] A glycative stress inhibitor containing one or more selected from the group consisting of glycyrrhizins, allantoins, azulenes, pyridoxine hydrochloride, hinokitiols, copper chlorophylls, lysozyme chloride, and epsilon-aminocaproic acid. [2] An AGE (Advanced Glycation End Products) production inhibitor containing one or more members selected from the group consisting of glycyrrhizinic acids, allantoins, azulenes, pyridoxine hydrochloride, hinokitiols, copper chlorophylls, lysozyme chloride, and epsilon-aminocaproic acid. [3] The AGE production inhibitor according to [2], wherein the AGEs are derived from collagen. [4] The AGE production inhibitor according to [2] or [3], wherein the AGEs are derived from collagen and glyceraldehyde. [5] A glycation reaction inhibitor containing one or more members selected from the group consisting of glycyrrhizins, allantoins, azulenes, pyridoxine hydrochloride, hinokitiols, copper chlorophylls, lysozyme chloride, and epsilon-aminocaproic acid. [6] The glycation reaction inhibitor according to [5], which inhibits the glycation reaction of collagen. [7] The glycation reaction inhibitor according to [5] or [6], which inhibits the glycation reaction of collagen caused by glyceraldehyde. [8] An inflammatory response inhibitor containing one or more members selected from the group consisting of glycyrrhizins, allantoins, azulenes, pyridoxine hydrochloride, hinokitiols, copper chlorophylls, lysozyme chloride, and epsilon-aminocaproic acid, which suppresses inflammatory responses caused by AGEs. [9] An external preparation comprising the agent according to any one of [1] to [8].

[10] An oral preparation comprising the agent according to any one of [1] to [8].

[11] The oral preparation according to

[10] , which is a gum care agent.

[12] The oral preparation according to

[10] , which is a periodontal disease preventive agent. [Effects of the Invention]

[0010] The present invention provides a glycative stress inhibitor that suppresses glycative stress occurring in the body, an AGE production inhibitor that has activity in inhibiting AGE production reactions, a glycation reaction inhibitor that suppresses glycation reactions (Maillard reaction), and an inflammatory response inhibitor that suppresses AGE-induced inflammatory reactions. These agents can be used as external preparations, oral preparations, gum care agents, periodontal disease preventatives, etc. [Brief explanation of the drawings]

[0011] [Figure 1] FIG. 1 shows the results of IL-6 concentration in Test Example 1. [Figure 2] FIG. 2 shows the effect of suppressing inflammation caused by AGEs in Test Example 3. DETAILED DESCRIPTION OF THE INVENTION

[0012] [1. Glycative stress inhibitor, AGE production inhibitor, glycation reaction inhibitor, inflammatory reaction inhibitor] [1.1 Active ingredient] In the present invention, the active ingredient is one or more selected from the group consisting of glycyrrhizins, allantoins, pyridoxine hydrochloride, hinokitiols, copper chlorophylls, lysozyme chloride, and epsilon aminocaproic acid.

[0013] <Glycyrrhizic acid> In this specification, glycyrrhizic acids refer to glycyrrhizin, glycyrrhizic acid, its hydrolysates, derivatives thereof, and plants and plant extracts containing them. Examples of glycyrrhizic acids include glycyrrhizinate salts such as dipotassium glycyrrhizinate and monoammonium glycyrrhizinate, glycyrrhetinic acid which is an acid hydrolysis product of glycyrrhizinic acid, glycyrrhetinic acid derivatives such as stearyl glycyrrhetinate, licorice (for example, root), and licorice extract, and dipotassium glycyrrhizinate, glycyrrhetinic acid, and stearyl glycyrrhetinate are preferred.

[0014] <Allantoin> In this specification, allantoins refer to allantoin, its derivatives, and salts thereof. Examples of allantoins include allantoin, allantoin chlorohydroxyaluminum, allantoin dihydroxyaluminum, and combinations of two or more thereof, with allantoin being preferred.

[0015] <Azulene> In this specification, azulenes refer to azulene, its derivatives, and salts thereof. Examples of azulenes include sodium azulene sulfonate, azulene, 1,4-dimethyl-7-isopropylazulene, 4,8-dimethyl-2-isopropylazulene, and combinations of two or more thereof, with sodium azulene sulfonate being preferred.

[0016] <Pyridoxine hydrochloride> Pyridoxine hydrochloride is the hydrochloride salt of pyridoxine.

[0017] <Hinokitiols> In this specification, hinokitiols refer to hinokitiol, its derivatives, and salts thereof (such as sodium salts). Methods for producing hinokitiol include, but are not limited to, obtaining it from oil obtained by distilling the wood or roots of Taiwan cypress or Aomori cypress. Examples of hinokitiols include hinokitiol, its salts, metal complexes, and combinations of two or more selected from these. From the viewpoint of achieving the desired effects of the present invention, hinokitiol is preferred. Hinokitiol is known to have 1) antibacterial and 2) anti-inflammatory effects.

[0018] <Copper chlorophylls> In this specification, copper chlorophylls refer to copper chlorophyllin and its salts. Methods for producing copper chlorophyllin and its salts include, but are not limited to, a method of stabilizing the chlorophyll by substituting magnesium in the chlorophyll molecules of plants with copper. Examples of copper chlorophylls include copper chlorophyllin, copper chlorophyllin salts (e.g., alkali metal salts such as sodium salt, potassium salt, calcium salt, and alkaline earth metal salts), and combinations of two or more of these. From the viewpoint of more efficiently achieving the desired effects of the present invention, copper chlorophyllin sodium is preferred as the copper chlorophyllin.

[0019] <Lysozyme chloride> Lysozyme chloride is a component contained in chicken eggs, and its production method may be any known method without particular limitation. As the lysozyme chloride, lysozyme chloride described in the Japanese Pharmacopoeia or the Standards for Cosmetic Ingredients can be used.

[0020] <ε-aminocaproic acid> As the epsilon aminocaproic acid, epsilon aminocaproic acid described in the Japanese Pharmacopoeia or the Standards for Cosmetic Ingredients can be used.

[0021] The active ingredient preferably contains at least one selected from glycyrrhizinic acids, pyridoxine hydrochloride, allantoin, hinokitiol, and epsilon aminocaproic acid, and more preferably glycyrrhizinic acid, glycyrrhizinate, allantoin, hinokitiol, or pyridoxine hydrochloride.

[0022] [1.2 Effective dose] The effective amount of each active ingredient can be determined appropriately depending on the subject of application, but the amount used per administration for humans is usually 0.00001 mg or more, preferably 0.001 mg or more, and more preferably 0.01 mg or more. The upper limit is usually 1 g or less, preferably 0.1 g or less. In particular, when the administration method is local administration (external application), the lower limit of the effective concentration in the administered formulation is preferably 0.0001 mass % or more, more preferably 0.001 mass %. The upper limit is preferably 10 mass % or less, and more preferably 1 mass % or less.

[0023] [1.3 Effect] The active ingredients can control glycative stress. As used herein, glycative stress encompasses all of the reactions that occur from the binding of sugars and proteins (glycation reaction; Maillard reaction) to the production of AGEs (AGE production reaction) to the action of AGEs on cells (inflammatory reaction). The inflammatory reaction caused by AGEs is a reaction in which AGEs bind to RAGE (Receptor for AGEs) expressed on the cell membranes of monocytes / macrophages, vascular endothelial cells, fibroblasts, and other cells, transmitting a signal into the cells and causing inflammation through the activation of AGE-induced oxidative stress, NF-κB activity, mitogen-activated protein kinase (MAPK), and protein kinase C (PKC). The active ingredients can suppress at least a portion of glycative stress and can be used, for example, as glycative stress inhibitors, AGE production inhibitors (which can also be referred to as glycation product production inhibitors, AGE formation inhibitors, glycation product formation inhibitors, etc.), and glycation reaction (Maillard reaction) inhibitors (which can also be referred to as anti-glycation agents, glycation inhibitors, etc.). The agent of the present invention is preferably used for the purposes of inhibiting skin aging, gum care, preventing periodontal disease, dry eye, cataracts, retinopathy, and presbyopia.

[0024] [1.4 Optional components] The agents of the present invention (glycative stress inhibitor, AGE production inhibitor, glycation reaction inhibitor, and AGE-induced inflammatory reaction inhibitor) are used in various dosage forms as described below. The agents may consist of the components described above, or may contain other optional components in addition to these. Examples of optional components include surfactants, buffers, diluents, binders, thickeners, preservatives, excipients, dispersants, disintegrants, solvents, antioxidants, stabilizers, sweeteners, colorants, flavoring agents, antiseptics, emulsifiers, and combinations of two or more selected from these. However, there are no particular limitations on the optional components as long as they are additives used in the field of each dosage form.

[0025] [1.5 Dosage Form] The dosage form of the glycative stress inhibitor, AGE production inhibitor, glycation reaction inhibitor, and AGE-induced inflammatory reaction inhibitor may be, for example, liquid, gel, cream (paste), powder (dispersed), granule, tablet, film, or sheet, and is not particularly limited.

[0026] [2.Application] The above-mentioned glycative stress inhibitor, AGE production inhibitor, glycation reaction inhibitor, and AGE-induced inflammatory reaction inhibitor can be used as quasi-drugs, medicines, cosmetics, and foods.

[0027] [2.1 Target and area of ​​application] The agent of the present invention can be applied to any animal including humans, and is usually mammals, birds, and fish, preferably mammals, more preferably humans.The agent can be applied to healthy individuals, elderly individuals, infected individuals, or individuals suspected of being infected.Except for humans, animals include, for example, mice, rats, hamsters, dogs, cats, sheep, goats, cattle, pigs, and monkeys.

[0028] The agent of the present invention may be applied either locally (to the oral cavity, ophthalmic mucosa, etc., to the skin, or to the hair) or systemically (by internal administration).

[0029] 2.2 Dosage Form Examples of dosage forms of the agent include oral administration (internal administration) and parenteral administration (e.g., transdermal transmucosal administration, intravenous administration, intramuscular administration, subcutaneous administration, nasal administration, and pulmonary administration). Among these, less invasive dosage forms are preferred, and transdermal transmucosal administration (external application) and oral administration are more preferred.

[0030] 2.3 Dosage Form The agent of the present invention can be used as a pharmaceutical, a quasi-drug, a cosmetic, a functional food, etc. The dosage form is not particularly limited, and examples thereof include a liquid, a spray, a solid, a semi-solid, a powder, a granule, and a sheet.

[0031] 2.4 Use in Pharmaceutical Formulations The agent of the present invention can be prepared in the form of various preparations such as topical preparations or oral preparations containing, as an active ingredient, one or more glycative stress inhibitors, AGE production inhibitors, glycative reaction inhibitors, or inflammatory reaction inhibitors caused by AGEs selected from the group consisting of glycyrrhizins, allantoins, azulenes, pyridoxine hydrochloride, hinokitiols, copper chlorophylls, lysozyme chloride, and epsilon-aminocaproic acid.

[0032] Examples of external preparations include preparations for mucous membranes, skin, and hair, with mucous membrane preparations being preferred, oral preparations being more preferred, and those for gum care and periodontal disease prevention being particularly preferred.

[0033] Oral preparations include dentifrices (e.g., toothpaste, gel toothpaste, moisturizing toothpaste, liquid toothpaste), gum care agents, mouthwashes, tongue polishes, oral sprays, oral tablets, gums, mouth fresheners, gargle tablets, oral pastes, oral gels, and oral ointments.

[0034] Examples of skin preparations (external skin preparations) include gels, ointments, creams, external liquids, lotions, sprays, and packs.

[0035] Examples of oral dosage forms (pharmaceuticals, functional foods) include oral liquids, syrups, creams, jellies, pastes, tablets, granules, fine granules, and capsules (soft capsules, hard capsules).

[0036] Cosmetics can be used in the form of, for example, creams, emulsions, packs, gels, aerosols, sheets, etc. Specific examples include skin cosmetics such as lotions, beauty serums, whitening agents, moisturizers, face masks, emulsions, foundations, eye shadows, mascaras, eyebrow pencils, eyeliners, blush powders, lipsticks, lip balms, packs, and soaps; and hair cosmetics such as hair rinses, hair conditioners, hair treatments, hair lotions, hair tonics, hair packs, hair creams, conditioning mousses, hair mousses, hair sprays, shampoos, leave-on treatments, hair dyes, and hair styling products.

[0037] Examples of foods (food compositions) include food compositions with uses such as health foods, functional foods, health foods, health supplements (supplements), nutritional supplements, foods for specified health uses, functional nutritional foods, medical foods, foods for the sick, foods for infants, foods for nursing care, and foods for the elderly.

[0038] [2.5 Content of agent] The content of the active ingredients of the present invention (the respective contents of azulenes, allantoins, glycyrrhizins, pyridoxine hydrochloride, copper chlorophylls, hinokitiols, lysozyme chloride, and epsilon-aminocaproic acid) in each of the above uses is not particularly limited. For example, it is preferably 0.0001% by mass or more, more preferably 0.001% by mass or more, based on the total mass of the agent. The upper limit is preferably 10% by mass or less, more preferably 2% by mass or less, and particularly preferably 1% by mass or less. The content of azulenes and allantoins is preferably 2% by mass or less, more preferably 1% by mass. Therefore, from the viewpoint of achieving the desired effects of the present invention, it is preferably 0.0001 to 10% by mass, more preferably 0.0001 to 2% by mass, and particularly preferably 0.001 to 1% by mass, based on the total mass of the agent. When licorice extract is used as the glycyrrhizinic acid compound, the content is calculated as the total amount of glycyrrhizinic acid, glycyrrhetinic acid, and salts thereof.

[0039] [2.6 Optional components] When the agent of the present invention is in the form of a so-called composition containing other optional ingredients, the other ingredients include, for example, ingredients other than the active ingredients, such as medicinal ingredients, buffers, solubilizers, isotonicity agents, stabilizers, chelating agents, pH adjusters, preservatives, oily ingredients, excipients, disintegrants, binders, lubricants, coating agents, colorants, color formers, flavoring agents (acidulants, fragrances, sweeteners), antioxidants, strengthening agents, leavening agents, thickeners, surfactants, abrasives, humectants, moisturizers, cooling agents, abrasives, binders, astringents, plant extracts, UV absorbers, aqueous solvents, preservatives, seasonings, food ingredients (including food additives), etc. The type and content of the optional ingredients can be selected depending on the intended use of the pharmaceutical, quasi-drug, food composition, or cosmetic, and / or the dosage form, administration method, etc., and may be one kind or a combination of two or more kinds.

[0040] <Medicinal ingredients> Examples of medicinal ingredients include enzymes such as dextranase, mutanase, amylase, protease, and Liteque enzyme; fluorides such as sodium fluoride, sodium monofluorophosphate, and stannous fluoride; anti-inflammatory agents such as tranexamic acid and dihydrocholesterol; metal salts such as zinc salts, copper salts, and tin salts; anti-tartar agents such as condensed phosphates and ethanehydroxydiphosphonate; blood flow promoters such as vitamin E (e.g., tocopherol acetate); hypersensitivity suppressants such as potassium nitrate, aluminum lactate, and strontium chloride; coating agents such as hydroxyethylcellulose dimethyldiallylammonium chloride; astringents such as vitamin C (e.g., ascorbic acid or its salt) and sodium chloride; water-soluble copper compounds such as copper gluconate; anti-tartar agents; amino acids such as alanine, glycine, and proline; plant extracts such as thyme, Scutellaria Root, clove, and witch hazel; callopeptide; and polyvinylpyrrolidone. Other examples include decongestants, anti-inflammatory agents, astringents, antihistamines, vitamins, amino acids, disinfectants, local anesthetics, ingredients other than the active ingredients of the present invention that have an AGE production inhibitory or anti-inflammatory effect, and combinations of two or more selected from these. Decongestants include, for example, naphazoline hydrochloride, tetrahydrozoline hydrochloride, phenylephrine hydrochloride, epinephrine, epinephrine hydrochloride, ephedrine hydrochloride, dl-methylephedrine hydrochloride, tetrahydrozoline nitrate, and naphazoline nitrate. Anti-inflammatory and astringent agents include, for example, neostigmine methylsulfate, berberine chloride, berberine sulfate, zinc sulfate, zinc lactate, bromelain, chamomile, and sodium cromoglycate. Examples of antihistamines include iproheptine hydrochloride, diphenhydramine hydrochloride, diphenhydramine, isothipendyl hydrochloride, and chlorpheniramine maleate. Examples of vitamins include flavin adenine dinucleotide sodium, cyanocobalamin, vitamin A (e.g., retinol acetate, retinol palmitate), and vitamin E (tocopherol acetate (e.g., d-α-tocopherol acetate)). Examples of amino acids include potassium L-aspartate, magnesium L-aspartate, aminoethylsulfonic acid, and sodium chondroitin sulfate.Examples of disinfectants include cetylpyridinium chloride, chlorhexidine gluconate, dequalinium chloride, benzalkonium chloride, benzethrium chloride, iodine, potassium iodide, sulfamethoxazole, sulfamethoxazole sodium, sulfisoxazole, sulfisomidine sodium, and isopropylmethylphenol. Examples of local anesthetics include lidocaine, lidocaine hydrochloride, dibucaine hydrochloride, and chlorobutanol. Each active ingredient may be used alone or in combination of two or more. The content of the active ingredient can be appropriately determined as an effective amount according to conventional methods.

[0041] <Surfactant> The surfactant may be an anionic surfactant, a nonionic surfactant, or an amphoteric surfactant.

[0042] Examples of anionic surfactants include alkyl sulfates, acylamino acid salts, acyltaurine salts, α-olefin sulfonates, hydrogenated coconut fatty acid monoglyceride monosulfates, and lauryl sulfoacetates. The alkyl and acyl groups may be linear or branched, saturated or unsaturated, and typically contain 10 to 20 carbon atoms, preferably 12 to 18, and more preferably 12 to 14 carbon atoms. The salts may be selected from pharmacologically acceptable salts. Examples of pharmacologically acceptable salts include base addition salts and amino acid salts. Specific examples include inorganic base salts such as sodium salts, potassium salts, calcium salts, magnesium salts, and ammonium salts; organic base salts such as triethylammonium salts, triethanolammonium salts, pyridinium salts, and diisopropylammonium salts; and basic amino acid salts such as arginine salts. Among these, inorganic base salts are preferred, with alkali metal salts (e.g., sodium salts and potassium salts) and ammonium salts being more preferred, and sodium salts being even more preferred.

[0043] Examples of alkyl sulfates include lauryl sulfate (sodium lauryl sulfate) and myristyl sulfate. Examples of acylamino acid salts include acyl glutamates such as lauroyl glutamate, myristoyl glutamate, palmitoyl glutamate, etc.; acyl glycine salts such as N-lauroyl-N-methyl glycine salt and cocoyl glycine salt; acyl alanine salts such as N-lauroyl-β-alanine salt, N-myristyl-β-alanine salt, N-cocoyl-β-alanine salt, N-lauroyl-N-methyl-β-alanine salt, N-myristoyl-N-methyl-β-alanine salt, and N-methyl-N-acylalanine salt; and acyl aspartates such as lauroyl aspartate. Examples of acyltaurine salts include lauroyl methyl taurine salt, N-methyl-N-acyltaurine salt, and N-cocoyl methyl taurine salt. Examples of α-olefin sulfonates include α-olefin sulfonates having 12 to 18 carbon atoms, such as tetradecene sulfonate. Other examples of anionic surfactants include hydrogenated coconut fatty acid monoglyceride sodium monosulfate and sodium lauryl sulfoacetate.

[0044] Examples of nonionic surfactants include polyoxyethylene alkyl ethers, polyoxyethylene hydrogenated castor oil, sorbitan fatty acid esters, polyoxyethylene sorbitan fatty acid esters (e.g., polyoxyethylene sorbitan monostearate), alkylolamides, polyoxyethylene fatty acid esters, polyoxyethylene alkenyl ethers, polyglycerin fatty acid esters, sucrose fatty acid esters (e.g., maltose fatty acid esters), sugar alcohol fatty acid esters (e.g., maltitol fatty acid esters, lactitol fatty acid esters), fatty acid diethanolamides (e.g., lauric acid mono- or diethanolamide), polyoxyethylene polyoxypropylene copolymers, and polyoxyethylene polyoxypropylene fatty acid esters. The number of carbon atoms in the alkyl chain of the polyoxyethylene alkyl ether is typically 14 to 18, and the average number of moles of ethylene oxide added is typically 5 to 30. The average number of moles of ethylene oxide added in polyoxyethylene hydrogenated castor oil is typically 20 to 100 moles, preferably 20 to 60 moles. The number of carbon atoms in the fatty acid of the sorbitan fatty acid ester is usually 12 to 18. The number of carbon atoms in the fatty acid of the polyoxyethylene sorbitan fatty acid ester is usually 16 to 18, and the average number of moles of ethylene oxide added is usually 10 to 40. The number of carbon atoms in the alkyl chain of the alkylolamide is usually 12 to 14.

[0045] Examples of amphoteric surfactants include betaine-type amphoteric surfactants such as alkyl dimethylamino acetic acid betaine (e.g., lauryl dimethylamino acetic acid betaine) and fatty acid amidopropyl dimethylamino acetic acid betaine (e.g., cocamidopropyl betaine); imidazoline-type amphoteric surfactants such as N-fatty acid acyl-N-carboxymethyl-N-hydroxyethyl ethylenediamine salts (e.g., N-coconut oil fatty acid acyl-N-carboxymethyl-N-hydroxyethyl imidazolinium betaine), coconut oil fatty acid imidazolinium betaine, and 2-alkyl-N-carboxymethyl-N-hydroxyethyl imidazolinium betaine; and alkyl betaines such as lauryl dimethylamino acetic acid betaine.

[0046] When a surfactant is contained, the content of each of the anionic, nonionic and amphoteric surfactants is usually 0.01 to 10% by mass, preferably 0.1 to 5% by mass, more preferably 0.2 to 3% by mass of the total agent.

[0047] <Abrasives> The abrasive may be either inorganic or organic. Examples of inorganic abrasives include abrasive silicas such as precipitated silica, aluminosilicate, zirconosilicate, crystalline zirconium silicate, and titanium-bonded silica; calcium phosphate compounds such as dibasic calcium phosphate dihydrate or anhydrate, monobasic calcium phosphate, tribasic calcium phosphate, and calcium pyrophosphate; calcium carbonate abrasives such as calcium carbonate; calcium abrasives other than carbonates or phosphates such as calcium hydroxide and calcium sulfate; aluminum-based materials such as aluminum oxide, aluminum hydroxide, and alumina; silicate-based materials such as anhydrous silicic acid, zeolite, and zirconium silicate; magnesium-based materials such as magnesium carbonate and tribasic magnesium phosphate; apatite-based materials such as hydroxyapatite, fluoroapatite, and calcium-deficient apatite; titanium-based materials such as titanium dioxide, titanium mica, and titanium oxide; and minerals such as bentonite. Examples of organic abrasives include polymethyl methacrylate and synthetic resin-based abrasives. Among these, abrasive silica and calcium phosphate compounds are preferred, and silicic anhydride is more preferred. The amount of the abrasive is preferably 50% by mass or less, more preferably 8 to 50% by mass, based on the total mass of the agent.

[0048] <Wetting agent> Examples of humectants include sugar alcohols and polyhydric alcohols other than sugar alcohols. Examples of sugar alcohols include sorbitol (sorbitol), erythritol, maltitol, lactitol, and xylitol. Examples of polyhydric alcohols other than sugar alcohols include glycerin; glycols such as ethylene glycol, propylene glycol, dipropylene glycol, butylene glycol, and polyethylene glycol (PEG); and reduced starch saccharification products. Examples of polyethylene glycols are, for example, polyethylene glycols with an average molecular weight of 150 to 6,000, and polyethylene glycols with an average molecular weight of 190 to 630 (PEG200, PEG300, PEG400, PEG600). The average molecular weight is the average molecular weight specified in the Quasi-drug Raw Materials Standards 2006. The content of the humectant is usually 40% by mass or less, preferably 1 to 30% by mass, of the total agent.

[0049] <Binding agent> Examples of binders include any suitable organic binders known in the art, such as polysaccharides, cellulose-based binders (e.g., carboxymethyl cellulose (CMC), hydroxyethyl cellulose, hydroxypropyl cellulose, hydroxypropylmethyl cellulose, methyl cellulose, and cationized cellulose), other polysaccharide thickeners (e.g., xanthan gum, guar gum, gellan gum, tragacanth gum, karaya gum, gum arabic, locust bean gum, carrageenan, and sodium alginate), and synthetic water-soluble polymers (e.g., sodium polyacrylate, carboxyvinyl polymer, polyvinylpyrrolidone, polyvinyl alcohol, and propylene glycol alginate). Furthermore, inorganic binders such as thickening silica and aluminum silicate may also be added. The content of the organic binder is preferably 0 to 3% by mass, and more preferably 0.1 to 2% by mass, based on the total mass of the agent. The content of the inorganic binder is preferably 0 to 10% by mass, and more preferably 1 to 8% by mass.

[0050] <Buffering agent> Examples of buffering agents include citric acid or a salt thereof (e.g., sodium citrate), phosphoric acid or a salt thereof (e.g., sodium hydrogen phosphate, sodium dihydrogen phosphate, dipotassium phosphate, potassium dihydrogen phosphate), tartaric acid or a salt thereof (e.g., sodium tartrate), gluconic acid or a salt thereof (e.g., sodium gluconate), acetic acid or a salt thereof (e.g., sodium acetate), carbonic acid or a salt thereof (e.g., sodium bicarbonate), trometamol, amino acids (e.g., potassium aspartate, aminoethylsulfonic acid, glutamic acid, sodium glutamate), and combinations of two or more thereof. Examples of solubilizing agents include polyoxyethylene higher fatty acid esters such as polyoxyethylene (e.g., p=60) hydrogenated castor oil, polyoxyethylene sorbitan higher fatty acid esters such as polyoxyethylene (e.g., p=20) sorbitan monooleate, propylene glycol, polyethylene glycol, and combinations of two or more thereof. Examples of isotonic agents include sodium chloride, potassium chloride, glycerin, and combinations of two or more thereof. Examples of stabilizers include sodium edetate, cyclodextrin, sulfites, citric acid or its salts, dibutylhydroxytoluene, ascorbic acid, and combinations of two or more thereof. Examples of chelating agents include sodium edetate, sodium citrate, and combinations of two or more thereof. Examples of pH adjusters include sodium hydroxide, potassium hydroxide, hydrochloric acid, and combinations of two or more thereof. Examples of preservatives include parahydroxybenzoic acid esters such as methylparaben, ethylparaben, propylparaben, and butylparaben, alcohol derivatives such as phenylethyl alcohol, benzyl alcohol, phenol, and acrinol, sorbic acid and its salts (potassium sorbate, etc.), benzalkonium chloride, benzethonium chloride, chlorhexidine gluconate, cetylpyridinium chloride, alkylpolyaminoethylglycine, and combinations of two or more thereof.

[0051] <Moisturizer> Examples of moisturizing agents include glycerin, concentrated glycerin, sugar alcohols (for example, sorbitol, xylitol, maltitol, mannitol, reduced starch syrup, reduced palatinose, erythritol, lactitol, and isomalt), and combinations of two or more of these.

[0052] <Flavoring agent> Examples of flavoring agents include sweeteners (e.g., saccharin sodium, aspartame, stevia, stevioside, para-methoxycinnamic aldehyde, neohesperidin dihydrochalcone, perillartine, aspartyl phenylalanine methyl ester, thaumatin, acesulfame potassium, sucralose, maltitol, sorbitol, mannitol, reduced starch syrup, reduced palatinose, xylitol, erythritol, lactitol and other artificial sweeteners), flavorings (e.g., anise oil, cassia), Natural essential oils such as citric acid, citrus oil, citrus limonene ... Fragrance components contained in the above natural essential oils, such as thall, methyl salicylate, eugenol, linalool, limonene, menthone, menthyl acetate, citral, decanal, camphor, borneol, pinene, spilanthol, n-decyl alcohol, citronellol, α-terpineol, citronellyl acetate, ethyl linalool, and vanillin; ethyl acetate, ethyl butyrate, isoamyl acetate, hexanal, hexenal, methyl anthranilate, and ethyl methylphenyl. Flavoring ingredients such as glycidyl glycidate, benzaldehyde, vanillin, ethyl vanillin, furaneol, N-ethyl-p-menthane-3-carboxamide, menthyl lactate, and ethylene glycol-l-menthyl carbonate; and various blended flavors such as mint, fruit, and herb flavors that combine several flavoring ingredients and natural essential oils (e.g., Peppermint Micron X-8277-T, Dry Coat Matcha #421), acidulants (e.g., citric acid, tartaric acid, malic acid), and green tea powder.

[0053] <Oily ingredients> Examples of oily components include fatty acid esters (e.g., glycerin fatty acid esters), hydrocarbons (e.g., paraffin, liquid paraffin, ceresin, squalane, petrolatum, microcrystalline wax), higher fatty acids (e.g., fatty acids having 8 to 22 carbon atoms such as lauric acid, myristic acid, oleic acid, and isostearic acid), higher alcohols (e.g., alcohols having 8 to 22 carbon atoms such as lauryl alcohol, cetyl alcohol, cetostearyl alcohol, oleyl alcohol, and isostearyl alcohol), vegetable oils (e.g., vegetable oils such as olive oil, castor oil, and coconut oil; fatty acid esters such as isopropyl myristate), beeswax, and combinations of two or more of these.

[0054] <Preservatives> Examples of preservatives include parahydroxybenzoic acid esters (eg, methyl parahydroxybenzoate, ethyl parahydroxybenzoate, butyl parahydroxybenzoate), sodium benzoate, and combinations of two or more thereof.

[0055] <Wetting agent> Examples of humectants include sugar alcohols and polyhydric alcohols other than sugar alcohols. Examples of sugar alcohols include sorbitol (sorbitol), erythritol, maltitol, lactitol, and xylitol. Examples of polyhydric alcohols other than sugar alcohols include glycerin; glycols such as ethylene glycol, propylene glycol, dipropylene glycol, butylene glycol, and polyethylene glycol (PEG); and reduced starch saccharification products. Examples of polyethylene glycols are, for example, polyethylene glycols with an average molecular weight of 150 to 6,000, and polyethylene glycols with an average molecular weight of 190 to 630 (PEG200, PEG300, PEG400, PEG600). The average molecular weight is the average molecular weight specified in the Quasi-drug Raw Materials Standards 2006. The amount of humectant is usually 40% by mass or less, preferably 1 to 30% by mass, of the total agent.

[0056] <Coloring agent> Examples of colorants include natural colorants such as safflower red pigment, gardenia yellow pigment, gardenia blue pigment, perilla pigment, red kojic pigment, red cabbage pigment, carrot pigment, hibiscus pigment, cocoa pigment, spirulina blue pigment, tamarind pigment, etc., legal colorants such as Red No. 2, Red No. 3, Red No. 104, Red No. 105, Red No. 106, Red No. 227, Yellow No. 4, Yellow No. 5, Green No. 3, Blue No. 1, etc., riboflavin, titanium dioxide, and the like. When a colorant is included, its content is preferably 0.00001 to 3% by mass based on the whole agent.

[0057] <pH adjuster> Examples of pH adjusters include organic acids such as phthalic acid, citric acid, succinic acid, acetic acid, fumaric acid, malic acid, and lactic acid or their salts (sodium citrate), inorganic acids such as phosphoric acid (orthophosphoric acid) or their salts (e.g., potassium salts, sodium salts, and ammonium salts), hydroxides such as sodium hydroxide and potassium hydroxide. Examples of inorganic acid salts include disodium hydrogen phosphate and sodium dihydrogen phosphate. The content of the pH adjuster can usually be an amount such that the pH of the agent after addition is 5 to 9, preferably 6 to 8.5. In this specification, the pH value usually refers to the value 25 °C and 3 minutes after the start of measurement. The pH value can be measured, for example, using a pH meter (model number Hm-30S) manufactured by Toa Denpa Kogyo Co., Ltd.

[0058] <Solvent> Examples of solvents include water (purified water) and ethanol, with water being preferred. The solvent may be used alone or in combination of two or more.

[0059] <Excipient> Examples of excipients include celluloses such as hydroxypropyl cellulose, hydroxypropylmethyl cellulose, methyl cellulose, crystalline cellulose, ethyl cellulose, methylethyl cellulose, and low-substituted hydroxypropyl cellulose, and pharmacologically acceptable derivatives thereof; synthetic polymers such as polyvinylpyrrolidone and partially saponified polyvinyl alcohol; polysaccharides such as gelatin, powdered gum arabic, pullulan, agar, alginic acid, sodium alginate, and xanthan gum; corn starch, potato starch, pregelatinized starch, and hydroxypropyl starch. starches and pharmacologically acceptable derivatives thereof; lactose, lactose granules, fructose, glucose, sucrose, granulated sugar, hydrous glucose, trehalose, palatinose, mannitol, sorbitol, erythritol, xylitol, maltotetraose, lactitol, isomalt, reduced palatinose, reduced starch syrup, powdered reduced maltose starch syrup, maltitol; inorganic excipients such as magnesium carbonate, calcium carbonate, light anhydrous silicic acid, silicon dioxide (also known as anhydrous silicic acid, fine silicon dioxide), titanium oxide, aluminum hydroxide gel, etc.; and combinations of two or more of these.

[0060] <Disintegrant> Examples of disintegrants include crospovidone, carmellose calcium, croscarmellose sodium, low-substituted hydroxypropyl cellulose, carboxymethyl cellulose, carboxymethyl starch sodium, croscarmellose sodium, cross-linked insoluble polyvinylpyrrolidone, hydroxypropyl starch, partially pregelatinized starch, corn starch, and combinations of two or more thereof.

[0061] <Binder> Examples of binders include hydroxypropyl cellulose, hydroxypropylmethyl cellulose, methyl cellulose, ethyl cellulose, polyvinyl alcohol, polyvinylpyrrolidone, gelatin, dextrin, starch, pregelatinized starch, and combinations of two or more thereof.

[0062] <Lubricant> Examples of lubricants include calcium stearate, magnesium stearate, sucrose fatty acid esters, light anhydrous silicic acid, sodium stearyl fumarate, polyethylene glycol, talc, stearic acid, and combinations of two or more thereof.

[0063] <Other optional ingredients> Examples of optional components other than those mentioned above include polyisobutylene, polybutadiene, urethane, silicone, and natural rubber. The content of these optional components can be appropriately set within a range that does not impair the effects of the present invention.

[0064] [3. Manufacturing method] The manufacturing method for oral preparations, topical preparations, and other preparations using glycative stress inhibitors, AGE production inhibitors, glycation reaction inhibitors, or AGE-induced inflammatory reaction inhibitors may be determined depending on the dosage form, intended use, and application site. For example, when used as a toothpaste, a method is exemplified in which the solvent-soluble components are prepared, followed by mixing with other insoluble components and optionally degassing (e.g., by reducing pressure). Another example is a method in which the active ingredient and other optional components are dispersed and dissolved in an aqueous solvent (e.g., purified water, sterilized water, etc.) to prepare a composition, which is then filled into an appropriate container (e.g., glass or resin). Examples of containers for oral preparations include laminated tubes, and materials that can be used include resins such as polyethylene, polypropylene, polyethylene terephthalate, and nylon. In the case of a spray formulation, a container equipped with a spraying device (e.g., a trigger-type, pump-type, or aerosol-type container) can be selected. The resulting toothpaste can then be filled into a container to produce a finished product. The shape and material of the container are not particularly limited, and any container that is commonly used for oral compositions can be used.

[0065] [4. How to use] Oral preparations, topical preparations, and other agents that suppress glycative stress, AGE production, glycation reaction, or AGE-induced inflammatory reaction can be used, for example, by administering the agent to the application site. The number of times administered per day is not particularly limited, but may be, for example, 1 to 6 times or more. In the case of oral preparations, a method of applying an appropriate amount of the agent to a toothbrush, brushing the surface of the teeth, and rinsing with water after use (dentifrice), or a method of holding an appropriate amount in the mouth, gargling, and then spitting it out (mouthwash) may be used. [Example]

[0066] [Test Example 1] Evaluation of gingival inflammation caused by glycation (reference example) Human gingival fibroblasts were seeded in multiwell plates and cultured in α-MEM (ThermoFischer) supplemented with 10% bovine serum until subconfluent. The medium was then removed and replaced with medium supplemented with 0, 250, 500, or 1000 μg / mL of Control-BSA (hereafter referred to as BSA, BioVision) or 0, 250, 500, or 1000 μg / mL of AGEs-BSA (glycosylated form of Control-BSA, hereafter referred to as AGEs, BioVision). After 24 hours, the culture supernatants were collected, and the IL-6 protein levels in the supernatants were assessed by ELISA (R&D Systems) (Figure 1). Compared to the supernatant supplemented with BSA, the IL-6 concentration was higher in the supernatant supplemented with AGEs. Furthermore, the IL-6 concentration increased depending on the AGE concentration. The results of this experiment demonstrate that AGEs promote inflammation in a concentration-dependent manner.

[0067] [Test Example 2] AGE production inhibitory activity test (Examples 1 to 8 and Comparative Example 1) <Preparation of collagen gel> The ice-cold neutralized collagen solution was dispensed into a 96-well black plate at 50 μL / well and left to stand overnight in a 37° C. incubator under humid conditions.

[0068] <Preparation of sample solution> Sodium azulenesulfonic acid (manufactured by Tokyo Chemical Industry Co., Ltd.), allantoin (manufactured by Fujifilm Wako Pure Chemical Corporation), dipotassium glycyrrhizinate (manufactured by Alps Chemical Industry Co., Ltd.), pyridoxine hydrochloride (manufactured by DSM), sodium copper chlorophyllin (manufactured by Tama Biochemical Co., Ltd.), hinokitiol (manufactured by Fujifilm Wako Pure Chemical Corporation), lysozyme chloride (manufactured by Fujifilm Wako Pure Chemical Corporation), and epsilon-aminocaproic acid (manufactured by Nippon Rikagaku Yakuhin Co., Ltd., Example 8) were each diluted with phosphate buffer (PBS) so that each component had a predetermined concentration and prepared. Tocopherol acetate (manufactured by Fujifilm Wako Pure Chemical Corporation) was dissolved in ethanol and then diluted with PBS so that the final ethanol concentration was 1% or less. In addition, the same commercially available products as in this test example were used in the following test examples for the samples.

[0069] <Preparation of Test Substances> 40 μL of the sample solution group and 10 μL of a 500 mM glyceraldehyde solution were mixed on a previously prepared collagen gel. Then, it was allowed to stand at 37 °C for 24 hours under wet conditions. Then, the collagen gel was irradiated with excitation light at 370 nm, and the fluorescence at 440 nm generated was measured. The result obtained from this measurement was designated as Measurement Value A.

[0070] <Preparation of blank> The blank was prepared as follows. 40 μL of PBS and 10 μL of a 500 mM glyceraldehyde solution were mixed on a previously prepared collagen gel. Then, it was allowed to stand at 37 °C for 24 hours under wet conditions. Then, the collagen gel was irradiated with excitation light at 370 nm, and the fluorescence at 440 nm generated was measured. The result obtained from this measurement was designated as Measurement Value B.

[0071] <Test Results> The test results were calculated as AGEs production inhibition rate (%) = 100 - {(Measurement Value A / Measurement Value B) × 100}. Each value is the average value of N = 3.

[0072]

Table 1

[0073] In Comparative Example 1, the AGE production inhibition rate was 0% or less, whereas in Examples 1 to 8, the AGE production inhibition rate exceeded 0%, and particularly, when the solution concentrations were 0.001, 0.01, and 1%, the production inhibition rate was high, at 20% or more. These results indicate that each compound of the Examples inhibits glycation reactions and acts as a glycation reaction inhibitor. Furthermore, as a result, AGE production is inhibited, indicating that the compound acts as an AGE production inhibitor.

[0074] It is known that oxidative stress is involved in glycation reactions, and tocopherol acetate (Comparative Example 1) is an antioxidant and was expected to suppress glycation reactions. However, in this example, tocopherol acetate did not inhibit AGE production (Table 1), and the results of this test example indicate that substances with antioxidant activity do not necessarily have AGE production inhibitory activity.

[0075] [Test Example 3] Evaluation of inflammatory response inhibition by AGEs <Preparation of sample solution> Components 1 to 7 shown in Table 2 were added to α-MEM (manufactured by ThermoFischer) to prepare a sample solution. [Table 2]

[0076] <Evaluation of inflammatory response in sample solutions> Human gingival fibroblasts were seeded onto multiwell plates and cultured in medium containing 10% bovine serum until subconfluent. The medium was then removed and replaced with sample solution. After 6 hours, the human gingival fibroblasts were harvested, RNA was extracted, cDNA was prepared, and gene expression analysis of IL-6 levels was performed by RT-PCR. IL-6 gene expression levels were normalized using the gene expression level of GAPDH, an endogenous control, and the IL-6 gene expression ratio was calculated with the negative control set at 1 (Figure 2).

[0077] The IL-6 gene expression rate at 250 μg / mL of AGEs (positive control: Comparative Example 3) was approximately 2.5 times higher than that at 250 μg / mL of BSA (negative control: Comparative Example 2). In a sample solution containing 250 μg / mL of AGEs and 0.1% each of allantoin, dipotassium glycyrrhizinate, pyridoxine hydrochloride, hinokitiol, and epsilon-aminocaproic acid, the IL-6 gene expression rate was lower than that of the positive control (Examples 9 to 13).

[0078] The results of this test demonstrate that AGEs induce inflammatory responses and that the agent of the present invention can suppress inflammatory responses caused by AGEs.

[0079] From the above examples, it is clear that each active ingredient has the activity of suppressing glycative stress occurring in the body, the AGE production reaction that causes inflammatory reactions, and the activity of suppressing the inflammatory reaction caused by AGEs itself, and can suppress inflammation caused by AGEs.

Claims

1. An AGEs (Advanced Glycation End Products) production inhibitor containing one or more members selected from the group consisting of azulene, sodium azulene sulfonate, 1,4-dimethyl-7-isopropylazulene, and 4,8-dimethyl-2-isopropylazulene.

2. 2. The AGE production inhibitor according to claim 1, wherein the AGEs are derived from collagen.

3. 3. The AGEs production inhibitor according to claim 1 or 2, wherein the AGEs are derived from collagen and glyceraldehyde.

4. A glycation reaction inhibitor comprising at least one member selected from the group consisting of azulene, sodium azulene sulfonate, 1,4-dimethyl-7-isopropylazulene, and 4,8-dimethyl-2-isopropylazulene.

5. The glycation reaction inhibitor according to claim 4, which inhibits the glycation reaction of collagen.

6. The glycation reaction inhibitor according to claim 4 or 5, which inhibits the glycation reaction of collagen caused by glyceraldehyde.

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