Intermedicine inhibitors and oral compositions

Abstract

The goal is to inhibit intermedicine using materials that are easier to obtain and can be applied to a wide range of dosage forms. [Solution] The intermedicine inhibitor contains at least one selected from the group consisting of loquat leaf extract, geranium extract, peony extract, copper chlorophyllin sodium, cherry bark extract, St. John's wort extract, black tea extract, peony extract, Houttuynia cordata extract, birch extract, oolong tea extract, hawthorn extract, linden extract, tea extract, witch hazel extract, saxifrage extract, Sanguisorba officinalis extract, and catechu extract.

Description

Technical Field

[0001] The present invention relates to an intermedilysin inhibitor and an oral composition.

Background Art

[0002] As a bacterium that normally resides in the oral cavity, Streptococcus intermedius is known. This bacterium is known as a pathogenic bacterium that causes abscess formation in deep organs such as pneumonia, lung abscess, empyema, brain abscess, and liver abscess. Regarding the pathogenicity of this bacterium, there is a report that the ability to avoid phagocytosis from immune cells is partly due to the production of intermedilysin (hereinafter also referred to as "ILY"), which is a cholesterol-dependent cytolysin.

[0003] For example, Patent Document 1 discloses the use of empty liposomes for the treatment and prevention of bacterial infections. The use of empty liposomes containing cholesterol and sphingomyelin, and mixtures of empty liposomes containing either cholesterol and sphingomyelin or phosphatidylcholine and sphingomyelin with other empty liposomes having a defined lipid composition is disclosed. ILY is disclosed as a toxin that causes bacterial infections.

Prior Art Documents

Patent Documents

[0004]

Patent Document 1

Summary of the Invention

Problems to be Solved by the Invention

[0005] However, empty liposomes are not readily available because they require multiple preparation steps. Furthermore, the stability of liposomes is compromised depending on the type and amount of surfactants used, or the presence of ethanol, making it difficult to incorporate them into a wide range of dosage forms. Therefore, they are not easily used for the treatment and prevention of bacterial infections. There is a need for an intermedicine inhibitor that is more readily available and can inhibit ILY using a material that can be applied to a wide range of dosage forms. [Means for solving the problem]

[0006] The essence of the intermedicine inhibitor for solving the above problems is that it contains at least one selected from the group consisting of loquat leaf extract, geranium extract, peony extract, copper chlorophyllin sodium, cherry bark extract, St. John's wort extract, black tea extract, peony extract, Houttuynia cordata extract, birch extract, oolong tea extract, hawthorn extract, linden extract, tea extract, witch hazel extract, saxifrage extract, Sanguisorba officinalis extract, and catechu extract.

[0007] The oral composition for solving the above problems contains at least one selected from the group consisting of loquat leaf extract, geranium extract, peony extract, copper chlorophyllin sodium, cherry bark extract, St. John's wort extract, black tea extract, peony extract, Houttuynia cordata extract, birch extract, oolong tea extract, hawthorn extract, linden extract, tea extract, witch hazel extract, saxifrage extract, Sanguisorba officinalis extract, and catechu extract, and is intended to inhibit intermedicine. [Effects of the Invention]

[0008] According to the intermedicine inhibitor of the present invention, ILY can be inhibited using materials that are easier to obtain and can be applied to a wide range of dosage forms. [Modes for carrying out the invention]

[0009] Embodiments of the intermedicine inhibitor (hereinafter also referred to as "ILY inhibitor") of the present invention will be described below. The ILY inhibitor contains at least one selected from the group consisting of loquat leaf extract, geranium extract, peony extract, copper chlorophyllin sodium, cherry bark extract, St. John's wort extract, black tea extract, peony extract, Houttuynia cordata extract, birch extract, oolong tea extract, hawthorn extract, linden extract, tea extract, witch hazel extract, saxifrage extract, Sanguisorba officinalis extract, and catechu extract.

[0010] <Loquat leaf extract> Loquat leaf extract is an extract derived from the leaves of the loquat tree, which belongs to the rose family. <Geranium thunbergii extract> Geranium extract is an extract obtained from the flowers, leaves, and stems of Geranium thunbergii, a perennial plant belonging to the genus Geranium in the family Geraniaceae.

[0011] <Peony extract> Peony extract is an extract obtained from the roots of the peony plant, which belongs to the Paeoniaceae family. <Sodium copper chlorophyllin> Sodium copper chlorophyllin is a compound in which magnesium, a coordinating element of chlorophyll naturally present in plants, is replaced with copper. It is commonly used as a green coloring agent.

[0012] <Cherry bark extract> Cherry bark extract is an extract obtained from the bark of trees belonging to the genus Prunus, such as the mountain cherry (Prunus jamasakura), which are part of the Rosaceae family. <Hypericum extract> St. John's wort extract is an extract derived from the flowers, leaves, and stems of St. John's wort, a perennial plant belonging to the genus St. John's wort in the family Hypericaceae.

[0013] <Black tea extract> Black tea extract is an extract derived from Assam tea leaves. <Peony extract> The peony extract is an extract obtained from the flowers and roots of the peony, a perennial plant of the Paeoniaceae family.

[0014] <Houttuynia cordata extract> Houttuynia cordata extract is an extract obtained from the flowers, leaves, and stems of Houttuynia cordata of the Saururaceae family. <Birch extract> Birch extract is an extract obtained from the bark and xylem of the birch of the Betulaceae family.

[0015] <Oolong tea extract> Oolong tea extract is an extract obtained from oolong tea produced from the leaves of Camellia sinensis.

[0016] <Japanese quince extract> Japanese quince extract is an extract obtained from the fruits of Japanese quince of the Rosaceae family. <Machilus thunbergii extract> Machilus thunbergii extract is an extract obtained from the flowers and leaves of Machilus thunbergii, Machilus philippensis, Machilus nanmu, etc. of the Machilus genus.

[0017] <Tea extract> Tea extract is an extract obtained from green tea. <Hamamelis extract> Hamamelis extract is an extract obtained from the leaves of Hamamelis virginiana of the Hamamelidaceae family.

[0018] <Saxifraga stolonifera extract> Saxifraga stolonifera extract is an extract obtained from the leaves and stems of Saxifraga stolonifera of the Saxifragaceae family.

[0019] <Rubus parvifolius extract> Rubus parvifolius extract is an extract obtained from the roots and stems of Rubus parvifolius of the Rosaceae family. <Morinda citrifolia extract> Morinda citrifolia extract is an extract obtained from the leaves and stems of Morinda citrifolia of the Rubiaceae family.

[0020] The ILY inhibitor of the present invention may contain only one of the above-mentioned extracts or solutions, or it may contain two or more in appropriate combinations. The extraction method for each of the above extracts and extractants is not particularly limited. For example, known extraction methods such as solvent extraction, ultrasonic extraction, pressing, and supercritical extraction can be employed.

[0021] Solvent extraction is performed using solvents such as water, ethanol, propylene glycol, glycerin, and 1,3-butylene glycol. For example, extraction may be performed using an aqueous solution of the above solvent, such as a 50% by volume aqueous solution of ethanol.

[0022] Each of the above extracts can be obtained by extracting with the above solvent and then concentrating the extracted extract under conditions that prevent volatilization. The concentrated liquid obtained may be further concentrated to a syrup-like consistency to prepare a soft extract, or the concentrated liquid may be cooled and dried, then pulverized to prepare a dry extract.

[0023] In the following explanation, the above-mentioned extracts, extract solutions, and sodium copper chlorophyllin will be collectively referred to as "extracts." The ILY inhibitors of the present invention can use commercially available products of the above-mentioned extracts.

[0024] <Other ingredients> ILY inhibitors may contain other components besides those mentioned above, depending on the intended use, form, and application. Examples of other components include surfactants, flavoring agents, sweeteners, humectants, binders, preservatives, colorants, pH adjusters, chelating agents, pharmacoactive ingredients, bases, abrasives, and stabilizers. Other components that are known to be incorporated into ILY inhibitors may be used. ILY inhibitors may contain only one of each of the above-mentioned other components, or they may contain two or more in appropriate combinations.

[0025] (Surfactants) Examples of surfactants include nonionic surfactants, anionic surfactants, and amphoteric surfactants.

[0026] Specific examples of nonionic surfactants include sugar fatty acid esters such as sucrose fatty acid esters, maltose fatty acid esters, and lactose fatty acid esters; fatty acid alkanolamides; glycerin fatty acid esters; sorbitan fatty acid esters; fatty acid monoglycerides; polyoxyethylene alkyl ethers with a polyoxyethylene addition coefficient of 8 to 10 and 13 to 15 carbon atoms in the alkyl group; polyoxyethylene alkylphenyl ethers with a polyoxyethylene addition coefficient of 10 to 18 and 9 carbon atoms in the alkyl group; diethyl sebacate; polyoxyethylene hydrogenated castor oil; and fatty acid polyoxyethylene sorbitan.

[0027] Specific examples of anionic surfactants include, for example, sulfate ester salts such as sodium polyoxyethylene lauryl ether sulfate; sulfosuccinates such as sodium lauryl sulfosuccinate and sodium polyoxyethylene lauryl ether sulfosuccinate; acyl amino acid salts such as sodium cocoyl sarcosinate and sodium lauroyl methylalanine; and sodium cocoyl methyl taurate.

[0028] Specific examples of amphoteric surfactants include, for example, betaine-type surfactants such as lauryldimethylaminoacetic acid betaine and coconut oil fatty acid amidopropyldimethylaminoacetic acid betaine; imidazoline-type surfactants such as N-cocoyl-N-carboxymethyl-N-hydroxyethylethylenediamine sodium; and amino acid-type surfactants such as N-lauryldiaminoethylglycine.

[0029] (Flavoring agent) Specific examples of flavoring agents include menthol, carboxylic acids, anethole, eugenol, methyl salicylate, limonene, ocimene, n-decyl alcohol, citronellol, α-terpineol, methyl acetate, citronenyl acetate, methyl eugenol, cineole, thymol, spearmint oil, peppermint oil, lemon oil, orange oil, sage oil, rosemary oil, perilla oil, wintergreen oil, clove oil, eucalyptus oil, pimento oil, d-camphor, d-borneol, fennel oil, cinnamon oil, cinnamaldehyde, peppermint oil, vanillin, and others.

[0030] (Sweetener) Specific examples of sweeteners include, for instance, sodium saccharin, potassium acesulfamethamphetamine, stevioside, neohesperidyl dihydrochalcone, perillartin, thaumatin, aspartylphenylalanyl methyl ester, and p-methoxycinnamic aldehyde.

[0031] (Humectant) Specific examples of wetting agents include, for example, sorbitol, ethylene glycol, propylene glycol, glycerin, 1,3-butylene glycol, polypropylene glycol, xylitol, maltitol, lactitol, and polyoxyethylene glycol.

[0032] (Binding agent) Specific examples of binders include, for example, cellulose derivatives such as sodium carboxymethylcellulose, carboxymethyl ethylcellulose salt, hydroxyethylcellulose, hydroxypropylcellulose, hydroxypropyl methylcellulose, methylcellulose, ethylcellulose, crystalline cellulose, and crystalline cellulose-carmellose sodium; microbially produced polymers such as xanthan gum; natural polymers or natural rubbers such as tragacanth gum, karaya gum, arabic gum, carrageenan, dextrin, agar, pectin, pullulan, gellan gum, locust bean gum, and sodium alginate; synthetic polymers such as polyvinyl alcohol, polyvinylpyrrolidone, carboxyvinyl polymer, polyvinyl methyl ether, and sodium polyacrylate; inorganic binders such as thickening silica and bee gum; and cationic binders such as O-[2-hydroxy-3-(trimethylammonio)propyl]hydroxyethylcellulose chloride.

[0033] (Preservative) Specific examples of preservatives include parabens such as methylparaben, ethylparaben, propylparaben, and butylparaben, as well as sodium benzoate, phenoxyethanol, and alkyldiaminoethylglycine hydrochloride.

[0034] (Coloring agent) Specific examples of colorants include legally approved pigments such as Blue No. 1, Yellow No. 4, Red No. 202, and Green No. 3, mineral pigments such as ultramarine, enhanced ultramarine, and Prussian blue, and titanium dioxide.

[0035] (pH adjuster) Specific examples of pH adjusters include, for example, citric acid, phosphoric acid, lactic acid, tartaric acid, glycerophosphate, acetic acid, nitric acid, or chemically possible salts thereof, or sodium hydroxide.

[0036] (Chelating agent) Specific examples of chelating agents include, for example, EDTA, sodium EDTA, potassium EDTA, and phytic acid.

[0037] (Medicinal ingredients) Specific examples of active ingredients include, for example, vitamin E compounds such as dl-α-tocopherol acetate, tocopherol succinate, or tocopherol nicotinate; vitamin C compounds such as ascorbic acid, sodium ascorbate, or magnesium ascorbyl phosphate; vitamin B6 compounds such as pyridoxine hydrochloride; amphoteric disinfectants such as dodecyldiaminoethylglycine; nonionic disinfectants such as triclosan and isopropylmethylphenol; anionic disinfectants such as sorbic acid; and cetylpyridinium chloride, chlorhexidine hydrochloride, benzalkonium chloride, and benzethonium chloride. Examples include cationic disinfectants, enzymes such as dextranase, amylase, protease, mutanase, lysozyme, and lytic enzymes (Litec enzyme), alkali metal monofluorophosphates such as sodium monofluorophosphate and potassium monofluorophosphate, fluorides such as sodium fluoride and stannous fluoride, tranexamic acid and epsilon-aminocaproic acid, aluminum chlorohydroxyl allantoin, dihydrocholesterol, chlorophyll, sodium chloride, caropeptide, allantoin, carbazochrome, potassium nitrate, and palatinite.

[0038] (Base) Specific examples of bases include alcohols, silicones, apatite, white petrolatum, paraffin, liquid paraffin, microcrystalline wax, squalane, and Plastibase.

[0039] Specific examples of alcohols include ethyl alcohol, lauryl alcohol, myristyl alcohol, and others. (Abrasive) Specific examples of abrasives include calcium carbonate, magnesium carbonate, dicalcium phosphate, tricalcium phosphate, magnesium phosphate, silica, zeolite, sodium metaphosphate, aluminum hydroxide, magnesium hydroxide, calcium pyrophosphate, red iron oxide, calcium sulfate, and anhydrous silicic acid.

[0040] (Stabilizer) Specific examples of the stabilizer include, for example, sodium thiosulfate, sodium sulfite, calcium lactate, lanolin, triacetin, castor oil, magnesium sulfate, and the like.

[0041] <Content in the ILY inhibitor> The content of each of the above extracts in the ILY inhibitor is not particularly limited. It can be appropriately set within the range where the effects as an ILY inhibitor are exhibited.

[0042] Assuming the total volume of the ILY inhibitor is 100%, the content of each of the above extracts is preferably 0.1% by volume or less, more preferably 0.05% by volume or less, and even more preferably 0.02% by volume or less. Also, preferably, it is 0.001% by volume or more.

[0043] <Application form, use, and dosage form> The application form of the ILY inhibitor is not particularly limited. For example, it can be used as food, cosmetics, pharmaceuticals, and quasi-drugs. Also, it can be used as an oral composition, a throat composition, an oral composition, and a pharmaceutical composition for the lungs and bronchi containing the ILY inhibitor.

[0044] When used as an oral composition or a throat composition, for example, it can be applied to sprays, dentifrices, liquid dentifrices, mouthwashes, gargles, etc. Examples of sprays include nasal sprays, throat sprays, oral sprays, etc.

[0045] When used as an oral composition, for example, it can be applied to candies, troches, tablets, gums, gummies, granules, powders, jellies, syrups, beverages, etc. When used as a pharmaceutical composition for the lungs and bronchi, for example, it can be applied as an inhalation drug for inhalers. It can be applied to inhalation drugs for aspiration pneumonia, etc.

[0046] <Actions and effects of this embodiment> The actions and effects of this embodiment will be described. (1) The ILY inhibitor contains at least one selected from the group consisting of loquat leaf extract, geranium extract, peony extract, copper chlorophyllin sodium, cherry bark extract, St. John's wort extract, black tea extract, peony extract, Houttuynia cordata extract, birch extract, oolong tea extract, hawthorn extract, linden extract, tea extract, witch hazel extract, saxifrage extract, Sanguisorba officinalis extract, and catechu extract.

[0047] According to the ILY inhibitor of this embodiment, ILY can be inhibited by the above-mentioned extracts binding to ILY or masking the surface of ILY. Therefore, ILY can be inhibited using materials that are easier to obtain and can be applied to a wide range of dosage forms.

[0048] (2) The oral composition contains at least one selected from the group consisting of loquat leaf extract, geranium extract, peony extract, copper chlorophyllin sodium, cherry bark extract, St. John's wort extract, black tea extract, peony extract, Houttuynia cordata extract, birch extract, oolong tea extract, hawthorn extract, linden extract, tea extract, witch hazel extract, saxifrage extract, Sanguisorba officinalis extract, and catechu extract, and is for ILY inhibition. Therefore, an oral composition for ILY inhibition can be obtained using materials that are easier to obtain and can be applied to a wide range of dosage forms. [Examples]

[0049] The ILY inhibitor of this embodiment will be described in more detail based on the following examples. Note that the ILY inhibitor is not limited to the configuration described in the Examples section. <Preparation of sample solution> Sample solutions for Examples 1-30 and Comparative Examples 1-4 were prepared. The types of materials contained in each sample solution are shown in Table 1. Commercially available materials were used for each sample.

[0050] The preparation method for each sample solution involved mixing each extract with phosphate-buffered saline (hereinafter also referred to as "PBS") and the toxin so that the concentration of the extract of each material was 1.25 times the concentration shown in Table 1. For example, in Example 1, a commercially available loquat leaf extract with a concentration of 1.5 vol% was prepared, and a sample solution of 0.0125 vol% was prepared using PBS. Subsequently, it was mixed with the toxin solution in a microcentrifuge tube. A sample solution with a final loquat leaf extract concentration of 0.01 vol% in the test system was prepared. For each sample solution, 40 μL of PBS and 50 μL of a 10 μg / mL solution of toxin were mixed. The toxin used was ILY prepared by the laboratory of Atsushi Tabata, Department of Bioresource Industry, Graduate School of Social and Industrial Science and Technology, Tokushima University.

[0051] Comparative Example 1 involved mixing only PBS and toxin without adding any extract. Comparative Example 2 used Phellodendron amurense bark extract, extracted from the bark of the Rutaceae family. Comparative Example 3 used Citrus unshiu peel extract, extracted from the fruit peel of the Rutaceae family. Comparative Example 4 used Coptis japonica extract, extracted from the rhizome of Coptis japonica, a perennial herb belonging to the Ranunculaceae family. Copper chlorophyllin sodium in Examples 6 and 7 means that the sample solution contained 1.66 mM and 166 μM of copper chlorophyllin sodium, respectively. Cherry bark extract in Example 8 means that the mass ratio of cherry bark extract to the volume of the final test system was 0.00167% by mass.

[0052] [Table 1]

[0053] <Method for measuring hemolysis rate> The hemolysis rate was measured for each sample solution. The hemolysis rate was measured by first keeping a mixture of each sample solution and ILY solution in a 37°C incubator for 15 minutes. Next, a mixture was prepared by mixing each sample solution with a 25% by volume red blood cell solution obtained by fractionating red blood cells from human blood. The mixture was prepared so that the red blood cell solution content in the mixture was 0.5% by volume. Next, the mixture was kept in a 37°C incubator for 1 hour. After that, the mixture was centrifuged with an 800g centrifugal force for 5 minutes. The supernatant after centrifugation was collected in a commercially available 96-well plate. The absorbance at 540 nm was measured using a cell imaging plate reader (Agilent BioTek Cytation 5, Agilent Technologies, Inc.). The hemolysis rate was calculated from the following formula (1). Absorbance measurements were performed with n=2, and the hemolysis rate was calculated from the average absorbance.

[0054]

number

[0055] In equation (1), sampleOD 540 This represents the absorbance of the above mixture at 540 nm. NCOD 540 This is the absorbance at 540 nm, measured without mixing the above ILY solution with each sample solution, as a negative control.

[0056] PCOD 540 The absorbance at 540 nm is the absorbance of a solution prepared by forcibly destroying red blood cells in the above mixture, which was obtained by mixing a 25% by volume red blood cell solution with each sample solution, as a positive control. The nonionic surfactant used was Triton ×-100, a polyoxyethylene alkylphenyl ether (Triton®)-based surfactant.

[0057] The denominator of equation (1) above is the absorbance of the solution in which red blood cells were forcibly destroyed, minus the absorbance of the background material. The numerator of equation (1) above is the absorbance of each sample solution, minus the absorbance of the background material. If the inhibitory effect on ILY is small, red blood cells are destroyed by ILY, and hemoglobin, the pigment in red blood cells, is more easily eluted. This makes the absorbance relatively larger. Therefore, the hemolysis rate value obtained from equation (1) above means that the closer it is to 100%, the smaller the inhibitory effect, and the closer it is to 0%, the larger the inhibitory effect. The method of evaluating the inhibitory effect by measuring the above hemolysis rate is generally called the hemolytic activity method or the cellular hemolysis method.

[0058] <Rating> The measured hemolysis rate was evaluated based on the following criteria. The results are shown in Table 1.

[0059] • Evaluation criteria for ILY inhibitors ◎ (Excellent): Hemolysis rate is 5% or less. ○ (Good): Hemolysis rate is greater than 5% but less than or equal to 20%.

[0060] × (Not acceptable): Hemolysis rate exceeds 20%. As shown in Table 1, Comparative Examples 1-4 all showed hemolysis rates exceeding 20%, confirming that the inhibitory effect on ILY was small. In contrast, Examples 1-30 all showed hemolysis rates of 20% or less, confirming that they could suitably inhibit ILY. In particular, in Examples 3, 5, 8, 10, 12, 15, 17, 20, 22, 24, 26, and 28, excellent effects were confirmed despite the low concentration of the extract in the sample solution being 0.001% by volume or 0.00167% by mass.

Claims

1. An intermedicine inhibitor characterized by containing at least one selected from the group consisting of loquat leaf extract, geranium extract, peony extract, copper chlorophyllin sodium, cherry bark extract, St. John's wort extract, black tea extract, peony extract, Houttuynia cordata extract, birch extract, oolong tea extract, hawthorn extract, linden extract, tea extract, witch hazel extract, saxifrage extract, Sanguisorba officinalis extract, and catechu extract.

2. An oral composition characterized by containing at least one selected from the group consisting of loquat leaf extract, geranium extract, peony extract, copper chlorophyllin sodium, cherry bark extract, St. John's wort extract, black tea extract, peony extract, Houttuynia cordata extract, birch extract, oolong tea extract, hawthorn extract, linden extract, tea extract, witch hazel extract, saxifrage extract, Sanguisorba officinalis extract, and catechu extract, and being used for intermedicine inhibition.

Images