Fusobacterium-sterilizing composition

Abstract

To provide a formulation that effectively sterilizes Fusobacterium within a short time.SOLUTION: A Fusobacterium-sterilizing composition contains cetylpyridinium chloride, DL-pyrrolidonecarboxylic acid salt of N-cocoyl-L-arginine ethyl ester, citric acid, and / or disodium edetate.SELECTED DRAWING: None

Description

Technical Field

[0001] The present disclosure relates to a composition for killing Fusobacterium, etc. The contents of all documents described in this specification are incorporated herein by reference.

Background Art

[0002] Dental plaque is a biofilm formed by the aggregation of oral microorganisms and is considered to be a cause of dental caries and periodontal disease.

[0003] Generally speaking, dental plaque is formed as follows. That is, first, a thin film of protein derived from saliva and physiological gingival sulcus exudate called "pellicle" is formed on the tooth surface, and facultative anaerobic bacteria such as Streptococcus attach to the tooth surface through the pellicle (initial adherent bacteria). Mediating bacteria such as Fusobacterium, which co-aggregate with various oral bacteria to these initial adherent bacteria, attach, and further, late adherent bacteria such as Porphyromonas gingivalis and Treponema denticola, which are anaerobic bacteria, attach and aggregate, and dental plaque matures. In particular, late adherent bacteria are known to be a cause of periodontal disease and are directly or indirectly related to the destruction of periodontal tissues. Therefore, suppressing the formation of dental plaque on the tooth surface, particularly suppressing the attachment and aggregation of late adherent bacteria and the maturation of dental plaque, is considered important for preventing periodontal disease (Non-Patent Document 1).

[0004] Fusobacterium is known as a bacterium related to periodontal disease. Since the presence of Fusobacterium is said to be directly or indirectly related to the destruction of periodontal tissues, it is necessary to effectively kill this Fusobacterium in order to prevent periodontal disease.

[0005] In addition, Patent Document 2 shows that an oral composition containing copper chlorophyllin salt and cocooyl arginine ethyl salt suppresses the activity of gingipain, which is one of the typical pathogenic factors produced by Porphyromonas gingivalis, a late colonizing bacterium.

Prior Art Documents

Patent Documents

[0006]

Non-Patent Document 1

Patent Document 2

Summary of the Invention

Problems to be Solved by the Invention

[0007] The present inventors focused on Fusobacterium, a mediating bacterium, and conducted studies. If the function of Fusobacterium, a mediating bacterium capable of coaggregation with various bacteria, can be suppressed, it is considered possible to suppress the adhesion and aggregation of late colonizing bacteria to the tooth surface, and ultimately suppress the maturation of plaque formation.

[0008] Effectively sterilizing Fusobacterium, a mediating bacterium at the previous stage, so as not to adhere and aggregate late colonizing bacteria that cause periodontal disease, ultimately leads to the prevention of periodontal disease. However, since conventional techniques for sterilizing Porphyromonas gingivalis, a known late colonizing bacterium, cannot sterilize Fusobacterium, a sterilization technique suitable for Fusobacterium is required.

[0009] In addition, many bactericidal agents are already known, but since many of them are cationic, there are concerns about irritation. Therefore, it is necessary to reduce the formulation of the bactericidal agent as much as possible and effectively sterilize Fusobacterium in as short a time as possible when used orally as a composition for sterilizing Fusobacterium.

[0010] Therefore, paying attention to N-ethyl-L-arginine DL-pyrrolidone carboxylate, which is one of the cationic surfactants, a method for effectively sterilizing Fusobacterium in a short time was studied while the blending amount of N-ethyl-L-arginine DL-pyrrolidone carboxylate is relatively small.

Means for Solving the Problems

[0011] The present inventor conducted studies focusing on the mediating bacterium Fusobacterium. If the function of Fusobacterium, a mediating bacterium capable of coaggregation with various bacteria, can be suppressed, it is considered that the late adherent bacteria can be suppressed from adhering and aggregating on the tooth surface, and thus the destruction of the periodontal tissue can be prevented. This is because.

[0012] Also, in Patent Document 1, it is described that N-ethyl-L-arginine DL-pyrrolidone carboxylate, which is a kind of ethyl cocoyl arginate, and sodium copper chlorophyllin are used, and depending on the blending amount of sodium copper chlorophyllin, an inhibitory effect on gingipain activity is shown in the range of 0.4% to 0.8% by mass of the concentration of N-ethyl-L-arginine DL-pyrrolidone carboxylate.

[0013] In the present invention, reduction of the blending amount of the cationic surfactant was studied to reduce irritation. As a result, it was found that even when the blending amount of N-ethyl-L-arginine DL-pyrrolidone carboxylate is 0.01% by mass, a high bactericidal effect is obtained by using cetylpyridinium chloride, citric acid, and disodium edetate.

[0014] The present disclosure includes, for example, the subject matters described in the following items. Item 1. A composition for sterilizing Fusobacterium containing the following A, B, C, and / or D. A: Cetylpyridinium chloride B: N-Yashi oil fatty acid acyl-L-arginine ethyl DL-pyrrolidone carboxylate C: Citric acid and / or its salts D: Disodium edetate Item 2. A method for sterilizing Fusobacterium using an oral composition containing the following A, B, C, and / or D. A: Cetylpyridinium chloride B: N-Yashi oil fatty acid acyl-L-arginine ethyl DL-pyrrolidone carboxylate C: Citric acid and / or its salts D: Disodium edetate

Advantages of the Invention

[0015] By providing a method for efficiently sterilizing Fusobacterium, it becomes possible to prevent periodontal disease.

Modes for Carrying Out the Invention

[0016] Hereinafter, embodiments according to the present invention will be described.

[0017] The composition for sterilizing Fusobacterium included in the present disclosure contains cetylpyridinium chloride, N-Yashi oil fatty acid acyl-L-arginine ethyl DL-pyrrolidone carboxylate, citric acid, and / or disodium edetate. The composition for sterilizing Fusobacterium may be referred to as "the composition for sterilizing Fusobacterium of the present disclosure".

[0018] Regarding the content of cetylpyridinium chloride in the composition of the present disclosure, the effect is achieved There is no particular limitation as long as it is within a range. For example, about 0.01 to 0.5 mass% can be mentioned. Note that the upper limit or lower limit of the said range can be, for example, 0.02, 0.03, 0.04, 0.05, 0.06, 0.07, 0.08, 0.09, 0.1, 0.11, 0.12, 0.13, 0.14, 0.15, 0.16, 0.17, 0.18, 0.19, 0.2, 0.21, 0.22, 0.23, 0.24, 0.25, 0.26, 0.27, 0.28, 0.29, 0.3, 0.31, 0.32, 0.33, 0.34, 0.35, 0.36, 0.37, 0.38, 0.39, 0.4, 0.41, 0.42, 0.43, 0.44, 0.45, 0.46, 0.47, 0.48, or 0.49 mass%. For example, the said range may be about 0.02 to 0.3 mass%, or about 0.03 to 0.1 mass%. When the content of cetylpyridinium chloride is 0.5 mass% or more, it may cause strong irritation and is thus not preferable.

[0019] N-coconut oil fatty acid acyl-L-arginine ethyl·DL-pyrrolidone carboxylate is the salt of the ethyl ester of coconut oil fatty acid and L-arginine and pyrrolidone carboxylic acid, and is an amino acid-based cationic surfactant. N-coconut oil fatty acid acyl-L-arginine e thyl·DL-pyrrolidone carboxylate is also referred to as cocoyl arginine ethyl PCA. As N-coconut oil fatty acid acyl-L-arginine ethyl·DL-pyrrolidone carboxylate, for example, "CAE" manufactured by Ajinomoto Health Supply Co., Ltd. can be used.

[0020] Examples of the fatty acids contained in the coconut oil fatty acids that constitute N-coconut oil fatty acyl-L-arginine ethyl·DL-pyrrolidone carboxylate include fatty acids having 8 carbon atoms (more specifically, caprylic acid), fatty acids having 10 carbon atoms (more specifically, capric acid), fatty acids having 12 carbon atoms (more specifically, lauric acid), fatty acids having 14 carbon atoms (more specifically, myristic acid), fatty acids having 16 carbon atoms (more specifically, palmitic acid), and the like. These may be contained alone or in combination of two or more. Among them, it is preferable to contain a fatty acid having 12 carbon atoms (more specifically, lauric acid) and a fatty acid having 14 carbon atoms (more specifically, myristic acid), and it is more preferable to contain a fatty acid having 12 carbon atoms (more specifically, lauric acid). The content of the fatty acid having 12 carbon atoms (more specifically, lauric acid) per fatty acid contained in the coconut oil fatty acids that constitute N-coconut oil fatty acyl-L-arginine ethyl·DL-pyrrolidone carboxylate may be, for example, about 40 to 70% by mass. The upper or lower limit of the range may be, for example, 45, 50, 55, 60, or 65% by mass. For example, the range may be about 45 to 65% by mass. The content of the fatty acid having 14 carbon atoms (more specifically, myristic acid) per fatty acid contained in the coconut oil fatty acids that constitute N-coconut oil fatty acyl-L-arginine ethyl·DL-pyrrolidone carboxylate may be, for example, about 15 to 35% by mass. The upper or lower limit of the range may be, for example, 20, 25, or 30% by mass. For example, the range may be about 20 to 30% by mass.

[0021] In the composition of the present disclosure, the content of N-coconut oil fatty acid acyl-L-arginine ethyl · DL-pyrrolidone carboxylate is not particularly limited as long as the effect is achieved. For example, it is about 0.005 to 0.5% by mass. The upper or lower limit of this range may be, for example, 0.005, 0.006, 0.007, 0.008, 0.009, 0.01, 0.015, 0.02, 0.03, 0.04, 0.05, 0.06, 0.07, 0.08, 0.09, 0.1, 0.11, 0.12, 0.13, 0.14, 0.15, 0.16, 0.17, 0.18, 0.19, 0.2, 0.21, 0.22, 0.23, 0.24, 0.25, 0.26, 0.27, 0.28, 0.29, 0.3, 0.31, 32, 0.33, 0.34, 0.35, 0.36, 0.37, 0.38, 0.39, 0.4, 0.41, 0.42, 0.43, 0.44, 0.45, 0.46, 0.47, 0.48, or 0.49% by mass.

[0022] It is preferable that N-coconut oil fatty acid acyl-L-arginine ethyl · DL-pyrrolidone carboxylate is contained in the composition for killing Fusobacterium in an amount of 0.005 to 0.3% by mass. It is more preferable that N-coconut oil fatty acid acyl-L-arginine ethyl · DL-pyrrolidone carboxylate is contained in the composition for killing Fusobacterium in an amount of 0.007 to 0.1% by mass. It is even more preferable that N-coconut oil fatty acid acyl-L-arginine ethyl · DL-pyrrolidone carboxylate is contained in the composition for killing Fusobacterium in an amount of 0.008 to 0.015% by mass.

[0023] The content ratio of N-coconut oil fatty acid acyl-L-arginine ethyl-DL-pyrrolidone carboxylate and cetylpyridinium chloride in the composition of the present disclosure is, for example, preferably about 1 to 100 parts by mass of cetylpyridinium chloride per 10 parts by mass of N-coconut oil fatty acid acyl-L-arginine ethyl-DL-pyrrolidone carboxylate. The upper or lower limit of the range is, for example, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99 It may be 1, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99 parts by mass. For example, the range may be about 2 to 90 parts by mass, or about 5 to 50 parts by mass.

[0024] The composition of the present disclosure contains cetylpyridinium chloride, N-coconut oil fatty acid acyl-L-arginine ethyl-DL-pyrrolidone carboxylate, citric acid, and disodium edetate, and thus exhibits excellent anti-Fusobacterial effect.

[0025] In this case, Fusobacterium is classified into early and late adherent bacteria in plaque formation. There is no particular limitation as long as it is a bacterium of the genus Fusobacterium, which is a mediator between the bacteria and the host, but Fus Fusobacterium nucleatum is preferred. Examples are given below.

[0026] The composition for Fusobacterium bactericidal of the present disclosure can be, for example, a solid composition or a liquid composition. The composition for Fusobacterium bactericidal can be used, for example, as pharmaceuticals, quasi-drugs, etc. Further, the form of the composition for Fusobacterium bactericidal of the present disclosure is not particularly limited, but according to a conventional method, for example, it can be made into forms (dosage forms) such as ointments, pastes, pastas, gels, liquids, sprays, mouthwashes, liquid dentifrices, dentifrices, gums, etc. Among them, mouthwashes, liquid dentifrices, dentifrices, ointments, pastes, liquids, gels are preferred.

[0027] The composition for Fusobacterium bactericidal of the present disclosure may further contain, alone or in combination of two or more, any components that can be formulated in the composition for Fusobacterium bactericidal, as long as the effects are not impaired.

[0028] For example, as the surfactant, a nonionic surfactant, an anionic surfactant or an amphoteric surfactant can be blended. Specifically, for example, as the nonionic surfactant, sugar fatty acid esters such as sucrose fatty acid ester, maltose fatty acid ester, lactose fatty acid ester; fatty acid alkanolamides; sorbitan fatty acid esters; fatty acid monoglycerides; polyoxyethylene alkyl ethers with a polyoxyethylene addition coefficient of 8 to 10 and an alkyl group having 13 to 15 carbon atoms; polyoxyethylene alkyl phenyl ethers with a polyoxyethylene addition coefficient of 10 to 18 and an alkyl group having 9 carbon atoms; diethyl sebacate; polyoxyethylene hydrogenated castor oil; fatty acid polyoxyethylene sorbitan and the like can be mentioned. As the anionic surfactant, sulfate esters such as sodium lauryl sulfate, sodium polyoxyethylene lauryl ether sulfate; sulfosuccinates such as sodium lauryl sulfosuccinate, sodium polyoxyethylene lauryl ether sulfosuccinate; acyl amino acid salts such as sodium cocoyl sarcosinate, sodium lauroyl methyl alaninate; sodium cocoyl methyl taurine and the like can be mentioned. As the amphoteric ion surfactant, betaine type surfactants such as lauryldimethylaminoacetic acid betaine, coconut oil fatty acid amide propyldimethylaminoacetic acid betaine; imidazoline type surfactants such as N-cocoyl-N-carboxymethyl-N-hydroxyethyl ethylenediamine sodium; amino acid type surfactants such as N-lauryl diaminoethyl glycine and the like can be mentioned. These surfactants can be blended alone or in combination of two or more. The blending amount can usually be, for example, about 0.1 to 5% by mass based on the total amount of the composition.

[0029] As flavoring agents, for example, menthol, carboxylic acid, anethole, eugenol, methyl salicylate, limonene, ocimene, n-decyl alcohol, citronellal, α-terpineol, methyl acetate, citronellyl acetate, methyl eugenol, cineole, linalool, ethyl linalool, thymol, spearmint oil, peppermint oil, lemon oil, orange oil, sage oil, rosemary oil, perilla oil, perilla oil, wintergreen oil, clove oil, eucalyptus oil, pimento oil, d-camphor, d-borneol, star anise oil, cinnamon oil, cinnamaldehyde, mint oil, vanilla, etc. can be used. These can be blended alone or in combination of two or more. The blending amount can be, for example, about 0.001 to 1.5% by mass based on the total amount of the composition.

[0030] As sweetening agents, for example, sodium saccharin, acesulfame potassium, stevioside, neohesperidin dihydrochalcone, perillartine, thaumatin, aspartyl phenylalanyl methyl ester, p-methoxycinnamic aldehyde, etc. can be used. These can be blended alone or in combination of two or more. The blending amount can be, for example, about 0.01 to 1% by mass based on the total amount of the composition.

[0031] As wetting agents, sorbitol, ethylene glycol, propylene glycol, glycerin, 1,3-butylene glycol, polypropylene glycol, xylitol, maltitol, lactitol, polyoxyethylene glycol, etc. can be blended alone or in combination of two or more.

[0032] As preservatives, parabens such as methyl paraben, ethyl paraben, propyl paraben, butyl paraben, sodium benzoate, phenoxyethanol, alkyldiaminoethyl glycine hydrochloride, etc. can be blended. These can be blended alone or in combination of two or more.

[0033] As colorants, legal dyes such as Food Blue No. 1, Food Yellow No. 4, Food Red No. 202, and Green No. 3, mineral dyes such as ultramarine, fortified ultramarine, and navy blue, and titanium oxide may be blended. These can be blended alone or in combination of two or more.

[0034] As a pH adjuster, phosphoric acid, malic acid, pyrophosphoric acid, lactic acid, tartaric acid, glycerophosphoric acid, acetic acid, nitric acid, or chemically possible salts thereof, sodium hydroxide, etc. may be blended. These can be blended alone or in combination of two or more so that the pH of the composition is in the range of 4 to 8, preferably 5 to 7. The blending amount of the pH adjuster may be, for example, about 0.01 to 2% by weight.

[0035] In the composition for killing Fusobacterium of the present disclosure, further, as medicinal components, vitamin Es such as dl-α-tocopherol acetate, succinic acid tocopherol, or nicotinic acid tocopherol, amphoteric bactericides such as dodecyldiaminoethyl glycine, nonionic bactericides such as triclosan, isopropylmethylphenol, and hinokitiol, anionic bactericides such as sodium lauroyl sarcosinate, cationic bactericides such as chlorhexidine hydrochloride and benzethonium chloride, enzymes such as dextranase, amylase, protease, mutanase, lysozyme, and lysing enzyme (Lytech 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 chlorhydroxyl allantoin, dihydrocholesterol, glycyrrhetinic acid, glycyrrhizic acid, sodium copper chlorophyllin, glycerophosphate, chlorophyll, sodium chloride, caropeptide, allantoin, carbazochrom, hinokitiol, potassium nitrate, paratinit, etc. can be blended alone or in combination of two or more.

[0036] As a base, it is also possible to add alcohols, silicon, apatite, white petrolatum, paraffin, liquid paraffin, microcrystalline wax, squalane, plastic base, etc. These can be blended alone or in combination of two or more kinds.

[0037] Moreover, the composition for killing Fusobacterium of the present disclosure can be prepared by a known method or a method easily conceivable from a known method. For example, it can be prepared by appropriately mixing cetylpyridinium chloride, N-coconut oil fatty acyl-L-arginine ethyl·DL-pyrrolidone carboxylate, citric acid, disodium edetate, and other components as necessary.

Examples

[0038] The content of the present disclosure will be specifically described using the following experimental examples and examples. However, the present disclosure is not limited to these in any way. In the following, unless otherwise specified, the experiments are carried out under atmospheric pressure and normal temperature conditions. Also, unless otherwise specified, “%” means “mass %”. Further, the blending amount values of each component described in each table also indicate “mass %” unless otherwise specified.

[0039] Cetylpyridinium chloride (CPC), N-coconut oil fatty acyl-L-arginine ethyl ·DL-pyrrolidone carboxylate (CAE), citric acid, and disodium edetate were dissolved in water so that each component had each concentration shown in Table 1, and the pH was adjusted to 6.5 - 7.0 using 1N NaOH. This was used as the bactericidal liquid.

[0040]

Table 1

[0041]

Table 2

[0042] Fusobacterium nucleatum ssp. nucleatum (ATCC: 25586) was used as the test bacterium.

[0043] The test bacterium was inoculated into 5 ml of modified GAM medium (Nissui Pharmaceutical Co., Ltd.) and cultured at 37°C for 2 days under anaerobic conditions. The resulting culture solution was used as the test bacterial solution.

[0044] 200 μl of the culture solution was added to each well of a 96-well plate, and the turbidity was measured using a spectrophotometer. Then, it was diluted using modified GAM medium (Nissui Pharmaceutical Co., Ltd.) to adjust the optical density to OD660 = 0.1.

[0045] 1.0 g of Tween 80, 0.14 g of lecithin, and 8.34 g of modified GAM medium (Nissui Pharmaceutical Co., Ltd.) were dissolved in 200 ml of distilled water, and sterilization treatment was performed using an autoclave at 121°C for 20 minutes to prepare an inactivated modified GAM medium.

[0046] 20 μl of the test bacterial solution was mixed with 200 μl of the bactericide solution. 20 μl of the mixture was sampled 30 and 90 seconds after the start of mixing, and each was mixed with 200 μl of the inactivated modified GAM medium to inactivate the bactericidal action of the bactericide.

[0047] Each mixed solution inactivated in

[0046] was cultured at 37°C for 2 days under anaerobic conditions. For the solution after culture, OD660nm was measured using a spectrophotometer, and the viability of various test bacteria was determined from the turbidity.

[0048] As the method for determining viability, using the OD660nm of 220 μl of the inactivated GAM medium as a blank, those with an OD660nm value in the bacterial solution after culture greater than that of the blank were determined to have growing bacteria, and those of the same level as the blank were determined to have dead bacteria. By this determination method, the treatment time when the turbidity value in the bactericide - test bacterium mixed solution was of the same level as the blank, that is, the time required for sterilization, was measured.

[0049] The results are shown in Table 2. The evaluation results were as follows. ○: The turbidity value is about the same as the blank, indicating that the bacteria are in a state of being killed. ×: Since the turbidity value is equal to or higher than the blank, it indicates that the bacteria are in a state of survival.

[0050]

Table 3

[0051] For Examples 1 to 3, the bacteria were killed 30 seconds after the start of mixing, and it was confirmed that they showed a high bactericidal power in a short time. As shown in Comparative Examples 1 to 12, when CPC, CAE, citric acid, and disodium edetate were used alone or in combination, they were not sterilized under the test conditions of this time if they were not in the combination as in the examples, or even if they were sterilized, the bacteria did not die in 30 seconds and it was found that it took time.

Claims

1. A composition for killing Fusobacterium, comprising the following A, B, C and / or D. A: Cetylpyridinium chloride B: N-coconut oil fatty acyl-L-arginine ethyl DL-pyrrolidone carboxylate C: Citric acid and / or its salts D: Disodium edetate

2. A method for killing Fusobacterium using an oral composition comprising the following A, B, C and / or D. A: Cetylpyridinium chloride B: N-coconut oil fatty acyl-L-arginine ethyl DL-pyrrolidone carboxylate C: Citric acid and / or its salts D: Disodium edetate

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