JP2003128580A - Oral composition for plaque control - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain an oral cavity composition which can control proliferation or pathogenicity of bacterial plaque forming bacteria by controlling intercellular information transmission of oral cavity bacteria and is effective for preventing disease of oral cavity such as dental caries, periodontal disease, foul breath. SOLUTION: This oral cavity composition for controlling bacterial plaque comprises a substance for controlling intercellular information transmission mechanism of bacterial plaque forming microorganism.

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to a composition for the oral cavity using a novel method for suppressing plaque by utilizing the information transmission mechanism of oral microorganisms, and it relates to prevention of dental caries, prevention of bad breath, and periodontal disease. The present invention relates to an oral composition for plaque control which is useful for prevention.

[0002]

2. Description of the Related Art Certain microorganisms control the expression of various traits by detecting the density of themselves and other bacterial species in the environment through information mediators produced by themselves. It is known. These phenomena have been termed Quorum sensing by Greenberg et al. (ASM News 1997 63 371).
-377).

Further, microorganisms naturally form a so-called biofilm, adapt to the environment and exert pathogenicity. It has been clarified from experiments using Pseudomonas aeruginosa that the above-mentioned information-transmitting substances are involved in the formation of these biofilms, and acyl homoserine lactone (AHL) is a P. aeruginosa has been reported to be involved in biofilm formation (Davis DG et al. Science 1998 2
80 295-298). In addition to AHL, it has been reported by Basler et al. That there is an information transduction mechanism between bacteria (Pros).
c. Natl. Acad. Sci. USA 1999
96 1639-1644).

In the oral cavity, it is well known that microorganisms form plaque on the tooth surface and the tongue surface and cause oral diseases such as dental caries, bad breath and periodontal disease. A number of methods for suppressing dental plaque for the purpose of preventing these oral diseases have been disclosed so far. However, the conventional method for suppressing plaque is not efficient because the permeation and diffusion of the active ingredient into the plaque is poor, and a more effective method is desired.

[0005]

Means for Solving the Problems and Modes for Carrying Out the Invention In order to meet the above-mentioned demands, the present inventors have conducted various studies to suppress plaque forming a biofilm, and as a result, have been known so far. In addition to existing antibacterial agents, antibacterial agents, and other antiplaque agents, there are effective methods to suppress dental plaque, and in particular, the active substance is present in the low-molecular weight fraction of the culture solution of oral bacteria. It was confirmed. That is, the inventors have found the possibility of suppressing the plaque by controlling the above-mentioned information transfer substance between bacteria and have completed the present invention.

Therefore, the present invention provides an oral composition for suppressing plaque, which is characterized by containing a substance which controls the inter-bacterial information transmission mechanism of plaque-forming microorganisms.

It has been known that microorganisms produce an antibacterial substance called bacteriocin and suppress the growth of coexisting bacteria. In addition, it is also possible to blend these bacteriocins and enzymes derived from bacteria into oral compositions to suppress dental plaque, for example, JP-A-52-44246, JP-A-7-502038, and JP-A-49-7439. It is described in the official gazette. The substance of the present invention, in that its molecular weight and the ability to suppress the growth and pathogenicity of microorganisms of the same genus and the same species,
It is fundamentally different from the aforementioned bacteriocin and the like.

The present invention will be described in more detail below.
The composition for oral cavity of the present invention contains a substance that controls the intercellular information transfer mechanism of plaque-forming microorganisms.

The information transmission control substance used in the present invention is
It can be obtained from the culture supernatant of the microorganisms constituting plaque. In this case, the molecular weight of the organic solvent extracted from the culture supernatant of the microorganism is 5,000 or less, preferably 300 to 3,000.

Here, the microorganisms (oral bacteria) belong to, for example, Streptococcus, Actinomyces, Lactobacillus, Bayonella, Fusobacterium, Porphyromonas, Prevotella, Bacteroides, Actinobacillus, etc. Microorganisms can be mentioned and can be obtained from the culture supernatant.

The culture supernatant of these oral bacteria is cultivated in a commonly used medium from the logarithmic growth phase to the stationary phase, and then the culture supernatant is filtered with a 0.22 μm filter or the like to remove mixed bacterial cells. A completely removed product can be used. Further, a medium-derived substance and a polymer substance produced by a microorganism, which are usually present in a mixed state, are optionally removed, and further purified products can be suitably used.

That is, as a method for removing a polymer substance from the culture supernatant, gel filtration or ultrafiltration after lyophilization can be preferably used, and as a method for extracting and desalting a target substance, an ion exchange chromatography is used. , Hydrophobic chromatography, and solvent extraction can be appropriately combined. The organic solvent that can be used here is not particularly limited, but a lower alcohol such as n-hexane, dichloromethane, chloroform, acetone, ethyl acetate, methanol, ethanol, propanol, or the like, or a mixture thereof in an arbitrary ratio is preferable. Can be used for

Further, substances other than the culture supernatant of the bacteria derived from the oral cavity can be applied to the present invention, and the organic solvent extracts of animals and plants which can control the above-mentioned information transfer substance between bacteria can also be preferably used.

The oral composition of the present invention can be applied to various dosage forms such as dentifrice such as toothpaste, mouthwash, gingival massage cream, topical application agent, troche and chewing gum. The compounding amount of the above information transfer controlling substance in the composition of the present invention is 0.0001 to 1 based on the entire composition.
0% (mass percentage, the same below), especially 0.001 to 3%
Is preferred.

In this case, the oral composition for suppressing plaque of the present invention can be used in combination with various base components used in ordinary oral compositions, in addition to the above-mentioned components. For example,
Various abrasives, thickeners, binders, surfactants, sweeteners, flavors, colorants, preservatives, active ingredients and the like can be added.

As the abrasive, silica-based abrasives such as precipitated silica, silica gel, aluminosilicate, zirconosilicate, dicalcium phosphate dihydrate and anhydride, calcium pyrophosphate, calcium carbonate, aluminum hydroxide, alumina. , Magnesium carbonate, tribasic magnesium phosphate, insoluble sodium metaphosphate, insoluble potassium metaphosphate, titanium oxide, zeolite, aluminum silicate, zirconium silicate, synthetic resin-based abrasives, and the like.

Examples of the thickener include glycerin, sorbitol, xylitol, propylene glycol, polyethylene glycol and the like.

As the binder, carrageenan, sodium alginate, propylene glycol alginate, xanthan gum, polyacrylic acid, sodium polyacrylate, carboxyvinyl polymer, tragacanth gum, guar gum, locust bean gum, sodium carboxymethyl cellulose, hydroxyethyl cellulose, hydroxypropyl. Cellulose, methyl cellulose, gellan gum, gelatin, curdlan, gum arabic, agar, pectin, polyvinyl alcohol, polyvinylpyrrolidone, pullulan and the like can be mentioned.

Examples of the surface active agent include anionic surface active agents, cationic surface active agents, nonionic surface active agents, and the like. Specific examples include sodium lauryl sulfate, sodium N-lauroyl sarcosine, and α-olefin sulfone. Sodium acid, N-acyl glutamate, 2-alkyl-N-carboxymethyl-N-hydroxyethyl imidazolinium betaine, N-acyl taurate, sucrose fatty acid ester, alkylol amide, polyoxyethylene hydrogenated castor oil, polyglycerin Fatty acid ester,
Pluronic, polyoxyethylene sorbitan monostearate and the like can be mentioned.

As the sweetener, saccharin sodium,
Examples thereof include stevioside, stevia extract, paramethoxycinnamic aldehyde, neohesperidyl dihydrochalcone, perillartine and the like.

Examples of the fragrance include terpenes such as 1-menthol, carvone, anethole and limonene, and their derivatives. Examples of colorants include Blue No. 1, Yellow No. 4, titanium dioxide and the like.

Further, in the present invention, cationic fungicides such as chlorhexidine, benzethonium chloride, benzalkonium chloride, cetylpyridinium chloride and decalinium chloride, phenolic compounds such as triclosan, hinokitiol and isopropylmethylphenol, dextranase. , Mutanase, lysozyme, amylase, protease, lytic enzyme, enzymes such as SOD, sodium monofluorophosphate, alkali metal monofluorophosphate such as potassium monofluorophosphate, sodium fluoride,
Fluoride such as stannous fluoride, tranexamic acid, epsilon aminocaproic acid, allantoin, dihydrocholestanol, glycyrrhizinic acid, glycyrrhetinic acid, glycerophosphate, chlorophyll, sodium chloride, zinc chloride, water-soluble inorganic phosphate compound, vitamin A ,
One or more known active ingredients such as vitamins such as vitamin B group, vitamin C and vitamin E and derivatives thereof can be used in combination.

[0023]

EXAMPLES The present invention will be specifically described below by showing experimental examples and examples, but the present invention is not limited to the following examples.

[Experimental example] (Preparation of test substance) Streptococcus sanguinis
Brain Heart Infusion (ATCC10556)
[BHI]), the cells were cultured at 37 ° C. for 24 hours, and the culture supernatant was collected by centrifugation. The culture solution was treated with a 0.22 μm filter to completely remove the mixed bacterial cells, and freeze-dried to obtain a culture supernatant fraction. After drying, it is dissolved in a small amount of distilled water and subjected to gel filtration (Biogel P100:
BioRad) was used to fractionate a fraction having an average molecular weight of 5,000 or less, extract with a chloroform / methanol mixture (2: 1), and evaporate the solvent to remove S. A sanguinis culture supernatant extract was obtained. Porphyromonas gingivalis ( Porphyrom
onas gingivalis ) 381 strain with hemin,
Todd Hebit Broth (Todd) with Menadione
It was cultured in Hewitt Broth [THB]) at 37 ° C. under anaerobic conditions for 48 hours, and the culture supernatant after the culture was collected by centrifugation. Use this culture solution with a pore size of 0.22μ
Then, the mixed bacterial cells were completely removed. The polymer was removed from this culture solution by ultrafiltration (Ultracell PLCC membrane: Amicon), and a low molecular weight fraction was obtained by freeze-drying. This product was dissolved in distilled water, extracted twice with dichloromethane, and dried under reduced pressure to give P. gingivalis culture supernatant extract was obtained.

(Model plaque formation suppressing effect) Diameter 20 mm
The hydroxyapatite (HA) plate in Example 1 was treated with human non-stimulative saliva filtered through a 0.45 μm filter for 4 hours,
After washing, soak in THB, P. gingivalis 3
81 strains or S. sanguinis ATCC 1055
Co-cultured with 6 strains. During the culturing period, a liquid medium to which the above-mentioned culture supernatant extract is added at various concentrations is continuously supplied,
Continuous culture was carried out for 5 days. After culturing for 5 days, the model plaque formed on the HA plate was dispersed by ultrasonic waves, subjected to a 10-fold serial dilution, spread on a blood agar plate, and the grown colonies were measured to measure the viable bacteria. I asked for the number. The results are shown in Table 1.

[0026]

[Table 1]

[0027] (trypsin-like enzyme activity inhibitory effect of P.gingivalis) P. of 5 days of culture of the above-mentioned ging
ivalis model plaque is taken out and washed, and then P. g
THB medium (2 ml) supplemented with various concentrations of the culture supernatant extract of Invivalis was newly added, and static culture was carried out for 24 hours. After the completion of the culture, the trypsin-like activity in the culture supernatant was measured using N-benzoyl-arginine-2-naphthylamide as a substrate, and the enzyme activity per concentration of the culture at 550 nm was calculated to obtain the enzyme activity for the control. I asked. The results are shown in Table 2.

[0028]

[Table 2]

Examples will be shown below. In the following examples, the culture supernatant extract of each bacterium was prepared by the same method as in the preparation of the above-mentioned test substance. [Example 1] Toothpaste Aluminum hydroxide 45.0% Gelling silica 2.0 Solbit 25.0 Sodium carboxymethylcellulose 1.0 Sucrose monopalmitate 1.0 Sodium lauryl sulfate 1.5 Sodium saccharin 0.2 Ethanol 0 1 Sodium benzoate 0.1 Streptococcus culture supernatant extract 0.001 Perfume 1.0 Water Residual amount 100.0%

[Example 2] Mouthwash Modified ethanol 18.0% Polyoxyethylene hydrogenated castor oil 2.0 Glycerin 10.0 Palmitoyl sarcosine sodium 0.1 Citric acid 0.01 Trisodium citrate 0.3 Actinomyces Genus culture supernatant extract 0.02 Perfume 0.5 0.1% Green No. 201 0.8 Water balance 100.0%

Example 3 Oral Pasta Hydroxyethyl Cellulose 3.0% Carrageenan 1.0 Sorbit 40.0 Sucrose Palmitic Acid Monoester 2.0 Lauroyl Sarcosine Sodium 0.3 Aluminum Lactate 0.8 Saccharin Sodium 0.1 Fuzobacterium Genus culture supernatant extract 0.05 Porphyromonas genus culture supernatant extract 0.05 Perfume 1.0 Water balance 100.0%

Example 4 Chewing gum gum base 20.0% sugar 15.0 isomaltose 20.0 palatinose 10.0 xylitol 10.0 corn syrup 12.0 starch syrup 12.3 Streptococcus culture supernatant extract 0.05 Bayonera culture supernatant extract 0.05 Perfume 0.6 Total 100.0%

[0033]

INDUSTRIAL APPLICABILITY According to the present invention, it is possible to suppress the proliferation or pathogenicity of plaque-forming bacteria by controlling the intercellular information transfer of oral bacteria, and to prevent oral diseases such as dental caries, periodontal disease and halitosis. It is valid.

Continued front page    F-term (reference) 4C083 AA031 AA032 AB172 AB222                       AC102 AC122 AC132 AC302                       AC312 AC432 AC662 AC782                       AC862 AD222 AD272 CC41                       EE32 EE33 EE36                 4C084 AA17 MA16 MA28 MA47 MA57                       NA14 ZB35 ZC41                 4C087 AA01 AA02 BC30 BC61 CA10                       CA11 CA24 CA25 CA26 CA27                       CA28 CA29 CA30 MA16 MA28                       MA57 NA14 ZB35 ZC41

Claims (2)

[Claims] 1. A composition for oral cavity for suppressing plaque, which is blended with a substance which controls a bacterial information transmission mechanism of a plaque-constituting microorganism. 2. A substance that regulates a mechanism of information transfer between bacteria,
Molecular weight of organic solvent extracted from culture supernatant of microorganisms 5,0
The oral composition for suppressing plaque according to claim 1, wherein the composition is 00 or less.