KR20140061349A - Oral bacteria proliferation inhibitor - Google Patents

Abstract

Providing a substance that inhibits the proliferation of bacteria in the oral cavity and is free from ingestion or harm. Especially, the usefulness of the combination of sugar and sugar alcohol has not yet been clarified, and it is intended to provide a combination of sugar and sugar alcohol which inhibits the growth of bacteria in the oral cavity. The agent for inhibiting the growth of bacteria in the oral cavity, which comprises galactose and sorbitol. The agent for inhibiting the growth of bacteria in the oral cavity according to the present invention is effective for the improvement and prevention of periodontal disease.

Description

ORAL BACTERIA PROLIFERATION INHIBITOR [0002]

The present invention relates to an agent for inhibiting the growth of bacteria in the oral cavity.

Hundreds of bacteria are known to reproduce in human oral cavity. Among these are many bacteria that cause periodontal disease or caries by causing irritation to gums and periodontal disease, or by producing acids to dissolve bones and teeth.

Fusobacterium nucleatum (hereinafter referred to as F. nucleatum) is a bacterium species that is frequently detected in plaques of both normal and diseased parts in humans (Non-Patent Documents 1 and 2). It is thought that this bacterium contributes to the onset and progression of periodontal disease in that the proportion thereof increases with the progress of periodontal disease (Non-patent Documents 3 to 5). It has also been known that it has the ability to adhere to many other bacteria and plays a role of bridging between early colony forming fungi and late colony forming fungi including periodontal pathogens in dental plaques (Non-Patent Document 6). Furthermore, it is also a producing species of volatile sulfur compounds that cause halitosis (Non-Patent Document 7). Therefore, the antimicrobial agent exhibiting activity against F. nucleatum is considered to be effective for prevention of periodontal disease, reduction of bacteria in the oral cavity, and prevention and reduction of bad breath.

Materials for activating or inhibiting the activity of these bacteria have also been studied. For example, it is widely known that saccharides are causative agents of caries causing bacteria. On the other hand, it is known that sugar alcohols have a proliferation inhibitory action on carnivorous microorganisms, and the proliferation inhibitory action against periodontal pathogens is also recently reported.

 Papapanou, P. N., Sellen, A., Wennstroem, J. L. and Dahlen, G., An analysis of the subgingival microflora in randomly selected subjects, Oral microbiology and immunology, 8, 24-29 (1993).  Moore, W.E. and Moore, L. V., The bacteria of periodontal diseases, Periodontology 2000, 5, 66-77 (1994).  Dzink, J. L., Socransky, S. S. and Haffajee, A. D., The predominant cultivable microbiota of active and inactive lesions of destructive periodontal diseases, Journal of Clinical Periodontology, 15, 316-323 (1988).  Socransky, S. S., Haffajee, A. D. and Dzink, J. L., Relationship of subgingival microbial complexes to clinical features at the sampled sites, Journal of Clinical Periodontology, 15, 440-444 (1988).  Tanner, A. and Bouldin, H., The microbiota of early periodontitis lesions in adults, Journal of clinical periodontology, 16, 467-471 (1989).  Kleenbrander, P. E., Andersen, R. N., Blehert, D. S., Egland, P. G., Foster, J. S. and Palmer, R. J. Jr., Communication among oral bacteria, Microbiology and molecular biology reviews, 66, 486-505 (2002).  Krespi, Y. P., Shrime, M. G. and Kacker, A., The relationship between oral malodor and volatile sulfur compound-producing bacteria, Otolaryngology-Head and Neck Surgery, 135, 671-676 (2006).  Bolstad, A. I., Jensen, H. B. and Bakken, V., Taxonomy, Biology, and Periodontal Aspects of Fusobacterium nucleatum, Clinical Microbiology Reviews, 9, 55-71 (1996).  G., Los, T., Lykidis, A. and Overbeek, R., et al., Genome Sequence and Analysis of the Oral Bacterium Fusobacterium nucleatum Strain ATCC 25586, Journal of Bacteriology, 184, 2005-2018 (2002).  Kapatral, V., Ivanova, N., Anderson, I., Reznik, G., Bhattacharyya A., Gardner, WL and Kyrpides, N., et al., Genome Analysis of F. nucleatum subspp vincentii and Its Comparison With Genome of F. nucleastum ATCC25586, Genome Research, 6A, 1180-1189 (2003).

As a periodontal pathogen or a caries pathogen, a plurality of oral bacteria are known, and it is required to provide a substance that inhibits the proliferation of the bacteria. It is preferable that such a substance is a substance which is harmless even if taken into the body, in view of its action in the oral cavity.

In view of the above problems, it is an object of the present invention to provide a substance that inhibits the proliferation of bacteria in the oral cavity and has no harmful effect on ingestion. Particularly, the usefulness of a combination of a sugar and a sugar alcohol has not yet been clarified. It is an object of the present invention to provide a combination of a sugar and a sugar alcohol which inhibits the growth of bacteria in the oral cavity.

As a result of intensive studies, the inventors of the present invention have found that proliferation of bacterial F. nucleatum in oral cavity is remarkably inhibited in a culture medium containing galactose and sorbitol, thereby completing the present invention. That is, the present invention relates to an agent for inhibiting the growth of bacteria in the oral cavity, which comprises galactose and sorbitol.

The agent for inhibiting the growth of bacteria in the oral cavity according to the present invention can be safely used or ingested on a routine basis by being contained in a food or oral composition and can effectively reduce the bacteria in the oral cavity and also improve and prevent periodontal disease, It can also prevent and reduce bad breath.

Fig. 1 is a graph showing OD values obtained for each medium in Example 2. Fig.
2 is a graph showing OD values obtained for each medium in Example 3. Fig.
Fig. 3 is a graph showing OD values obtained for each medium in Example 4. Fig.

The agent for inhibiting the growth of bacteria in oral cavity containing galactose and sorbitol of the present invention has an effect of synergistically inhibiting the proliferation of F. nucleatum. Because F. nucleatum causes dental plaque formation by adhering to many other bacteria, it can inhibit the growth of other bacteria in the oral cavity, which causes periodontal disease and caries, by inhibiting the proliferation of this bacterium. In addition, since F. nucleatum is one of pathogenic bacteria of periodontal disease, the present invention is useful for improvement and prevention of periodontal disease. Furthermore, this bacterium produces volatile sulfur compounds that cause bad breath, so inhibiting the proliferation of F. nucleatum is considered to be effective for prevention and reduction of bad breath.

The total molar concentration of galactose and sorbitol is preferably 0.5 M or more and 0.9 M or less. The molar concentration of galactose and sorbitol is preferably in a molar ratio of galactose to sorbitol of 2: 1 to 1: 3.

As the galactose in the present invention, it is preferable to use? -D-galactose. As the sorbitol in the present invention, D-sorbitol (D-sorbitol) is preferably used.

In addition, the proliferation inhibitor of the present invention can be used for oral compositions such as a moisturizer, a toothpaste, an inhalant and a troche, a confectionery such as a chewing gum, a candy, a jelly, a jelly, a chocolate, a biscuit and a snack, It can be used in combination with beverages such as beverages, breads, hot cakes, dairy products, ham and sausage products, fish meat products such as fish cake and chikuwa, whole foods, puddings, soups or jams.

Example 1

sample

D (+) - galactose (Wako Pure Chemical Industries, Ltd.) and D (-) - sorbitol (Wako Pure Chemical Industries, Ltd.) were used.

Published strain

F. nucleatum JCM12990 strain was used as oral bacteria.

badge

Yeast extract (3.0 g / ℓ) , Hemin (5.0 ㎎ / ℓ), Menadione (0.5 ㎎ / ℓ) a Tripticase soy broth (after TSB medium), anaerobic conditions, and (10% CO ₂ at 37 ℃ using the addition of They were cultured in _{10% H 2, 80% N} 2). For medium containing galactose or sorbitol, or galactose or sorbitol, TSB medium in which galactose and sorbitol were dissolved was prepared so that the final concentration when 3.0 ml was added to 7.0 ml of TSB medium was the set concentration. All the media were sterilized by a filter (ADVANTEC, pore size 0.20 mu m).

Bacterium  Measure

The pre-culture of oral bacteria F. nucleatum JCM12990 was added to 7.0 ml of TSB medium and cultured to logarithmic growth phase. The same amount of the culture medium as that of the previous culture medium was removed, and once the volume of the culture medium was adjusted to 7.0 ml, 3.0 ml of a TSB medium containing galactose, sorbitol or galactose and sorbitol prepared so as to have a predetermined concentration in 10.0 ml was added. At that time, 3.0 ml of a normal medium containing neither galactose or sorbitol was added to control the medium. After culturing them in the anaerobic condition for 1 hour, the culture tube was transferred into a small shaking culture apparatus (Biophotoreater TVS062CA, Advanetek TYOYANG Co., Ltd.), and cultured at 37 ° C for 24 hours. Optical density at OD 660 nm ) (HITACHI U-3900H Spectrophotometer).

Example 2

In Example 1, a TSB medium supplemented with galactose containing 0.1 M or 0.6 M of galactose, a TSB supplemented with sorbitol containing 0.2 M, 0.3 M or 0.6 M sorbitol, 0.4 M of galactose and 0.2 M of sorbitol, Mixed TSB medium containing 0.3 M of galactose and 0.3 M of sorbitol, 0.3 M of galactose and 0.6 M of sorbitol, and control were prepared and tested. The OD values obtained for each TSB medium are shown in Fig. When cultured in TSB medium containing 0.1 M or 0.6 M of galactose for 24 hours, the OD value exceeded the control OD value. It has been reported that F. nucleatum metabolizes galactose (Non-patent Documents 8-10), and it is presumed that the effect of F. nucleatum is promoted by the effect of F. nucleatum. When cultured in TSB medium containing 0.2 M, 0.3 M or 0.6 M sorbitol for 24 hours each, OD value exceeding control at 0.2 M, OD value at the same level as control at 0.3 M, control at 0.6 M The lower OD value was confirmed.

Here, an OD value lower than the control was confirmed by culturing for 24 hours in a mixed-addition TSB medium containing 0.4 M of galactose, which is expected to promote proliferation, and 0.2 M of sorbitol, which has been confirmed to promote proliferation. Likewise, the OD value under control was also confirmed for medium containing 0.3 M galactose, 0.3 M sorbitol, 0.3 M galactose and 0.6 M sorbitol, and this value was found to be 0.3 M sorbitol, 0.6 M sorbitol Value.

From the above results, it was confirmed that the proliferation of F. nucleatum was inhibited by containing galactose and sorbitol in the medium.

Example 3

From Example 2, it was confirmed that when galactose and sorbitol were added together to the medium, the proliferation inhibitory effect of F. nucleatum was confirmed, it was examined whether the same proliferation inhibitory effect was confirmed by changing the ratio of galactose and sorbitol in which this effect was confirmed . The total molar concentration of galactose and sorbitol was set to 0.6 M. The OD values obtained for each TSB medium are shown in Fig.

As a result, the OD value under control was confirmed under the conditions of the ratio of galactose: sorbitol = 2: 1, 1: 1, 1: 2, and 1: The degree of inhibition of proliferation was not dependent on any concentration of galactose or sorbitol, and the strongest inhibitory effect was confirmed under the conditions of galactose: sorbitol = 1: 2 (galactose: sorbitol = 0.2 M: 0.4 M).

Example 4

The same test was conducted under the conditions of galactose: sorbitol = 1: 2, in which the strongest proliferation inhibitory effect was confirmed in Example 3, while changing the total molar concentration while maintaining the ratio. The OD values obtained for each TSB medium are shown in Fig.

As a result, OD values exceeding the control were confirmed for the conditions in which the total molar concentrations were 0.3 M and 0.4 M, respectively. On the other hand, for the conditions in which the total molar concentrations were 0.5 M, 0.6 M, and 0.9 M, OD values lower than the control were confirmed. The degree of inhibition of proliferation was not dependent on the total molar concentration, and the strongest proliferation inhibitory effect was confirmed under the conditions of 0.6 M (galactose: sorbitol = 0.2 M: 0.4 M).

Example 5

Examples Hereinafter, typical formulation examples according to the present invention are shown, but the present invention is not limited thereto.

The hydraulic agent was prepared according to the following formulation.

2.0% by weight of ethanol

Spice 1.0

Galactose 3.7

Sorbitol 7.4

Water rest

100.0

Example 6

The toothpaste was prepared according to the following prescription.

Calcium carbonate 40.0 wt%

Glycerin 10.0

Carboxymethylcellulose 2.0

Sodium lauryl sulfate 2.0

Spice 1.0

Saccharin 0.1

Chlorhexidine 0.01

Galactose 5.5

Sorbitol 11.0

Water rest

100.0

Example 7

A breath spray was prepared according to the following prescription.

Ethanol 10.0 wt%

Glycerin 5.0

Fragrance 0.05

Coloring matter 0.001

Galactose 5.5

Sorbitol 11.0

Water rest

100.0

Example 8

Troches were prepared according to the following prescription.

Arabic gum 6.0 wt%

Paramount 75.8

Spice 1.0

Galactose 5.5

Sorbitol 11.0

Sodium monofluorophosphate         0.7

100.0

Example 9

The gum was prepared according to the following prescription.

Gum base 20.0 wt%

Paratunit 61.5

Galactose 5.5

Sorbitol 11.0

Spice 2.0

100.0

Example 10

Candies were prepared according to the following prescription.

Reduced starch syrup 34.0 wt%

Paratiunit 33.5

Citric acid 2.0

Fragrance 0.2

Galactose 5.5

Sorbitol 11.0

Water rest

100.0

Example 11

The tablets were prepared according to the following prescription.

0.2% by weight of sucrose fatty acid ester

Paratyite 78.6

Fragrance 0.2

Galactose 5.5

Sorbitol 11.0

Water 4.5

100.0

Example 12

Gumi jelly was prepared according to the following prescription.

Gelatin 60.0 wt%

Reduced starch syrup 21.6

Plant Maintenance 4.5

Malic acid 2.0

Galactose 3.8

Sorbitol 7.6

Fragrance 0.5

100.0

Example 13

The beverage was prepared according to the following prescription.

Orange juice 30.0 wt%

Citric acid 0.1

Vitamin C 0.04

Fragrance 0.1

Galactose 5.0

Sorbitol 10.0

Water rest

100.0

This application claims priority from Japanese Patent Application No. 2011-145737, filed on June 30, 2011, which is hereby incorporated by reference in its entirety.

Claims (7)

The agent for inhibiting the growth of bacteria in the oral cavity, which comprises galactose and sorbitol. The method according to claim 1,
Galactose, and sorbitol in a concentration of 0.5 M to 0.9 M in total. 3. The method according to claim 1 or 2,
Wherein the ratio of galactose to sorbitol is in the range of 2: 1 to 1: 3. 4. The method according to any one of claims 1 to 3,
The agent for inhibiting the growth of bacteria in the oral cavity, wherein the bacteria in the oral cavity are periodontal pathogens. 5. The method according to any one of claims 1 to 4,
Wherein the bacterium in the oral cavity is Fusobacterium nucleatum. The oral composition according to any one of claims 1 to 5, which contains the agent for inhibiting the growth of bacteria in the oral cavity. A food characterized by containing the agent for inhibiting the growth of bacteria in the oral cavity according to any one of claims 1 to 5.

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