JPH05271085A - Agent for preventing periodontosis - Google Patents

Abstract

PURPOSE:To provide the periodontosis-preventing agent containing the hydrophobic extract of the white-bark of a mulberry tree and having an antibacterial activity against gram-negative anaerobic pathogenic bacteria. CONSTITUTION:The objective preventing agent contains 0.0005-10wt.% of the hydrophobic extract of the mulberry white bark obtained by extracting the root barks of Morus alba L, Morus bombycis koids, etc., with an organic solvent and 0.005-5% of 1-menthol and/or 1-carvone.

Description

Detailed Description of the Invention

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a preventive agent for periodontal disease, which suppresses pathogenic bacteria of periodontal disease and is useful for preventing and treating periodontal disease and suppressing bad breath.

[0002]

2. Description of the Related Art Periodontal disease is one of the two major diseases in the oral cavity comparable to caries, and is a general term for diseases that occur in periodontal tissues such as gingivitis and periodontitis. While caries is caused by the demineralization of enamel by the acid produced by Gram-positive facultative anaerobic pathogens (Streptococcus mutans, etc.) that have settled on the tooth surface, periodontal disease is localized in the periodontal disease. It is a disease that occurs in the periodontal tissue due to a Gram-negative anaerobic pathogenic bacterium represented by Porphyromonas gingivalis, and these two diseases have completely different symptoms and causes.

The organic acids produced by the bacteria causing periodontal disease also cause bad breath. Thus, periodontal disease is an infection caused by certain Gram-negative anaerobic bacteria, and these bacteria are one of the means for preventing or treating periodontal disease from the viewpoint that they are deeply associated with the development of halitosis. Attempts have been made to suppress pathogenic bacteria with antibacterial agents, and hitherto, synthetic antibacterial agents such as chlorhexidine and cetylpyridinium chloride have been mixed in toothpaste and mouthwash, and their effects have been confirmed.

However, when considering the long-term use of these synthetic antibacterial agents and further application to the mouth, it is preferable to use naturally derived antibacterial agents from the viewpoint of safety.
However, until now, studies on naturally-occurring antibacterial agents against pathogens of periodontal disease have not been sufficiently conducted.

[0005] On the other hand, some studies have hitherto been made on the antibacterial activity in mulberry plants. For example, an inhibitory effect on growth of caries fungi by a hydrophilic organic solvent extract of mulberry bark (Japanese Examined Patent Publication No. 63-24489) and a quannon compound effective against Gram-positive bacteria (Japanese Examined Patent Publication No. 1-49152 and Japanese Examined Patent Publication No. 2-1824). However, all of them only show effectiveness against aerobic or facultative anaerobic gram-positive bacteria, and no report on periodontopathic bacteria which are gram-negative anaerobic bacteria is found.

[0006]

In view of these circumstances, the present inventors have conducted various studies in order to find a substance that suppresses the growth of periodontal disease-causing bacteria from various natural extracts, and as a result, mulberry Hydrophobic extract of skin shows very strong antibacterial activity against Porphyromonas gingivalis, a periodontopathic bacterium which is a gram-negative anaerobic bacterium, and a small amount of l-menthol and // Or, by adding l-carvone, its antibacterial activity is significantly increased,
The inventors have found that not only Porphyromonas gingivalis, but also Fusobacterium nucleatum, which is a causative bacterium of halitosis, and Actinomyces viscosus, which is a causative bacterium of gingivitis, have strong antibacterial activity, and the present invention has been completed.

That is, the present invention comprises a periodontal disease preventive agent characterized by containing a mulberry bark hydrophobic extract, and further the mulberry bark hydrophobic extract and l-menthol and / or l-carvone. The present invention provides a preventive agent for periodontal disease. The agent for preventing periodontal disease of the present invention remarkably suppresses the causative bacteria of periodontal disease, and is very useful for the prevention and treatment of periodontal disease and the suppression of halitosis associated with these causative bacteria.

The mulberry bark hydrophobic extract used as an antibacterial component in the preventive agent for periodontal disease of the present invention is a mulberry of the Moraceae family or its homologous plants, for example, mulberry (Morus alba).
L. ), Yamaguchi (Morus bombycis koidz), and Rosou (Morus Lhou koidz) root bark or commercial mulberry bark hydrophobic extract.

The hydrophobic extract can be obtained by extracting mulberry bark with an organic solvent according to a method known per se. Examples of the organic solvent used include organic solvents having a small polarity such as n-hexane and benzene, fatty acid esters such as methyl acetate, ethyl acetate and butyl acetate, ethers such as methyl ether and ethyl ether, anhydrous methanol and anhydrous ethanol. Be done.

In the present invention, the mulberry bark hydrophobic extract is used as an extract dry weight based on the total amount of the periodontal disease preventive agent.
0.0005-10% by weight, preferably 0.005-5
A desired effect can be obtained by blending the composition in a weight percentage.

Further, in the present invention, as described above,
It is preferable to add l-menthol and / or l-carvone, which increases the antibacterial activity of the mulberry bark hydrophobic extract. L-menthol and l-carvone were each added to 0.005 to 5 with respect to the total periodontal disease preventive agent.
A desired antibacterial activity-increasing effect is exhibited by blending it in a weight percentage. It is not preferable to add an excessive amount because it affects the flavor of the periodontal disease preventive agent.

The preventive agent for periodontal disease of the present invention is prepared according to a conventional method.
It may be in the form of toothpaste, mouthwash, pasta, chewing gum, candy or the like, and the other compounding ingredients are not particularly limited, and those normally compounded in this type of composition can be used.

[0013]

EXAMPLES Next, the present invention will be described in more detail with reference to examples and test examples. Example 1 Preparation of Extract A 10 L of n-hexane was added to 1 kg of commercially available powder of mulberry bark, and the mixture was extracted under reflux for 5 hours. The obtained extract was concentrated under reduced pressure and dried, 10 l of ethyl acetate was added to the residue, and extraction was carried out for 5 hours. The ethyl acetate extract was concentrated under reduced pressure and dried to obtain about 20 g of Extract A.

Example 2 Preparation of Extract B 2 kg of roots of mulberry (Morus alba L.) were ground and n
-Hexane: absolute ethanol (2: 1) mixed solvent 10
L was added and extraction was carried out for 2 hours under reflux. The extract was concentrated under reduced pressure and dried to obtain about 40 g of Extract B.

Example 3 Toothpaste Toothpaste containing mulberry bark hydrophobic extract A and l-carvone was prepared according to the following formulation. Ingredients Amount (wt%) Silicic acid 15 Carrageenan 0.5 Gelatin 0.3 Propylene glycol 3.0 Sorbit solution 60.5 Sodium lauryl sulfate 1.2 Lauric acid diethanolamide 1.5 Sodium saccharin 0.1 Mulberry peel hydrophobic Sex extract A 0.1 l-Carvone 0.5 Purified water Residual total 100.0%

Example 4 Mouthwash A mousewash containing mulberry bark hydrophobic extract B and 1-menthol was prepared according to the following formulation. Ingredients (wt%) Ethyl alcohol 15.0 Sorbit 10.0 Citric acid 0.05 Sodium citrate 0.2 Sodium benzoate 0.2 Sodium lauryl sulphate 0.2 Sodium saccharin 0.05 Blue No. 1 0.001 Mulberry White skin hydrophobic extract B 0.02 l-Menthol 0.05 Purified water Residual total 100.0%

Example 5 Chewing gum A chewing gum having the following composition was prepared by a conventional method. Ingredients Amount (% by weight) Chewing gum base 20 Powdered sugar 51.15 Glucose 10 Syrup candy 18 Mulberry bark hydrophobic extract B 0.05 l-Menthol 0.8 combined 100.0%

Example 6 Hard Candy A hard candy having the following composition was prepared by a conventional method. Ingredients Amount (wt%) Granulated sugar 46.8 Water syrup 52.34 Citric acid 0.3 Mulberry bark hydrophobic extract A 0.03 Mulberry bark hydrophobic extract B 0.03 l-Carvone 0.5 Total 100.0%

Test Example 1 The minimum inhibitory concentration (MIC) against the three periodontal pathogens of mulberry bark hydrophobic extracts A and B, l-menthol and l-carvone extracted in Examples 1 and 2 was determined as follows. Heart infusion broth (BHI broth)
Was determined by the liquid medium dilution method using. As a positive control, cetylpyridinium chloride, which is an antibacterial agent, was used.
The bacterial suspension prepared to about 10 ⁸ cells / ml and each test drug prepared to various concentrations were mixed in BHI broth,
Anaerobic culture (H ₂ 10%, CO ₂ 5%, N ₂ 8)
5%), the presence or absence of bacterial growth was visually determined. MIC (μg / m
l). The results are shown in Table 1.

[0020]

[Table 1] Minimum inhibitory concentration (MIC) (μg / ml) against periodontal pathogens Control Subject Strain Cetyl chloride Mulberry bark Hydrophobic l-Men l-Cal Pyridinium extract Tall bomb A AB Porphyromonas gingivalis 381 0.78 3.9 2.0> 800> 800 Azobacterium Nucleatum 0.78 7.8 3.9> 800> 800 ATCC 23726 Actinomyces viscosus T14V 0.39 15.6 7.8> 800> 800

Both the mulberry bark hydrophobic extracts A and B exhibited antibacterial activity in the range of 2.0 to 15.6 μg / ml against the tested strains. Especially for Porphyromonas gingivalis, which is a gram-negative anaerobic bacterium, 2.
It had a MIC of 0 to 3.9 μg / ml and showed excellent antibacterial activity. However, its activity was lower than that of cetylpyridinium chloride. On the other hand, l-menthol and l-
Carvone showed no antibacterial activity in the concentration range used.

Test Example 2 In the presence of perfume ingredients such as 1-menthol and 1-carvone,
The antibacterial activity of the mulberry bark hydrophobic extract A and B against periodontal pathogens was examined. 100μg / ml or 500μ
BHI broth containing g / ml of a fragrance component was added with various concentrations of the mulberry bark hydrophobic extract to inoculate the same as in Test Example 1.
After culturing, the presence or absence of growth was determined. The results are shown in Table 2. The results are shown as the minimum concentration of the mulberry bark hydrophobic extract, which exhibits antibacterial activity in the presence of each fragrance component, in μg / ml.

[0023]

[Table 2] Antibacterial activity of mulberry bark hydrophobic extract with addition of fragrance ingredients Porphyromonas fusobacterium gingivalis nucleatum 381 ATCC23726 Fragrance ingredients extract A extract B extract A extract B no addition 3.9 2.0 7.8 3.9 l-menthol 100 μg / ml 0.5 0.25 1.0 0.5 500 μg / ml 0.25 0.125 0.5 0.25 l-carvone 100 μg / ml 1.0 0.5 2. 0 1.0 500 μg / ml 0.5 0.25 1.0 0.5 anisaldehyde 500 μg / ml 3.9 2.0 7.8 3.9 cinnamic aldehyde 500 μg / ml 3.9 2.0 7. 8 3.9 Isoamyl acetate 500 μg / ml 3.9 2.0 7.8 3.9 Vanillin 500 μg / ml 3.9 2.0 7.8 3. 9 Minimum concentration of mulberry bark extract showing antibacterial activity: μg / ml

As shown in Table 2, l-menthol and l-
Due to the presence of carvone, mulberry bark hydrophobic extracts A and B
The antibacterial activity of the was significantly increased. On the other hand, the other fragrance ingredients did not affect the antibacterial activity of the mulberry bark hydrophobic extract.

Test Example 3 According to the formulation of Example 4, a mouthwash (test product) containing mulberry bark hydrophobic extract B and 1-menthol was prepared. On the other hand, instead of l-menthol, a mouse wash containing vanillin at the same concentration (control product 1) and a mouse wash containing no mulberry bark hydrophobic extract B (control product 2) were prepared and their antibacterial activities were compared. BHI agar medium was sterilized by heating and cooled to 50 ° C., to which was added Actinomyces viscosus T14V bacterial solution preliminarily cultivated in BHI broth to a concentration of about 10 ⁷ cells / ml, and a diameter of 90 m immediately.
10 ml each was poured into a sterile petri dish of m to be solidified. A cylinder having an inner diameter of 8 mm and a height of 10 mm was erected on this flat plate, filled with the above-mentioned three types of mouthwashes, and 37 ° C.
After anaerobic culture for 48 hours, the diameter of the zone (inhibition circle) in which bacteria did not grow was measured to determine the antibacterial activity. The results are shown in Table 3.
Shown in.

[0026]

[Table 3] Test product Control product 1 Control product 2 Diameter of blocking circle (mm) 15 8 0

As shown in Table 3, no inhibition circle was observed in the reference product 2 containing no mulberry bark hydrophobic extract B. On the other hand, in the control product 1, an inhibition circle was observed only in the inner diameter portion of the cylinder, whereas in the test product containing both the mulberry bark hydrophobic extract B and l-menthol, a large inhibition circle was formed and a strong antibacterial effect was observed. It showed activity.

[0028]

INDUSTRIAL APPLICABILITY According to the present invention, a prophylactic agent for periodontal disease having a strong antibacterial activity against Gram-negative anaerobic periodontal pathogenic bacteria is further blended with a hydrophobic extract of mulberry bark. By adding l-menthol and / or l-carvone, its antibacterial activity is enhanced, and a periodontal disease preventive agent having antibacterial activity against bacteria causing bad breath is obtained.

─────────────────────────────────────────────────── ─── Continuation of the front page (51) Int.Cl. ⁵ Identification code Internal reference number FI technical display location // (A61K 35/78 31: 045) 8413-4C (A61K 35/78 31:12) 8413- 4C

Claims (2)

[Claims] 1. A preventive agent for periodontal disease, which comprises a hydrophobic extract of mulberry bark. 2. The periodontal disease preventive agent according to claim 1, which comprises a hydrophobic extract of mulberry bark and l-menthol and / or l-carvone.