JPS6317819A - Oral composition effective against dental caries and periodontosis - Google Patents

Abstract

PURPOSE:An oral composition, containing an extract essence of a honeybee nest, capable of selectively suppressing multiplication of causative bacteria of dental caries and periodontosis and effective in preventing and treating the dental caries and periodontosis and having high safety. CONSTITUTION:An oral composition obtained by blending an extract essence of a honeybee nest (extract solution obtained by extracting a honeybee nest with a water-miscible organic solvent, preferably methanol, ethanol, acetone, etc., or a blend thereof with water, concentrated essence or dried powder thereof) in an amount of >=0.05wt% based on the total weight of the above-mentioned composition expressed in terms of the concentrated essence in an oral composition, e.g. dentifrice, gargle, chewing gum, ointment, troche, strips, etc., having selective antibacterial action on causative bacteria of dental caries, e.g. Streptococcus mutans, etc., causative germs of periodontosis, e.g. bacterial cluster of the genus Bacteroides, etc., and effective in preventing and treating dental caries and periodontosis.

Description

DETAILED DESCRIPTION OF THE INVENTION Field of Industrial Application The present invention relates to an oral composition, and more particularly to a composition prepared from honey bee honeycomb which has selective antibacterial activity against caries-causing bacteria and periodontal disease-causing bacteria. The present invention relates to an oral composition effective for preventing and treating caries and periodontal disease, which contains an extract as an essential ingredient.

BACKGROUND OF THE INVENTION Caries and periodontal disease are two major diseases in the field of dentistry, and it is no exaggeration to say that almost all causes of tooth loss are attributable to one or the other disease.

In recent years, as a result of microbiological research into the etiology of these diseases, it has become clear that these are all bacterial infections caused by oral cavity. That is, oral bacteria, especially Streptococcus 0 muutans (S
Treptococcus mutans (hereinafter referred to as S. mutans) converts sucrose in food into sticky polysaccharides (glucan), and the glucan adheres to the tooth surface to form bacterial aggregates, that is, plaque. . When this plaque comes out, organic acids such as lactic acid produced by the proliferation of bacteria demineralize the teeth, causing caries to develop and progress.

The thick plaque deposited on the teeth also creates an anaerobic environment, allowing anaerobic bacteria to breed. Anaerobic bacteria actively proliferate in the more anaerobic subgingival region, causing gingival inflammation and forming periodontal pockets (cul-de-sacs). As the disease progresses, the periodontal pockets become deeper, the inflammation reaches from the gingiva to the alveolar bone, and the alveolar bone resorbs, eventually leading to the mountain becoming unstable and falling off. Periodontal disease is a process like this. This is a general term for inflammatory diseases that occur in the periodontal tissues.

In recent years, efforts have been made to search for periodontal disease-causing bacteria, and currently only Bacteroides
Bacteria, Actinobacillus
s) Bacterial group, Capnocytophaga
F usob
acterium), Eikenella
Durham-negative anaerobic bacteria such as the lla) group are attracting attention as causative bacteria. In addition to this, Actinomyces (Ac
tinomyces) and spirochetes (Spir□
It is said that bacteria such as P. chetes may also be causative bacteria, but in any case, it is a well-established theory that periodontal disease is caused by a limited number of types of bacteria, rather than by non-specific bacteria in plaque. It has become.

As described above, dental caries is an infectious disease caused by S. mutans, and periodontal disease is also an infectious disease caused by a limited number of types of bacteria. Therefore, suppressing the growth of pathogenic bacteria is considered to be the most effective means for preventing or treating these diseases. However, continuous use of antibiotics and antibacterial agents for a long period of time is undesirable because it may cause side effects, the appearance of resistant bacteria, and bacterial replacement due to disturbance of bacterial flora.

Therefore, the present inventors conducted a search for a safer antibacterial substance that is highly selective for the bacteria that cause caries and periodontal disease.As a result, we found that honey bee honeycomb extract was found to be the cause of the above-mentioned diseases. The present inventors have discovered that the proliferation of these cells can be selectively inhibited, and have completed the present invention.

That is, the present invention provides an oral composition for preventing and treating caries and periodontal disease, which contains honey bee honeycomb extract as an essential ingredient.

An example of blending a honeycomb extract as an oral composition is disclosed in Japanese Patent Publication No. 109516/1983. However, the invention disclosed in this publication focuses on the anti-inflammatory effect of extracts from the honeycombs of paper wasps, wasps, and black hornets, and has developed an effective treatment for the prevention and treatment of gingivitis containing extracts from these honeycombs. The present invention aims to provide an oral composition. All of these bees belong to the family Vespidae, and are taxonomically and evolutionary phylogenetically different from honey bees at the family level. Furthermore, this publication does not mention at all the antibacterial effects of these honeycomb extracts, especially the selective antibacterial effects.

The oral composition of the present invention can be produced by mixing the honeycomb extract with various carriers and/or adjuvants commonly used in the production of oral compositions in accordance with a conventional method. .

Honeybee hive extract is made by crushing the hive properly.
It can be obtained by extracting this with a suitable solvent. The honey bee nest is Apis, an ancient Japanese species that is distributed all over Japan and kept in Japan for honey collection.
Indica japonica (Apis 1ndica j
aponica) and the original species of this species, Apis indica (Ap), which is widely distributed or domesticated in Asia.
In addition to Apis 1ndica) and its subspecies, varieties, or regional types, there is also a different species of Western honey bee, Apis melihuera (Apis +1l), which is distributed and kept in Europe and America.
ellH'era) and its subspecies, varieties, or local forms can be used. That is, in this specification, honey bees refer to species belonging to the family Apidae and the genus Apis.

As the extraction solvent, it is preferable to use water-miscible organic solvents such as methanol, ethanol, acetone, or a mixture of water and these organic solvents.

In this specification, the honeycomb extract means a honeycomb extract obtained by the above method, a concentrated extract thereof, or a dry powder thereof, unless otherwise specified.

The carrier and auxiliary agent to be mixed with the above extracted extract should be appropriately selected from known ones depending on the type of oral composition (e.g., dentifrice, deodorant, chewing gum, ointment, troche, strips, etc.). Although there are no particular limitations, particularly preferred examples include dibasic calcium phosphate, glycerin, gelatin, vinyl acetate resin, carrageenan, sodium lauryl sulfate, sodium lauroyl sarcosine, polyisobutylene, butyl hydroxybenzoate, sorbitol, mannitol, Carbopol, macrogol, carboxin methylcellulose sodium, white petrolatum,
Plastinobase, polyethylene glycol, silicic anhydride, calcium carbonate, magnesium stearate, talc, hydroxypropyl cellulose, liquid paraffin,
These include ethanol, methylparaben, and saccharin.

Dexamethacin, which has an anti-inflammatory effect, may also be used as needed.
Indomethacin, water-soluble azulene, etc. can also be blended.

The content of the extracted extract in the oral composition varies depending on the type of the above-mentioned preparation, but it is usually 0.05% by weight or more of the aphrodisiac extract based on the total weight of the composition.

The present invention will be explained in more detail with reference to Examples below.

Example 1 Production of extract extract An extract was obtained using ethanol from the dried powder of beehives after collecting honey and beeswax.

That is, after crushing a beehive and collecting honey by centrifuging it, when the beehive is put into Q7A, the beeswax floats to the surface. After collecting and removing this as beeswax, the nest residue was dried and crushed to obtain a dry powder. 500 inches of ethanol was added to this dry powder 1009, and the mixture was refluxed for 3 hours in a cooling tube while being heated using a mantle heater. After cooling, the filtrate obtained by filtration was compressed under reduced pressure to obtain extract 209.

Test Example 1 An ethanol solution (100 m
9/x+2), create a 2-fold dilution series with ethanol,
An ethanol solution of g/aQ was prepared. A sterilized plain heart infusion (BHI) agar medium (or CAM agar medium) 9 , 9 zQ was kept at 50° C., and 1 ml of an ethanol solution of the above extract was added and mixed to prepare an agar plate. Separately, an extract-free agar plate was prepared to which 0.1 x 12 ethanol was added. By the above operations, the beehive extract was prepared at a final concentration of 1000.500.250.125.
A BHI agar medium (or CAM agar medium) containing 62.5, θμ9/YuQ and 1% ethanol was prepared.

A loopful of the overnight culture of the 61 strains to be tested was applied to this agar plate, and cultured at 37°C for 1 to 3 days under the conditions listed in Table 1-1. ) was determined for each bacterial strain. The breakdown of the 61 bacterial strains tested was 18 caries-causing bacteria (S, mutans) and 15 periodontal disease-causing bacteria.
strains, enteric lactic acid bacteria 12 special, other Durham-positive bacteria 10 strains,
There are 6 other strains of Durham-negative bacteria. The results are shown in Tables 1-1 to 4.
and shown in Table 2.

As is clear from Table 1-1, the extract according to the present invention exhibited antibacterial activity against most strains of cariogenic bacteria, S. mutans, at a concentration of 250 μ9 IRQ or less. In particular, all MICs for strains of human-derived serotypes c, d, e, r, and g were 250μ9/1tQ or less.

Table 1-2 shows the MIC for periodontal disease-causing bacteria.
The MIC for all strains was below 250μ9IRQ. In particular, Bacteroides gingivalis, which is attracting the most attention as a periodontal disease-causing bacterium.
It has strong antibacterial activity against S gingivalis),
In both of the two strains tested, no bacterial growth was observed even at the lowest concentration tested, 62°5μ9/sQ.

On the other hand, the MIC for any strain of intestinal lactic acid bacteria shown in Table 1-3 is higher than 500 μ9/2 (!
Furthermore, as shown in Table 1-4, no antibacterial activity was shown against Durham-negative aerobic bacteria even at 1000μ9/RQ.

In this way, honey bee hive extract is effective against caries-causing bacteria.
Although it exhibits strong antibacterial activity against periodontal disease-causing bacteria, it has unique properties that make it rarely effective against intestinal bacteria and lactic acid bacteria. Therefore, the extract of the present invention can be used for a long period of time for the prevention and treatment of dental caries and periodontal disease without causing disturbance of intestinal flora or bacterial replacement.

Examples of formulations of oral compositions containing the honeybee honeycomb extract of the present invention as an essential ingredient are listed below.

Toothpaste Dibasic Calcium Phosphate 42 Glycerin
18 Karaginano
0.9 Sodium lauryl sulfate 1.
2 Sabcalin 1 Honeybee's nest extract 1 Paraoxyamne 9, butyl fragrant 0.1 Fragrance
0.1 water
Total remaining amount 100 (wt%) Manufactured according to a conventional method for manufacturing toothpaste. That is, the prescribed amounts of water, glycerin, carrageenan, saccharin, honeybee's nest extract, butyl paraoquin benzoate, and fragrance are mixed together to swell the binder, and then dibasic calcium phosphate and lauryl are added. Add sodium sulfate,
Mix well.

After thoroughly defoaming and mixing, fill into a tube. To use, apply 1 amount on a toothbrush in the same way as regular dentifrice, and brush the teeth and gums with 5. Sing.

Formulation example 2 Stimulating agent ethanol (90%) 20 Saccharin 0.3 Sodium lauroyl sarcosine 05 Gelatin
0.5 Beehive extract
l fragrance l
Water Total remaining amount 100 (wt%) A bedding agent having the above formulation is manufactured according to a conventional method.

To use it, dilute it about 5 times with water, put an appropriate amount of it in your mouth, hold it for a few minutes, then spit it out and rinse with water.

Formulation example 3 Chewing gum vinyl acetate 23 polyisobutylene 3 calcium carbonate
2 sorbit
60 Mannito 10 Beehive extract l Fragrance
lTotal amount
100 (wt%) A chewing gum having the above formulation is manufactured according to a conventional method.

Formulation example 4 Gingival pine surge ointment Macrogol 4000 4 s, s Macrogol 400 48.5 Carbobol 934
0.5 Fragrance 1.5 Beehive extract 0.5 Water-soluble azulene
0.5 total! l
100 (wt%) The above prescription ointment is manufactured according to a conventional method. To use, take an appropriate amount on your finger or toothbrush and massage the gums.

Formulation Example 5 Oral Ointment Sodium Carboxymethyl Cellulose 31.5 White Petrolatum 11.5 Plastibase
23 Beehive extract
05 Indomethacin 0.5 Liquid paraffin Total remaining amount
100 (wt%) An ointment having the above formulation is manufactured according to a conventional method. To use, apply an appropriate amount to the gums.

Formulation examples Lozenge polyethylene glycol 6000 2 Silicic anhydride l Magnesium stearate 06 Talc 0
, 3 Hydroxypropyl cellulose 0.7 Beehive extract 05 Mannito
Full amount remaining
l 00 (wt%) A lozenge having the above formulation is manufactured according to a conventional method.

Formulation Example 7 Gingival Pocket Insert Agent (Strips) Methylparaben 0,1 Dexamethacin
0.1 Beehive extract
0.5 Hydroquine Propyl Cellulose 99
．． 3 Total amount +00 (weight%)
The above formulation is uniformly mixed with 9 times the amount of water, a film is prepared by a casting method, and the film is cut to an appropriate size to produce strithopus. To use it, cut it according to the width and depth of the periodontal pocket and insert it into the periodontal pocket.

Procedural amendments Commissioner of the Patent Office Miyazaki January 29, 1982 2 Name of the invention Oral composition effective for caries and periodontal disease 3 Relationship to the case of the person making the amendment Patent applicant address Osaka City, Osaka Prefecture 1-8-1 Tatsunishi, Ikuno-ku Name Rohto Pharmaceutical Co., Ltd. 4 Agent address 2-1-61 Mi, Higashi-ku, Osaka City, Osaka Prefecture, 540
Issue Tweet: /21 MID Tower Publication Telephone (06)949
-12617, Correction details (1) Page 8, line 11, “Ethanol solution in Example 1” is corrected as follows: “Extraction of honey bee nest obtained in Example 1 The antibacterial activity of the extract was investigated using the dilution method.

Ethanol solution of the above extracted extract.'' (2) On page 10, two lines from the bottom, insert the following sentence between ``Ru.'' and one line from the bottom, ``Below'': ``Test Example 2 below.'' 3 explains the in vivo effect of the honeybee hive extract of the present invention on dental caries and periodontal disease.

Test Example 2 The effectiveness of the extract according to the present invention against dental caries in rats was investigated.

Twenty-four 16-day-old SD male rats were fed commercially available powdered feed C.
E-2 (Nippon Talea) and penicillin G (4000 units/j10-containing water) were reared for 4 days to suppress the resident bacterial flora.
They were randomly divided into groups (8 animals each). S. mutans MT8 was found in the rat oral cavity of the infected control group and the extract administration group.
1.18R strain (streptomycin resistant) overnight culture suspension (number of bacteria: 8 x 10'/, IC) at 0, 2 x
The infection was caused by inoculation with Q for 5 days. From the start date of strain inoculation, the non-infected control group and infected control group were on a diet.
2000, and the extract-administered group was fed Diet 2000 with feed containing 1% (w/w) of this extract.

Diet 2000 is a waxy caries-inducing feed that is usually used in this type of experiment and contains 56% sucrose, 28% skim milk powder, 6% wheat flour, 4% brewer's yeast, 3% alfalfa powder, and 1%.
Consists of liver end, 2% salt.

The status of weekly colonization of the inoculated bacteria in the oral cavity of each rat was examined once a week by the method described below. After swabbing the tooth surface of each rat with a sterile cotton swab, it was placed in 1612 sterile physiological saline and stirred well. This was diluted stepwise 10 times, and 0 and 1 mQ of this solution was added to streptomycin (500μ9I).
RQ) containing Mitis salivarius agar plate (Difc)
S mutans MT81 in the physiological saline containing the original cotton swab, based on the number of colonies that grew after application to S. mutans MT81.
The number of 48R bacteria was determined. When the number of bacteria is 0/tenji, the score is 0.1 to 102/mQ, the score is 1.102 to 1.
03f[U/i(, the score is 2, and the following is 1.
0'~10' pieces/I score 4.105 pieces/1
In the above, the score of each individual was determined as a score of 5.

When they reached 72 days of age, all rats were decapitated and Reg
olati and Hotz method (Helv, 0dont
o1. Acta, 16:13, 1972) and the method of Keyes (J, Den.
The caries score was evaluated using the following method.

Table 3 shows the results of the bacterial colonization test. In the group treated with this extract, bacterial colonization was completely suppressed for two weeks after inoculation, and the number of bacteria gradually increased thereafter, but the number of bacteria remained significantly lower than that of the infected control group until 7 weeks after being sacrificed. . In the non-infected control group, no inoculated bacteria were detected throughout the experimental period. Table 4 shows the plaque index and caries score. In both cases, the group administered with this extract showed a significant suppressive effect (risk rate of 0.1% or less) compared to the control group. As mentioned above, this extract was used in rat animal experiments to inoculate S.

It has been shown that it inhibits the colonization of P. mutans and plaque adhesion, and exhibits a caries preventive effect.

The effectiveness of the extract according to the present invention against experimental periodontal disease in hamsters was investigated.

Twelve 4-week-old male Golden Syrian hamsters were reared for 4 days on solid commercial feed CE-2 (Nippon Talea) and water containing penicillin G (4000 units/II (1)) to eliminate the resident bacterial flora. After suppression, the hamsters were randomly divided into three groups (4 animals each): a non-infected control group, an infected control group, and an extract-administered group. Actinomyces viscosus ATCC 15987, which is known to induce
Infection was carried out by inoculating a 2-day culture suspension of the strain (number of bacteria: 2.5 x to 8 cells/Shoulder Q) at a dose of 0.25πQ for 5 days. From the start date of bacterial inoculation, the non-infected control group and infected control group received Diet 2050, and the extract administration group received Diet 2.
050 was fed with feed containing 1% (V/W) of this extract. In addition, Diet2050 is Test Example 2
The 56% sucrose in the caries-inducing feed Diet 2000 described in 2000 was replaced with 28% sucrose and 28% corn starch to create a periodontal disease-inducing feed that does not cause caries.

Eight weeks after infection, the salivary occult blood of all the animals was evaluated under anesthesia using floor occult blood test paper (Hemastyx ■, Miles Sankyo) according to a colorimetric table in 5 grades from 1 to 1.

The gingivitis index was determined according to Rosenberg's criteria (J, Pe
riodontol, 37:208.1966) visually evaluated on a scale of 0 to 3. Next, all animals were decapitated, and the plaque index was measured by the method of Rogolati and Hotz (described above) after staining the teeth with Red Coat (Patola). After measuring the plaque index, the jawbone was extracted, and the left jaw was used as a bone specimen by removing the soft tissue. After one hour, a photograph of the molar region was taken from the west side using a stereomicroscope. The distance from the alveolar bone crest to the cement-enamel junction was measured on the photograph, and the average alveolar bone resorption distance per tooth root was determined for each individual.
), 750μF or more is evaluated as (-)7. On the other hand, the right jaw was fixed in formalin, dehydrated, decalcified, and embedded in paraffin, then sections were prepared, stained with hematoxylin and eosin, and histologically observed under a microscope.

As shown in Table 5, the results are not shown as the average value for each group due to the small number of individuals, but the measured values for each individual are shown as they are. Salivary occult blood, which indicates bleeding from the gums, gingivitis index, which indicates the degree of gingival inflammation, and plaque index, which indicates the degree of plaque adhesion, as well as alveolar bone resorption, which is the most important characteristic of periodontal disease.
The infection in the extract-administered group was mild compared to the control group, indicating that the onset of periodontal disease was suppressed. Also,
Although not shown in the table, histological observations showed that in the infected control group, significant plaque accumulation and fugue migration of polymorphonuclear leukocytes were observed, indicating the formation of periodontal disease pathology. In the group they were mild. On the other hand, in the non-infected control group, almost no signs of periodontal disease were observed in any of the measurement items. (3) Insert the following table on page 25, line 1, before "Patent application k": "The values in the colonization test table for inoculated bacteria in animal experiments on caries-1 are the average of the bacterial index of each group." Values in the Table of Plaque Suppression and Caries Preventive Effects of Beehive Extract are mean ± standard error N5 Results of hamster periodontal disease animal experiment Oral Composition Effective for Erosion and Periodontal Disease 3, Relationship with the Amendment Case Patent Applicant Address 1-8-1 Tatsunishi, Ikuno-ku, Osaka-shi, Osaka Name Rohto Pharmaceutical Co., Ltd. 4 Agent Address 〒 540 2-1-61 Higashi District Mi, Osaka City, Osaka Prefecture
Issue Tweet'/21 Inside MID Tower Telephone (06)949
-12616, Subject of amendment: ``Detailed Description of the Invention,'' Part 7 of the description, Contents of the amendment By the procedural amendment dated January 29, 1988, 1 of the description
Regarding the text inserted between lines 19 and 20 on page 0, please do the following 2.
Amendments will be made in the following places: 1) r1612j (in the written amendment, page 3, line 14) shall be changed to “
Corrected to "l'IQ".

2) “Rogolat i J (in the same procedural amendment, 6
page 13) was corrected to r Regolat i J.

that's all

Claims (1)

[Scope of Claims] 1. An oral composition for preventing and treating dental caries and periodontal disease, which is characterized by containing an extract of honeybee honeycomb. 2. The oral composition according to item 1, wherein the content of the extracted extract is 0.05% by weight or more of the concentrated extract based on the total weight of the composition.