JPH03261716A - Composition for oral cavity application - Google Patents

Abstract

PURPOSE:To provide a composition for oral cavity application, containing protamine, effective in suppressing the adhesion of oral bacteria to the tooth and the periodontium and useful for the prevention and remedy of dental caries and periodontosis. CONSTITUTION:The objective composition contains 0.001-5wt.%, especially 0.01-0.5wt.% of protamine. Protamine has an effect to suppress the adhesion of oral bacteria such as Bacteroides gingivalis. Protamine is a kind of strongly basic simple proteins, separable from the sperm nucleus of fish and classified to monoprotamine (e.g. salmine and clupeine), a diprotamine (e.g. dipurinine) and triprotamine (e.g. sturine). The composition is used as dentifrice and medicine and, in the case of using as a medicine, the dose of the peptide for adult is 5-50mg per day for oral administration and 1 to several tens mg at a time for external use.

Description

[Detailed description of the invention]

INDUSTRIAL APPLICATION FIELD The present invention relates to an oral composition that has the effect of suppressing the adhesion of oral cavity spores to teeth and periodontal tissue, and has preventive and therapeutic effects on dental caries and periodontal disease. . Conventional techniques and challenges It is clear that dental caries and periodontal disease are caused by certain types of oral hygiene products, such as Bacteroides gingivalis.
Periodontal pathogenic bacteria such as S) adhere to and settle on teeth and periodontal tissue, and the enzymes and endotoxins produced by these bacteria destroy the periodontal tissue, causing periodontal disease. Therefore, methods to suppress the adhesion of Bacteroides gingivalis have been investigated to prevent the onset of periodontal disease. For example, arginine and lysine, which are amino acids, suppress the adhesion of Bacteroides gingivalis to buccal mucous epithelial cells. There are complaints such as doing so (Journal of the Japanese Society of Hygiene 38:590-591% 1988)
However, these methods have problems in that the target cells are not derived from periodontal tissues and their effects are weak. In view of these circumstances, the present inventors investigated various drugs effective for preventing and treating dental caries and periodontal disease, and found that protamine is suitable for use outside of the kiln. I was able to complete it. Means for Solving the Problems The present invention provides an oral composition characterized by containing protamine. Since protamine effectively inhibits the adhesion of bacteria such as Bacteroides gingivalis to teeth and periodontal tissue, the oral composition of the present invention is useful as a dentifrice or pharmaceutical for the prevention and treatment of caries and periodontal disease. be. Protamine is a type of strong basic simple protein that has been isolated from fish sperm nuclei, etc., and can be classified into monoprotamines such as salmin, crupein, and sfunpurin, diprotamines such as diprinin, and triprotamines such as sullin, and is commercially available. In the present invention, these can be used alone or in combination of two or more. The protamine content in the composition is 0.001 to 5% by weight,
Especially 0. 1 to 0.5% by weight of O is preferred. The composition for use in the present invention can be prepared in accordance with conventional methods such as toothpaste, toothpaste, liquid toothpaste, mouthwash, chewing gum, gargle, troche, pasta, cream, ointment, patch, etc.
It can be a toothpaste such as a tablet or a medicine. Other ingredients are not particularly limited, and ingredients that are normally used in this type of composition can be included. For example, in the case of toothpaste, abrasives, binders, wetting agents, sweeteners, fragrances, preservatives, other medicinal agents, etc. are appropriately added. In the case of pharmaceuticals, the amount of peptide used for oral administration is usually 5 to 50 mg per adult molar, and for external use, the amount of peptide per dose ranges from 1 to several tens of grams at a time, without harmful side effects. It can be used to prevent and treat periodontal disease. When using toothpaste, take these dosages into consideration and use it according to the usual method. Effects: Hereinafter, the effects of protamine used in the present invention will be specifically shown through experiments. (Experiment 1) Bacteroides gingivalis 381 to gingival epithelial cells
Effect on strain attachment: Bacteroides gingivalis strain 381, which had been cultured for 48 hours under anaerobic conditions, was incubated with 10 μg containing 0.15 M NaCl.
After thoroughly washing with 1 M phosphate buffer (pH 7.0, PBS), it was suspended in PBS. On the other hand, human gingival epithelial cells were collected from an adult without periodontitis, thoroughly washed, and then suspended in PBS. 1 m 12 f of skeletal cell suspension (I
1 L was incubated at 37° C. for 30 minutes with shaking to allow the adhesion reaction of Bacteroides gingivalis strain 381 to denture epithelial cells. After the reaction was completed, epithelial cells were collected using a membrane filter with 1112 μm holes. The collected epithelial cells were suspended in PBS, smeared on a glass slide, air-dried, flame-fixed, and stained with gentian bioleft. The number of somites attached to the epithelial cell is
Thirty epithelial cells were randomly selected under an optical microscope (magnification: 10×40), and the number of adherent bacteria per 1 m of epithelial cells was calculated. In addition, as a negative control, epithelial cells were reacted with PBS only, and the number of spores originally attached to the epithelial cells was determined and used as a negative control value. The value obtained by subtracting the negative control value was used. The effect of peptides on the adhesion of Bacteroides gingivalis to epithelial MPM was investigated in the same manner as described above, after Bacteroides gingivalis was reacted with these substances for 15 minutes at room temperature. The results are shown in Table 1 below. As shown in Table 1, adhesion was slightly inhibited by the amino acids arginine and lysine, but almost no inhibition was observed by glycine or bradykinin potentiator C. On the other hand, basic peptides (protamine) such as salmin and crupein significantly inhibited cell adhesion to epithelial cells. (Experiment 2) Effect on the adhesion of Bacteroides gingivalis strain 381 onto saliva-coated hydroxyapatite (HAP) beads.
5- for 1 hour at room temperature. Visit beads with 0.05M KCI% 1+mM KHz-PO
,% 1mM CaC15 and 1mM MgC1,
(This buffer is a model for salivary inorganic components) l−, pH 6.0. The beads were then incubated for 1 hour at room temperature with a 0.5-p) sample solution of 17.0 and washed twice with the buffer 1aff. Next, in 0.5 ml of the buffer solution,

[3H] A suspension containing 5.0×10 thymidine bacteria (Bacteroides gingivalis) was added to the beads and incubated at room temperature for 1 hour. After washing three times with the above buffer solution 1-, the beads were transferred to a vial and radioactivity was counted using a liquid synthesis counter. On the other hand, we counted the autopsies of known [''H]I[llI cells using the same method, and created a calibration curve for bacteria.
Shown in the table. As shown in Table 2, protamines such as salmin and crupein significantly inhibited the adhesion to the saliva-covered HAP of the narrow mouth. (Mitsubishi No. 11113) Concentration dependence of protamine in inhibiting the adhesion of Bacteroides gingivalis Using salmin and kulperin from various epidermises, we investigated the effect of protamine on inhibiting the adhesion of Bacteroides gingivalis to gingival epithelial cells in the same manner as in Experiment 1. The results are shown in Table 3 below. As shown in Table 3, the adhesion inhibitory effect of protamine was observed even at a concentration of 0.001+*M. (Experiment 4) Protamine localized to hamster periodontal Effect of inhibiting colonization of Bacteroides gingivalis Ten 5-week-old germ-free hamsters were divided into two groups. On the first day of the experiment, a 0.1-M salmine hydrochloride solution was applied to the mandibular molars of the first group of hamsters using a cotton swab. PBS was applied to the surface of the second group as a control.Subsequently, a ligature was applied to the target molars, and a
Bacteroides gingivalis ESOI prepared in 12
32 strains of sodium bicarbonate solution were administered. Thereafter, the aqueous salmine solution and the PBS solution were applied once a day every day, and the sodium bicarbonate solution was administered twice a week for 6 weeks. One week after the final sodium hydroxide administration, the degree of gingival inflammation at the test site was observed, and after the ligature was removed, plaque from the site was collected with a curette and suspended in Ringer's solution maintained anaerobically. , Bacteroides gingivalis in this sample was counted by the maim method. The results are shown in Table 4 below. As shown in Table 4, in the Salmino aqueous solution treated group, colonization of Bacteroides gingivalis in the periodontal area was significantly suppressed and the degree of gingival inflammation was also mild compared to the control group. Next, the present invention will be explained in more detail with reference to Examples. In the Examples, all "%" means % by weight. Example 1 (Toothpaste) A toothpaste was manufactured according to a conventional method using the following formulation. Ingredients Amount (% by weight) Calcium diphosphate 45 Rubit dihydrate (70% solution) 10 Glycerin 10 Sodium lauryl sulfate 1.5 Carboxymethyl cellulose l Carrageenan
0.5 Salmine
0.05 Saccharin Sodium 0.1 Preservatives and Fragrances 1.2 Distilled Water
Adjustment to 100% Example 2 (Toothpaste) A toothpaste was manufactured according to a conventional method using the following formulation. Ingredients Amount (wt%) Silicic anhydride 15 Sodium lauryl sulfate l Polyacrylic acid l
Nrubit (70% solution) 60 Carrageenan 0.2 Kurupein 0.01 Salmine
0. Ol saccharin sodium
0.2 Preservatives and fragrances
1.2 Distilled water adjusted to 100%*
Example 3 (Mouthwash) A mouthwash was manufactured according to a conventional method using the following formulation. Ingredients Amount (wt%) Ethanol 5 N Rubit (70% solution) 15 Polyoxyethylene (60 EO)
0.5 Linghua castor oil polyoxyethyl monooleate 0.5 (20EO
) Sorbitan Clupein 0.05 Saccharin Sodium Preservatives and Flavors 0.8 Distilled Water
100% Preparation Example 4 (Mouthwash) A mouthwash was manufactured according to a conventional method using the following formulation. Ingredients Amount (wt%) Ethanol 15 Glycerin
15 Polyoxyethylene (60EO) 1 Hydrogenated castor oil salmine 0.05 Cetylpyridinium chloride 0.05 Saccharin sodium 0.1 Fragrance
0.5 Distilled water to 100% Preparation Example 5 (Pasta in the Paste) Pasta in the Pasta was produced according to the conventional method according to the following recipe. Ingredients Amount (wt%) White petrolatum 1060 Stearin alcohol 8.0 Propylene glycol
5.6 Sodium lauryl sulfate 0.6 Ethyl paraoxybenzoate 0.01 Distilled water
16.1 Salmin
1.0 Carboxymethylcellulose Adjusted to 100% Sodium Effects of the Invention According to the present invention, a composition useful for the prevention and treatment of dental caries and periodontal disease is obtained.

Claims (1)

[Claims] (1) An oral composition characterized by containing protamine.