Stomatological composition

Abstract

A stomatological composition is provided. The stomatological composition is capable of stably compounding an antibody obtained from a hen egg yolk, and preventing diseases in an oral cavity such as odontonecrosis and periodontal disease from occurring, or improving such diseases in the oral cavity. The stomatological composition includes at least one selected from the group consisting of lipase, protease and the antibody obtained from the hen egg yolk, citric acid and a metal salt of citric acid. A total amount of the lipase, the protease and the antibody obtained from the hen egg yolk is in the range of 0.001 to 10 wt %. When an amount of the citric acid is defined as “A” [wt %] and an amount of the metal salt of citric acid is defined as “B” [wt %], the following relation is satisfied: 0.005≦A/B≦1. Further, the stomatological composition of the present invention includes a polyglyceryl fatty acid ester or an amino acid-based ampholytic surfactant as a surfactant. Further, the stomatological composition of the present invention includes a collagen.

Claims

1. A stomatological composition comprising: (C) at least one selected from a group consisting of lipase, protease and an antibody obtained from a hen egg yolk; (A) citric acid; and (B) sodium citrate, wherein (A) and (B), by weight percent (wt %) of each in the composition, are present in the composition in a ratio of 0.01≦A/B≦0.5, where (A) is present in the composition in a range of 0.005≦A≦0.08 and (B) is present in the composition in a range of 0.2≦B≦1 so that the composition can contain the lipase, the protease, and the antibody obtained from the hen egg yolk stably therein.

2. The stomatological composition as claimed in claim 1, wherein a total amount of the lipase, the protease and the antibody obtained from the hen egg yolk is in a range of 0.001 to 10 wt %.

3. The stomatological composition as claimed in claim 2, further comprising a polyglyceryl fatty acid ester or an amino acid-based ampholytic surfactant as a surfactant.

4. The stomatological composition as claimed in claim 1, wherein (A) and (C), by weight percent (wt %) of each in the composition, are present in the composition in a ratio of 0.0005≦A/C≦10000.

5. The stomatological composition as claimed in claim 4, wherein (B) and (C), by weight percent (wt %) of each in the composition, are present in the composition in a ratio of 0.005≦B/C≦10000.

6. The stomatological composition as claimed in claim 5, further comprising a polyglyceryl fatty acid ester or an amino acid-based ampholytic surfactant as a surfactant.

7. The stomatological composition as claimed in claim 6, further comprising a collagen.

8. The stomatological composition as claimed in claim 7, wherein a salt concentration in the stomatological composition is in a range of 0.1 to 20%.

9. The stomatological composition as claimed in claim 1, wherein (B) and (C), by weight percent (wt %) of each in the composition, are present in the composition in a ratio of 0.005≦B/C≦10000.

10. The stomatological composition as claimed in claim 1, further comprising a collagen.

11. The stomatological composition as claimed in claim 1, further comprising a polyglyceryl fatty acid ester or an amino acid-based ampholytic surfactant as a surfactant.

12. The stomatological composition as claimed in claim 1, wherein a salt concentration in the stomatological composition is in a range of 0.1 to 20%.

Description

EXAMPLES

(1) Next, description will be made on concrete examples of the invention.

Example 1

(2) First, a toothpaste agent of the stomatological composition was produced by using the following components 1 to 11 (unit: wt %).

(3) In this regard, an immune globulin was used as the antibody obtained from the hen egg yolk. Such an immune globulin was prepared from an egg which was produced by hen and in which the membrane-bound glucosyltransferase of Streptococcus mutans was immunized. The toothpaste agent was produced in the usual manner, except that the antibody obtained from the hen egg yolk was added to water with a hydrolysis collagen to dissolve them. 1. Antibody obtained from the hen egg yolk: 1.0 2. Citric acid: 0.08 3. Sodium citrate: 1.0 4. Hydrolysis collagen: 0.5 5. Polyglyceryl monolaurate: 0.2 6. Xylitol: 1.0 7. Concentrated glycerin: 20.0 8. Sorbitol: 40.0 9. Carboxymethylcellulose sodium: 2.7 10. Perfume material: 1.0 11. Distilled water: residue

Examples 2 to 10

(4) In each of the Examples 2 to 10, a toothpaste agent was produced in the same manner as the Example 1, except that an amount of each of the antibody obtained from the hen egg yolk, the citric acid and the metal salt of citric acid was changed as shown in Table 1.

Example 11

(5) A toothpaste agent was produced in the same manner as the Example 1, except that the immune globulin as the antibody obtained from the hen egg yolk was changed to an immune globulin prepared from an egg which was produced by hen and in which the gingipains of Porphyromonas gingivalis was immunized, the hydrolysis collagen was changed gelatin, and 2-alykyl-N-carboxymethyl-N-hydroxyethyl imidazolinium betaine as a surfactant instead of polyglyceryl monolaurate was contained in an amount shown in Table 1.

Example 12

(6) A toothpaste agent of the stomatological composition was produced by using the following components 1 to 13 (unit: wt %).

(7) In this regard, an immune globulin was used as the antibody obtained from the hen egg yolk. Such an immune globulin was prepared from an egg which was produced by hen and in which the membrane-bound glucosyltransferase of Streptococcus mutans was immunized. The toothpaste agent was produced in the usual manner, except that the antibody obtained from the hen egg yolk was added to water with a hydrolysis collagen to dissolve them. 1. Antibody obtained from the hen egg yolk: 1.0 2. Citric acid: 0.08 3. Sodium citrate: 1.0 4. Hydrolysis collagen: 1.0 5. Polyglyceryl monolaurate: 2.0 6. Calcium hydrogen phosphate: 25.0 7. Silicic anhydride: 7.0 8. Xylitol: 1.0 9. Concentrated glycerin: 25.0 10. Carboxymethylcellulose sodium: 1.0 11. Polyethyelene glycol 400: 2.0 12. Perfume material: 1.0 13. Distilled water: residue

Example 13

(8) A toothpaste agent of the stomatological composition was produced by using the following components 1 to 11 (unit: wt %).

(9) In this regard, the toothpaste agent was produced in the usual manner, except that lipase was added to water with a hydrolysis collagen to dissolve them. 1. Lipase (derived genus Pseudomonas): 1.0 2. Citric acid: 0.08 3. Sodium citrate: 1.0 4. Hydrolysis collagen: 0.5 5. Polyglyceryl monolaurate: 0.2 6. Xylitol: 1.0 7. Concentrated glycerin: 20.0 8. Sorbitol: 40.0 9. Carboxymethylcellulose sodium: 2.7 10. Perfume material: 1.0 11. Distilled water: residue

Examples 14 to 22

(10) In each of the Examples 14 to 22, a toothpaste agent was produced in the same manner as the Example 13, except that an amount of each of the lipase, the citric acid and the metal salt of citric acid was changed as shown in Table 2.

Example 23

(11) A toothpaste agent was produced by using the following components 1 to 11 (unit: wt %).

(12) In this regard, the toothpaste agent was produced in the usual manner, except that protease was added to water with a hydrolysis collagen to dissolve them. 1. Protease (papain): 1.0 2. Citric acid: 0.08 3. Sodium citrate: 1.0 4. Hydrolysis collagen: 0.5 5. Polyglyceryl monolaurate: 0.2 6. Xylitol: 1.0 7. Concentrated glycerin: 20.0 8. Sorbitol: 40.0 9. Carboxymethylcellulose sodium: 2.7 10. Perfume material: 1.0 11. Distilled water: residue

Examples 24 to 32

(13) In each of the Examples 24 to 32, a toothpaste agent was produced in the same manner as the Example 23, except that an amount of each of the protease, the citric acid and the metal salt of citric acid was changed as shown in Table 2.

Comparative Example 1

(14) A toothpaste agent was produced in the same manner as the Example 1, except that no citric acid was contained in the toothpaste agent.

Comparative Example 2

(15) A toothpaste agent was produced in the same manner as the Example 1, except that no sodium citrate was contained in the toothpaste agent and the surfactant shown in Table 1 was used.

Comparative Example 3

(16) A toothpaste agent was produced in the same manner as the Example 1, except that both the citric acid and the sodium citrate were not contained in the toothpaste agent and the surfactant shown in Table 1 was used.

Comparative Example 4

(17) A toothpaste agent was produced in the same manner as the Example 13, except that no citric acid was contained in the toothpaste agent.

Comparative Example 5

(18) A toothpaste agent was produced in the same manner as the Example 13, except that no sodium citrate was contained in the toothpaste agent and the surfactant shown in Table 2 was used.

Comparative Example 6

(19) A toothpaste agent was produced in the same manner as the Example 13, except that both the citric acid and the sodium citrate were not contained in the toothpaste agent and the surfactant shown in Table 2 was used.

Comparative Example 7

(20) A toothpaste agent was produced in the same manner as the Example 23, except that no citric acid was contained in the toothpaste agent.

Comparative Example 8

(21) A toothpaste agent was produced in the same manner as the Example 23, except that no sodium citrate was contained in the toothpaste agent and the surfactant shown in Table 2 was used.

Comparative Example 9

(22) A toothpaste agent was produced in the same manner as the Example 23, except that both the citric acid and the sodium citrate were not contained in the toothpaste agent and the surfactant shown in Table 2 was used.

(23) In each of the Examples and the Comparative Examples, the kind and the amount of the antibody obtained from the hen egg yolk or the enzyme, the amount of each of the citric acid, the sodium citrate and the sodium chloride, the kind and the amount of each of the collagen and the surfactant, and the like were shown in Table 1 and Table 2. Further, in Tables, indicated were the antibody obtained from the hen egg yolk as the immune globulin prepared from the egg, which was produced by hen and in which the membrane-bound glucosyltransferase of Streptococcus mutans was immunized, as “X”, the antibody obtained from the hen egg yolk as the immune globulin prepared from the egg, which was produced by hen and in which the gingipains of Porphyromonas gingivalis was immunized, as “Y”, the lipase as “L”, the protease as “P”, the hydrolysis collagen as “a”, the gelatin as “b”, the polyglyceryl monolaurate as “A”, the sodium lauroampho acetate as “B”, and the 2-alkyl-N-carboxymethy-N-hydroxyethyl imidazolinium betaine as “C”.

(24) TABLE-US-00001 TABLE 1 Toothpaste agents Antibodies Metal obtained salts of Evalua- from hen Citric citric Salt tions of egg yolk acids acid Collagens Surfactants concen- stable Amounts Amounts Amounts Amounts Amounts trations con- kinds C[wt %] A[wt %] B[wt %] kinds D[wt %] kinds E[wt %] A/B A/C B/C D/C E/C [%] taining Ex. 1 X 1 0.08 1 a 0.5 A 0.2 0.08 0.08 1 0.5 0.2 1 A Ex. 2 X 1 0.75 3 a 0.5 A 0.2 0.25 0.75 3 0.5 0.2 3 B Ex. 3 X 1 0.03 6 a 0.5 A 0.2 0.005 0.08 6 0.5 0.2 6 C Ex. 4 X 1 0.01 0.5 a 0.5 A 0.2 0.02 0.01 0.5 0.5 0.2 0.5 A Ex. 5 X 1 0.02 2 a 0.5 A 0.2 0.01 0.02 2 0.5 0.2 2 B Ex. 6 X 1 0.6 1 a 0.5 A 0.2 0.6 0.6 1 0.5 0.2 1 B Ex. 7 X 1 0.009 1 a 0.5 A 0.2 0.009 0.009 1 0.5 0.2 1 C Ex. 8 X 0.1 0.08 1 a 0.5 A 0.2 0.08 0.8 10 5 2 1 A Ex. 9 X 0.1 0.75 3 a 0.5 A 0.2 0.25 7.5 30 5 2 3 B Ex. 10 X 0.1 0.03 6 a 0.5 A 0.2 0.006 0.3 60 5 2 6 C Ex. 11 Y 1 0.08 1 b 0.5 C 0.2 0.08 0.08 1 0.5 0.2 1 A Ex. 12 X 1 0.08 1 a 1.0 A 2.0 0.08 0.08 1 1 2 1 A Com. X 1 — 1 a 0.5 A 0.2 — — 1 0.5 0.2 1 D Ex. 1 Com. X 1 0.08 — a 0.5 B 0.2 — 0.08 — 0.5 0.2 — D Ex. 2 Com. X 1 — — a 1.5 B 1.2 — — — 1.5 1.2 — D Ex. 3

(25) TABLE-US-00002 TABLE 2 Toothpaste agents Metal salts of Evalua- Citric citric Salt tions of Enzymes acids acid Collagens Surfactants concen- stable Amounts Amounts Amounts Amounts Amounts trations con- kinds C[wt %] A[wt %] B[wt %] Kinds D[wt %] kinds E[wt %] A/B A/C B/C D/C E/C [%] taining Ex. 13 L 1 0.08 1 a 0.5 A 0.2 0.08 0.08 1 0.5 0.2 1 A Ex. 14 L 1 0.75 3 a 0.5 A 0.2 0.25 0.75 3 0.5 0.2 3 B Ex. 15 L 1 0.03 6 a 0.5 A 0.2 0.005 0.03 6 0.5 0.2 6 C Ex. 16 L 1 0.01 0.5 a 0.5 A 0.2 0.02 0.01 0.5 0.5 0.2 0.5 A Ex. 17 L 1 0.02 2 a 0.5 A 0.2 0.01 0.02 2 0.5 0.2 2 B Ex. 18 L 1 0.6 1 a 0.5 A 0.2 0.6 0.6 1 0.5 0.2 1 B Ex. 19 L 1 0.009 1 a 0.5 A 0.2 0.009 0.009 1 0.5 0.2 1 C Ex. 20 L 0.1 0.08 1 a 0.5 A 0.2 0.08 0.8 10 5 2 1 A Ex. 21 L 0.1 0.75 3 a 0.5 A 0.2 0.25 7.5 30 5 2 3 B Ex. 22 L 0.1 0.03 6 a 0.5 A 0.2 0.005 0.3 60 5 2 6 C Ex. 23 P 1 0.08 1 a 0.5 A 0.2 0.08 0.08 1 0.5 0.2 1 A Ex. 24 P 1 0.75 3 a 0.5 A 0.2 0.25 0.75 3 0.5 0.2 3 B Ex. 25 P 1 0.03 6 a 0.5 A 0.2 0.005 0.03 6 0.5 0.2 6 C Ex. 26 P 1 0.01 0.5 a 0.5 A 0.2 0.02 0.01 0.5 0.5 0.2 0.5 A Ex. 27 P 1 0.02 2 a 0.5 A 0.2 0.01 0.02 2 0.5 0.2 2 B Ex. 28 P 1 0.6 1 a 0.5 A 0.2 0.6 0.6 1 0.5 0.2 1 B Ex. 29 P 1 0.009 1 a 0.5 A 0.2 0.009 0.009 1 0.5 0.2 1 C Ex. 30 P 0.1 0.08 1 a 0.5 A 0.2 0.08 0.8 10 5 2 1 A Ex. 31 P 0.1 0.75 3 a 0.5 A 0.2 0.25 7.5 30 5 2 3 B Ex. 32 P 0.1 0.03 6 a 0.5 A 0.2 0.005 0.3 60 5 2 6 C Com. L 1 — 1 a 0.5 A 0.2 — — 1 0.5 0.2 1 D Ex. 4 Com. L 1 0.08 — a 0.5 B 0.2 — 0.08 — 0.5 0.2 — D Ex. 5 Com. L 1 — — a 1.5 B 1.2 — — — 1.5 1.2 — D Ex. 6 Com. P 1 — 1 a 0.5 A 0.3 — — 1 0.5 0.2 1 D Ex. 7 Com. P 1 0.08 — a 0.5 B 0.2 — 0.08 — 0.5 0.2 — D Ex. 8 Com. P 1 — — a 1.5 B 1.2 — — — 1.5 1.2 — D Ex. 9

(26) Evaluations

(27) [Evaluation of Stable Containing of Antibody Obtained from the Hen Egg Yolk]

(28) In the toothpaste agent obtained in each of the Examples 1 to 12 and the Comparative Examples 1 to 3, the toothpaste agent of 1 g just after the production thereof and the tooth agent of 1 g after it was stored at a temperature of 40° C. for 6 months were suspended in a phosphate buffered saline (PBS) of 9 mL to obtain a solution. The solution was incubated in a water bath of 37° C. for 15 minutes. Chloroform of 10 mL was added into the solution to obtain a mixture. The mixture was incubated in a water bath of 37° C. for 15 minutes. Thereafter, the mixture was subjected to a centrifugal separation for 20 minutes to obtain a water phase part. Thus, the water phase part was used as a sample solution. The sample solution was added into an ELISA plate, on which a solid phase antigen was adsorbed, at a ratio of 100 μL/well. The ELISA plate was left at 25° C. for 1 hour to carry out an antigen-antibody reaction. After the completion of the reaction, the ELISA plate was washed with PBS-T five times. Moreover, 100 μL of a peroxidase bonding anti-chicken IgG antibody as a secondary antibody was added to the ELISA plate to carry out the antigen-antibody reaction at 25° C. for 30 minutes. Thereafter, the ELISA plate was washed with the PBS-T six times. Next, 100 μL of a solution, in which phenylenediamine of 20 mg of a substrate and hydrogen peroxide of 10 μL were dissolved, was added into 50 mL of a 0.2 M disodium phosphate-0.1 M citric acid buffer solution (pH 5.0). Thus, a mixture was obtained. Thereafter, the mixture was reacted at 25° C. for 20 minutes. The reaction stoppage was performed by adding 100 μL of a 3N sulfuric acid solution into the mixture. After the completion of the reaction, an absorbance (OD490) of the obtained reaction solution was measured to obtain an antibody value. An amount of the antibody value obtained by using the toothpaste agent stored at 40° C. for 6 months with respect to an antibody value obtained by using the toothpaste agent just after the production was obtained. The amount was evaluated according to the following four criteria as a residual ratio of the antibody obtained from the hen egg yolk.

(29) A: A residual ratio of the antibody obtained from the hen egg yolk is 80% or more.

(30) B: A residual ratio of the antibody obtained from the hen egg yolk is 70% or more but lower than 80%.

(31) C: A residual ratio of the antibody obtained from the hen egg yolk is 50% or more but lower than 70%.

(32) D: A residual ratio of the antibody obtained from the hen egg yolk is lower than 50%.

(33) [Evaluation of Stable Containing of Lipase]

(34) In the toothpaste agent obtained in each of the Examples 13 to 22 and the Comparative Examples 4 to 6, the toothpaste agent of 5.0 g just after the production thereof and the toothpaste agent of 5.0 g after it was stored at a temperature of 40° C. for 6 months were suspended in a 0.1 M phosphate buffer solution (pH 7.0) of 10 mL to obtain a suspension liquid. Next, the suspension liquid was subjected to a centrifugal separation for 5 minutes to obtain a centrifugal supernatant. 2 mL of a 0.1 M phosphate buffer solution (pH 7.0) was added into 1 mL of the centrifugal supernatant to obtain a mixture A. The mixture A was heated for 5 minutes in a constant temperature reservoir of 37° C. Further, 3 mL of an olive oil emulsified liquid heated at 37° C. for 5 minutes in advance was added into the mixture A to obtain a mixture B. The mixture B was stirred at 37° C. for 30 minutes. Thereafter, 20 mL of a mixture of equal parts of acetone and ethanol was added into the mixture B to obtain a sample solution.

(35) Aside from this, in the toothpaste agent obtained in each of the Examples 13 to 22 and the Comparative Examples 4 to 6, the toothpaste agent of 5.0 g just after the production thereof and the toothpaste agent of 5.0 g after it was stored at a temperature of 40° C. for 6 months were suspended in 10 mL of a 0.1 M phosphate buffer solution (pH 7.0) to obtain a suspension liquid. Next, the suspension liquid was subjected to a centrifugal separation for 5 minutes to obtain a centrifugal supernatant. 2 mL of a 0.1 M phosphate buffer solution (pH 7.0) of 2 mL was added into 1 mL of the centrifugal supernatant to obtain a mixture C. Furthermore, 20 mL of a mixture of equal parts of acetone and ethanol was added into the mixture C, and then it was mixed sufficiently. Thereafter, 3 mL of an olive oil emulsified liquid was added into the mixture C to obtain a mixture D. The mixture D was stirred at 37° C. for 30 minutes to obtain a control solution.

(36) Three droplets of phenolphthalein were added into the sample solution and the control solution, respectively, to titrate with a 0.1 N sodium hydroxide solution. A lipase titer (unit/g) was calculated by the following formula. An amount of the lipase titer obtained by using the toothpaste agent stored at 40° C. for 6 months with respect to a lipase titer obtained by using the toothpaste agent just after the production was obtained. The amount was evaluated according to the following four criteria as a residual ratio of the lipase.
Lipase titer [unit/g]=[400×(a−b)]/3

(37) where “a” represents a titer (mL) of the sample solution and “b” represents a titer (mL) of the control solution.

(38) A: A residual ratio of the lipase is 80% or more.

(39) B: A residual ratio of the lipase is 70% or more but lower than 80%.

(40) C: A residual ratio of the lipase is 50% or more but lower than 70%.

(41) D: A residual ratio of the lipase is lower than 50%.

(42) [Evaluation of Stable Containing of Protease]

(43) In the toothpaste agent obtained in each of the Examples 23 to 32 and the Comparative Examples 7 to 9, 20 mL of water was accurately added into the toothpaste agent of 0.25 g just after the production thereof and the toothpaste agent of 0.25 g after it was stored at a temperature of 40° C. for 6 months. They were mixed and suspended to obtain suspension liquids, respectively. Thereafter, the suspension liquid was subjected to a centrifugal separation for 5 minutes to obtain a centrifugal supernatant. 4 mL of a 0.1 mol/L phosphate buffer solution was added into 1 mL of the centrifugal supernatant to obtain a mixture E. The mixture E was heated for 5 minutes in a constant temperature reservoir of 30° C. Further, 4 mL of a 0.1 mol/L p-tosyl arginine methylester hydrochloride heated at 30° C. for 5 minutes in advance was added into the mixture E to obtain a mixture F. The mixture F was heated at 30° C. for 30 minutes, and then was filtered immediately (a membrane filter having a pore size of 0.45 μm). Then, 0.4 mL of a trichloroacetic acid solution (15.fwdarw.100) was added into 0.6 mL of the obtained filtrate to stop an enzyme reaction. Thus, a mixture G was obtained. Next, 0.2 mL of a potassium permanganate solution (2.fwdarw.100) was added into the mixture G to obtain a mixture H. The mixture H was stirred and left for 1 minute. The contents thereof were oxidized, and thereafter, 0.2 mL of a sodium acid sulfite solution (1.fwdarw.10) was added into the mixture H to obtain a mixture I. The mixture I was stirred and excess potassium permanganate was reduced. Next, 8 mL of a 0.4% chromotropic acid solution was added into the mixture I to obtain a mixture J. The mixture J was stirred well and heated in a boiling-water bath for 15 minutes accurately. Thus, the mixture J was colored. The mixture J was left at room temperature for 40 minutes. Thereafter, an absorbance in a wavelength of 580 nm of the mixture J was measured by using a blank solution of water. Similarly, a blank test was carried out, and a protease titer [unit/g] was obtained by the following formula. An amount of the protease titer obtained by using the toothpaste agent stored at 40° C. for 6 months with respect to a protease titer obtained by using the toothpaste agent just after the production was obtained. The amount was evaluated according to the following four criteria as a residual ratio of the protease.
Protease titer [unit/g]=(a−b)×188

(44) where “a” represents an absorbance of the sample solution and “b” represents an absorbance of the sample solution for the blank test.

(45) A: A residual ratio of the protease is 80% or more.

(46) B: A residual ratio of the protease is 70% or more but lower than 80%.

(47) C: A residual ratio of the protease is 50% or more but lower than 70%.

(48) D: A residual ratio of the protease is lower than 50%.

(49) The results evaluated as described above are shown in Table 1 and Table 2.

(50) As shown in Table 1 and Table 2 clearly, the toothpaste agent obtained in each of the Examples could contain the lipase, the protease and the antibody obtained from the hen egg yolk stably.

(51) In contrast, the toothpaste agent obtained in each of the Comparative Examples could obtain insufficient results.

INDUSTRIAL APPLICABILITY

(52) The present invention provides with the stomatological composition comprising at least one selected from the group consisting of the lipase, the protease and the antibody obtained from the hen egg yolk, citric acid, and the metal salt of citric acid. According to the present invention, it is possible to provide with the stomatological composition that is capable of containing the antibody obtained from the hen egg yolk and the enzyme stably, and preventing or improving the disease in the oral cavity such as odontonecrosis or periodontal disease. Accordingly, the present invention has industrial applicability.