JPH0130803B2 - - Google Patents

Description

[Detailed description of the invention]

(Industrial Application Field) The present invention relates to a dentifrice composition for preventing dental caries, and more particularly to a dentifrice composition containing hydroxyapatite with fixed dextranase. (Prior art) Dextranase has a plaque-dissolving effect due to its ability to decompose dextran, and it is known that toothpaste compositions containing this have a good caries prevention effect (Patent No. 782154). . However, dextranase has relatively poor stability, and dextranase in toothpaste products that contain it tends to decompose and deactivate over time. To keep it stable, it is necessary to use an appropriate stabilizer. It is said that there is
Various ideas have been proposed. Anionic active agents used in dentifrice compositions, for example as foaming agents, promote the degradation of dextranase. To prevent this, attempts have been made to use terpene-based hydrocarbons and aliphatic alcohol flavorings in combination (Patent No. 9242229), and methods to use carvone and l-menthol together in a specific ratio (Japanese Patent Laid-Open No. 110609/1982) , or a method using aluminum oxide as an abrasive (Japanese Unexamined Patent Publication No. 56-63915) has been proposed. On the other hand, since toothpaste products are used in the oral cavity, they are required to be safe and have an excellent usability. Therefore, it is important to achieve both the stability of dextranase, the excellent feeling of use in the oral cavity, and the safety of dextranase, which are required for dentifrice products containing dextranase. It can be said that difficulties remain. (Problems to be Solved by the Invention) The present invention solves the problems that arise in achieving both stability and usability due to the use of additives in conventional dextranase-containing dentifrice products, and stabilizes dextranase. An object of the present invention is to provide a dextranase-containing dentifrice composition that does not require the addition of a curing agent and has an excellent caries-preventing effect. (Means for solving the problem) It is known that hydroxyapatite, which has a hardness close to that of enamel teeth, is suitable as an abrasive used in the preparation of toothpaste compositions from the viewpoint of cleaning and preserving teeth. (Japanese Unexamined Patent Publication No. 56-73014). Therefore, if it is possible to stably immobilize dextranase, which has been recognized as effective in oral hygiene compositions, on hydroxyapatite, then the hydroxyapatite prepared in this way can be used as a compounding ingredient. As a result of various studies based on the idea that by doing so, a toothpaste product that meets the purpose can be obtained, the present invention was completed. Recently, much knowledge has been obtained regarding enzyme immobilization. For example, it is known that enzymes have characteristics such that they are easily adsorbed by proteins, and that cross-linking occurs between enzymes and proteins due to bifunctional substances, making the enzyme insolubilized and stabilized. On the other hand, when hydroxyapatite is made into fine particles, it exhibits strong adsorption properties. Focusing on this matter, we have previously filed a patent application for a method for preparing hydroxyapatite in which dextranase is stably immobilized as a result of various studies. That is, in a low-temperature aqueous medium, proteins selected from lysozyme, casein, albumin, cytochrome C, etc.
Hydroxyapatite with immobilized dextranase is obtained by gradually adding a bifunctional substance such as glutaraldehyde to dextranase and hydroxyapatite while stirring. The hydroxyapatite used is normally used in toothpaste compositions, and the dextranase and protein are used in approximately equal amounts.
Use 10 to 100 times more hydroxyapatite than total protein. The pH of the water medium used is 5.6 to 9.0
The amount of glutaraldehyde added is preferably as small as possible since the enzyme will be inactivated if the amount is too large. According to this method, hydroxyapatite powder with immobilized dextranase can be easily obtained, and this can be used as an abrasive and a product prepared according to the formulation of ordinary toothpaste compositions, which can be used as an excellent preventive agent for tooth decay. You can get effective toothpaste products. (Function) A toothpaste product that uses dextranase-immobilized hydroxyapatite prepared using the method for which a patent application was previously applied is different from conventional toothpaste products that contain dextranase and a stabilizer. Since dextranase is immobilized on the abrasive hydroxyapatite, it has excellent stability and can prevent decomposition and deactivation of dextranase during product storage. It is also stable in the presence of other formulation ingredients, such as blowing agents such as anionic active agents. Furthermore,
When the product is used in the oral cavity, the decomposition of dextranase due to various factors such as ions and pH is prevented, and at the same time, the effects of dextranase continue for a relatively long period of time. That is, in the mouth, the ends of the hydroxyapatite particles and/or
Alternatively, it is presumed that the terminal end of dextranase is adsorbed to the tooth surface enamel layer, and the dextranase stays in the oral cavity for a relatively long period of time, thereby making the plaque decomposition effect permanent. Dextranase-immobilized hydroxyapatite can be prepared by the method described below. No special technique is required to prepare the dentifrice composition of the present invention. That is, in the formulation of conventional dentifrice compositions, at least all or part of the abrasive used may be replaced with dextranase-immobilized hydroxyapatite prepared according to the method described below. (Examples) The present invention will be explained with reference to the following examples, in which the compositions are expressed in parts by weight. Example 1 Toothpaste dextranase fixed hydroxyapatite
13.2 Calcium phosphate 25.0 CMC sodium salt 0.3 Carrageenan 1.2 Glycerin 10.0 Sorbit 15.0 Sodium lauryl sulfate 2.0 Flavoring 1.2 Sodium saccharin 0.1 Silicon dioxide 2.0 Water 30.0 Example 2 Toothpaste dextranase-fixed hydroxyapatite
7.2 Calcium phosphate 10.0 Calcium pyrophosphate 20.0 CMC sodium salt 1.0 Sodium alginate 0.1 Glycerin 10.0 Sorbit 10.0 Sodium lauryl sulfate 1.5 Sodium lauroyl sarcosinate 0.5 Flavor 0.5 Sodium saccharin 0.1 Silicon dioxide 2.5 Sodium phosphate 1.0 Water 35.0 Example 3 toothpaste dextranase fixed hydroxyapatite
20.0 Calcium pyrophosphate 10.0 CMC sodium salt 0.5 Carrageenan 0.6 Glycerin 20.0 Sorbit 10.0 Sodium lauryl sulfate 2.0 Flavoring 1.0 Satucharin sodium 0.1 Silicon dioxide 2.0 KCl 2.0 MgCl ₂ 0.3 Sodium phosphate 0.5 Water 30.0 Example 4 Toothpaste dextraner Ze-fixed hydroxyapatite
35.0 CMC sodium salt 1.0 Carrageenan 0.3 Glycerin 35.0 Sodium lauryl sulfate 2.0 Flavoring 1.0 Satucalin sodium 0.1 Silicon dioxide 2.5 NaCl 2.0 MgCl ₂ 0.1 KCl 1.0 Water 20.0 Example 5 Toothpaste dextranase-fixed hydroxyapatite
90.8 Sodium lauryl sulfate 2.0 Flavoring 1.5 Satucharin sodium 0.2 NaCl 5.0 MgCl ₂ 0.5 Example 6 Toothpaste dextranase-fixed hydroxyapatite
38.0 Calcium pyrophosphate 50.0 Silicon dioxide 5.0 Sodium lauryl sulfate 2.0 Flavoring 2.0 Satucharin sodium 0.3 NaCl 1.8 MgCl ₂ 0.2 Potassium phosphate 0.7 Example 7 Moisturizing toothpaste dextranase-fixed hydroxyapatite
63.0 Calcium phosphate 10.0 Sorbit 10.0 Sodium lauryl sulfate 2.0 Flavor 1.5 NaCl 3.3 MgCl ₂ 0.08 Satucalin 0.12 Water 10.0 The dextranase-immobilized hydroxyapatite used in the examples was prepared as follows. Preparation of dextranase-immobilized hydroxyapatite. Hydroxyapatite used in each example was prepared by the method described below. That is, the hydroxyapatite of Example 1 was produced by dry synthesis method,
6CaHPO ₄ +4CaCO ₃ →Ca ₁₀ (PO ₄ ) ₆ (OH) ₂ , Example 2
and that of Example 3 is a hydrothermal synthesis method, _CaHPO4・
2H ₂ O→Ca ₁₀ (PO ₄ ) ₆ (OH) ₂ , that of Examples 4 and 5 is a wet synthesis method, CaCl ₂ +H ₃ PO ₄ →Ca ₁₀ (PO ₄ ) ₆
(OH) ₂ , that of Examples 6 and 7, is a wet synthesis method,
It was synthesized by the reaction Ca(OH) ₂ +H ₃ PO ₄ →Ca ₁₀ (PO ₄ ) ₆ (OH) ₂ , dried and ground, and used as hydroxyapatite for immobilization. Mix 50 mg of dextranase, 50 mg of lysozyme, and 5 g of hydroxyapatite powder, add 500 ml of 0.05 molar concentration PH6.8 potassium phosphate buffer solution, and keep stirring at 4°C with 0.2% glutaraldehyde aqueous solution.
500 μl was gradually added dropwise, and stirring was continued for 5 hours at 4°C. The reaction product was collected and washed three times with 100 ml of the above buffer solution to remove unreacted material, and the residue was lyophilized to obtain 5.06 g. Weigh out approximately 1 g of this accurately, add 1 molar concentration PH6.8 potassium phosphate buffer solution 20
ml was added, stirred for 3 hours, centrifuged, the precipitate was washed and combined with the liquid, and protein was measured by the Lowry method, and it was found that 10.9 mg of protein was adsorbed per g of dry hydroxyapatite. As a result of measuring the dextranase activity of this protein, it was confirmed that 0.513 g of dextran/g of protein was degraded in 2 hours. The dextranase activity of the obtained dextranase-immobilized hydroxyapatite increases with time from immediately after production, reaches the highest activity for a certain period of time, and then gradually decreases. Dextranase-immobilized hydroxyapatite can be obtained similarly to lysozyme by using albumin, casein, or cytochrome C instead of lysozyme. Changes over time in dextranase activity Instead of 35 g of dextranase-immobilized hydroxyapatite in Example 4, a toothpaste containing 35 g of hydroxyapatite, 350 mg of dextranase, and other reagents in equal amounts and water to make 100 g was prepared. , served as a control. Weigh 2g of each toothpaste accurately, and check the pH
Add 10 ml of a solution of 1% dextran dissolved in a 0.05 molar potassium phosphate buffer solution of 7.0, stir at 35°C for 2 hours, centrifuge, and wash the residue with the same buffer solution.
The amount of glucose produced by glucose oxidase in combination with the centrifuged solution was measured over time, and the following results were obtained.

[Table] (Effects of the Invention) By incorporating hydroxyapatite with immobilized dextranase into toothpaste, it is possible to obtain a toothpaste with stable dextranase efficacy and which does not require special consideration in its formulation.

Claims (1)

[Claims] 1. A dentifrice composition containing hydroxyapatite with immobilized dextranase.