Enzymatic dentifrices

© Start of patent duration: 1 p. July 1972 Longest possible duration:

© Published on: 26.March 1973 © Inventors: Hendrik Hoogendoorn in Krimpen, Rutger Matthijsen and Huibert Cornelis Theüs Moelker in Oss (the Netherlands) © Dependency:

© Pamphlets considered to delineate the prior art:

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Nr.306243

The invention relates to a novel enzyme-containing dentifrice, such as toothpaste, mouthwash, or a tablet, which is dissolved in water to a mouthwash.

Tooth decay, especially tooth decay, is caused by a number of factors. It is known that on the surface of the tooth there is usually a deposit consisting of food and bacteria, which is called a sheet. The bacteria found in this deposit lead to the rot of the food, whereby acids are formed. At the surface of the tooth the p_HValue of the saliva, which generally has a value of 7.0 to 7.5, by this formation of acids. Depending on the type of food consumed and the frequency of food intake, the py value can reach different values ​​and also the time necessary to return to normal p_HFor example, it has been shown that the intake of sugar in the form of viscous masses, such as toffees, a much lower p ^ value is achieved than when these sugars in one Product with a fibrous structure, such as an apple, are included. In addition, the regeneration time in the first case is much longer. Further, it has been found that the repeated consumption of sweets or other acid-forming products gradually increases the regeneration time. Depending on the type of food, the p ^ value may drop to 5.5 to 4.5 and sometimes even lower. The region below the limits of 5.5 is often referred to as a danger zone, since in such acidic conditions the calcium compounds of the tooth dissolve in the saliva, which leads to the development of tooth decay. It is thus clear that the lower the p + value and the longer the regeneration time, the greater the risk that the teeth will be attacked.

Besides these two factors, there is at least one other factor that plays an important role in tooth decay, including: between the thickness of the coating. It is a fact that if the coating is very thick, the acids formed in it by bacterial decomposition are very difficult to penetrate to the surface, resulting in a lower pi value and a longer regeneration time.

In connection with this latter factor, it has long been common practice to add grinding and / or polishing materials to the toothpastes to reduce or remove the coating. That's why eating apples is recommended.

It is also known that the toothpastes u.ähnl. Dentifrices to add enzymes, for example proteases, such as pepsin, pancreatin, trypsin u.ähnl., Or amylases. All of the enzymes used hitherto belong to the group of hydrolases, such as the enzymes that can disrupt peptide or glycoside bonds, thereby converting macromolecular compounds into oligomeric or monomeric products that are easier to dissolve in saliva and so can be removed.

The invention now relates to a new dentifrice, which is characterized in that it optionally contains in admixture with other enzymes an oxido-reductase, such as glucose oxidase, which is the degradation of a substrate which is already present in the oral cavity or the dentifrice is hydrogen peroxide provides. Two important effects are achieved in the use of such enzyme systems in dentifrices: the plaque is attacked and, most importantly, hydrogen peroxide is formed upon enzymatic decomposition, which has been found to normalize the oral flora, thereby decreasing the bacterial Decomposition and consequently no or only a slight formation of harmful acids occurs.

An essential feature of the invention is that, under the influence of a suitable oxido-reductase, a substrate, which is usually present in the mouth, especially in the plaque, is degraded to form hydrogen peroxide, whereby the hydrogen peroxide normalizes the oral flora and forms a low pH. Value and consequently the tooth rot. This means that it is usually sufficient to add an oxydo reductase to the toothpaste, since a substrate in the mouth is already mostly available for this enzyme, as stated above.

However, depending on the choice of oxydo reductase, it may also be the case that the substrate is not or only insufficiently available in the mouth. In this case there are two possibilities:

a) addition of a hydrolase enzyme (which differs substantially from an oxydo reductase enzyme), which degrades a compound present in the mouth and gives a substrate for the applied oxydo reductase, or

b) add the substrate as such to the toothpaste.

In principle, any oxydo reductase which forms hydrogen peroxide can be used to prepare the agents of this invention, generally any oxydo reductase which will decompose the substrate with oxygen as the acceptor.

As examples of such enzymes may be mentioned the oxidases acting on the CHOH, aldehyde or keto group of the substrate, the lactate oxidase, malate oxidase, glucose oxidase, hexose oxidase, galactose oxidase, pyruvate oxidase and oxalate oxidase , Oxydases on the CH-NH₂Group of the substrate, the L-amino acid oxidase, D-amino acid oxidase, tyraminase and histaminase, and other oxidases which attack at another point, such as xanthine oxidase, sarcosine oxidase, N-methylamino acid oxidase and sulfite. oxidase.

The amount which must be used by the above-mentioned enzyme may vary widely, but they

No. 306243 is generally between 0.01 and 500 units / g or milliliter of the dentifrice.

By an enzyme unit is meant the amount of an enzyme containing 1 μ mol substrate / min at 30 ° C and a p_HValue of 6 is oxidized under standard conditions.

It has also been found advantageous to add one or more other enzymes, especially those enzymes which can provide a substrate for the oxidase enzyme system used, such as enzymes belonging to the group of hydrolases. The hydrolases can eg Carbohydrases, proteases and lipases such as os-amylase, β-amylase, glucoamylase or amyloglucosidase, cellulase, dextranase, invertase, α- and β-glucosidase, α- and β-galactosidase, pepsin, trypsin, chymotrypsin, papain and proteases of bacterial origin ,

These hydrolases are measured by known methods and their activities are reported in units according to the recommendations of the International Union of Biochemistry (Report of the Commission in Enzymes of the IUB, Pergamon Press, Oxford [1961]).

Instead of or simultaneously with one or more hydrolases, it is also possible to add to the agent the substrate of the oxidase used, such as a sugar, eg galactose or glucose, a lactate or malate, an amino acid, a pyruvate or oxalate or the like. admit.

The addition of glucose oxidase together with amyloglucosidase has also proved to be a very beneficial combination.

To show the surprising effect of these enzymes, the pjj value on the surface of the tooth in the coating was measured. The μ-antimony electrode used for this purpose was developed by F. Clarence Thompson et al. in Journal of Dental Res. Bd.33 [1954], p.849. The measurements were made on the buccal surfaces of the first and second molars at least 1 h after the last food intake. Thus, a total of four measuring points per person were obtained, each in the order:

m₁sd-m₂sd-m₁ss-m₂ss.

On the first day, the subjects rinsed their mouths with 10 ml of a 70% sucrose solution for 4 minutes, whereupon the p_H- Value was measured. After these measurements, they rinsed with 10 ml of mouthwash in which the enzymes were dissolved, which of course the comparators did not do.

The examinations were carried out so that a gap of at least 1 h was observed between the last rinsing and the next food intake.

On the second day, the procedure was repeated with 10 ml of a 70% sucrose solution, whereupon the p_HValue was measured.

The next day, the good influence of the mouthwash with enzyme on the ppj value could be clearly demonstrated by changing the p_HValue measured on the surface of the tooth after rinsing with sucrose.

In order to avoid a mechanical attack on this coating, both the sucrose and the enzymes were used in liquid form.

The experiments were carried out with groups of 15 to 30 children (boys and girls) between 12 and 14 years.

The following table shows the results of the p ^ measurements. The children were divided into three groups:

a) those whose p ^ value was higher than 6, 8

b) those whose pjj value was between 5.6 and 6.8

c) those whose py value was less than 5.6.

<td rowspan="2"></td><td colspan="3">Percentage of children with a pp value of</td>

<td>&gt; 6.8</td><td>5,6 to 6, 8</td><td>&lt;5.6</td>

<td>Equalizers I-Day</td><td>23</td><td>53</td><td>24</td>

<td>2 day</td><td>19</td><td>58</td><td>23</td>

<td>30 U amyloglucosidase / 10 ml l.Tag</td><td>33</td><td>39</td><td>28</td>

<td>2 day</td><td>28</td><td>47</td><td>25</td>

<td>6 E glucose oxidase / 10 ml l.Tag</td><td>25</td><td>55</td><td>20</td>

<td>2 day</td><td>31</td><td>60</td><td>9</td>

<td>30 E amyloglucosidase + 6 E glucose oxidase / 10 ml l.Tag</td><td>21</td><td>59</td><td>20</td>

<td>2 day</td><td>60</td><td>40</td><td>0</td>

Nr.306243

From this table, it can be seen that amyloglucosidase alone has no significant effect on the percentage of children in the danger zone below 5.6, and that glucose oxidase, and to an even greater extent glucose oxidase along with amyloglucosidase, greatly increases this percentage or down to zero.

Another important factor was that the coating became less solid by treatment with the enzyme mixture, so that in some cases it was easily lost.

The dentifrices of the invention may be in various forms, for example in the form of toothpaste, mouthwash, tablets dissolved in water to form a mouthwash, or other conventional forms. In addition to the enzymes of the invention and, where present, a substrate for the oxidase used, these dentifrices contain the usual substances. To contain toothpastes, eg Abrasive and / or polishing substances such as calcium carbonate, dicalcium phosphate, calcium phosphate, calcium sulfate or silicon compounds, thickeners such as carboxymethyl cellulose, tragacanth or guar gum, water, flavorings and / or natural or synthetic sweeteners. Further, fluorine compounds such as sodium or potassium monofluorophosphate or sodium fluoride may be added,

The invention is further illustrated by the following examples.

Example 1: Toothpaste

Precipitated silica (eg neosyl) paraffin

Ethyl p-hydroxybenzoate

methylcellulose

Naarden 17391 flavor substance Glucose oxidase (20 U / g) Amylglucosidase (30 U / g) Distilled water

Example 2: Toothpaste

calcium carbonate

tricalcium phosphate

Sorbitol (70% solution)

glycerin

tragacanth

Naarden ZD 00037-flavor substance glucose oxidase (10 U / g)

Dextranase (10 U / g) Ethyl p-hydroxybenzoate Distilled water about 23%

0.2% 1.8% 2% to 100%%

%% 20% 2%

0.8%

0.1% to 100%

Example 3: Toothpaste

Aluminum hydroxide

Na-fluoride

Sorbitol (70% solution)

glycerin

eucalyptol

Na alginate

Propyl p-hydroxybenzoate

saccharin

Glucose oxidase (5 U / g)

Invertase (25 E / g)

water

Example 4: Tooth powder

Eucalyptol and menthol (1: 1)

Na cyclamate

Detergent (Texapon L 100 from the Fa.Dehydag) galactose oxidase (4 U / g) glucose oxidase (4 U / g) £ -galactosidase (30 U / g)

Calcium phosphate%

0.1%%

%

1.2% 1% 0.1% 0.25% to 100%% 0.5% 1% to 100%

Nr.306243

Example 5: Mouthwash

Methylcellulose (low viscous) Naarden 17391 flavor substance Ethyl p-hydroxybenzoate Disodium phosphate 0 aq. Citric acid 1 aq.

Glucose oxidase (3 U / ml) Amyloglucosidase (5 U / ml) Distilled water

Example 6: Tooth powder

Na cyclamate

eucalyptol

L-amino acid oxidase (95 U / g) glycine

tricalcium phosphate

Example 7: Tooth powder

Naarden 17391-flavor substance Na-fluoride

Detergent (Texapon L 100 of the Fa Dehydag) xanthine

Xanthine oxidase (5 U / g)

Na-saccharin

Microcrystalline aluminum hydroxide