RU2407512C1 - Nonabrasive tooth paste containing papain, dextranase, alpha-amilase, potassium or ammonium thiocyanate, invertase or glucoamylase, glucose oxidase, lactoperoxidase, lysozyme or lactoferrin and lactulose enzymes - Google Patents

Abstract

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to medicine, namely to dentistry. For the purpose of ensuring an improved cleansing, antimicrobial and anti-inflammatory action, natural oral microflora restoration, there is developed a nonabrasive tooth paste formulation containing the following ingredients, wt %: papain 0.01-5.00; dextranase 0.01-5.00; alpha-amylase 0.01-5.00; potassium or ammonium thiocyanate 0.01-5.00; invertase or glucoamylase 0.01-5.00; glucose oxidase 0.01-5.00; lactoperoxidase 0.01-5.00; lysozyme or lactoferrin 0.01-5.00; lactulose 2-4; sodium carboxymethyl cellulose 0.5-0.7; emulsifying agent 0.01-5; preserving agent 0.01-1.0; aromatiser 0.01-1.00; 70% sorbitol 30-40; demineralised water to 8.30-67.39.

EFFECT: tooth paste have good organoleptic property, a pleasant odour and good taste properties, and allows to reduce plaque and oral inflammations without no abrasive ingredients added.

9 tbl, 2 ex

Description

The present invention relates to dentistry, namely to the non-abrasive toothpaste for cleaning the oral cavity comprising the following components: papain, dextranase, alpha-amylase, potassium or ammonium thiocyanate, invertase and glucoamylase, glucose oxidase, lactoperoxidase, lysozyme, or lactoferrin, lactulose, sodium carboxymethylcellulose , emulsifier, preservative, flavor, sorbitol 70%, demineralized water.

The invention allows to reduce without the use of abrasive materials such as calcium carbonate, calcium pyrophosphate, dicalcium phosphate, silicon dioxide, aluminum hydroxide, the formation of plaque on the surface of the teeth by 70% due to the enzymatic destruction of plaque, restoration of the natural microflora of the oral cavity and the destruction of pathogenic microflora , reduce inflammation in the oral cavity by 90%. Toothpaste does not contain abrasive components and has good organoleptic properties: it is white in color, with a pleasant smell and good taste.

The invention relates to medicine, namely to dentistry.

There are various medicinal and preventive toothpastes used for the prevention of inflammatory periodontal diseases, containing anti-inflammatory components that are not enzymes, for example extracts of medicinal plants and vitamins (medical and preventive toothpaste "Silca Herb plus" (Germany)), tea tree oil (tooth paste "Natural whitening toothpaste with tea tree oil" (USA)). However, this paste has a low cleaning ability, because it does not contain components that directionally dissolve plaque.

Known toothpaste with an enzyme-containing composition, where the enzyme is selected from the group of proteinases, including papain (Rembrandt whitening toothpaste (USA)), as well as a paste containing lysozyme, glucose oxidase, lactoperoxidase and lactoferrin (Biotene toothpaste (Finland) )), which is used to treat xerostomia. The described toothpaste does not contain lactulose, which has the ability to normalize the microflora of the oral cavity and stimulate the formation of endogenous lysozyme, and therefore cannot be used to restore the microflora of the oral cavity.

From RU 2355420, publ. 05/20/2009 (examples, claims) a toothpaste is known, including: an abrasive component - silicon dioxide, lactulose - 2.0-6.0 wt.%, A combination of enzymes: papain - 0.2-1.0 wt.% And lysozyme 0.01-0.4 wt.%. Toothpaste also contains: sodium carboxymethyl cellulose - 0.5-0.7 wt.%, Preservatives (sodium methylparaben and sodium propylparpaben) in a total amount of 0.2-0.8 wt.%, Sodium lauryl sulfate and sodium lauroyl sarcosinate in total - 0.6-3.0 wt.%, Flavoring - 0.05-2.0 wt.% And demineralized water. In RU 2355420, publ. 05/20/2009 (examples, claims), it is also indicated that the introduction of lactulose in the composition of the toothpaste helps to restore the natural microflora of the oral cavity.

From US 4150113, publ. 04/17/1979 (the entire document, in particular the description column 5, examples, claims), dentifrices are known, including toothpastes, mouthwashes, chewable tablets and gums, the main active ingredient of which is glucose oxidase. It is indicated that in addition to glucose oxidase, it is advisable to include carbohydrase, for example, alpha-amylase, glucoamylase (amyloglucosidase), dextranase, and invertase.

From US 4578265, publ. 03/25/1986 (examples, claims), a dentifrice with antibacterial properties is known, including glucose oxidase, lactoperoxidase and potassium or ammonium thiocyanate.

However, these agents do not contain proteases, in particular papain.

From RU 2333768, publ. September 20, 2008 (see the Enzymes section), it is also known that it is advisable to include carbonic anhydrases such as glucoamylase, alpha-amylase, dextranase, and proteases such as papain in the composition for caring for the oral cavity. Moreover, the specified combination of enzymes can be supplemented with lysozyme. However, these compositions do not contain lactulose.

Toothpastes containing papain enzymes; dextranase; alpha amylase; invertase or glucoamylase; glucose oxidase; lactoperoxidase; lysozyme or lactoferrin, lactulose, for topical use to restore the microflora of the oral cavity and the prevention of inflammatory periodontal diseases are not described.

Thus, an object of the present invention is to provide a toothpaste containing papain enzymes; dextranase; alpha amylase; invertase or glucoamylase; glucose oxidase; lactoperoxidase; lysozyme or lactoferrin, potassium or ammonium thiocyanate and lactulose and not containing abrasive components, which can be used to restore the microflora of the oral cavity and to prevent inflammatory periodontal diseases caused by cariogenic microorganisms such as S. mutans, S. aureus and others.

According to the present invention, there is provided a toothpaste containing 0.01-5.0% by weight of papain as an active ingredient; 0.01-5.0 wt.% Dextranase; 0.01-5.0 wt.% Alpha-amylase; 0.01-5.0 wt.% Invertase or glucoamylase; 0.01-5.0 wt.% Glucose oxidase; 0.01-5.0 wt.% Lactoperoxidase; 0.01-5.0 wt.% Lysozyme or lactoferrin; 0.01-5.0 wt.% Potassium or ammonium thiocyanate; 2.0-4.0 wt.% Lactulose, and cosmetically acceptable carriers or excipients.

As a preferred embodiment of the claimed invention, the toothpaste indicated above, in the following ratio of components, wt.%:

Papain 0.01-5.00 Dextranase 0.01-5.00 Alpha amylase 0.01-5.00 Potassium or ammonium thiocyanate 0.01-5.00 Invertase or glucoamylase 0.01-5.00 Glucose oxidase 0.01-5.00 Lactoperoxidase 0.01-5.00 Lysozyme or lactoferrin 0.01-5.00 Lactulose 2.00-4.00 Sodium carboxymethyl cellulose 0.50-0.70 Emulsifier 0.01-5.00 Preservative 0.01-1.00 Flavoring 0.01-1.00 Sorbitol 70% 30.00-40.00 Demineralized water 8.30-67.39

The active ingredient of the claimed non-abrasive paste are: papain, dextranase, alpha-amylase, potassium or ammonium thiocyanate, invertase or glucoamylase, glucose oxidase, lactoperoxidase, lysozyme and lactulose. Papain is capable of breaking peptide bonds from sulfur-containing amino acid residues. Dextranase hydrolyzes dextrans and dextrins to oligosaccharides. Alpha-amylase hydrolyzes starch and amylopectin to dextrans and dextrins. Invertase and glucoamylase hydrolyze sucrose into fructose and glucose. Glucose oxidase oxidizes glucose with the formation of hydrogen peroxide, together with lactoperoxidase, potassium or ammonium thiocyanate form the so-called lactoperoxidase system, the principle of which is as follows: hydrogen peroxide formed under the influence of glucose oxidase oxidizes thiocyanate cells to . Lactoferrin has the specific ability to bind the last vital trace elements that are part of the respiratory chain cytochromes, catalases, peroxidases and superoxide dismutases through the binding of iron and other metals of variable valency from the surface structures of the shells of microorganisms, and also reduces their resistance to the toxic effects of chemical reactive oxygen derivatives . Thus, the integrated use of the above active components leads to the following result: modification of plaque with the formation of easily removable water-soluble compounds, activation of the lactoperoxidase system with the aim of antimicrobial and anti-inflammatory protection against pathogenic microorganisms. To prepare a non-abrasive paste, the substances used are: Papain, Dextranase, Alpha Amylase, Potassium or Ammonium Thiocyanate, Invertase or Glucoamylase, Glucose Oxidase, Lactoperoxidase, Lysozyme or Lactoferrin, ". Lactoferrin and lactoperoxidase are obtained from whey, papain from papaya, by extraction from crushed raw materials, followed by purification and freeze drying. Freeze-dried powder “Papain” is used in medicine together with lysozyme for the treatment of intervertebral hernia, in the form of powders and lotions in dentistry for gingivitis and periodontitis and other inflammatory conditions of the oral cavity. Freeze-dried powder "Lactoferrin" is used for acne. Freeze-dried dextranase is used to remove plaque.

The non-abrasive paste of the present invention is usually made by conventional methods using solid or liquid cosmetically acceptable carriers or excipients selected from emulsifiers, dispersants, preservatives, flavorings, pH adjusters, polymer carriers and other excipients that are suitable for the preparation of compositions for topical application.

Additional advantages of this composition are the ability to restore the natural microflora of the oral cavity.

The proposed non-abrasive paste reduces plaque formation by 70% and is completely harmless and safe for prolonged use.

The significant differences of the proposed non-abrasive paste are as follows.

The claimed cosmetic composition has stability with respect to the activity of the enzymes included in its composition during prolonged storage, which is achieved by immobilization of enzymes on polymer carriers contained in the composition. As these polymeric carriers, any polymers capable of binding to enzymes to form enzyme-carrier complexes can be used. Examples of such polymeric carriers are chitosan polyethylene glycol ether (Chitoxy-20 from Aquion CJSC) and sodium carboxymethyl cellulose (Hercules).

The activity of the claimed composition is due to the activity of its constituent enzymes. It was found that a topical cosmetic composition containing: 0.01-5.0 wt.% Papain, 0.01-5.0 wt.% Dextranase, 0.01-5.0 wt.% Alpha-amylase; 0.01-5.0 wt.% Invertase or glucoamylase; 0.01-5.0 wt.% Glucose oxidase; 0.01-5.0 wt.% Lactoperoxidase, 0.01-5.0 wt.% Lysozyme, 0.01-5.0 wt.% Potassium or ammonium thiocyanate; 2.0-4.0 wt.% Lactulose and cosmetically acceptable carriers or excipients, is stable in relation to the activity of enzymes during its long-term storage, while the activity of these enzymes, prepared in the form of a solution, rapidly decreases. Therefore, the claimed composition can be used for the effective prevention of inflammatory periodontal diseases.

The present invention is illustrated by the following examples.

EXAMPLE 1

A cosmetic composition in the form of a non-abrasive paste based on papain, dextranase, alpha-amylase, invertase, glucose oxidase, lactoperoxidase, lysozyme, potassium thiocyanate, lactulose and cosmetically acceptable carriers or excipients has the following composition:

Papain 0.01 Dextranase 0.01 Alpha amylase 0.01 Potassium thiocyanate 0.01 Invertase 0.01 Glucose oxidase 0.01 Lactoperoxidase 0.01 Lysozyme 0.01 Lactulose 2.00 Sodium carboxymethyl cellulose 0.50 Emulsifier 0.01 Preservative 0.01 Flavoring 0.01 Sorbitol 70% 30.00 Demineralized water 67.39

The specified composition is prepared according to the following method. All components except sodium carboxymethyl cellulose are successively dissolved in demineralized water, then sodium carboxymethyl cellulose is added and mixed until a gel forms. The resulting gel is stirred, vacuum and homogenized.

EXAMPLE 2

A cosmetic composition in the form of a non-abrasive paste based on papain, dextranase, alpha-amylase, invertase, glucose oxidase, lactoperoxidase, lysozyme, potassium thiocyanate, lactulose and cosmetically acceptable carriers or excipients has the following composition:

Papain 5,0 Dextranase 5,0 Alpha amylase 5,0 Potassium thiocyanate 5,0 Invertase 5,0 Glucose oxidase 5,0 Lactoperoxidase 5,0 Lysozyme 5,0 Lactulose 4.0 Sodium carboxymethyl cellulose 0.70 Emulsifier 5,0 Preservative 1,0 Flavoring 1,0 Sorbitol 70% 40.00 Demineralized water 8.30

The specified composition is prepared as in EXAMPLE 1.

Evaluation of the stability of enzymes in topical compositions

Since the main disadvantage of enzymes is their instability during long-term storage of finished forms based on them during the shelf life, a comparative assessment was made of the preservation of the activity of the enzyme during storage in the form of a solution and in the form of a composition for local use.

Papain activity was determined by the following methods.

To determine the proteolytic activity of papain, a modified Kunitz method was used. Such a quantity of enzyme is taken as one proteolytic unit, upon exposure to which casein on the substrate for 10 minutes under standard conditions, such a quantity of hydrolysis products is released that leads to an increase in optical density at a wavelength of 280 nm per unit.

To quantify papain, the Lowry method was used. The determination method is based on the fact that proteins with copper salts in an alkaline medium restore Follin's reagent. A Coomassie G-250 dye solution was prepared as follows: 100 mg of the dye was dissolved in 100 ml of phosphoric acid, diluted with water 5 times, then filtered through a thick paper filter. To determine the concentration in the sample, an equal volume of dye was added, mixed and the absorbance of the solution was measured at a wavelength of 595 nm.

The results are shown in the following table.

Table 1 Test samples Main ingredients Storage stability one 2 3 Pasta No. 1 According to Example 1 Activity 95% for papain within 1 year of observation Pasta No. 2 According to Example 2 Activity 82% for papain within 1 year of observation Native enzyme solution - 10% activity for papain for 7 days

As can be seen from the table 1, the introduction of enzymes in the composition for topical application allows you to get stable cosmetic compositions for topical use in the prevention of inflammatory periodontal diseases. Despite the fact that upon the introduction of enzymes into the composition of such compositions, a slight loss of enzymatic activity occurs, upon further storage for 1 year, the enzymes do not change their activity due to the fixation of the labile structure of the enzyme by the carrier polymer.

The antimicrobial activity of lactulose was determined by serial dilution. As a nutrient medium used meat-peptone broth; as test microorganisms, pure cultures: Staphylococcus aureus, Bacillus cereus, Esherichia coli, Candida albicans. A one-day culture of 1000 cells / ml was added to each tube. Cell cultures: Staphylococcus aureus, Bacillus cereus, Esherichia coli were grown in test tubes in a medium with meat and peptone broth at 37 ° C for 24 hours, and the Candida albicans culture was grown on Saburo agar medium at 24 ° C for two days. Titration was started at a concentration of 10 mg / ml (1/10 dilution). As a control, a test tube was used, into which only nutrient medium and microorganism were introduced. The concentration in the last tube with no growth was taken as the minimum inhibitory concentration. To assess the bactericidal effect of the tubes with a lack of growth, reseeding was done on plates with meat and peptone broth. The cups were thermostated at 37 ° C for a day. Bactericidal action was noted in the absence of growth on a solid nutrient medium.

table 2 Name of ingredient Staphylococcus aureus Bacillus cereus Esherichia coli Candida albicans MIC, mg / ml MCC, mg / ml MIC, mg / ml MCC, mg / ml MIC, mg / ml MCC, mg / ml MIC, mg / ml MCC, mg / ml Lactulose twenty 40 twenty 40 twenty 40 thirty fifty

As can be seen from table 2, the optimal inhibitory concentration for lactulose is: 20 mg / ml in the case of Staphylococcus aureus, Bacillus cereus, Escherichia coli; 30 mg / ml for Candida albicans; bactericidal concentration for lactulose - 40 mg / ml - in the case of Staphylococcus aureus, Bacillus cereus, Escherichia coli; 50 mg / ml - for Candida albicans.

Clinical trials of the proposed toothpaste were carried out for one month on 30 volunteers aged 21-24 years old, clinically healthy, without deformation of the teeth and jaws. To determine the cleaning effect of the paste, controlled and uncontrolled brushing of the teeth by the proposed paste was carried out. The controlled brushing of the teeth was carried out according to the method proposed in the “Methodological guidelines for experimental (pharmacological) and clinical trials of preventive and therapeutic toothpastes” (Moscow, Sofia, 1983), for 3 minutes on the 1st, 7th, 14- th, 21st and 28th day. On the remaining days, probants brushed their teeth with the same brush and paste, morning and evening, at home without control. Before and after controlled cleaning, the Green-Vermillion Hygiene Index was determined.

Quantitative assessment was carried out according to a 4-point system:

- staining of the entire surface of the tooth crown was evaluated at 3 points;

- staining 2/3 of the surface of the tooth crown was evaluated at 2 points;

- staining 1/3 of the surface of the tooth crown was evaluated at 1 point;

- the absence of staining of the surfaces of the teeth was evaluated at 0 points.

The calculation of the hygiene index indicators according to Green-Vermillion was carried out according to the formula:

K _av = K / n, where

To _cf - the averaged digital index of the hygiene index for all teeth in which the index indicators were determined;

K is the total sum of the index values for all teeth in which the indicators were evaluated;

n is the number of teeth that were evaluated.

The cleaning effectiveness of toothpaste was calculated by the formula:

Purifying efficiency = [(IG _do1 -IG _DOX) / IG _do1] × 100 where

IG _do1 - an indicator of the hygiene index obtained during the initial examination, before brushing your teeth;

IG _doX is an indicator of the hygiene index obtained during a subsequent examination (7, 14, 21, 28 days of examination), before brushing your teeth.

Plaque reduction during a single brushing was determined by the formula for each controlled brushing:

Reduction = [(IG _doX- IG _afterX ) / IG _doX ] × 100, where

IG _doX - an indicator of the hygiene index obtained during examination before the start of controlled brushing (1, 7, 14, 21, 28 days);

IG _afterX is an indicator of the hygiene index obtained during examination after conducting controlled brushing of teeth (1, 7, 14, 21, 28 days).

From table 1, 2 and 3 it can be seen that the proposed toothpaste has an increased cleaning ability, and therefore anti-caries effect, due to a more pronounced removal of dental plaque and provides removal of 58.4% of plaque.

To determine the anti-inflammatory properties of the proposed toothpaste, all the probants (the same 30 people) carried out controlled brushing for one month.

The anti-inflammatory effect of the paste was evaluated by changing the state of the gums attached to the teeth using the PMA index according to the formula:

PMA index = [Sum of indicators / (3 × number of teeth)] × 100, where

P - inflammation of the gingival papilla, which is estimated at 1 point;

M - inflammation of the gingival margin, which is estimated at 2 points;

A - inflammation of the attached gum of the alveolar process, which is estimated at 3 points.

Evaluation of the anti-inflammatory effect of toothpaste was carried out on the basis of an analysis of the dynamics of the PMA index, which was determined during repeated examinations on the 1st, 7th, 14th, 21st and 28th days. Anti-inflammatory efficacy was calculated by the formula:

Anti-inflammatory efficacy = [(PMA ₁ -PMA _X ) / PMA ₁ ] × 100, where

PMA ₁ - an indicator of the PMA index obtained during the initial examination on the 1st day of the study;

RMA _X - an indicator of the RMA index obtained during a subsequent examination (7, 14, 21 and 28 days).

The data are given in Tables 4 and 5. It is clearly seen from them that after using this paste, the condition of periodontal tissues improved significantly: 100% of the probants disappeared from catarrhal gingivitis, and the PMA index decreased from 48.3 to 6.2, which corresponded to 87 , 16% anti-inflammatory efficacy of this paste.

Assessment of hemostatic effect was carried out according to the index of hemorrhage Kechke:

I degree - rarely, estimated at 2 points;

II degree - during brushing, estimated at 4 points;

III degree - with meals and spontaneously, is estimated at 8 points.

Hemostatic effectiveness was determined by the formula:

Hemostatic effectiveness = [(IK ₁ -IR _X ) / IK ₁ ] × 100, where

IR ₁ - bleeding index at the beginning of the study;

IR _X - bleeding index at the end of the experiment.

The data are given in Tables 6 and 7. From these, it is seen that after using this paste, the condition of periodontal tissues improves and bleeding of the gums decreases from 2.6 to 0.7 points, which corresponds to 73.08% of the hemostatic effectiveness of this paste.

Thus, the proposed toothpaste has a higher cleansing (58.4% - cleansing effectiveness), anti-inflammatory (87.16% - anti-inflammatory effect) and hemostatic (73.08% - hemostatic effectiveness) effect.

Table 1 condition OHI-S Green-Vermillion 1st day 7th day 14th day 21st day 28th day Before cleaning 2.5 2.0 1,6 1.23 1,04 After cleaning 1.3 1,2 1,1 0.92 0.78

Table 2 Plaque reduction with a single use of toothpaste 1st day 7th day 14th day 21st day 28th day 48% 40% 31.25% 25.20% 25.0%

Table 3 The cleaning effectiveness of the paste (compared with the 1st day) 7th day 14th day 21st day 28th day twenty% 36% 50.8% 58.4%

Table 4 RMA Index 1st day 7th day 14th day 21st day 28th day 48.3% 27.5% 15.8% 10.09% 6.2%

Table 5 Anti-inflammatory efficacy 7th day 14th day 21st day 28th day 43.06% 67.29% 79.11% 87.16%

Table 6 Kechka bleeding index 1st day 7th day 14th day 21st day 28th day 2.6 2.2 1.9 1.4 0.7

Table 7 Hemostatic effectiveness 7th day 14th day 21st day 28th day 15.38% 26.92% 46.15% 73.08%

Claims (1)

A non-abrasive toothpaste containing enzyme preparations, a polymeric carrier, water, a preservative, emulsifier and other acceptable excipients, characterized in that it contains: papain, dextranase, alpha-amylase, potassium or ammonium thiocyanate, invertase or glucoamylase, glucose oxidase, lactoperoxidase, lactoplase or lactoferrin, lactulose, as a carrier, sodium carboxymethyl cellulose, emulsifier, preservative, flavoring agent, sorbitol 70%, demineralized water in the following ratio of components, wt.%: papain 0.01-5.00; dextranase 0.01-5.00; alpha amylase 0.01-5.00; potassium or ammonium thiocyanate 0.01-5.00; invertase or glucoamylase 0.01-5.00; glucose oxidase 0.01-5.00; lactoperoxidase 0.01-5.00; lysozyme or lactoferrin 0.01-5.00; lactulose 2-4; sodium carboxymethyl cellulose 0.5-0.7; emulsifier 0.01-5; preservative 0.01-1.0; flavoring 0.01-1.00; sorbitol 70% 30-40; demineralized water to 8.30-67.39.