RU2527343C1 - Toothpaste containing papain, lactoperoxidase and lactulose enzymes - Google Patents

Abstract

FIELD: chemistry.

SUBSTANCE: invention provides toothpaste containing enzyme preparations, a polymer carrier, water, a preservative, an emulsifier and other acceptable excipients, the toothpaste being characterised by that it contains: as active ingredients - papain, lactoperoxidase, lactulose, hydroxyapatite, lysine ursolate, oxyproline oleanolate, amino fluoride; as a carrier - silicone quaternium-16, or guar hydroxypropyltrimonium chloride, or polyquaternium, or palmitoyl hydroxypropyltrimonium amylopectin/glycerine crosspolymer, or xanthane gum, or carrageenate, or modified acrylic thickener, or hydroxypropyltrimonium maltodextrin crosspolymer, hydroxypropyl-beta-cyclodextrin, excipients - emulsifier, preservative, aromatising agent, sorbitol 70%, demineralised water. The invention enables to produce toothpaste which enables to remineralise hard dental tissue, reduce formation of dental plaque by 62.96% owing to selective decomposition of its protein base and reduce mouth inflammatory phenomena by 85.11% owing to restoration of the natural microflora of the mouth and destruction of pathogenic microflora.

EFFECT: absence of highly abrasive components in the paste and good organoleptic properties.

2 cl, 8 ex, 11 tbl

Description

The present invention relates to medicine, namely to dentistry. In order to increase the cleansing, antimicrobial, anti-inflammatory and hemostatic and remineralizing effects, restore the natural microflora of the oral cavity, a toothpaste formulation has been developed containing the following components, wt.%: Papain 0.01-5.00; lactoperoxidase 0.01-5.00; lactulose 2.00-4.00; hydroxyapatite 5.00-25.00; aminofluoride 0.05-1.95; silicone quaternium-16, or guar hydroxypropyltrimmonium chloride, or polyquaternium, or palmitoylhydroxypropyltrimmonium amylopectin / glycerin crosspolymer, or xanthan gum, or carrageenate, or a modified acrylic thickener, or hydroxyprosopropoltrimmonium 5; lysine ursolate 0.001-5.00; hydroxypropyl beta-cyclodextrin 0.10-5.00; oxyproline oleanolate 0.001-5.00; emulsifier 2.00-5.00; preservative 0.50-1.00; flavoring 0.05-1.00; sorbitol 70% 30.00-40.00; demineralized water - the rest. The proposed toothpaste allows you to remineralize the hard tissues of the teeth, reduce the formation of plaque by 62.96%, due to the selective destruction of its protein base and reduce inflammation in the oral cavity by 85.11% due to the restoration of the natural microflora of the oral cavity and the destruction of pathogenic microflora due to exposure lactulose and lactoperoxidase. Toothpaste does not contain highly 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.

Known compositions containing sodium ursolate and bisabolol as anti-inflammatory components (Internet link http://web.archive.org/web/20071221022251; http://www.biocosm.ru/NatuRoyle.htm).

There are various therapeutic 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 toothpaste "Natural whitening toothpaste with tea tree oil" (USA). However, these pastes have a low cleansing ability, because they do not contain components that specifically dissolve plaque.

A toothpaste with an enzyme-containing composition is known, 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 Ia oral microflora.

From RU 2006135434, a toothpaste is known containing enzyme preparations, a polymeric carrier, water, a preservative, emulsifier and other acceptable excipients, the paste containing in wt.%: Papain 0.20-0.40; lactoperoxidase 0.20-0.40; lactulose 2.00-4.00; silica 20.00-25.00; sodium carboxymethyl cellulose 0.50-0.70; sodium ursolate 0.01-0.02; bisabolol 0.30-0.50; emulsifier 2.00-5.00; preservative 0.50-1.00; flavoring agent 0.05-0.10; sorbitol 70% 30.00-40.00; demineralized water "up to 71.99-77.52" (ie, the amount of demineralized water in the range 71.99-77.52, which must be added so that the total content of the components is 100%.).

The claimed toothpaste, in contrast to RU 2006135434, contains quaternium-16 silicone, or guar hydroxypropyltrimmonium chloride, or polyquaternium, or palmitoylhydroxypropyltrimmonium amylopectin / glycerin, crosspolymer, or xanthan gum, hydroxymethyl, hydroxymethyl, hydroxymethyl, hydroxymethyl, hydroxybenzene, or modified carboxymean has the ability to remineralize hard tissue of teeth.

From RU 2355420, publ. 05/20/2009 (examples, claims) Known toothpaste, including: the abrasive component is 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 propylparaben) 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. The difference between the claimed claims is that it contains lactoperoxidase.

From US 4150113, publ. 04/17/1979 (the entire document, in particular, description column 5, examples, claims), dentifrices are known, including toothpastes, mouthwashes, chewing 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.

An oral composition is known from RU 2292872, which provides oral hygiene with improved properties, containing proteolytic enzymes: papain, bromelain, chymotrypsin.

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. 09/20/2008 (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. Toothpaste "Dent Check-Up foam" (Japan) is also known - a liquid soap for cleaning the oral cavity. (Internet link: http://www.fukuoka.ru/zubnaya_bezabrazivnaya pena s ftorom dlya chistki zubov chec k up foam dent 100 g)

Toothpaste containing papain enzymes is known from RU 2407512; dextranase; alpha amylase; invertase or glucoamylase; glucose oxidase; lactoperoxidase; lysozyme or lactoferrin, lactulose to restore the microflora of the oral cavity and the prevention of inflammatory periodontal diseases.

From RU 2220713 C1 and from RU 2158582, methods for treating periodontitis patients with drugs containing lactulose are known.

Toothpastes containing papain, lactoperoxidase, lactulose, hydroxyapatite, aminofluoride, silicone quaternium-16, or guar hydroxypropyltrimmonium chloride, or polyquaternium, or palmitoylhydroxypropyltrimmonium amylopectrin or carboxymerylcriminodimethylcriminolithromylodergene gumperinacetamide, or glycerol acrylamidecrylamide or camrylated acrylate lysine ursolate, hydroxypropyl-beta-cyclodextrin, oxyproline oleanolate for remineralization of hard tissues of teeth, restoration of microflora olosti mouth and prophylaxis of inflammatory periodontal diseases are not described.

Thus, it is an object of the present invention to provide a toothpaste containing papain, lactoperoxidase, lactulose, hydroxyapatite, aminofluoride, silicone quaternium-16, or guar hydroxypropyltrimmonium chloride, or polyquaternium, palmitoylhydroxypropperimonimonium, or palmitoylhydroxypropperimonimonium, or gamma hydroxypropperimonimonium, carrageenate, or a modified acrylic thickener, or hydroxypropyltrimmonium maltodextrin cross-polymer, lysine ursolate, hydroxypropyl beta-cyclodextrin, oxyproline oleanolat which it may be used for remineralization of dental hard tissues, oral cavity microflora recovery and prevention of inflammatory periodontal disease caused by cariogenic microorganisms, such as S. mutans, S. aureus and others.

According to the present invention, there is provided a toothpaste containing in wt.%: Papain 0.01-5.00; lactoperoxidase 0.01-5.00; lactulose 2.00-4.00; hydroxyapatite 5.00-25.00; aminofluoride 0.05-1.95; silicone quaternium-16, or guarhydroxypropyltrimmonium chloride, or polyquaternium, or palmitoylhydroxypropyltrimmonium amylopectin / glycerin crosspolymer, or xanthan gum, or carrageenate, or a modified acrylic thickener, or hydroxyprosopropoltrimmonium 5; hydroxypropyl beta-cyclodextrin 0.10-5.00 and acceptable excipients.

As a preferred embodiment of the claimed invention, there is provided the toothpaste indicated above, further comprising one or more anti-inflammatory ingredients.

Most preferred is a toothpaste containing papain, lactoperoxidase, lactulose, hydroxyapatite, aminofluoride, quaternium-16 silicone, or guarhydroxypropyltrimmonium chloride, or polyquaternium, or palmitoylhydroxypropyltrimmonium amylopropermene glycerylpropyltrimethylene glycerolpropylene or maltodextrin cross-polymer, lysine ursolate, hydroxypropyl beta-cyclodextrin, oxyproline oleanolate and excipients, in the following Ocean, wt.%:

Papain 0.01-5.00 Lactoperoxidase 0.01-5.00 Lactulose 2.00-4.00 Hydroxyapatite 5.00-25.00 Aminofluoride 0.05-1.95 Silicone Quaternium-16, or guar hydroxypropyltrimmonium chloride, or polyquaternium, or palmitoylhydroxypropyltrimmonium amylopectin / glycerin crosspolymer, or xanthan gum, or carrageenate, or modified acrylic thickener, or hydroxypropyltrimmonium maltodextrin crosspolymer 0.05-5.00 Hydroxypropyl Beta Cyclodextrin 0.10-5.00 Lysine ursolate 0.001-5.00 Oxyproline oleanolate 0.001-5.00 Emulsifier 2.00-5.00 Preservative 0.50-1.00 Flavoring 0.05-0.10 Sorbitol 70% 30.00-40.00 Demineralized water rest

The active ingredients of the claimed toothpaste are: papain, lactoperoxidase, lactulose, hydroxyapatite, lysine ursolate, oxyproline oleanolate. Papain is capable of breaking peptide bonds from sulfur-containing amino acid residues. Lactoperoxidase has the specific ability to oxidize thiocyanate ions to hypothiocyanate in the presence of hydrogen peroxide, delaying the growth of pathogenic bacteria. For the preparation of toothpaste, the enzyme substances Lactoperoxidase and Papain are used. Lactoperoxidase was obtained from horseradish rhizomes, papain from papaya fruits 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. Lyophilized dried powder “Lactoperoxidase” is used for acne. Ursolate lysine and oxyproline oleanolate obtained from sea buckthorn fruits have anti-inflammatory and antibacterial effects. Aminofluoride is fixed on the surface of tooth enamel and creates a depot of fluoride ion, exerting an anticaries effect. Hydroxyapatite (English hydroxyapatite, hydroxylapatite, HA) is a mineral of Ca ₁₀ (PO ₄ ) ₆ (OH) ₂ , a type of calcium apatite. It is the main mineral component of bones (about 50% of the total bone weight) and teeth (96% in enamel). In medicine, synthetic hydroxyapatite is used as a filler to replace parts of lost bone, and as a coating for implants, which helps to build up new bone. (Found from the Internet: http://en.wikipedia.org/wiki/%D0%93%D0%B8%D0%B4%D1%80%D0%BE%D0%BA%D1%81%D0%B8% D0% B0% D0% BF% D0% B0% D1% 82% D0% B8% D1% 82)

The toothpaste of the present invention is usually made by conventional methods using solid or liquid 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 use.

Additional advantages of this composition are the ability to remineralize the hard tissues of the teeth, restore the natural microflora of the oral cavity.

The proposed toothpaste reduces the formation of plaque by 62.96%, reduces inflammation in the oral cavity by 85.11%, reduces bleeding gums by 72.34% and is completely harmless and safe for prolonged use.

The significant differences of the proposed toothpaste 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 immobilizing the enzymes on polymer carriers contained in the specified 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 silicone quaternium-16, or guarhydroxypropyltrimmonium chloride, or polyquaternium or palmitoylhydroxypropyltrimmonium amylopectin / glycerin crosspolymer, or xanthan gum, or carrageenate, or modified acrylic hydroxymethyl thickener, or modified acrylic hydroxymethylpropylene or molybrosimetol, or thickener. The advantages of xanthan gum and carrageenate are its high compatibility with calcium salts and biological tissues. The advantages of polymer carriers such as quaternium-16, guar-hydroxypropyltrimmonium chloride, polyquaternium, palmitoylhydroxypropyltrimmonium amylopectin / glycerin crosspolymer, modified acrylic thickener, hydroxypropyltrimmonium maltodextrin cross-linked polymer, the presence of a positively charged ammonium polymer. This determines the remineralizing and antimicrobial properties of these polymers, determines the high affinity for negatively charged hard tissues of the teeth, the ability to fix calcium ions and obstruct the dentinal tubules, as well as increase the stability of the complex with enzymes. An additional carrier is hydroxypropyl beta-cyclodextrin, which helps to increase the solubility of oxyproline oleanolate and ursolate lysine.

The activity of the claimed composition is due to the activity of the functional components included in its composition. It was found that a toothpaste containing papain 0.01-5.00 wt.% And lactoperoxidase 0.01-5.00 wt.%, Is stable in relation to the activity of enzymes during its long-term storage, while the activity of these enzymes cooked as a solution, drops rapidly. 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 toothpaste based on a mixture of lactulose, papain and lactoperoxidase has the following composition:

Papain 0.40 Lactoperoxidase 0.40 Lactulose 4.00 Hydroxyapatite 25.00 Aminofluoride 0.05 Palmitoylhydroxypropyltrimmonium amylopectin / glycerin crosspolymer 0.70 Hydroxypropyl Beta Cyclodextrin 0.70 Lysine ursolate 0.001 Oxyproline oleanolate 0.50 Emulsifier 4.30 Preservative 1.00 Flavoring 0.10 Sorbitol 70% 40.00 Demineralized water rest

The specified composition is prepared according to the following method. In demineralized water, palmitoylhydroxypropyltrimmonium amylopectin / glycerol crosspolymer and lactulose are dissolved, the resulting gel is mixed, vacuum and homogenized. Papain, lactoperoxidase, hydroxyapatite, aminofluoride, hydroxypropyl-beta-cyclodextrin, lysine ursolate, oxyproline oleanolate, emulsifier, preservative, flavoring agent, sorbitol 70% are successively added to the resulting gel. Toothpaste is homogenized and vacuum.

BUT,

Toothpaste containing a mixture of lactulose, papain and lactoperoxidase based on guarhydroxypropyltrimmonium chloride has the following composition:

Papain 0.20 Lactoperoxidase 0.20 Lactulose 2.00 Hydroxyapatite 20.00 Aminofluoride 1.95 Hydroxypropyltrimmonium Chloride 0.50 Hydroxypropyl Beta Cyclodextrin 0.50 Lysine ursolate 0.01 Oxyproline oleanolate 0.001 Emulsifier 2.00 Preservative 0.50 Flavoring 0.05 Sorbitol 70% 30.00 Demineralized water rest

The specified composition is prepared according to the following method. Guarhydroxypropyltrimmonium chloride and lactulose are dissolved in demineralized water, the resulting gel is stirred, vacuum and homogenized. Papain, lactoperoxidase, hydroxyapatite, aminofluoride, hydroxypropyl-beta-cyclodextrin, lysine ursolate, oxyproline oleanolate, emulsifier, preservative, flavoring agent, sorbitol 70% are successively added to the resulting gel. Toothpaste is homogenized and vacuum.

EXAMPLE 3

Toothpaste containing a mixture of lactulose, papain and lactoperoxidase based on hydroxypropyltrimmonium maltodextrin cross-polymer has the following composition:

Papain 0.25 Lactoperoxidase 0.25 Lactulose 4.00 Hydroxyapatite 25.00 Aminofluoride 1.95

Hydroxypropyltrimmonium maltodextrin

crosspolymer 0.50 Hydroxypropyl Beta Cyclodextrin 1.00 Lysine ursolate 0.015 Oxyproline oleanolate 0.50 Emulsifier 5.00 Preservative 1.00 Flavoring 0.10 Sorbitol 70% 35.00 Demineralized water rest

The specified composition is prepared according to the following method. Hydroxypropyltrimmonium maltodextrin cross-polymer and lactulose are dissolved in demineralized water, the resulting gel is mixed, vacuum and homogenized. Papain, lactoperoxidase, hydroxyapatite, aminofluoride, hydroxypropyl-beta-cyclodextrin, lysine ursolate, oxyproline oleanolate, emulsifier, preservative, flavoring agent, sorbitol 70% are successively added to the resulting gel. Toothpaste is homogenized and vacuum.

EXAMPLE 4

Toothpaste containing a mixture of lactulose, papain and lactoperoxidase based on silicone quaternium-16, has the following composition:

Papain 3.25 Lactoperoxidase 3.25 Lactulose 3,50 Hydroxyapatite 22.00 Aminofluoride 1.95 Silicone Quaternium-16 3,50 Hydroxypropyl Beta Cyclodextrin 1.00 Lysine ursolate 0.015 Oxyproline oleanolate 0.50 Emulsifier 5.00 Preservative 1.00 Flavoring 0.10 Sorbitol 70% 35.00 Demineralized water rest

The specified composition is prepared according to the following method. Quaternium-16 silicone and lactulose are dissolved in demineralized water, the resulting gel is mixed, vacuum and homogenized. Papain, lactoperoxidase, hydroxyapatite, aminofluoride, hydroxypropyl-beta-cyclodextrin, lysine ursolate, oxyproline oleanolate, emulsifier, preservative, flavoring agent, sorbitol 70% are successively added to the resulting gel. Toothpaste is homogenized and vacuum.

EXAMPLE 5

Toothpaste containing a mixture of lactulose, papain and lactoperoxidase based on a modified acrylic thickener (Sepin EMT 10), has the following composition:

Papain 2,50 Lactoperoxidase 2,50 Lactulose 2,50 Hydroxyapatite 22.00 Aminofluoride 1.95 Sepinov EMT 10 1,50 Hydroxypropyl Beta Cyclodextrin 1.00 Lysine ursolate 0.015 Oxyproline oleanolate 0.50 Emulsifier 5.00 Preservative 1.00 Flavoring 0.10 Sorbitol 70% 30.00 Demineralized water rest

The specified composition is prepared according to the following method. Modified acrylic thickener (Sepinov EMT 10) and lactulose are dissolved in demineralized water, the resulting gel is mixed, vacuum and homogenized. Papain, lactoperoxidase, hydroxyapatite, aminofluoride, hydroxypropyl-beta-cyclodextrin, lysine ursolate, oxyproline oleanolate, emulsifier, preservative, flavoring agent, sorbitol 70% are successively added to the resulting gel. Toothpaste is homogenized and vacuum.

EXAMPLE 6

Toothpaste containing a mixture of lactulose, papain and lactoperoxidase based on carrageenate has the following composition:

Papain 2,50 Lactoperoxidase 2,50 Lactulose 2,50 Hydroxyapatite 22.00 Aminofluoride 1.95 Carragenate 1,50 Hydroxypropyl Beta Cyclodextrin 1.00 Lysine ursolate 0.015 Oxyproline oleanolate 0.50 Emulsifier 5.00 Preservative 1.00 Flavoring 0.10 Sorbitol 70% 30.00 Demineralized water rest

The specified composition is prepared according to the following method. Carrageenate and lactulose are dissolved in demineralized water, the resulting gel is mixed, vacuum and homogenized. Papain, lactoperoxidase, hydroxyapatite, aminofluoride, hydroxypropyl-beta-cyclodextrin, lysine ursolate, oxyproline oleanolate, emulsifier, preservative, flavoring agent, sorbitol 70% are successively added to the resulting gel. Toothpaste is homogenized and vacuum.

EXAMPLE 7

Toothpaste containing a mixture of lactulose, papain and lactoperoxidase based on xanthan gum, has the following composition:

Papain 4,50 Lactoperoxidase 4,50 Lactulose 2.20 Hydroxyapatite 24.00 Aminofluoride 1.95 Xanthan gum 1,50 Hydroxypropyl Beta Cyclodextrin 5.00 Lysine ursolate 0.015 Oxyproline oleanolate 0.50 Emulsifier 4.00 Preservative 1.00 Flavoring 0.10 Sorbitol 70% 30.00 Demineralized water rest

The specified composition is prepared according to the following method. Xanthan gum and lactulose are dissolved in demineralized water, the resulting gel is mixed, vacuum and homogenized. Papain, lactoperoxidase, hydroxyapatite, aminofluoride, hydroxypropyl-beta-cyclodextrin, lysine ursolate, oxyproline oleanolate, emulsifier, preservative, flavoring agent, sorbitol 70% are successively added to the resulting gel. Toothpaste is homogenized and vacuum.

EXAMPLE 8

A toothpaste containing a mixture of lactulose, papain and lactoperoxidase based on polyquaternium has the following composition:

Papain 0.20 Lactoperoxidase 0.20 Lactulose 2.00 Hydroxyapatite 20.00 Aminofluoride 1.95 Polyquaternium 0.50 Lysine ursolate 0.01 Oxyproline oleanolate 0.30 Hydroxypropyl Beta Cyclodextrin 5.00 Emulsifier 2.00 Preservative 0.50 Flavoring 0.05 Sorbitol 70% 30.00 Demineralized water rest

The specified composition is prepared according to the following method. Polyquaternium and lactulose are dissolved in demineralized water, the resulting gel is mixed, vacuum and homogenized. Papain, lactoperoxidase, hydroxyapatite, aminofluoride, hydroxypropyl-beta-cyclodextrin, lysine ursolate, oxyproline oleanolate, emulsifier, preservative, flavoring agent, sorbitol 70% are successively added to the resulting gel. Toothpaste is homogenized and vacuum.

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 expiration date, a comparative assessment was made of the preservation of enzyme activity during storage in the form of a solution and in the form of a composition for local use.

The activity of papain and lactoperoxidase was determined by the following methods.

To determine the proteolytic activity of papain, a modified Kunitz method was used. Such a quantity of an 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 determine the activity of lactoperoxidase, the Birden method was used. A Coomassie G-250 dye solution was prepared as follows: 100 mg of the dye was dissolved in 100 ml of phosphoric acid, diluted 5 times with water, 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 Toothpaste No. 1 According to Example 1 Activity 95% for papain and 80% for lactoperoxidase within 1 year of observation Toothpaste No. 2 According to Example 2 Activity 82% for papain and 80% for lactoperoxidase within 1 year of observation Native enzyme solution - 40% activity for papain and 10% lactoperoxidase for 1 day

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 lactoperoxidase, sodium ursolate and 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, and Esherichia coli were grown in test tubes in a medium with meat and peptone broth at 37 ° C for 24 hours, and a Candida albicans culture on Saburo agarized seldure at 24 ° C for two days. Titration began with a concentration of 1 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 Lactoperoxidase 10 twenty 10 twenty 10 twenty twenty thirty Ursolate Lysine one 2 one 2 one 2 one 2 Hydroxyproline Oleanolate one 2 one 2 one 2 one 2

As can be seen from table 2, the optimal inhibitory concentration is: for lactulose: 20 mg / ml in the case of Staphylococcus aureus, Bacillus cereus, Escherichia coli; 30 mg / ml for Candida albicans; lactoperoxidase - 10 mg / ml in the case of Staphylococcus aureus, Bacillus cereus, Escherichia coli; 20 mg / ml - for Candida albicans; ursolate lysine and oxyproline oleanolate-1 mg / ml in the case of Staphylococcus aureus, Bacillus cereus, Escherichia coli, 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; for latoperoxidase, 20 mg / ml in the case of Staphylococcus aureus, Bacillus cereus, Escherichia coli; 30 mg / ml for Candida albicans; ursolate lysine and oxyproline oleanolate - 2 mg / ml - in the case of Staphylococcus aureus, Bacillus cereus, Escherichia coli, 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, 14th th, 21st and 28-1 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, a 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 _cf = K / n, where

K _cf - the averaged digital indicator 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 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 up to _1- IG up to _X ) / IG up to ₁ ] × 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 _{beforeX —} IG _afterX ) / IG _beforeX ] × 100, where

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

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

From table 3, 4 and 5 it can be seen that the proposed toothpaste has an increased cleaning ability, and therefore the anti-caries effect due to a more pronounced removal of dental plaque and provides the removal of 62.96% 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;

РМА _x - indicator of the РМА index, obtained during the subsequent examination (7, 14, 21 and 28 days).

The data are given in Tables 6 and 7. It is clearly seen from them that after using this paste, the condition of periodontal tissues improved significantly: 100% of the probans disappeared from catarrhal gingivitis, and the PMA index decreased from 57.1 to 8.5, which corresponded to 85 , 11% anti-inflammatory efficacy of this paste.

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

I degree - the gum bleeds rarely, is estimated at 2 points;

II degree - the gum bleeds during brushing, is estimated at 4 points;

III degree - the gum bleeds with food and spontaneously, is estimated at 8 points.

Hemostatic effectiveness was determined by the formula:

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

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

IR _x is the bleeding index at the end of the experiment.

The data are given in Tables 8 and 9. It can be seen from them that after using this paste, the condition of periodontal tissues improves and bleeding of the gums decreases from 4.7 to 1.3 points, which corresponds to 72.34% of the hemostatic effectiveness of this paste.

Thus, the proposed toothpaste has a higher cleansing (62.96% - cleansing effectiveness), anti-inflammatory (85.11% - anti-inflammatory effect) and hemostatic (72.34% - hemostatic effectiveness) effect.

Table3 condition OHI-S Green-Vermillion 1st day 7th day 14th day 21st day 28th day Before cleaning 2.7 2.0 1,5 1,2 1.00 After cleaning 1.3 1,2 1,1 0.92 0.78

Table 4 Plaque reduction with a single use of toothpaste 1st day 7th day 14th day 21st day 28th day 51.85% 60% 73.33% 76.66% 78.00%

Table 5 The cleaning effectiveness of the paste (compared with the 1st day) 7th day 14th day 21st day 28th day 25.9% 44.44% 55.55% 62.96%

Table 6 RMA Index 1st day 7th day 14th day 21st day 28th day 57.1% 47.7% 35.6% 18.7% 8.5%

Table 7 Anti-inflammatory efficacy 7th day 14th day 21st day 28th day 16.46% 37.65% 67.25% 85.11%

Table 8 Kechka bleeding index 1st day 7th day 14th day 21st day 28th day 4.7 3,1 2.7 2.2 1.3

Table 9 Hemostatic effectiveness 7th day 14th day 21st day 28th day 34.04% 42.55% 53.19% 72.34%

Table 10 Changing the state of remineralization of teeth according to the TER test TER test Survey period Start 1st week 2nd week 3rd week 4th week 60.0 55.0 55.0 50,0 45.0

Table 11 summarizes the results of determining the remineralizing efficacy of the TER test.

Table 11 Change in remineralizing efficiency according to the TER test Remineralizing efficacy by TER test Survey period 1st week 2nd week 3rd week 4th week 8.3% 8.3% 16.7% 25.00%

The proposed means of oral hygiene also has a remineralizing effect due to the fact that there is a more active cleaning of the surfaces of the teeth and the mineral components of the oral fluid are able to more effectively penetrate the surface layer of enamel and form a protective layer of CaF ₂ .

Claims (2)

1. Toothpaste containing enzyme preparations, a polymeric carrier, water, preservative, emulsifier and other acceptable excipients,
characterized in that it contains: as active ingredients - papain, lactoperoxidase, lactulose, hydroxyapatite, lysine ursolate, oxyproline oleanolate, aminofluoride; as a carrier - silicone quaternium-16, or guarhydroxypropyltrimmonium chloride, or polyquaternium, or palmitoylhydroxypropyltrimmonium amylopectin / glycerin, crosspolymer, or xanthan gum, or carrageenate, or modified acrylic thickener, hydroxypertroxymethrocyrmethroxymethroxymethroxymethroxy-hydroxypropyl malin hydroxypropylene preservative, flavoring, sorbitol 70%, demineralized water. 2. Toothpaste according to claim 1, having the following composition:
Papain 0.01-5.00 Lactoperoxidase 0.01-5.00 Lactulose 2.00-4.00 Hydroxyapatite 5.00-25.00 Aminofluoride 0.05-1.95 Silicone Quaternium-16, or guar hydroxypropyltrimmonium chloride, or polyquaternium, or palmitoylhydroxypropyltrimmonium amylopectin / glycerin crosspolymer, or xanthan gum, or carrageenate, or modified acrylic thickener, or hydroxypropyltrimmonium maltodextrin crosspolymer 0.05-5.00 Hydroxypropyl Beta Cyclodextrin 0.10-5.00 Lysine ursolate 0.001-5.00 Oxyproline oleanolate 0.001-5.00 Emulsifier 2.00-5.00 Preservative 0.50-1.00 Flavoring 0.05-0.10 Sorbitol 70% 30.00-40.00 Demineralized water rest