RU2527692C1 - Tooth paste containing complex of hydrolases - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: invention represents a tooth paste containing papain, lysozyme, collagenase, chemopsin, trisamine, urea, mannitol, polyethylene glycol chitosan, D-panthenol, characterised by the fact that the following ingredients are used for preparing it, wt %: papain 0.01-5.00; abrasive 2.00-20.00; humidifying agent 5.00-10.00; collagenase 0.01-5.00; chemopsin 0.01-5.00; mannitol 1.00-10.00; trisamine 0.001-4.00; urea 1.00-10.00; D-panthenol 0.001-5.00; polyethylene glycol chitosan 0.001-5.00; hydrocolloid 0.10-2.00; surfactant 0.10-1.00; sodium fluoride 0.05-0.32; lysozyme 0.01-5.00; tetrasodium pyrophosphate 1.00-5.00; chlorhexidine bigluconate 0.05-0.20; flavouring agent 0.10-1.50; demineralised water - the rest.

EFFECT: preparing the composition which can be used for oral hygiene, gingival pocket cleaning from purulo-necrotic mass, higher saliva lysozyme, treatment and prevention of dentoalveolar diseases.

2 ex, 9 tbl

Description

The present invention relates to medicine, namely to dentistry. In order to increase the cleaning, antimicrobial and anti-inflammatory effect, restore the natural pH level of tissues, a toothpaste containing a complex of hydrolases has been developed: papain, lysozyme, collagenase, chymopsin, trisamine, urea, mannitol, polyethylene glycol ether of chitosan, d-panthenol and acceptable carriers and excipients . The proposed toothpaste can reduce inflammation and accelerate the regeneration processes in the tissues of the dentition in inflammatory diseases. Toothpaste has good organoleptic properties: it is white in color, with a pleasant smell and good taste.

2. The level of technology

Compositions for the oral cavity, such as toothpastes, gels, elixirs, balms, rinses are intended mainly for removing plaque and / or preventing its formation with regular use. Dental plaque is a multi-component, constantly forming biotope of the oral cavity, the basis of which is almost all representatives of the microflora of the oral cavity in a conglomerate with proteins and polysaccharides. In the process of plaque formation, a marked change in the composition of microflora occurs. In the first phase, lasting 2-4 hours, an early plaque is formed with a predominance of aerobic and facultative anaerobic bacteria - streptococci, staphylococci, neisseria and lactobacillus. The total bacterial content is not more than 100-1000 in 1 g. In the second phase (day 4-5), aerobic and facultative anaerobic microflora shift towards anaerobic leptotrichia and fusobacteria. The total bacterial content increases to 1-10 million in 1 g. In the third phase (days 6-7 and further), the formation of a qualitative composition of dental plaque ends - the content of aerobes and facultative anaerobes sharply decreases and obligate anaerobes (bacteroids, fusobacteria, veilonella prevail) , actinomycetes, peptostreptococci), emitting a complex of toxic substances that cause the destruction of Ig A and Ig G, a sharp shift in pH to the acid side, which leads to the progression of microbial invasion, the development of caries and periodontal diseases.

Various therapeutic and prophylactic toothpastes used for the prevention of inflammatory periodontal diseases, containing anti-inflammatory and antimicrobial components, for example, extracts of medicinal plants and vitamins (Silca Herb plus (Germany) and tea tree oil, are known in the art) Toothpaste "Natural whitening toothpaste with tea tree oil" ((USA). However, this paste has a low cleaning ability, because it does not contain components that specifically dissolve plaque.

Known toothpaste "Audrey-Dent" for the elimination and prevention of various inflammatory processes in the oral cavity, containing liposomes, antiseptic - dioxidine, urea, Maclea extract, vitamins C and P. (http://liderm.audrie.ru/audrie/audrie_catalog /audrie_dentifrice/34-zubnaya-pasta-odri-dent.html)

Known toothpaste Lacalut Brilliant White Menta®, which removes plaque due to highly formative compounds.

Known desensitizing paste Colgate (Sensitive Pro-Relief ™, containing arginine and calcium carbonate to reduce tooth sensitivity. (Http://colgateprofessional.ru/products/5938a27a02315210b27a0231521028016e0aRCRD/specifics)

Known toothpaste with an enzyme-containing composition, where the enzyme is selected from the group of proteinases, including papain, which destroys the protein base of plaque (Whitening toothpaste "Rembrandt" ((USA), as well as a paste containing lysozyme, glucose oxidase, lactoperoxidase and lactoferrinrin (Biotene® toothpaste (Finland), which is used to treat xerostomia. The described toothpaste generates active forms of oxygen during toothbrushing and can only affect aerobic and facultative anaerobic bacteria.

Toothpaste containing papain enzymes is known from RU 2407512 C1; dextranase; alpha amylase; invertase or glucoamylase; glucose oxidase; lactoperoxidase; lysozyme or lactoferrin, to restore the microflora of the oral cavity and the prevention of inflammatory periodontal diseases.

From RU 2333768 C2, enzyme-containing compositions with cetylpyridinium chloride with improved hygienic properties against plaque are known.

Known chymopsin is an enzymatic preparation of proteolytic action, which breaks down necrotic tissue without affecting living tissue, due to the presence of specific antienzymes in it. It liquefies viscous secrets and exudates and is used for burns, bedsores, trophic ulcers, purulent wounds to cleanse purulent-necrotic masses, accelerate healing. (http://www.vidal.ru/poisk_preparatov/chymopsin.htm)

Collagenase is known - a proteolytic enzyme isolated from Clostridium histolyticum. It contains several other proteases as concomitant enzymes. Under the influence of the drug, the wound cleanses, which contributes to the more rapid development of granulation tissue. Does not affect intact epithelium, adipose and muscle tissue. (http://www.vidal.ru/poisk_preparatov/lact_269.htm)

Known drug "Lysozyme", produced in the form of hermetically sealed vials of 0.05; 0.1 and 0.15 g, which has a bacteriolytic effect and has the ability to stimulate the nonspecific reactivity of the body. Lysozyme catalyzes the hydrolysis of 1,4-bonds between N-acetylmuramic acid and 2-acetamido-2-deoxy-D-glucose (N-acetylglucosamine) contained in mucopolysaccharides, mucopeptides and chitin of the cell walls of bacteria, fungi and protozoa, it has a bright pronounced antimicrobial (destroys the cell walls of most gram-positive and some gram-negative bacteria), mycostatic (delays the development of microscopic fungi), as well as wound healing effect. (Found from the Internet: <URL: http: //provizor.Org/node/l1671)

The known enzyme is papain, a plant proteolytic enzyme of the hydrolase class, exhibiting broad substrate specificity: it catalyzes the hydrolysis of peptide bonds in proteins and peptides, and also hydrolyzes amides, esters and thioethers. 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.

(Found from the Internet:

<URL: http: //slovari.yandex.ru/%D0%BF%D0%B0%D0%BF%D0%B0%D0%B8%D0%BD/%D0%91%D0%A1%D0%AD /% D0% 9F% D0% B0% D0% BF% D0% B0% D0% B8% D0% BD /)

Known drug trisamine (tri- (oxymethyl) -aminomethane), which is an antacid of systemic action, has buffering properties, eliminates acidosis, including and intracellular, has an osmotic diuretic effect, is used for acute and chronic diseases, accompanied by metabolic and mixed acidosis. (Mashkovsky M.D. Medicines. - 15th ed., - M: New Wave, 2006. - p.686)

Known mannitol is an osmotic diuretic. Increasing the osmotic pressure of plasma and filtration without subsequent tubular reabsorption leads to the retention of water in the tubules and an increase in urine volume. Increasing the osmolarity of plasma, it causes the movement of fluid from tissues (in particular, the eyeball, brain) into the vascular bed, (http://www.vidal.ru/poisk_preparatov/act_633.htm)

Urea (urea) is a known chemical compound, carbonic diamide. White crystals soluble in polar solvents (water, ethanol, liquid ammonia). Urea forms inclusion complexes (clathrates) with many compounds, for example, with hydrogen peroxide CO (NH ₂ ) ₂ -H ₂ O ₂ , used as a convenient and safe form of “dry” hydrogen peroxide (hydroperite). (http://en.wikipedia.org/wiki/%CC%EE%F7%E5%E2%E8%ED%E0)

It is known to use enzyme preparations, in particular, proteolytic enzymes, such as trypsin, chymotrypsin, chymopsin, terrilithin, lysoamidase, papain, profesim, the drug iruxol containing clostridiopeptidase and chloramphenicol in case of ulcerative gingivitis, periodontitis, and especially during purulent carcinoma and parodontitis. (Tsepov L.M., Nikolaev A.I. Diagnostics and treatment of periodontal diseases. - M .: MEDpress-inform, 2002. - p. 94, 95, 98)

Known adhesive balm for the gums Asept® containing chlorhexidine bigluconate - an antiseptic that is effective against aerobic and facultative anaerobic bacteria and metronidazole - an antibacterial component that is effective against anaerobic bacteria and protozoa.

Known gel toothpaste Dentofam® containing a concentrate of bacteriophages, selective for species Streptococccus and Staphylococcus.

Chitoxy-20 (polyethylene glycol ether of chitosan) is known, which is a water-soluble form of chitosan. Chitosan is a saponification product of a natural carbohydrate-containing biopolymer - chitin, which, along with cellulose, is the most common organic substance in nature. The use of chitosan in cosmetic compositions is due to its following properties:

- biocompatibility with human cells;

- regenerative effect on the skin;

- antibacterial effect;

- antifungal effect;

- ability to retain moisture.

Chitoxy-20 is a viscous transparent liquid of light yellow color, odorless, easily soluble in water at any pH. The ideal solubility in water allows the use of Hitoxy-20 as a component in any percentage. The consistency of this form of chitosan can be regulated (taking the form that is most convenient for use in this type of cosmetics). The drug Chitoxy-20 preserves all the useful properties of the original chitosan and has new specific properties:

- pronounced hydrating properties; forms around itself a "fur coat" of water molecules, including tightly bound water;

- the ability to form semipermeable films that keep the skin hydrated by passing oxygen, i.e. allow the skin to breathe;

- the ability to stabilize other components of the composition, for example, vitamins.

(http://www.akvion.ru/catalog/skk/skkhitoksi.php)

Known dexpanthenol, which refers to the vitamins of group B and is a derivative of pantothenic acid. It plays an important role in the processes of acetylation and oxidation, participates in carbohydrate and fat metabolism, in the synthesis of acetylcholine, corticosteroids, porphyrins. It has a pronounced effect on the formation and function of epithelial tissue, has some anti-inflammatory activity, (http://www.vidal.ru/poisk_preparatov/act_303.htm)

Known medical and preventive toothpaste N-ZIM Prebio, produced by Artlife since 2009 (Medical and preventive toothpaste N-ZIM Prebio. 08/02/2011, Found on the Internet: <URL: http://www.artlife-chel.ru / main / 466-n-zim-prebio>), includes a lactobacillus lysate, which increases local immunity, thereby preventing the propagation of pathogenic microorganisms in the oral cavity and the preservation of beneficial microflora. N-ZIM Prebio Toothpaste also contains the enzyme papain, which effectively breaks down plaque without damaging tooth enamel.

According to Artlife company data, N-ZIM Prebio toothpaste contains: demineralized water, sorbitol, silicon dioxide, dicalcium phosphate dihydrate, polyethylene glycol, sodium lauroyl sarcosinate, sodium carboxymethyl cellulose, flavoring, D-panthenol, sodium lauryl sulfate, papa titan, lysine, lysine fluoride, aminofluoride, allantoin, saccharin, lactobacillus lysate, methyl paraben, propyl paraben, stevioside, aloe extract; while the mass fraction of fluoride is at least 0.1%.

The declared toothpaste differs from N-ZIM Prebio toothpaste in that it contains urea, mannitol, trisamine, chitosan polyethylene glycol ether, lysozyme, collagenase, chymopsin, d-panthenol.

Toothpaste compositions are known from RU 2355420 and RU 2381021, including enzymes, in particular lysozyme and papain.

The declared toothpaste differs from RU 2355420 and RU 2381021 in that it contains urea, mannitol, trisamine, polyethylene glycol chitosan ether, collagenase, chymopsin, d-panthenol.

Compositions for the oral cavity, such as toothpastes, gels, elixirs, rinses, for the manufacture of which are used: enzymes: papain, lysozyme, collagenase, chymopsin, polyethylene glycol ether of chitosan, trisamine, urea, mannitol, d-panthenol.

Thus, the objective of the present invention is to provide a composition for the oral cavity, the composition of the starting components of which includes: papain, lysozyme, collagenase, chymopsin, polyethylene glycol ether of chitosan, trisamine, urea, mannitol, d-panthenol, which can be used for oral hygiene, purification of periodontal pockets of purulent-necrotic masses, an increase in saliva lysozyme, treatment and prevention of inflammatory diseases of the dentition, caused by cariogenic microorganisms Anism, such as S. mutans, S. aureus and others.

SUMMARY OF THE INVENTION

According to the present invention, it was unexpectedly found that the introduction of toothpaste as the starting components: papain, lysozyme, collagenase, chymopsin, chitosan polyethylene glycol ether, trisamine, urea, mannitol, d-panthenol helps to purify periodontal pockets of purulent-necrotic masses, reduce inflammatory phenomena and accelerate the regeneration processes in the tissues of the dentition in inflammatory diseases.

Description of preferred embodiments

Urea, mannitol are introduced into toothpaste in a concentration of from 1.00 to 10.00 wt.%; chitosan d-panthenol and polyethylene glycol ether in a concentration of from 0.001 to 5.0 wt.%; trisamine in a concentration of from 0.001 to 4.0 wt.%.

Enzymes that can be used in the practice of the present invention are represented by proteases, hydrolyzing proteins and lysozyme. As proteolytic enzymes can be used proteases: papain, chymopsin and collagenase in a concentration of from 0.01 to 5.0 wt.% And, preferably, from 0.1 to 5.0 wt.%.

Lysozyme derived from egg white with a molecular weight of approximately 15,000 amu has an antimicrobial effect against gram-positive bacteria and to a lesser extent gram-negative, hydrolyzing fragments of the peptidoglycan wall of bacteria and can be used in a concentration of from 0.01 to 5.0 wt.% and, preferably, from 0.1 to 5.0 wt.% .

Toothpaste

When receiving toothpaste, chitosan polyethylene glycol ether, mannitol, d-panthenol and enzymes are mixed with the base of the toothpaste and get a product that helps purify periodontal pockets of purulent-necrotic masses, increase saliva lysozyme, reduce inflammation and accelerate the regeneration processes in the tissues of the dentition with inflammatory diseases.

Toothpaste base

Acceptable fillers used to make the toothpaste of the present invention are represented by a hydrophilic phase consisting of water and a humectant, which can be used propylene glycol, glycerin, sorbitol 70%, PEG with a molecular weight of from 200 to 400 amu, or mixtures thereof. The concentration of the humidifier is usually in the amount of from 5 to 10.00 wt.%. The water present in the toothpaste will be the amount remaining from 100 wt.%, After calculating the sum of the wt.% Of all components of the paste.

Of the abrasives, silica-based abrasives with an average particle size of 5 to 20 microns, for example Sorbosil AC 35; AC 39; TC 15 (Inneos Silicas), Zeodent 113: 163 (J.M. Huber) and others similar in characteristics. Other abrasives used in toothpastes are tricalcium phosphate, dicalcium phosphate dihydrate, and aluminum silicate. The concentration of abrasive is usually in the amount of from 2 to 20 wt.%.

Of the thickeners, natural and synthetic hydrocolloids can be used, such as carboxymethyl cellulose and its salts, hydroxyethyl cellulose, sodium alginate, sodium carrageenate, xanthan gum under various trade names. The concentration of thickener is usually in the amount of from 0.05 to 5.0 wt.%, And preferably from 0.1 to 2.0 wt.%.

Of the surfactants, various anionic, cationic and nonionic surfactants are allowed for use in oral hygiene products under various trade names. The concentration of surfactant is usually in the amount of from 0.1 to 2.0 wt.%, And preferably from 0.1 to 1.0 wt.%.

Of the fluorides, sodium fluoride, tin fluoride, sodium monofluorophosphate, aminofluoride with a fluoride ion concentration in the range from 0.05 to 0.15 wt.%, And preferably from 0.1 to 0.15 wt.%, Can be used.

Of the phosphates, sodium and / or potassium tetrapyrophosphates, disodium pyrophosphate, sodium polyphosphate in a concentration of from 0.5 to 5.0 wt.%, And preferably from 1.0 to 5.0 wt.%, Can be used.

Of the antibacterial substances, methyl and propyl parabens or their salts can be used in a concentration of from 0.15 to 0.4 wt.%, And preferably from 0.2 to 0.3 wt.%; chlorhexidine bigluconate in a concentration of from 0.05 to 0.2 wt.%.

Of the flavors, any flavoring additives permitted for use with a concentration in the range of from 0.05 to 1.5 wt.%), And preferably from 0.1 to 1.5 wt.%, Can be used.

Other components that can be incorporated into the composition of the toothpaste are desensitizing agents, whitening additives, dyes without significantly affecting the properties and characteristics of the toothpaste according to this invention.

Getting Toothpaste

The toothpaste according to this 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.

Examples

As an embodiment of the claimed invention, a toothpaste is proposed, the composition of the starting components of which includes, in wt.%: 0.01-5.0 papain, 0.01-5.00 lysozyme, 0.01-5.00 collagenase, 0.01 -5.0 chymopsin; trisamine 0.001-4.00, 1.00-10.00 urea; 1.00-10.00 mannitol; 0.001-5.00 d-panthenol; 0.001-5.00 polyethylene glycol ester of chitosan; 2.00-20.00 abrasive component; 5.00-10.00 humidifier; 0.05-5.0 hydrocolloid; 0.10-2.00 surfactant (surfactant); 0.10-0.32 sodium fluoride; 0.50-5.00 tetracal pyrophosphate; 0.15-0.40 preservative; 0.05-1.50 flavoring; water - the rest.

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

Papain 0.10-5.00 Abrasive component 2.00-20.00 Humidifier 5.00-10.00 Collagenase 0.1-5.00 Chymopsin 0.1-5.00 Trisamine 1.00-4.00 Mannitol 1.00-5.00 Urea 1.00-5.00 D-panthenol 1.00-5.00 Chitosan Polyethylene Glycol Ether 1.00-5.00 Hydrocolloid 0.10-2.00 Surfactant 0.10-1.00 Sodium Fluoride 0.05-0.32 Lysozyme 0.1-5.00 Tetracal pyrophosphate 1.00-5.00 Chlorhexidine bigluconate 0.05-0.20 Flavoring 0.10-1.50 Demineralized water Rest

The active ingredients of the claimed toothpaste are: papain, lysozyme, collagenase, chymopsin, mannitol, urea, chitosan polyethylene glycol ether, d-panthenol. Trisamine, mannitol and urea, having high osmolarity, contribute to the extraction of purulent-necrotic masses from the lesion. Trisamine, urea, chitosan polyethylene glycol ether and d-panthenol, due to their main properties, provide neutralization of acidic components of putrid masses and maintaining the optimum pH of the action of proteolytic enzymes. Papain hydrolyzes the peptide bonds of plaque, which leads to its softening and investment removal during brushing. Chymopsin and collagenase break down necrotic tissue and viscous secrets and exudates. D-panthenol accelerates the epithelization of damaged tissues.

Lysozyme derived from egg white with a molecular weight of approximately 15,000 amu is a factor of nonspecific immunity, has an antimicrobial effect against gram-positive bacteria and to a lesser extent gram-negative, hydrolyzing fragments of the peptidoglycan wall of bacteria.

Thus, the integrated use of the above active components leads to the following result: plaque modification with the formation of easily removable water-soluble compounds, normalization of the natural pH level and purification of damaged tissues from purulent-necrotic masses, activation of the immune system with the aim of antimicrobial and protection against pathogenic microorganisms, elimination of inflammatory phenomena and acceleration of tissue regeneration of the dentition. The following substances are used to prepare toothpaste: Papain, Collagenase, Himopsin, Lysozyme, Hitoxy-20, d-panthenol, trisamine, mannitol, urea with a basic substance content of at least 97%. An additional advantage of this composition is the ability to increase the content of lysozyme in saliva and oral fluid.

The proposed toothpaste reduces plaque formation by 61.53% and is completely harmless and safe for prolonged use.

The significant differences of the proposed toothpaste are as follows.

The claimed toothpaste has stability with respect to the activity of the enzymes included in its composition during long-term storage, which is achieved by immobilizing the 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 sodium alginate (FMC) and sodium carboxymethyl cellulose (Hercules).

The activity of the declared toothpaste is due to the activity of its constituents: papain, lysozyme, collagenase, chymopsin, mannitol, urea, polyethylene glycol ether of chitosan and d-panthenol. It was found that toothpaste, the composition of the starting components of which includes, in wt.%: 0.01-5.0 papain, 0.01-5.00 lysozyme, 0.01-5.00 collagenase, 0.01-5, 0 chymopsin; 1.00-10.00 urea; 0.001-4.00 trisamine; 1.00-10.00 mannitol; 0.001-5.00 d-panthenol; 0.001-5.00 polyethylene glycol ester of chitosan; 2.00-20.00 abrasive component; 5.00-10.00 humidifier; 0.05-5.0 hydrocolloid; 0.10-2.00 surfactant (surfactant); 0.10-0.32 sodium fluoride; 0.50-5.00 tetracal pyrophosphate; 0.15-0.40 preservative; 0.05-1.50 flavoring; water - the rest 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 diseases of the dentition.

The present invention is illustrated by the following examples.

EXAMPLE 1

Toothpaste for the treatment and prevention of inflammatory diseases of the dentition, for the manufacture of which papain, lysozyme, collagenase, chymopsin, trisamine, mannitol, urea, chitosan polyethylene glycol ether and d-panthenol and other cosmetically acceptable carriers or excipients are used:

Papain 0.10 Dicalcium Phosphate Dihydrate 2.00 Glycerol 5.00 Collagenase 0.1 Chymopsin 0.1 Mannitol 1.00 Trisamine 1.00 Urea 1.00 D-panthenol 1.00 Chitosan Polyethylene Glycol Ether 0.001 Sodium Carrageenate 0.50 Polysorbate 20 0.10 Sodium Fluoride 0.05 Lysozyme 0.1 Tetracal pyrophosphate 1.00 Chlorhexidine bigluconate 0.05 Flavoring 0.10 Demineralized water Rest

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

EXAMPLE 2

Toothpaste for the treatment and prevention of inflammatory diseases of the dentition, for the manufacture of which papain, lysozyme, collagenase, chymopsin, trisamine, mannitol, urea, chitosan polyethylene glycol ether and d-panthenol and other cosmetically acceptable carriers or excipients are used:

Papain 5.00 Silica 20.00 Sorbitol 70% 10.00 Collagenase 5.00 Chymopsin 5.00 Trisamine 4.00 Mannitol 5.00 Urea 5.00 D-panthenol 5.00 Chitosan Polyethylene Glycol Ether 5.00 Sodium CMC 2.00 PEG-40 hydrogenated castor oil 1.00 Sodium Fluoride 0.32 Lysozyme 5.00 Tetracal pyrophosphate 5.00 Methylparaben 0.15 Propylparaben 0.05 Flavoring 1,50 Demineralized water Rest

The specified composition is prepared according to the following method. In demineralized water, all components are sequentially dissolved except sodium CMC and silicon dioxide, then sodium CMC and silicon dioxide are added, mixed until a gel forms. The resulting toothpaste is mixed, vacuum and homogenized.

Assessment of the stability of enzymes in toothpaste

Since the main disadvantage of enzymes is their instability during long-term storage of ready-made forms based on them during the expiration date, a comparative assessment was made of the preservation of the activity of proteolytic enzymes: papain, chymopsin, collagenase during storage in the form of a solution and as a composition for local use.

The activity of proteolytic enzymes was determined by the following method.

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.

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; 94% chymopsin; 90% collagenase during 1 year of observation Pasta No. 2 According to Example 2 Activity: 82% for papain; 86% chymopsin; 80% collagenase within 1 year of observation Native enzyme solution Activity: 10% for papain; 12% chymopsin; 1% collagenase for 7 days

As can be seen from the table 1, the introduction of enzymes in toothpastes allows you to get stable cosmetic compositions for topical use in the prevention of inflammatory diseases of the dentition. 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 lysozyme was determined by the method of serial dilutions. 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 sarrel 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
ingredient Staphylococcus aureus Bacillus cereus Esherichia coli Candida albicans MIC, mg / ml mck, mg / ml MIC, mg / ml MCC, mg / ml MIC, mg / ml MCC, mg / ml MIC, mg / ml MCC, mg / ml Lysozyme 2 four 2 four twenty 40 thirty fifty

As can be seen from table 2, the optimal inhibitory concentration is: for lysozyme: 2 mg / ml in the case of Staphylococcus aureus, Bacillus cereus, Escherichia coli; 30 mg / ml for Candida albicans; bactericidal concentration for lysozyme - 4 mg / ml - in the case of Staphylococcus aureus, Bacillus cereus; 40 mg / ml for 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, 14th the 21st, 28th and 28th days. 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 _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 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 - hygiene index indicator 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 (1, 7, 14, 21, 28 days).

From tables 3, 4 and 5 it can be seen that the proposed toothpaste has an increased cleaning ability, and hence the anti-caries effect due to a more pronounced removal of dental plaque and provides removal of 61.53% 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 shown 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 probants disappeared from catarrhal gingivitis, and the PMA index decreased from 49.2 to 6.0, which corresponded to 87, 80% anti-inflammatory effectiveness 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 = [(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 2.6 to 1.0 points, which corresponds to 74.07% of the hemostatic effectiveness of this paste.

Thus, the proposed toothpaste has a higher cleansing (61.53% cleansing effectiveness), anti-inflammatory (87.80% anti-inflammatory effect) and hemostatic (76.92% hemostatic effectiveness) effect.

Table 3 condition OHI-S Green-Vermillion 1st day 7th day 14th day 21st day 28th day Before cleaning 2.6 2.0 1.7 1.12 1.00 After cleaning 1.3 1,1 1,0 0.83 0.77

Table 4 Plaque reduction with a single use of toothpaste 1st day 7th day 14th day 21st day 28-1 day 50.00% 45.0% 41.17% 25.89% 23.0%

Table 5 The cleaning effectiveness of the paste (compared with the 1st day) 7th day 14th day 21st day 28th day 23.00% 34.61% 56.92% 61.53%

Table 6 RMA Index 1st day 7th day 14th day 21st day 28th day 49.2% 27.8% 15.4% 10.00% 6.0%

Table 7 Anti-inflammatory efficacy 7th day 14th day 21st day 28th day 43.49% 68.69% 79.67% 87.80%

Table 8 Kechka bleeding index 1st day 7th day 14th day 21st day 28th day 2.7 2.1 1.7 1.4 0.7

Table 9 Hemostatic effectiveness 7th day 14th day 21st day 28th day 22.22% 37.03% 48.14% 74.07%

Claims (1)

Toothpaste containing papain, lysozyme, collagenase, chymopsin, trisamine, urea, mannitol, chitosan polyethylene glycol ether, d-panthenol, characterized in that the following components are used for its manufacture, wt.%:
Papain 0.01-5.00 Abrasive component 2.00-20.00 Humidifier 5.00-10.00 Collagenase 0.01-5.00 Chymopsin 0.01-5.00 Mannitol 1.00-10.00 Trisamine 0.001-4.00 Urea 1.00-10.00 D-panthenol 0.001-5.00 Chitosan Polyethylene Glycol Ether 0.001-5.00 Hydrocolloid 0.10-2.00 Surfactant 0.10-1.00 Sodium Fluoride 0.05-0.32 Lysozyme 0.01-5.00 Tetracal pyrophosphate 1.00-5.00 Chlorhexidine bigluconate 0.05-0.20 Flavoring 0.10-1.50 Demineralized water Rest