RU2824364C1 - Agent possessing anti-inflammatory and antimicrobial action for prevention and treatment of oral diseases - Google Patents

Abstract

FIELD: medicine; pharmaceutics.

SUBSTANCE: invention refers to pharmaceutics and medicine, particularly to dentistry, and can be used for treating and preventing dental diseases of the oral mucosa of various origins. Disclosed is an anti-inflammatory and antimicrobial agent in the form of a film obtained by dissolving 4.0 g of sodium carboxymethyl cellulose in 93.69–93.87 ml of purified water heated to temperature of 60 °C, with subsequent addition of 0.13 to 0.31 g of dry water extract of garden strawberry leaves and 2.0 g of glycerine into a film-forming base, thorough homogenisation for 10 minutes, uniform filling by 28.0 g taking into account the watering layer height of 5–7 mm on pre-weighed substrates and drying in a drying cabinet with a heater and forced air ventilation at temperature of 40–45 °C for 16 hours.

EFFECT: agent has independent antimicrobial and anti-inflammatory action, which provides its effectiveness in treating dental diseases of the oral mucosa.

1 cl, 2 tbl, 3 ex

Description

The invention relates to pharmaceuticals and medicine, in particular to dentistry, and can be used for the treatment and prevention of dental diseases of the oral mucosa of various origins.

One of the current problems of medical science and practical health care is the prevention and treatment of inflammatory diseases of the oral mucosa. This disease is characterized by all the signs of an inflammatory process: redness, pain, swelling, an increase in local and general body temperature. The inflammatory process can occur on the mucosa of any part of the oral cavity: on the gum, tongue, palatine mucosa, in the vestibule of the oral cavity and retromolar fossa, on the mucosa of the cheeks and lips. The disease can manifest itself in a variety of forms: it can be redness of the mucosa in a certain area or the appearance of ulcers in the oral cavity [1].

The importance of this problem is due to the frequency of occurrence and wide spread of the disease [2]. The problem of inflammatory diseases of the oral mucosa is one of the most pressing in dentistry. Inflammation accompanies many diseases of the oral mucosa and develops with any traumatic impact on the mucosa. In this regard, the search for and development of new effective and safe medicinal products and substances from plant materials that affect inflammation and activate regenerative processes in the treatment of diseases of the oral mucosa and have low toxicity are an urgent task in outpatient dental treatment of adults and children.

The choice of drugs for oral diseases depends on the severity of the inflammatory process, swelling, bleeding, the presence of epithelial defects, the stage of the process and destructive disorders. The inflammatory process in the oral cavity begins with its colonization by various microorganisms. By reducing the barrier function of the integumentary tissues, microbes contribute to the penetration and spread of toxic products, lytic enzymes [3]. In modern preventive dentistry, great importance is attached to creating a balance between the microflora and the natural defense systems of the oral cavity. In order to correct the composition of the microbial flora, broad-spectrum antiseptics have been proposed, most often chlorhexidine, triclosan, hexetidine, which are included in many hygiene products - rinses, gels, balms, toothpastes. Such products dominate and are widely advertised as effective in eliminating inflammation and regulating the composition of the microflora. However, despite the correction of the microbial composition as a result of the antiseptic, this group of drugs has an adverse effect on the tissues of the oral cavity and the composition of the oral fluid. Their long-term use leads to a violation of the resistant flora, the development of dysbiosis, taste sensitivity disorder and sometimes to allergic reactions, tissue staining.

For the normal functioning of the oral cavity defense mechanisms, preparations based on medicinal plant materials have been used since ancient times. Dentistry has accumulated experience in using medicinal plants to treat various types of pathology, since herbal medicine is highly effective, harmless, and rarely causes side effects and allergic reactions. Since plants contain biologically active substances such as vitamins, phytohormones, phytoncides, chlorophylls, microelements, essential and fatty oils, they affect metabolic processes, increase protective properties, and normalize homeostasis. Preparations of medicinal plants have anti-inflammatory, analgesic, immunomodulatory, hemostatic and wound-healing effects. At the same time, in terms of the effectiveness of antimicrobial and deodorizing effects, some of them are not inferior to synthetic agents [3]. In the presence of pathology (gingivitis, periodontitis, periodontosis), herbal preparations are an excellent addition to other remedies and are often used as components of complex therapy. Liquid hygiene products are used for therapeutic and prophylactic purposes. These include aqueous extracts from medicinal raw materials (infusions, decoctions), as well as tinctures and extracts in the composition of dental elixirs, rinses, balms, mouth fresheners. Many preparations can be used for rinsing the mouth, but rinses with tinctures of eucalyptus, sage, St. John's wort, peppermint, yarrow are considered the most effective [4]. The listed herbal products are suitable not only for rinsing the mouth, but also for applications, instillations, mouth baths. It is advisable to take these products as prototypes due to the presence of common pharmacological characteristics.

Prototypes (infusions, decoctions, tinctures) have the following disadvantages:

- lack of precise dosing;

- manifestation of short-term pharmacological activity (only during rinsing);

- inconvenience of use on the mucous membranes of the oral cavity due to the lack of adhesive properties of the infusion and, as a consequence, the impossibility of ensuring prolonged action of the active component on the area of the pathological focus due to regular washing with saliva;

- limited shelf life of aqueous extracts (2 days).

Effective means of local antimicrobial and anti-inflammatory therapy are films or self-absorbable plates. The basis of the self-absorbable plates "KP-Plast", "KPlast-phyto", "KP-Plast-vita", "KP-Plast-antimicrobial", "CM-1 Plates" is represented by natural polysaccharides. In addition, dental self-adhesive films "Diplen Denta" and collagen plates "Farmadont" are used. They are applied directly to the affected area of the mucosa, the release of medicinal components is prolonged, for different compositions lasts from 2 to 12 hours. The advantages of films are considered to be the accuracy of dosage, constant concentration of biologically active substances during the exposure time, strong fixation to wet and hard tissues, sufficient gas permeability for the normal course of reparative processes, ease of use, which allows independent use by patients. Good adhesion of films and plates is ensured by the polymer included in their composition (sodium carboxymethylcellulose, methylcellulose, gelatin, collagen, polyvinyl alcohol). The plates, in contact with wet gums, absorb saliva and gradually turn into a gel, fixing on the gum surface. However, the range of such drugs is very limited.

The aim of the present invention is to develop a composition of films with dry extract of garden strawberry leaves and to establish the anti-inflammatory and antimicrobial action of a new medicinal product based on plant raw materials.

The technical result of the claimed invention consists in achieving an increase in the effectiveness of treating inflammatory diseases of the oral mucosa by developing a composition of films with dry garden strawberry leaf extract and establishing anti-inflammatory and antimicrobial action in a new medicinal product based on plant raw materials that has a sufficient raw material base.

Garden strawberry leaves - Fragaria ananassa L. are used as a source of raw materials, from which a dry aqueous extract [5] is obtained at JSC Biokhimmash, Moscow, which serves as a substance for the development of a method for obtaining a new dosage form - a film. Dry extract of garden strawberry leaves is a powder from yellow-green to brownish-green in color with a faint odor and a bitter, slightly astringent taste. The content of flavonoids is 5.12 ± 0.17%, oxidizable substances - 82.3 ± 8.5%, ascorbic acid - 1.1 ± 0.12%, humidity - 3.5 ± 0.3% [6].

The said technical result is achieved by the fact that for the prevention and treatment of dental diseases of the oral cavity, a means is proposed in the form of films obtained by introducing dry garden strawberry leaf extract into a film-forming base with subsequent drying of the film mass until a film is formed. When introducing dry garden strawberry leaf extract into a watering solution, the "shrinkage" coefficient of the films is taken into account, which was 16. Based on this, the concentration of dry garden strawberry leaf extract in the watering solution was 0.13-0.31%, which allows the finished film to have a concentration of dry garden strawberry leaf extract within 2-5%. This concentration of dry garden strawberry leaf extract was introduced by us into a sodium carboxymethylcellulose solution, from which films with dry garden strawberry leaf extract were obtained by the watering method with the following ratio of components in wt.%:

Garden strawberry leaf dry extract 0.13-0.31

Sodium carboxymethylcellulose - 4.0

Glycerin - 2.0

Water purified to 100.0

Examples of specific compositions.

Example 1

Garden strawberry leaf dry extract 0.13

Sodium carboxymethylcellulose - 4.0

Glycerin - 2.0

Water purified to 100.0

Example 2

Garden strawberry leaf dry extract 0.22

Sodium carboxymethylcellulose - 4.0

Glycerin - 2.0

Water purified to 100.0

Example 3

Garden strawberry leaf dry extract 0.31

Sodium carboxymethylcellulose - 4.0

Glycerin - 2.0

Water purified to 100.0

In all the above examples, the films are obtained as follows:

93.70 - 93.87 ml of purified water are heated to a temperature of 60 °C. In parallel, 4.0 g of sodium carboxymethyl cellulose are weighed out. The water is transferred to a container for preparing a polymer solution with an anchor-type stirrer (speed 90-100 rpm). Then, while stirring, sodium carboxymethyl cellulose is poured onto the surface in small portions. The resulting solution is homogenized until the lumps disappear, after which 0.13-0.31 g of dry garden strawberry leaf extract is introduced into the polymer base, thoroughly homogenizing for 10 minutes. Lastly, 2.0 g of glycerol are introduced, homogenization is carried out for another 10-20 minutes. Deaeration of the polymer solution is carried out by settling the mixture for 30-40 minutes at room temperature. The polymer solution is evenly poured onto pre-weighed substrates. The substrates are filled with a polymer solution weighing 28 g. The height of the watering layer is 5-7 mm. Drying is carried out in a drying cabinet with a heater and forced air ventilation at a temperature of 40-45 ° C for 16 hours. The shrinkage coefficient was determined experimentally to be 16.

The study of antimicrobial activity was carried out in relation to the reference strains E. coli ATCC 25922, S. aureus ATCC 6538P, C. albicans 885-653 NCTC by the agar diffusion method in accordance with the principles of the State Pharmacopoeia XIV OFS 1.2.4.0010.18 [7].

Nutrient agar (State Research Center of Applied Microbiology and Biotechnology, Obolensk) was used to determine antibacterial activity; Sabouraud agar (State Research Center of Applied Microbiology and Biotechnology, Obolensk) was used to determine antifungal activity. Two-day cultures grown on Sabouraud agar were used to prepare a yeast culture suspension. Antibacterial activity was determined using typical daily cultures grown on nutrient agar. To prepare a working suspension of microorganisms, the grown culture was washed with an isotonic sodium chloride solution and the optical density of the microbial suspension was set at 0.5 McF. This suspension was sown on the corresponding nutrient media as a lawn. Sterile cylinders of uniform size and weight, (10±1) mm in height and (6.0±0.1) mm in internal diameter, made of stainless steel or aluminum, were placed on the surface of the seeded medium at equal distances from each other and from the edge of the dish.

The test samples of films of medicinal with extract of garden strawberry leaves dry with an area of about 1 sq. cm, moistened with isotonic solution of sodium chloride were introduced into the wells of each dish. The dishes were placed in a thermostat at a temperature of 36±1°C for 18-20 hours. The presence of antimicrobial activity was judged by measuring the diameter of the average zone of growth inhibition in millimeters. The results are given in Table 1.

Table 1
Antimicrobial activity of medicinal films with dry garden strawberry leaf extract Microorganism growth inhibition zone, mm Control (Calendula ointment) Films with extract concentration of 1% Films with extract concentration of 2% Films with extract concentration of 5% E. coli ATCC 25922 10.3 + 0.6* 10.3 + 0.6* 10.3 + 0.6* 10.3 + 0.6* S. aureus ATCC 6538P 10.3 + 0.6* 16.6 + 2.8* 18.0 + 2.0* 17.3 + 1.1* C. albicans 885-653 NCTC 10.3 + 0.6* 10.3 + 0.6* 10.3 + 0.6* 10.3 + 0.6* * - standard deviation

The results of the study of antimicrobial activity showed that this effect is most pronounced in films of medicinal products with a concentration of dry garden strawberry leaf extract of 2%. The degree of antimicrobial activity is most pronounced in relation to the studied strain of S. aureus. The antimicrobial activity of films with dry garden strawberry leaf extract is significantly higher than the activity of the ointment "Calendula" (p < 0.02).

Determination of anti-inflammatory activity is based on the assessment of the increase in the volume of the inflamed foot of rats, caused by the introduction of a phlogogenic agent.

The study was conducted on nonlinear rats of both sexes, sexually mature, weighing 200-250 g, of both sexes (the group included 6 animals) on the model of acute inflammatory edema caused by subplantar injection of 0.1 ml of 1% aqueous carrageenan solution into the hind paw of the rat. The increase in foot volume, indicating the development of edema, was assessed oncometrically [8] before the injection and 3 hours after the injection of carrageenan. The objects under study in the amount of 0.3 g were applied to the skin of the foot 0.5 hour before the injection of the phlogogenic agent. The animals that did not receive the drug served as a control. Statistical processing was performed using the Student's method. Based on the results obtained, the effect of inflammation inhibition was determined as a percentage of the control level. The presence of an anti-inflammatory effect was judged by the severity of the inhibition of the inflammatory reaction. If this indicator was greater than 30%, the result was considered positive. The study results are presented in Table 2.

Table 2
Anti-inflammatory activity of films with dry garden strawberry leaf extract Preparation Increase in foot volume
after 3 hours, % Inhibition of reaction
after 3 hours, % Films with strawberry extract (Gel - non-dried base) 38.2 ± 10.6 *
p < 0.01
p'> 0.05 50.1 Ointment "Calendula" 35.8 ± 4.4 *
p < 0.001 35.8 Indomethacin ointment 13.7 ± 1.7
p < 0.001 82.1 Control 76.6 ± 7.2 p - level of statistical significance of differences compared to the control;
p' - the level of statistical significance of differences in comparison with Calendula ointment;
* - level of statistical significance of differences in comparison with Indomethacin ointment

The studied dosage form showed anti-inflammatory activity in the model of carrageenan inflammation. Compared with the control results, the severity of inflammatory edema when using medicinal films decreases and is statistically significant. The anti-inflammatory activity of films with dry garden strawberry leaf extract does not significantly differ from the activity of the ointment "Calendula". At the same time, the ointment "Indomethacin" acts more actively than all the studied objects.

Thus, films with 2% dry garden strawberry leaf extract have anti-inflammatory activity. The anti-inflammatory activity of films with dry garden strawberry leaf extract is comparable to that of Calendula ointment.

As follows from the data provided, films with dry garden strawberry extract have independent antimicrobial and anti-inflammatory effects.

The proposed method for producing films, in comparison with known prototypes, allows obtaining a preparation of constant composition with antimicrobial and anti-inflammatory effect for use on the mucous membranes of the oral cavity, providing prolonged action of the active component on the area of the pathological focus; allows control over the technological stages of production and standardization of the target product; allows obtaining a preparation that is convenient for storage and transportation.

The above-mentioned advantages, combined with the proposed production scheme, contribute to a rational method of using medicinal plant raw materials, expanding the range of herbal medicines with antimicrobial and anti-inflammatory activity and open up the prospect of introducing this method into the pharmaceutical industry.

List of references.

1. Novikov O. O., Zhilyakova E. T., Tsimbalistov A. V., Trifonov B. V., Malyutina A. Yu., Pisarev D. I. A new look at the problem of prevention and treatment of periodontal diseases // Scientific results of biomedical research. - 2016. - V. 2. - No. 3. - P. 64-69.

2. Suerkulov E. S., Yuldashev, I. M., Mamyraliev, A. B., Toktosunova, S. A., Tsepeleva, A. S., Sooronbaev, A. A. Prevalence and structure of the incidence of stomatitis in children / / Bulletin of science and practice. - 2018. - T. 4. - No. 11. - pp. 91-96.

3. E. I. Goncharova Herbal remedies in the prevention and treatment of periodontal diseases // RUSSIAN DENTAL JOURNAL, No. 3, 2012. - P. 48-52.

4. State Register of Medicines: website “RLS® Medicine Directory” [Electronic resource] - access mode https://grls.rosminzdrav.ru/grls.aspx.

5. Oleshko G.I., Petukhova O.V., Yushkov V.V., Blinova O.A., Marchenko S.D. Method for obtaining a drug with diuretic and anti-inflammatory activity // Russian Federation Patent No. 2342945. Patent holder: State Educational Institution of Higher Professional Education PGFA of the Russian Health Ministry. 2009. Bulletin No. 1.

6. Yaborova O.V., Sosnina S.A., Turyshev A.Yu. Selection of the optimal technology for obtaining garden strawberry leaf extract / Perm State Pharmaceutical Academy of the Ministry of Health of the Russian Federation, Perm, Russian Federation // Medical and Pharmaceutical Journal "Pulse" - 2021.

7. State Pharmacopoeia of the Russian Federation [Electronic resource]. - 14th ed. - Moscow, 2018. - Volume 1, 2, 3, 4. - Access mode: http://www.femb.ru/feml

8. Guidelines for conducting preclinical studies of medicinal products / edited by Mironov A.N. - M.: Grif i K, 2012. - 944 p.

Claims (1)

A product for the treatment and prevention of dental diseases of the oral mucosa, with anti-inflammatory and antimicrobial action, in the form of a film obtained by dissolving 4.0 sodium carboxymethylcellulose in 93.7-93.87 ml of purified water heated to a temperature of 60°C, followed by the addition of 0.13 to 0.31 g of dry aqueous extract of garden strawberry leaves and 2.0 g of glycerin into a film-forming base, thoroughly homogenized for 10 minutes, uniformly poured into 28.0 g, taking into account the height of the watering layer of 5-7 mm, on pre-weighed substrates and dried in a drying cabinet with a heater and forced air ventilation at a temperature of 40-45°C for 16 hours.