RU2552330C1 - Multi-component melatonine-containing preparation for oral tissue regeneration and method for producing it - Google Patents

Abstract

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to pharmaceutical industry, namely to a method for producing a preparation applicable for oral tissue regeneration. A method for producing a multi-component melatonine-containing preparation for oral tissue regeneration, involving incubating eggs, cooling them, performing homogenisation and ultramicrofiltration; the incubation procedure is performed for 5-6 days; the ultramicrofiltration procedure is added with 0.1% sodium benzoate; the prepared substance of embryo-egg mass is added with a second substance containing medical gelatine, sage and St. John's wort tea, melatonine, eucalyptus oil, glycerol and prepared according to certain procedure that is followed by mixing in a homogeniser; the produced mass is bottled, placed in a thermostat, cooled until film hardened; the films are cut in strips 1 cm wide and packed into plastic package. The multi-component melatonine-containing preparation for oral tissue regeneration.

EFFECT: preparation prepared by the method described above is effective for oral tissue regeneration.

3 cl, 4 ex

Description

FIELD OF THE INVENTION

The present invention relates to biotechnology and medicine, and in particular to methods for producing a biologically active drug with a regenerative stimulating effect, and can be used in dentistry to accelerate tissue repair in inflammatory and inflammatory-destructive processes.

State of the art

The oral cavity is constantly exposed to adverse factors (physical, chemical, biological), which lead to dental diseases of both inflammatory and non-inflammatory nature. Features of the course, duration, outcome and success of treatment and preventive measures depend on the ratio of the degree of damage and the state of compensatory-adaptive processes, which include regeneration. Moreover, the level of regeneration can lag significantly even against the background of positive dynamics of the intensity of damage reactions, which is especially often observed in chronic pathological processes. In this regard, the struggle for a high regenerative potential and an adequate level of damage to the reparative activity of the tissues of the oral cavity today has become one of the central tasks of the dentist.

The mechanisms of the pathogenesis of damage and regeneration processes are unified and among them have recently been distinguished: immunological and metabolic. The leading links in the provision and regulation of these mechanisms is redox, stress-adaptive reactions, providing homeostasis, characterizing the morphofunctional usefulness of the tissue. At the same time, it is necessary to highlight the priority role of factors that primarily determine the high level of the proliferative activity of tissue elements (antioxidants, cellular metabolites, adaptogens and direct regulators of cell division). Therefore, to optimize the balance between damage and regeneration of the tissues of the oral cavity, it is advisable, if possible, to act simultaneously, if not all, then at least the vast majority of the above processes and factors of etiopathogenesis. For the full restoration of the morphofunctional usefulness of the tissues of the oral cavity, activation and significant acceleration of the repair, the use of regenerative stimulating agents is required. In this case, optimal results can be achieved either by combining various drugs with a multidirectional etiopathogenetic effect, or by using innovative complex etipatogenetically multicomponent drugs.

A known method of obtaining a dental gel with chlorhexidine and dibunol for the treatment of inflammatory diseases of periodontal and oral mucosa. The gel contains chlorhexidine gluconate in the form of a 20% solution and dibunol as active substances, a gel base including polyethylene glycol 400, glycerin, liquid paraffin, cremophor RH-40, aerosil and water purified at a certain ratio of components (see US Pat. 2 395 292 C1, publ. 06/10/2011). However, the drug is limited mainly by antibacterial and antioxidant effects. In addition, the use of a strong antiseptic chlorhexidine gluconate does not exclude the suppression of saprophytic microflora, which can lead to a decrease in the immunoresistance of oral tissues.

A known method of obtaining a gel composition for the treatment of dental diseases containing an active component, chlorhexidine gluconate, a mucoadhesive reagent, a gelling agent and a pharmaceutically acceptable diluent, characterized in that diclofenac is used as an active component in the form of a salt in the following components, wt. %: chlorhexidine gluconate (0.05-2.0), mucoadhesive reagent (1.0-20.0), diclofenac (1.0-5.0), gel-forming agent (0.2-7), a pharmaceutically acceptable solvent ( the rest) (see Pat. RU 2 288 699 C2, publ. 10.12.2006). The disadvantage of the drug, which has a powerful antiseptic and anti-inflammatory effect, along with the above is the lack of components covering other regenerative-stimulating mechanisms. In addition, the content of a strong anti-inflammatory component (diclofenac) does not exclude the side effect of the drug on the body.

The local use of melatonin in dentistry is known in the form of an application on the gum of a 0.05% chlorhexidine solution with the addition of a 0.1% solution of melaxen (melatonin) on turundas in periodontal pockets. The effect of the inclusion of melaxen is manifested by a mild suppression of inflammatory processes in the periodontium, antioxidant effect, immunological effect (Elbekyan K.S., Markarova G.V. Application of the drug melaxen in the treatment of inflammatory diseases of the oral cavity / Physicochemical Biology: Mat. International Internet Conference . - Stavropol, 2012 .-- S. 21-23). However, such use of the solution, despite the positive effect, is limited by the conditions of dental procedures, which exclude the possibility of its use by patients on their own and insufficient prolongation of action.

A known method of producing a film that provides prolonged contact with a pronounced anti-inflammatory, bacteriostatic, antioxidant, membrane-stabilizing and stimulating regeneration action, by triple extraction of cocoa leaves with spear-shaped 70% ethanol for 1.5 hours each time at 96-98 ° C at a ratio raw materials: extractant (1: 16-18) followed by combining the extracts, distillation of ethanol, evaporation and drying of the intermediate in a vacuum dryer, grinding the intermediate with a mixture of Tween 80-dime tilsulfoxide-glycerin in a ratio of 1: 1: 10 and then with a 5% solution of carboxymethyl cellulose in a ratio of 1: 5 and a 10% solution of gelatin in a ratio of 1: 5, pouring into a mold and drying 20-22 ° C (see Pat. RU 2395292 C1, publ. 07.27.2010).

This drug has a pronounced healing effect, good penetration of components into periodontal tissues and the oral mucosa, however, it has a rather complicated production technology.

The closest analogue of the claimed invention is a method for producing a biologically active substance from an embryonic egg mass for the preparation of an anti-burn plate and an anti-burn plate based on it having a multicomponent regenerating stimulating effect on integumentary tissues in violation of their integrity. The method includes homogenizing a chicken egg taken on the 9th day of development, followed by cooling for 6-7 days at a temperature of 2-4 ° C, and after homogenization, the homogenate is treated with ultrasound, then microfiltration is carried out through filters with a diameter of 0.2 μm, and before ultramicro-filtration to the obtained homogenate 1: 1 add a mixture of a 0.1% solution of sodium benzoic acid and a 0.1% solution of potassium sorbic acid in equal parts, in addition prepare a homogeneous mass containing medical gelatin, a decoction alendula and chamomile, antibiotic agents, anesthesin, amaranth oil, glycerin, brilliant green (see US Pat. RU 2433171 C1, publ. 10.11.2011).

However, this technology of obtaining a biologically active substance from the embryonic egg mass on the 9th day of incubation does not provide the maximum yield of biologically active substances, in particular alpha-fetoprotein, which seems to be significant for the regeneration of oral tissue and, due to its low molecular weight, will penetrate the mucous membrane the oral cavity. In addition, the maximum concentration of total protein capable of providing high-quality occlusion occurs at earlier incubation times.

Disclosure of invention

The problem to which the invention is directed, is to expand the arsenal of methods for producing drugs aimed at stimulating regeneration processes that ensure rapid restoration of the organotypicity of the tissues of the oral cavity. Its solution is to create a multicomponent melatonin-containing preparation for the regeneration of the oral cavity, which would maximally affect the main links of the regeneration mechanisms, was enclosed in an optimal form that ensures long-term contact and delivery of the active substances of the drug in the tissues of the oral cavity.

The technical result is achieved using the method of obtaining a multicomponent melatonin-containing preparation for the regeneration of oral tissue, including the preparation of gelatin films based on two obtained substances.

The first substance is prepared by selecting fresh, non-infected chicken eggs, incubated for 5-6 days in order to obtain an embryonic egg mass containing almost the entire complex of biologically active substances that positively affect most cells and tissues. The quantitative ratio used for further processing according to the method of V.P. Filatova of embryonic and extraembryonic tissues should be 1: 4, which leads to a high content of vitamins, amino acids, lipids, and high molecular weight proteins in extraembryonic tissues, which, being included in the surface layers of the film, thereby increasing occlusion, inhibit fluid removal and increase the permeability of mucous tissue shells of the oral cavity for other components that make up the film.

In order to include biogenic stimulants in the preparation, embryos incubated for up to 5-6 days are placed in unfavorable conditions for survival in the cold chamber at a temperature of 2-8 ° C for 6-7 days, followed by extraction and homogenization of all embryonic and extraembryonic tissues.

The second substance is prepared by dissolving in the broth of sage and St. John's wort medical gelatin, glycerin, eucalyptus oil and melatonin with thorough mixing to a homogeneous mass. The addition of melatonin to the substance is due to its properties as a stimulant of metabolic processes, a powerful natural antioxidant and immunomodulator, a substance with a good antitoxic effect. The structure and properties of melatonin allow it to easily penetrate through cell membranes, which suggests the presence of its intracellular effect. Its inclusion enhances the immunomodulatory, regenerative, stimulating and antioxidant effects of the embryonic egg mass. The inclusion of eucalyptus oil is justified by its pronounced antimicrobial, antiviral and fungicidal action, as well as anti-inflammatory, astringent, wound healing and analgesic effect when applied locally to the mucous membranes.

The addition of St. John's wort herb extract is justified by its multilateral pharmacological properties, among which, according to local action, anti-inflammatory, hemostatic, analgesic, antiseptic and wound healing are pronounced in the oral cavity. The inclusion of sage extract in the substrate has the additional function of a binder and tanning component.

The presence of a gelatin-glycerin composite in the preparation performs the structural function of creating a dosage form in the form of films with elasticity and adhesion. Gelatin also serves as a depot of natural healing substances, which determines their long-term and dosed intake into the tissues of the oral cavity.

Then, 48.0-50.0 ml of the first substance is added to 100 ml of the second substance and mixed in a homogenizer for 6-7 minutes. The resulting mass is poured into 15.0 ml in a special container with sides 10 cm and a height of 1 cm, placed in a thermostat for 3.5-4 hours at a temperature of 37-39 ° C, then cooled for 1-2 days until solidified in the form films, then cut the film into strips 1 cm wide and packaged in a plastic sheath. The advantages of gelatin drug films when applied to the mucous membrane of the oral cavity are their strong fixation in the pathology zone. Such contact provides targeted delivery of drugs and allows to achieve a therapeutic effect in doses 10-20 times lower than in traditional dosage forms. Gelatin drug films slowly dissolve in the secrets of the mucosa, releasing a drug substance within 3-5 hours.

The essence of the method of obtaining a multicomponent melatonin-containing preparation for oral tissue is as follows. Fresh uninfected chicken eggs are selected, then they are incubated for 5-6 days, followed by cooling for 6-7 days at a temperature of 2-4 ° C, and after homogenization ultramicrofiltration is carried out through filters with a diameter of 0.2 μm, while before ultramicro-filtration to the resulting homogenate add a 0.1% solution of sodium benzoate, thus obtaining a substance from the embryonic egg mass.

To obtain the second substance in the preparation of a multicomponent melatonin-containing film, take 3.5-4.0 g of medical gelatin, pour 1 to 1 ml of sage broth and St. John's wort broth, incubate for 55-60 minutes at a temperature of 20-25 ° C , then placed in a water bath at a temperature of 65-70 ° C until gelatin is completely dissolved, after which 1.5-2.0 g of melatonin, 0.9-1.0 ml of eucalyptus oil, 2.5- are added to the solution with gelatin 3.0 ml of glycerol, followed by stirring in a homogenizer for 4-5 minutes, then to the resulting homogeneous mass adj 48.0-50.0 ml of the substance from the embryo-egg mass are mixed and mixed in a homogenizer for 6-7 minutes, the resulting mass is poured into 15.0 ml into a special container with a side of 10 cm and a height of 1 cm, placed in a thermostat 3.5-4 hours at a temperature of 37-39 ° C, then cooled for 1 to 2 days to solidify in the form of films, then the films are cut into strips 1 cm wide and enclosed in a plastic bag.

Identified distinguishing features allow us to conclude that the proposed method for obtaining the criterion of "novelty."

The implementation of the invention

The method is illustrated by the following examples of the preparation of a multicomponent melatonin-containing preparation for stimulating the processes of tissue regeneration of the oral cavity.

Example 1. A selection of fresh uninfected chicken eggs is carried out, then they are incubated for 6 days, followed by cooling for 6-7 days at a temperature of 2-4 ° C, and after homogenization ultramicrofiltration is carried out through filters with a diameter of 0.2 μm, while Before ultramicro-filtration, a 0.1% sodium benzoate solution is added to the resulting homogenate, thereby obtaining a substance from the embryonic egg mass.

To obtain the second substance in the preparation of a multicomponent melatonin-containing film, take 2.0 g of medical gelatin, pour 1 to 1 ml of sage broth and St. John's wort broth, incubate for 45 minutes at a temperature of 20-25 ° C, then place in a water bath at a temperature of 65-70 ° C until gelatin is completely dissolved, after which 0.5 g of melatonin, 0.5 ml of eucalyptus oil, 1.5 ml of glycerin are added to the solution with gelatin, followed by stirring in a homogenizer for 3 minutes, then to the resulting 30.0 ml are added to a homogeneous mass substances from the embryo-egg mass and mixed in a homogenizer for 6-7 minutes, the resulting mass is poured 10.0 ml into special dishes with a side of 10 cm and a height of 1 cm, placed in a thermostat for 4 hours at a temperature of 37-39 ° C , then cooled for 1-2 days to solidify in the form of films, then the films are cut into strips 1 cm wide and enclosed in a plastic bag.

The films are stored in a dry place at a temperature of plus 2-4 ° C, but the films are too thin, easily torn.

Example 2. The preparation of the substance from the embryo-egg mass is carried out analogously to example 1. To obtain the second substance in the preparation of the drug, take 3.0 g of medical gelatin, pour broth of sage and broth of St. John's wort to 100 ml in a ratio of 1: 1, incubated for 55 minutes at a temperature of 20-25 ° C, then placed in a water bath at a temperature of 65-70 ° C until gelatin is completely dissolved, after which 1.0 g of melatonin, 0.7 ml of eucalyptus oil, 2.0 ml of glycerin are added to the solution with gelatin followed by stirring in a homogenizer for 3.5 minutes, then 45.0 ml of the substance from the embryo-egg mass is poured to the obtained homogeneous mass and mixed in the homogenizer for 6-7 minutes, the resulting mass is poured into 20 ml in special dishes with a side of 10 cm and a height of 1 cm, placed in a thermostat for 4 hours at a temperature of 37-39 ° C, then cooled for 1-2 days to solidify in the form of films, then the films are cut into strips 1 cm wide and enclosed in a plastic bag.

However, the resulting films are too thick, poorly solidify, easily break.

Example 3. The preparation of the substance from the embryo-egg mass is carried out analogously to example 1. To obtain a homogeneous mass in the preparation of a multicomponent melatonin-containing preparation, take 3.5 g of medical gelatin, pour sage broth and hypericum decoction to 100 ml in a ratio of 1: 1, incubated for 1 hour at a temperature of 20-25 ° C, then placed in a water bath at a temperature of 65-70 ° C until the gelatin is completely dissolved, after which 1.5 g of melatonin, 1.0 ml of eucalyptus oil, 3.0 are added to the solution with gelatin ml of glycerol followed by by stirring in the homogenizer for 5 minutes, then 50.0 ml of substance from the embryo-egg mass is poured to the resulting homogeneous mass and mixed in the homogenizer for 6-7 minutes, the resulting mass is poured into 15.0 ml into a special container with a side of 10 cm and a height of 1 cm, placed in a thermostat for 3.5 hours at a temperature of 37-39 ° C, then cooled for 1-2 days to solidify in the form of films, then the films are cut into strips 1 cm wide and enclosed in a plastic bag.

The obtained multicomponent melatonin-containing films have an optimal thickness and are convenient to apply to the oral mucosa.

Example 4. The preparation of the substance from the embryonic egg mass is carried out analogously to example 1. To prepare a multicomponent melatonin-containing preparation from the activated embryonic egg mass, 4.0 g of medical gelatin are taken, poured with sage broth and hypericum hypericum to 100 ml in a ratio of 1: 1, maintained for 1 hour at a temperature of 20-25 ° C, then placed in a water bath at a temperature of 65-70 ° C until gelatin is completely dissolved, after which 2.0 g of melatonin, 2.0 ml of eucalyptus oil are added to the solution with gelatin, 4 , 0 ml of glycerol with followed by stirring in the homogenizer for 5 minutes, then 70.0 ml of substance from the embryo-egg mass is poured to the resulting homogeneous mass and mixed in the homogenizer for 6-7 minutes, the resulting mass is poured into 15.0 ml into a special container with side 10 cm and a height of 1 cm, placed in a thermostat for 4 hours at a temperature of 37-39 ° C, then cooled for 1-2 days to solidify in the form of a film, then the films are cut into strips 1 cm wide and enclosed in a plastic bag.

However, the resulting preparation is more expensive, as it contains a large amount of fetal egg mass and all other components.

Thus, the most optimal is the cooking method, reflected in example 3.

Test results of a multicomponent melatonin-containing preparation prepared according to the technology of Example 3 on volunteers in a UTA-Dent clinic (Stavropol) The drug was tested in a UTA-Dent clinic in Stavropol on patients with various dental pathologies. An agreement of informed consent was concluded with patients with the UTA-Dent clinic. The following examples illustrate the present invention.

Clinical example 1

Patients: 18 people, age from 37 years to 62 years.

Diagnosis: secondary partial adentia complicated by periodontitis of moderate severity.

Treatment: after complete rehabilitation of the oral cavity, all patients. A 10-fold application of the drug was carried out to stimulate the regenerative potential of the tissues of the oral cavity for 10 days 1 time per day (for each patient). In the third procedure, hyperemia, discharge from the gingival groove, and halitosis disappeared. Inspection 30 days after the end of the procedures, the result is a stable remission.

Clinical example 2

Patients: 23 people, age from 32 years to 49 years.

Diagnosis: cervical caries, wedge-shaped defects of the frontal group of teeth of both jaws.

Treatment: during rehabilitation, a retraction thread was introduced, subgingival polishing was carried out, immediately after polishing, the application of a multicomponent melatonin-containing preparation to stimulate the process of tissue regeneration of the oral cavity. Re-apply the film at bedtime at home.

The result the next day - the gum is intact.

Clinical example 3

Patients: 12 people, age from 30 years to 70 years. Diagnosis: complete and partial loss of teeth.

Treatment: wearing plate and clasp prostheses with preliminary application to the mucous membrane of the gingival part of the drug from 5 to 12 days (once a day) during the addiction period.

Result: epithelization of decubital ulcers on the third day, on the fifth day (after correction of the prosthesis) the disappearance of hyperemia and swelling of the tissues of the prosthetic bed.

Thus, a multicomponent melatonin-containing drug has established itself with the following properties:

- anti-inflammatory and antimicrobial;

- local immunomodulatory and antioxidant;

- strengthens the walls of blood vessels and stops bleeding gums;

- restores metabolic processes in cells;

- Stimulates the division and differentiation of lost cells. Clinical results indicate the feasibility of using the proposed method for producing a multicomponent melatonin-containing preparation and its effectiveness in order to increase the potentials of damaged tissues of the oral cavity for regeneration, which can significantly increase efficiency and significantly reduce treatment time without performing unnecessary and often expensive procedures.

The proposed method is simple, easily reproducible, does not require significant material costs and the use of complex equipment. The multicomponent melatonin-containing preparation obtained according to this method is capable of providing reliable stimulation of the processes of tissue regeneration of the oral cavity and can be widely used in practical dentistry.

Claims (2)

1. A method of obtaining a multicomponent melatonin-containing preparation for the regeneration of oral tissue, including incubation of eggs with subsequent cooling for 6-7 days at a temperature of 2-4 ° C, homogenization and ultramicro-filtration, characterized in that the incubation of eggs is carried out for 5-6 days , before ultramicro-filtration, a 0.1% sodium benzoate solution is added to the homogenate, a second substance containing medical gelatin, a sage broth, and s is added to the obtained substance from the embryonic egg mass. Veroboy, melatonin, eucalyptus oil, glycerin, while to obtain a second substance, pre-medical gelatin in an amount of 3.5-4.0 g is poured with a sage and St. John's wort broth to 100 ml in a ratio of 1: 1, kept for 55-60 minutes at temperature of 20-25 ° C, then placed in a water bath at a temperature of 65-70 ° C until gelatin is completely dissolved, after which 1.5-2.0 g of melatonin, 0.9-1.0 ml of oil are added to the solution with gelatin eucalyptus, 2.5-3.0 ml of glycerol, followed by stirring in a homogenizer for 4-5 minutes, then to the resulting homogeneous mass, pr pour 48.0-50.0 ml of substance from the embryo-egg mass and mix in the homogenizer for 6-7 minutes, the resulting mass is poured into 15 ml into a container with sides 10 cm and a height of 1 cm, placed in a thermostat by 3.5 -4 hours at a temperature of 37-39 ° C, then cooled for 1-2 days to solidify in the form of films, then cut the film into strips 1 cm wide and Packed in a plastic bag. 2. A multicomponent melatonin-containing preparation for the regeneration of oral tissue, prepared by the method according to p. 1.