RU2693228C2 - Composite for accelerated healing of wounds of various aetiologies, use of composite as cosmetic agent and as a therapeutic agent in veterinary science, agent for skin regeneration based on composite - Google Patents

Abstract

FIELD: medicine; pharmaceuticals.SUBSTANCE: group of inventions relates to the pharmaceutical industry and is a composite for accelerated wound healing of various aetiology, containing ionol, carnosine, colloidal silver, lecithin, triethanolamine, carbopol and water, wherein the components in the composite are in a certain ratio, wt. %, as well as an agent for skin regeneration containing said composite.EFFECT: invention provides a wider range of therapeutic and preventive agents for accelerated wound healing of various aetiologies and for skin regeneration, does not cause allergic and irritant reactions, has compatibility with traditional means and methods of therapy and prevention and has the possibility of local application.4 cl, 9 ex, 6 tbl, 8 dwg

Description

The invention relates to the related fields of medicine and pharmacology, in particular, to the creation of means of accelerated healing of wounds of various etiologies using composites developed on a rational basis, as well as for use in veterinary medicine and medical cosmetics. The invention also relates to medicine, in particular to purulent surgery, and can be used to treat purulent wounds and burns.

It is known that during thermal lesions the skin and underlying tissues become a powerful source of free radicals, which in the first hours after injury leads to the excessive formation of reactive oxygen species and a sharp intensification of POL processes in the internal organs of the injured. Cytolysis and activation of POL in the areas of ischemia adjacent to the area of necrosis in the wound, cause the active release of the products of these reactions into the plasma. In the process of burn shock development, an excess of ROS and LPO products leads to the breakdown of the body’s antioxidant protection - the development of “oxidative stress, a sharp decrease in the level of endogenous antioxidants and disruption of the functioning of internal organs, primarily the kidneys, subjected to ischemia, reperfusion, toxic damage by the products of tissue breakdown and increased load due to violations of water and electrolyte balance and lungs, due to the increase in lung blood volume due to the high centralization of blood circulation.

It is also known that the local treatment of burns, despite the many existing methods, is still a difficult and not fully resolved problem. Therefore, the search for tools and methods intended for the local treatment of burns seems to be relevant and promising. The system of local treatment of burns plays a significant role in ensuring an uncomplicated course of burn disease in general and a burn wound in particular. Adequately selected drugs for the local treatment of burn wounds reduce the influence of factors contributing to the deepening of necrosis, such as tissue ischemia, infection of wounds, and create optimal conditions for regeneration. .

The objective of the invention is the creation of a new composite for the treatment of wounds of various origins, increasing efficiency and reducing the time of wound treatment.

The problem is solved by the proposed composite for accelerated healing of wounds of various etiologies.

The technical result of the invention is to expand the Arsenal of therapeutic and prophylactic agents for the integrated treatment and prevention of inflammatory skin diseases, such as burns, gunshot wounds, pressure sores, nonhealing wounds, etc., which do not cause allergic and irritating reactions compatible with traditional means and methods of therapy and prevention, and with the possibility of local application.

The above technical result is achieved by a composite, hereinafter referred to as “Leran”, containing three active components with high antioxidant and antiradical activities, with wound healing, antibacterial, antiseptic and anti-inflammatory activities, each component enhancing the effect of the other components of the composite, and the stability and uniformity of distribution of the components of the composite fixed gel base, with the following ratio of components, in percent:

-ionol 4,5-5,5

-L-carnosine - 1.5 -2.5

- colloidal silver - 2.5-3.5

- lecithin -1 - 1.5

- triethanolamine - 1 -1.5

- Carbopol - 50-51

- water - the rest.

The task is also solved by the use of the inventive composite as a cosmetic in cosmetic practice.

In addition, the problem is solved by using the composite as a therapeutic agent in veterinary medicine.

One aspect of the invention is a means for regeneration of the skin, containing the claimed composite.

Below are descriptions of the constituent components of the composite.

Ionol (Dibunol).

Physico-chemical characteristics of ionol. 2,6-di-tert-butyl-4-methylphenol (ionol, dibunol) is a synthetic sterically hindered phenol, a derivative of 2,6-di-tert-butyphenol, as a chemical substance known since 1921 and, by its basic biological effect, direct antioxidants. Antioxidant properties of the drug are associated with its ability to bind the active forms and compounds of oxygen to form a stable phenoxyl radical, not participating in the chain of oxidative transformations and interrupting the oxidation chain in the substrate. It is well absorbed from the surface of the skin, accumulates in tissues rich in lipids and is determined in the lesion in concentrations sufficient for the manifestation of a therapeutic effect - 32-86 mg / kg. Deeply penetrates the skin, protects it from the harmful effects of the external environment. Reduces wrinkles around eyes. Reduces puffiness of eyes, relieves fatigue. It has antimicrobial and wound healing effects. The tool has a combination of wound healing, bacteriostatic and immunostimulating properties, stimulating regenerative-preparative processes and accelerating the growth of healthy wound cells. Ionol is a classic synthetic fat-soluble antioxidant phenolic, which has a pronounced anti-radical activity and in most model systems is superior in efficiency to other spatially obstructed phenols.

Currently produced in many countries of the world and the volume of its production is constantly increasing [Molchanov VI, Popov VL, Kalmykov K.N. "Gunshot injuries and their forensic examination", L., Medicine, 1990]. Treats low-toxic antioxidants. When injected into an oil solution through a probe into the stomach, its LD ₁₀₀ is 2500 mg / kg for mice, 3500 mg / kg for rats, 2100 mg / kg for rabbits, and even 10700 mg / kg for guinea pigs. No teratogenic mutagenic effects with repeated administration of large doses of the compound (up to 1 g / kg) in different periods of pregnancy, as well as its effect on the reproduction of animals. It is an acceptor of reactive oxygen species that inhibits lipid peroxidation (POL) and proteins. Forms complexes with free fatty acids, creating conditions to prevent the oxidation of lipids of cell membranes. [.Varfolomeev V., N., Bogdanov G.N., “Mechanochemical processes in the pathogenesis of gunshot wounds and gel composite for their accelerated healing”, Institute of Problems of Chemical Physics, Russian Academy of Sciences, Yearbook, Chernogolovka, 2011, Vol. VII, P. 117-124.

Varfolomeev V.N. "Physico-chemical and biomedical aspects of the action of factors of military weapons", Diss. Doctor of Biological Sciences, M., 2003], contributes to the biosynthesis of macromolecules, ensuring the protection of cells from toxic effects [Varfolomeev VN, Composite on a gel basis for preventing the onset and development of secondary necrosis in gunshot wounds and treatment of gunshot wounds., III International Scientific and Practical Conference "High Technologies, Basic and Applied Research in Physiology and Medicine", April 26-28, 2012, p. 94, St. Petersburg].

L-carnosine.

One of the natural regulators of free radical cell processes is carnosine. This fairly simple dipeptide (its molecule consists of two amino acids - histidine and beta-alanine) acts as a regulator of many processes that occur with the participation of free radicals. Studies of the antioxidant properties of carnosine have shown that it can affect various stages of the process of lipid peroxidation, from the neutralization of reactive oxygen species to interaction with molecular products of free radical oxidation. Carnosine can serve as a trap for peroxyl and hydroxyl radicals, singlet oxygen and superoxide anion oxygen, as well as neutralize the hypochlorite anion, forming stable chloramine complexes with it.

As noted, carnosine is able to inhibit membrane lipid peroxidation, caused both by enzymatic and non-enzymatic, and neutralize the products of free radical reactions. These effects do not have tissue specificity and are accompanied by a membrane stabilizing effect, manifested in the preservation and restoration of the structure of intact cell membranes. The recently discovered immunomodulatory effect of carnosine on the production of reactive oxygen species by leukocytes carrying out phagocytosis makes it possible to explain the immunomodulatory effect of carnosine in the cellular immunity system. It has been shown that carnosine stimulates the healing of trophic ulcers, accelerates the healing of cervical erosion, promotes healing of corneal and lung wounds.

Another advantage of carnosine is its animal origin, since Most of the other antioxidants are of plant origin and are poorly included in human metabolism, therefore, their overdose and undesirable accumulation in the tissues are possible. Carnosine is highly soluble in water, easily formed in the form of tablets, ointments, creams. This compound is completely metabolized in the human body and does not accumulate in organs or tissues with prolonged use.

Membranoprotective effect of carnosine is associated not only with the antioxidant ability of the molecule. A similar protective effect of carnosine on cellular structures was also found under conditions of osmotic shock of cells, as well as during the growth of cell cultures. Carnosine served as a factor that increases resistance to adverse conditions, preventing cell death. In the literature, carnosine has been attributed to a wide variety of functions, including the role of pH buffer, a source of histidine, a chelator of heavy metal ions, a neurotransmitter, a wound healing agent, an immunostimulant, and an antioxidant. The metabolism of carnosine in cells and tissues produces products that have an important biological significance: β-alanine, which stimulates the formation of collagen, and histidine, which is involved in the biosynthesis of proteins and neurotransmitters. The combination of antioxidant and immunomodulatory activity appears to be the basis of the pharmacological effect of carnosine.

Currently, our country uses the method of industrial production of carnosine from cattle meat, based on column chromatography. Carnosine isolated by this technology is less toxic when used in vivo than most synthetic drugs. In terms of purity and low toxicity, preparative carnosine is not inferior to commercial synthetic carnosine produced by the Japanese company Zeria.

Colloidal silver.

The widespread use of antibiotics has revealed a number of negative factors. First, the emergence and rapid spread of antibiotic-resistant strains of microorganisms, which necessitates the development of new antibiotics. Secondly, antibiotics negatively affect the macroorganism as a whole, causing dysbacteriosis and reducing the immune status. Thirdly, antibiotics do not act on viruses.

The antimicrobial activity of silver and its preparations is associated with the complexing, biochemical, and catalytic action of silver ions on bacterial enzymes, proteins, and membrane structures. A positive point is the very large difference in the toxicity of silver compounds for lower life forms (single-celled, bacteria, viruses, etc.), and for higher organisms (animals, man), reaching 5-6 orders of magnitude (10 ⁵ -10 ⁶ times ). That is, the concentration of silver compounds, lethal to microorganisms, is practically harmless to animals and humans.

Silver has a broad spectrum of antimicrobial activity against aerobic and anaerobic microflora, including antibiotic-resistant; shows virucidal and fungicidal activity; has anti-inflammatory effect. Available in the form of a concentrated solution, used in the form of dilute aqueous solutions. Firstly, colloidal silver does not contain cations, which was the case in silver preparations of the previous generation. Cations have an irritating effect on the mucous membrane, which significantly reduced the positive effect of silver preparations of the previous generation. In colloidal silver, there is no cationic form, which makes colloidal silver to be absolutely safe substances (hazard class 4), there is no irritating effect on mucous membranes and this greatly expands the boundaries and effectiveness of colloidal silver, silver is a completely different product, with absolutely new properties and wide margin of application.

And one more not less important fact. Colloidal silver has a unique ability to inhibit the work of the enzyme. And another very important plus - silver of zero valence, unlike cationic silver (a strong oxidizing agent), is a reducing agent, that is, an antioxidant. According to these data, we can safely say that the colloidal using which the protozoa carry out the oxygen exchange. As a result of such action of silver, foreign microorganisms die, because in the presence of silver they cannot carry out oxygen exchange.

Carbopol - is a whole group of compounds of carboxyacrylic and carboxyvinyl polymers. Such polymers resemble latex - sap of a rubber tree. In the world of cosmetics, Carbopol plays the role of a gelling agent. Also, this substance has the ability to soften and moisturize the skin, while not making it sticky. No toxicity detected. Rare acrylic polymer. Highly effective associative thickener, suspending agent, stable at low input levels. When neutralized with alkali turns into a transparent gel. Used in creams, shampoos and alcohol-containing lotions. Compatible with electrolytes. The input rate is 0.1-0.25% to obtain a creamy consistency and up to 1% to obtain a dense gel. Carbomer is sensitive to pH in the formula. To prepare the gel, the carbomer must be completely dissolved in water. By increasing the pH to 7 (range 5-8), carbomer gives a gel structure. Usually neutralization is performed with inorganic bases (NaOH, KOH) or amines (triethanolamine).

Carbopol has the following properties: high viscosity gels at low polymer concentrations; thermal and microbiological resistance; stability and chemical stability during storage; compatibility with many medicinal substances; the possibility of obtaining gels with a wide pH range from 4 to 10; ease of controlling the viscosity of the resulting gels; excellent suspending properties; the ability to stabilize emulsions of the first and second kinds; hypoallergenic; thixotropy, facilitating their extrusion from tubes; ease of application and removal from the skin; high absorption of drugs.

In the claimed invention proposed composite of the following composition.

Table 2 shows the characteristics of the raw materials, materials and intermediates for the composite.

The preparation of the composite is carried out as follows.

1 g of lecithin is added to the porcelain mortar, then 5 g of ionol is added and the mixture is thoroughly ground to obtain a white homogeneous mass.

Then 10 g of a 1% solution of carbomer 974 is added to the mortar and the mixture is also thoroughly ground.

Then 2 g of 1% L-carnosine is applied and the mixture is thoroughly ground.

Then 40 g are added to the mortar. 1% solution of carbomer 974 and the mixture is thoroughly ground.

3 g. 0, 1% KND-SP is introduced into the mortar and the mixture is thoroughly ground.

In conclusion, 1 g of triethanolamine is added to the mortar and the mixture is thoroughly ground.

The result is a gel-based composite, convenient for applying to the surface to be treated. Further, the gel-based composite is conventionally called “Leran”.

CONDUCTING MEDICAL AND BIOLOGICAL TESTS OF COMPOSITE ON GEL BASIS "LERAN".

STUDY OF THE ANTIMICROBIAL PROPERTIES OF "LERAN".

To determine the intrinsic antimicrobial activity of the gel-based composite for skin regeneration, “Leran”, studies were conducted with respect to test strains of the following microorganism types: Escherichia coli ATCC 25922, Staphylococcus aureus ATCC 6538-P, Pseudomonas aeruginosa ATCC 9027, Bacillus cereus АТСС 9027, Bacillus cereus АТСС 9027, Bacillus cereus TC АТСС 9027, Pseudomonas aeruginosa АТСС 9027, Bacillus cereus АТСС 9027, Bacillus cereus TC АТСС 9027, Pseudomonas aeruginosa АТСС 9027, Bacillus cereus АТСС 9027, Bacillus cereus TC АТСС 9027; Candida albicans ATCC 885-653. Own antimicrobial activity was determined once.

Cultures of Escherichia coli, Staphylococcus aureus, Pseudomonas aeruginosa, Bacillus cereus were cultivated on meat-peptone broth at 37 ° C for 19 hours. Candida albicans was cultivated in Saburo's liquid medium at a temperature of 30 ° C for 48 hours. From each grown culture a suspension of microorganisms was prepared in a physiological solution in a 1: 1000 ratio. In 10 ml of a buffer solution consisting of mono-substituted potassium phosphate and disubstituted sodium phosphate in physiological solution, 1 ml was added from a microbial suspension and the first dilution of the test Leran skin. Served as control tubes with nutrient media and the corresponding test strains, in which instead of the tested cosmetic products made the same amount of distilled water. Crops were incubated at 38 ° C for 48 hours. After the expiration of the incubation period, they were sown on selective media: Escherichia coli - on Endo medium, Staphylococcus aureus - on yolk-salt agar, Pseudomonas aeruginosa - on nutrient agar for Ps.aeruginosa, Bacillus cereus - 5% blood agar, Candida albicans - chromogenic agar for fungi Candida.

The research results showed that the cream-gel for skin regeneration "Leran", exhibits an antimicrobial bacteriostatic effect (no growth) in relation to test strains of E.coli, St. Aureus and Ps.aeruginosa (Table 3).

The strains of Bacillus cereus and Candida albicans after treatment with Composite solution for skin regeneration Leran grew on nutrient media.

STUDY OF THE ANTI-INFLAMMATORY ACTIVITY OF THE COMPOSITE LERAN.

The definition of anti-inflammatory action was carried out according to the method of F.P. Trinusa et al., Creating a model of experimental inflammation by subplantary administration of a 1% formalin solution in an amount of 0.1 ml into one of the hind paws of rats. An increase in the volume of the paw was recorded in the dynamics every hour since the development of inflammation and the application of the studied agent within three hours and after 24 hours (day). The intact animals served as controls, and nothing was applied to the skin after the development of the inflammatory focus. The percentage of inhibition of inflammation was calculated according to the formula defined in this method:

,где V_k - объем отечной лапки в контроле минус исходный объем этой же лапки до отека

where V _k is the volume of the edematous paw in the control minus the initial volume of the same paw before edema

- объем отечной лапки в опыте минус исходный объем этой же лапки до отекаY _k

- the volume of the swollen paw in the experiment minus the initial volume of the same paw before the swelling

Experimental studies on the study of anti-inflammatory action were carried out on 20 white rats, males, line "Wistar", weighing 220-230 g, are on standard vivarium mode. The intact animals served as controls, which, like the experimental ones, were in standard vivarium mode, but they did not receive skin applications of the studied compositions.

In evaluating the anti-inflammatory effect, we noted that the cream-gel for skin regeneration rather actively inhibited the process of inflammation. The degree of effectiveness was almost directly proportional to the duration of its impact. So, after 1 hour after application, inflammation decreased by 28.4%, after 2 hours, the effect increased and amounted to 58.9%. After 3 hours, the anti-inflammatory effect continued to increase and reached 70.0% already. A day after its use, the anti-inflammatory effect was maximal, pronounced and amounted to 91.1% compared with the control. The anti-inflammatory activity of the inventive composite is shown in FIG. one.

One of the urgent problems of pharmaceutical science is the search for new and improvement of existing ointments and their introduction into medical practice.

Recently, skin diseases (dermatosis) have become much more frequent, which is explained by the influence of many factors, including sensitization, the organism, exposure to microorganisms, environmental pollution, the use of synthetic detergents, etc.

STUDY OF ANTISEPTIC ACTIVITY OF "LERAN".

- противогнилостные средства, предназначенные для предупреждения процессов разложения на поверхности открытых ран, например в ранах, образующихся после больших операций или ушибов, или для задержания уже начавшихся изменений в крови. Некоторые антисептики являются действительно гермицидными, способными уничтожать микробов, в то время как другие являются бактериостатическими и только предотвращают или подавляют их рост.

- protivognotnye means designed to prevent decomposition processes on the surface of open wounds, for example in wounds formed after large surgeries or bruises, or to arrest the already begun changes in the blood. Some antiseptics are truly germicidal, capable of killing germs, while others are bacteriostatic and only prevent or inhibit their growth.

Antiseptics are called broad-spectrum antimicrobial drugs that cause protein denaturation and do not have a selective effect. They are used externally for disinfecting the skin, mucous membranes. Antiseptics have a wide spectrum of action, affect bacteria, fungi, protozoa and large viruses. They can cause protein denaturation, violate the permeability of the cytoplasmic membrane, inhibit the activity of enzymes necessary for the vital activity of microorganisms.

Antiseptic ointments - medicines for external (local) use, which are prescribed for the prevention and treatment of purulent-inflammatory processes. These drugs are effective against most pathogens, i.e. possess a wide spectrum of action, without showing selectivity. Antiseptic ointments can be applied to the skin and mucous membranes. These drugs delay the development of microorganisms, affecting proteins, enzymatic systems of microbial cells, or causing their death. As a result, the infection is eliminated, the development of the inflammatory process stops or is prevented and the damage heals as soon as possible. The activity of antiseptic ointments depends on their concentration, duration of exposure, ambient temperature, the presence of organic substances in the treated medium, the sensitivity of infectious agents, etc. Unlike liquid antiseptics, antiseptic ointments are well absorbed and retained for a long time in the damaged tissues, for a long time affecting and not overdrying the treated surfaces.

An antiseptic is a complex of therapeutic and preventive measures aimed at the destruction of microbes in a wound or the body as a whole.

Recently, skin diseases (dermatoses) have become much more frequent, due to the influence of many factors, including sensitization, of the organism, exposure to microorganisms, environmental pollution, the use of synthetic detergents.

Due to the success of chemistry for the treatment of purulent wounds and infectious processes, a number of new antiseptics have been proposed, much less toxic to the tissues and the patient's body than carbolic acid.

Modern antiseptic preparations have a wide antibacterial spectrum and do not induce microbial resistance. Their use is a promising direction, as it allows you to change the nature of the impact on cellular factors of inflammation (Budevskaya TV. Justification of the choice and rational use of antiseptics for the treatment of apical periodontitis: Abstract. Dis. Cand. Medical Nauk, Minsk, 1993. - 120 with.;

Barer G.M., Grigoryan S.S., Surazhev B.Yu. Evaluation of the effectiveness of liniment 5% cycloferon complex therapy of periodontitis // Materials of the II scientific-practical conference dedicated to the memory of prof. HER. Platonov (MSMSU). M., 2004. - p. 14-16.

Elizarova V.A. Chlorophyllipt as an effective antiseptic in the treatment of periodontitis // Proceedings of the II International Congress of Dentists Tbilisi, 2000. - P. 109-111.

Grigoryan A.S., Grudyanov A.I., Rabukhina N.A. Periodontal diseases: pathogenesis, diagnosis, treatment. / / Guide for doctors. M., 2004. - 320 p.).

The dominant factors that determine the widespread clinical use of antiseptics are the enormous species and quantitative diversity of microorganisms, the slow production of resistant bacterial strains to antiseptics, and the rarely occurring allergic reactions in patients.

Previously, we noted the relationship between microbial seeding and the occurrence of purulent complications. The table shows that the use of the composite "Leran" for the treatment of wound surfaces in order to obtain a therapeutic effect reduces microbial seeding of wounds to extremely low amounts, which is accompanied by the complete absence of animals with purulent complications. The average time for wound cleansing is reduced by 2.8 times compared with the control, and complete wound healing with hair restoration occurs 10 days earlier than in the control (for 18 days).

Similar results can be explained by an additional contribution to the antiseptic activity of “Leran” of a solution of colloidal silver, which is part of the composite. A solution of colloidal silver has a disinfecting effect, at the same time, being a tool that has a high therapeutic effect. Influencing microorganisms, silver solution is not addictive and does not lead to the stability of viruses. Colloidal silver solution is a powerful antiseptic with a broad spectrum of activity that prevents the growth and reproduction of viruses, bacteria and fungi, unlike antibiotics, does not cause the formation of pathogenic microorganisms resistant to it

STUDYING THE HEATING EFFECTS OF LERAN.

Experimental studies of the wound-healing effect were carried out on 20 white rats, males, line "Wistar", weighing 220-230 g, are in standard vivarium mode.

The wound healing effect of the gel-based composite for skin regeneration was studied according to the method commonly used in pharmacology in experimental animals. Animals of the experimental group received daily, once a day, skin applications of the studied cosmetic, and in the control group wound healing (scar formation) occurred spontaneously. After applying the cosmetic, the animals were placed in individual metabolic cells for 1 hour, to prevent the drug from licking. Visual and planimetric assessment of wound healing was performed on the 3rd, 5th, 8th, 10th, 12th and 14th day before the formation of the scar, in a comparative aspect with the control group. A clinical examination of each animal was performed daily.

The results of experimental studies on the wound-healing effect showed that under the action of a gel-based composite for skin regeneration, the Leran wound healing process proceeded faster than in the control group. We noted that already on the 3rd day of observation the regenerating effect of the experimental group was 28.2% with 18.0% in the control group (a difference of 10.2%). On the 5th day, the difference in the wound-healing effect in a comparative aspect between the experimental and control groups was expressed at the same level and amounted to 10.3%. On the 8th day, the wound healing process was more pronounced in the experimental group, compared with the control also by 10.3%. By the 10th day, the wound healing process under the influence of Cream-gel for skin regeneration “Leran” almost came to an end and was faster by 3.9%, and by the 12th day it was fully completed - by 100.0%. A thin scar formed in the wound site, the color of normal healthy skin, whereas in the control group the wound healing process was only 96.1%, and complete regeneration in this group occurred on the 14th day (Fig. 2).

In the study of gunshot wounds [4], we drew attention to the fact that the differences in the therapeutic efficacy of drugs are higher, the heavier the initial damage. We conducted a series of experiments to determine the healing activity of Lera. In this case, the wound was additionally infected with a culture of staphylococcus. The rest of the procedure was carried out in accordance with previous experience. The results showed that by 21 days in the group of animals with the use of "Leran" there is a complete recovery of damage, while in the control group the wound surface of a sufficiently large size is maintained.

The introduction of "Leran" significantly increases in the wound discharge the number of macrophages, which for all periods of observation exceeds their content in the control by 4-5 times, which indicates an intensive process of cleansing wounds. The favorable direction of the wound process is also indicated by the significant excess in the number of polyblasts already in the early stages in the experimental series compared with the control groups. In addition, it was found that the content of young and mature fibroblasts progressively increased at the indicated observation times by 3-5 times, and fibroblasts by 2-4, while in the control series the number of fibroblasts remained low.

Studies over the past two decades have convincingly shown that a balance shift in the system of prooxidant-antioxidant balance accompanies many physiological and most pathological processes.

Used in the treatment of purulent wounds, ointment dosage forms in most cases can have a significant impact on free radical processes both in the wound and in the underlying healthy structures.

An increase in ROS production, the appearance of a pool of catalytically active Fe2 + ions and a drop in the activity of the endogenous system of CPP inhibitors cause the activation of CPP in cell membrane and lipoproteins observed in various pathological conditions.

This is a pathogenetic rationale for the use in the clinic of various natural or synthetic inhibitors of CPP as antioxidant therapy drugs.

Recently, a wide search and testing of the properties and activity of various new CPP inhibitors and their mixtures has been carried out.

One of the modern methods of diagnosing impaired prooxidant-antioxidant status is the measurement of parameters of luminol-dependent induced chemiluminescence, based on determining the number of produced radicals during the induction of free radical reactions by introducing hydrogen peroxide, while showing the maximum BVHL value that reflects the level of products.

The duration of the latent period (t) depends on the time to reach the so-called. "critical" concentration of divalent iron ions in the system, which in turn is determined by the contribution of substances that affect the oxidation and / or elimination (chelation) of Fe2 +. The intensity (H) or light sum of slow luminescence is determined by the rate of CPP flowing in the hydrophobic phase of membranes or lipoproteins, and the ability of inhibitors to intercept lipid radicals [Vladimirov Yu.A., Archakov AI (1972) Lipid peroxidation in biological membranes. M., Science. The LM oxidation is accompanied by the formation of the LM radical and ultimately leads to the formation of an excited oxidation product, which passes to the ground state with the emission of the quantum of light CL. The amount of released light quanta of CL is proportional to the amount of the resulting oxidation product and therefore is a measure of the degree of oxidation of the LM. Addition of substances capable of preventing the oxidation of LM (antioxidants) to this model system will lead to a decrease in the number of CHL light quanta. Measuring the kinetics of the intensity of CL in the Hb-H2O2-LM system without and in the presence of this substance, one can thus study the antioxidant properties of this substance [Teselkin Yu.O., Babenkova IV, Lyubitsky OB, Klebanov GI , Vladimirov Yu.A. (1997) Q. honey. chemistry 43, 87-92].

The applicant conducted a comparative study of a number of drugs used to repair various kinds of damage to the skin. The results are presented in FIG. 3 and 4. (The intensity of the chemiluminescence signal of a number of wound-healing drugs available in the pharmacy chain and Leran is shown in Fig. 3, Fig. 4 shows the light sum of a number of wound-healing drugs available in the pharmacy network and Leran). From the figures it is clear that the drugs can be divided into three groups according to their action:

- possessing a prooxidant effect, i.e. enhancing the formation of reactive oxygen species (Levomekol and Malavit);

- weakly affecting chemiluminescence indices (counter-tube, D-panthenol, argosulfan);

- causing a significant decrease in the studied parameters (nanovit, special grease, acriderm, radevit, and lerand, while it should be noted that Leran has two or more times the above listed ointments in its activity).

It can be assumed that the effect of the studied samples of wound-healing drugs available in the pharmacy chain is due to the interaction of inhibitors not so much with the initiator radicals that are formed when the reactions of the model system are triggered, but with the radicals formed in subsequent luminol oxidation reactions. One of them should be called superoxide anion radical (O ₂ ^- ), which is an intermediate product in the reaction of free radical oxidation of luminol. One of the arguments in favor of this assumption is the fact that the introduction of SOD into the model system caused a decrease in the amplitude of the CL without a significant change in the duration of the latent period. In this regard, it can be assumed that Mexidol, Emoxipin and Proxipin, water-soluble antioxidants that extinguish the CL model system, are able to intercept O2 * and thereby inhibit the initiation of CPP lipid peroxidation in vivo [Teselkin Yu.O., Babenkova I.V. , Lyubitsky OB, Klebanov G.I., Vladimirov Yu.A. (1997) Q. honey. chemistry 43, 87-92].

Thus, we can conclude that the antioxidant effect of the claimed composite is based on the ability to inhibit the stage of initiation of CPP of lipid peroxidation, largely due to the formation of reactive oxygen species and the appearance of catalytically active iron ions. This fact allows us to recommend the use as CPP inhibitors in the acute period of inflammation, when the effectiveness of the initiation stage is very high. This is especially true in the case of the use of CPP inhibitors in the treatment of wounds [Tolstoy PI, Krivihin V.T., Lutsevich EV, Kalinin MR, Dotsenko N.M. (1998) Laser radiation and antioxidants in the treatment of purulent-necrotic processes of the lower extremities in patients with diabetes mellitus. M, Orbita-M]. In this case, external use, in particular, “Lerana”, as having shown the greatest activity in the in vitro system, will allow creating the necessary concentration of inhibitors in the inflammation center (in the wound), which will reduce the production of ROS and thereby decrease the effect of lipid peroxidation of lipid peroxidation on subsequent wound healing stages.

The essence of the invention is characterized by the following examples.

Example 1. Studies on the anti-burn activity of the Leran composite versus the action of the drug Levomekol, which is the most popular among surgeons, and the often advertised drug D-panthenol (comparison groups 1 and 2, respectively).

Assessing the data obtained, attention is drawn to the fact that in the treatment of a purulent wound with the Leran composite in the purulent-necrotic phase of the wound process, noticeable stimulation of all links of the wound process occurs. Thus, the scab rejection in the main group of animals treated with "Leran" was observed approximately 7 days and 12 days earlier than in comparison groups 1 and 2, respectively. Analyzing the further dynamics of epithelialization, both in the control and in the main group, the significant advantages of the proposed composition were revealed. Thus, in animals receiving a new composition for treating burn wounds, on average, the completion of wound epithelization occurred 7 days and 21 days earlier than in animals treated with D-Panthenol ointment and Levomekol, respectively.

Below, FIG. 5, which depicts the wound surfaces of burn wounds on the 29th day after causing damage in the control, comparison groups and the main group. The data clearly demonstrates the advantage of using the Leran composite developed by us.

Example 2. Studies of the composite on pus-plicating complications.

The test group included 15 volunteer patients who underwent surgical interventions for melanomas of the skin and their metastases in regional lymph nodes. The indications for the use of the composite were purulent-septic complications from the surgical wound in the early postoperative period. The composite was applied to an open wound and evenly distributed over the surface. It was noted that the wound was cleared of purulent-necrotic raids, a decrease in the swelling of the surrounding tissues, skin hyperemia and pain syndrome already on the first day. Within 7-10 days the wounds were completely carried out with viable granulations, active epithelization was underway. Positive effects were recorded for all patients included in the study group.

Example 3. The composite was used in the treatment of severe purulent-septic processes, in particular after necrosis of skin grafts and subcutaneous tissue after Ducén's surgery, as well as pressure sores in patients on prolonged mechanical ventilation in the intensive care unit. As a result of the use of the composite, rapid wound cleansing, the development of granulations and wound healing by secondary tension with the formation of a tender scar are noted.

Thus, the gel-based composite "Leran" proved to be an active substance with high activity and can be used in purulent surgery to clean wounds and stimulate regenerative processes.

Efficiency assessment of the use of "Lerana" with a gunshot wound.

Studies were conducted on two groups of animals.

In the control group of animals, the inlet zone of the secondary necrosis extends up to 15 mm from the edge of the wound, the boundaries of the zones are fuzzy, of a mosaic nature, extensive hemorrhages into the muscle tissue are observed at a considerable distance from the wound canal. The wound is moderately swollen, with pronounced infiltration, especially in the area of the orifice. For wounds in the control is characterized by the presence of a significant amount of purulent discharge. At the opening of the wound canal, the fully preserved wound canal, made of necrotic tissues, shows a sizeable region of non-viable tissues. The mass of removed non-viable tissues is 9 ± 3 g. The healing of such wounds occurs 16-18 days after injury.

In the experimental group of animals, the wound was treated with Leran. The inlet is usually overgrown. The zone of secondary necrosis is practically absent. Hemorrhages are either absent or extend to a distance of 2-3 mm from the edge of the wound only into the subcutaneous tissue without penetration into the muscle mass. Discharge wounds is serous. There is a marginal epithelialization, the presence of mature granulation tissue. When a wound is dissected, an almost overgrown wound canal is seen, purulent discharge is absent, around the wound canal there are viable muscle tissues, which, when cut, contract and bleed. Wounds are overgrown at 9-10 days after injury.

Example 4. FIG. 6, 7. presents data on the treatment of trophic ulcers of the foot. Patient A. 72, trophic ulcer of the foot, 2 years of unsuccessful conservative treatment in the clinic (levomekol, streptotsid, sea buckthorn oil), is shown in FIG. 6. Type of ulcer (Fig. 7) after 1.5 months of treatment with the composite (applications were performed 2 times a week), 80% of the wound is epithelized.

Example 5. The composite was used after a knife wound of a finger. FIG. 8. treatment of a knife wound of a finger (little finger, right) with a gel-based composite for 2 days (A), for 5 days (B), for 10 days (C) and for 14 days (D) after injury.

The figures on the left show a light knife injury of the ring finger, received simultaneously. This wound has not been treated.

Despite significant quantitative and qualitative differences in the character of the wounds of the little finger and ring finger, by the 11th day the wounds look almost the same, and by the 14th day the wounds are completely overgrown.

The following are examples of the use of the composite on a gel basis "Leran" in veterinary medicine.

Example 6. Dog, small poodle, 14 years old. Diagnosis: epolis of the upper gum. size 7 × 5 cm. Removal of the neoplasm, five teeth and jaw bone section in healthy tissues + electrocoagulation, further antibiotic therapy + treatment of papilloma locally using a composite. The prognosis was unfavorable due to neglect of the process, extensiveness of the lesion, age of the animal and abundant dissemination of the microflora interventions, but after two weeks the mucous membranes of the oral cavity of the papilloma were completely washed out. In the future, metastasis was not observed.

Example 7. Dog, Drathar 5 years old, multiple injured pelvic limbs with a fraction. After extracting the fraction-antibiotic therapy + composite. After 7 days, there was a complete recovery.

Example 8. Dog, German Shepherd, 8 years. After the other dogs bites, three purulent-necrotic foci of skin and muscle tissue with a diameter of 15 cm developed. After stripping, a composite was applied locally. Recovery occurred in three weeks.

The gel-based composite was tested on wounds caused by thermal burns.

Example 9. The experiment was carried out on 30 outbred rats. Keeping animals and staging experiments was carried out in accordance with the requirements of orders No. 1179 of the USSR Ministry of Health on 11.10.1983 and No. 267 of the Ministry of Health of the Russian Federation of June 19, 2003, and in accordance with the requirements of the Helsinki Declaration on the humane treatment of animals [4] .. the first day after the application of burns, Levomekol ointment was applied daily to the burn scab to animals of the 2nd group (control), pectin films were applied to the 3rd group (experimental), the 1st group of animals was left without treatment. In all groups of animals, the beginning of the scab rejection occurred approximately at the same time — on 13.8 ± 0.39 days. It was established that, thanks to the “Levomekol” ointment, animals of the 2nd group had a scab in a more humid state. In the experimental group, a pectin film fixed to the surface of a scab did not significantly affect its properties.

Experimental animals were divided into 3 groups of 20 animals each: Group 1 - intact animals; 2nd group - experimental (animals with a full wound with application of a mixture consisting of nanoparticles of metals and chitosan); Group 3 - the comparison group, animals with a full wound, which was treated with 0.9% isotonic sodium chloride solution. The maintenance of animals, their nutrition and care for them was carried out according to the "Sanitary rules for the design, equipment and maintenance of experimental biological clinics (vivariums) of April 6, 1973"

Assessing the data obtained, attention is drawn to the fact that in the treatment of a purulent wound with the Leran composite in the purulent-necrotic phase of the wound process, noticeable stimulation of all links of the wound process occurs. So, the first islands of granulation tissue appeared in the main group of animals by the third day (2.9 ± 0.3 days) of treatment, and in the control group - by 4.5 ± 0.2 days. In the main group, complete wound cleansing from a purulent-necrotic discharge was observed on average by 5.1 ± 0.2 days, and in the control group - 7.4 ± 0.3 days. Analyzing the further dynamics of epithelialization, both in the control and in the main group, the significant advantages of the proposed composition were revealed. Thus, in animals receiving a new composition for the treatment of purulent wounds, on average, the appearance of marginal epithelialization occurred two days earlier than in animals treated with Levomekol ointment: 7.2 ± 0.19 and 9.5 ± 0.3, respectively. Complete epithelialization of the wound defect in the main group occurred at 11.3 ± 0.3 days, and in the control group - at 18.4 ± 0.3 days from the start of treatment. Clinical evaluation of the effectiveness of local treatment of purulent wounds is presented in table 5.

As a result of the research it was found that under the influence of thermal effects in rats of all experimental groups developed severe skin burns of III-IV degree. In the next 2-3 days after the burn injury, the animals were lethargic and adynamic, refused to feed and lost in weight to 10-15% of the initial body weight. Over the next 5-10 days after the application of the burn, its area in animals of the experimental groups treated with "Leran" D-panthenol was significantly less than in control rats and in the group of rats treated with Levomikol. It should be noted that the greatest differences with the control were recorded in rats fed "Leran". During the same periods of observation, the degree of necrotic changes in the burn surface in the animals of the experimental groups was different. They were the least pronounced in rats treated with Leran; then, in order of magnification, the group of rats treated with the “-D-panthenol” comparator was located. The largest necrotic changes were observed in the control animals and in the group of rats treated with “Levomikol”. In the subsequent follow-up periods ≈ for 15-20 days, healing of burns in rats treated with Leran was accelerated compared with the control. Healing of a burn wound was worse in rats treated with the D-panthenol comparator. Under the influence of Leran, on the 25th day after the application of the burn wound in individual rats there was a complete healing of the burns and the formation of a tender scar. When examining the skin at the site of a burn in control rats (a burn without treatment), tissue necrosis was observed on the 30th day after the application of the burn injury. Revealed the place of rejection of the scab and suppuration, the absence of the upper layers of the skin. There was visible damage to the styloid and basal layers of the integumentary epithelium. There were signs of damage to the follicles of the hair follicles, exocrine and apocrine glands. The pattern of wound defects healing in rats during the treatment with "Leran" on the 30th day after the application of the burn was significantly different from that described above in the control. In many rats epithelialization of wounds was noted. A moderate number of thin-walled vessels was detected in the granulation tissue. In rats treated with burns with D-panthenolone used as a reference drug, there were scab remains on the surface of wounds in the form of a fibrous-leukocyte layer, under which granulation tissue was arranged with an ordered course of weakly fuxinophilic collagen fibers. The granulation tissue of the healing burn wound was characterized by the predominance of cellular elements over the collagen fibers. By the effectiveness of the anti-burn effect, Leran is superior to the anti-inflammatory drug D-Panthenolone and the pharmacopoeial drug Levomekol when applied topically over the next 30 days of burn treatment. The mechanism of the anti-burn action of "Leran" seems to be related to its influence on the processes of regeneration of collagen fibers.

Drugs with antioxidant properties are characterized by a stimulating effect on regenerative processes, manifested by the acceleration of the time of epithelization of burns, the absence of cases of suppuration of burn wounds. The use of adequately selected drugs for the local treatment of wounds improves the results of treatment of patients with burns and their consequences.

Summing up the study of the effect of "Leran" on the healing of severe thermal skin burn in rats, it should be noted that applying the drug to a burn surface immediately after a burn injury and its daily single application over the next 30 days significantly weakens the development of severe necrotic processes and accelerates the healing of the burn. The effectiveness of the antiburn effect of the studied drug. "Leran" surpasses that used as reference preparations.

Thus, according to the results of planimetric studies in animals using the “Leran” composite, it can be concluded that it stimulates epithelial regeneration. Based on the foregoing, it is safe to say that the use of a composite on a gel basis "Leran" is a very promising physiologically active form of impact on the wound process.

Analysis of the obtained results gives grounds to draw the following conclusions:

The gel-based composite for skin regeneration “Leran” is effective according to the studied objective indicators of the functional state of the skin.

The gel-based composite for skin regeneration “Leran” has an anti-inflammatory effect in a directly proportional degree, depending on the duration of its effect, with the most pronounced value in one day - 91.1%;

The gel-based skin regeneration composite Leran has its own antimicrobial activity against microorganisms E.coli, St. Aureus, and Ps.aeruginosa.

The gel-based composite for skin regeneration “Leran” significantly speeds up the process of regeneration of the wound surface after its application for all periods of observation.

The gel-based composite for skin regeneration “Leran” can be recommended for wide practical use as an effective means for the studied parameters.

Claims (5)

1. Composite for accelerated healing of wounds of various etiologies, containing active components, characterized in that it contains three active components with wound healing, anti-inflammatory, antibacterial and antiseptic activities, due both to their antioxidant properties and the ability to directly influence the pathogenetic features of the wound process , in the following percentages by weight: ionol 4.5-5.5 carnosine 1.5-2.5 colloidal silver 2.5-3.5 lecithin 1.0-1.5 triethanolamine 1.0-1.5 carbopol   50-51 water rest 2. The use of a composite according to claim 1 as a cosmetic for skin regeneration in cosmetology practice. 3. The use of a composite according to claim 1 as a therapeutic agent in veterinary medicine for accelerated healing of wounds. 4. Means for regeneration of the skin, characterized in that it contains a composite according to claim 1.

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