RU2456013C2 - Wound-healing agent for wound preventiong and repair in diabetic patients - Google Patents

Abstract

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to medicine and concerns preventing and treating the complication of diabetes - diabetic foot infections (DFIs). A wound-healing agent for wound prevention and repair in the diabetic patients contains an alcoholic-aqueous extract of St. John's wort, sea-buckthorn oil, propolis infusion and as a base, PEO-400 and PEO-1500 alloy (1:1) in the following proportions, wt %: the alcoholic-aqueous extract of St. John's wort 5.0; sea-buckthorn oil 3.0; propolis infusion 2.0; the base - the rest.

EFFECT: agent promotes line skin wound healing with underlying experimental alloxan diabetes at all three stages of the wound process (exudation, proliferation, epithelisation).

6 dwg

Description

The invention relates to medicine and for the treatment of patients with diabetes mellitus. It can be used for the prevention and treatment of complications of diabetes mellitus - diabetic foot syndrome (SDS).

Damage to the skin of the feet for a healthy person will be practically invisible, but for a patient with diabetes mellitus, against the background of a weakened immune system, the slightest cut, wear on the legs is enough for infection to occur. This condition is usually accompanied by hyperthermia, leukocytosis. In such cases, urgent surgical intervention with necrectomy is necessary. With extensive purulent-necrotic lesions, amputation is performed. In the United States, more than 50,000 amputations are performed annually, each costing about $ 10,000, with total costs reaching $ 500 million (Komelyagina E.Yu., 1998). In Russia, about 11-12 thousand high amputations are performed annually, which is relatively more than in other countries (Dedov I.I., 2000). According to Russian researchers, after amputation of the lower extremities, postoperative complications are observed in 64% of patients, the average length of hospital stay is 58.2-65.7 days, and in every second life expectancy after surgery does not exceed two years (Briskin B.S. et al. ., 1999).

Over the past 20 years, the attitude towards amputations has changed in the direction of preserving the limbs as much as possible and preventing them.

The question of the most rational composition of the comprehensive treatment of CDS and the appropriateness of including some new methods in it remains relevant. At the same time, the use of local therapy is still an indispensable method of treatment, and in particular, ointments of various compositions are used. The selection of optimal agents for topical treatment of trophic disorders of the skin of the lower extremities with SDS is an urgent problem. Doctors from all over the world face it.

Local treatment should correspond to the stage of the wound process:

1) exudation;

2) proliferation;

3) epithelization.

In recent years, there have been reports of the treatment of flaccid granulating ulcers in CDS with the Kuriozin preparation, which is an associate of hyaluronic acid and zinc. The positive effect of this drug in patients with SDS was noted in the 1st phase of the wound healing process, and since patients with the 2-3rd phase of the wound healing process come to the hospital, Curiosin can only be used in the complex treatment of SDS.

The Levomekol ointment has proven itself well (V. Datsenko, T. Tamm. Study of a multicomponent ointment on a water-soluble basis with dioxidine. Klin. Khir., 1981, No. 1, p. 43-45). This is a combined drug (chloramphenicol and methyluracil) for external use, with an antibacterial, anti-inflammatory effect. But it is also indicated for use mainly in the 1st phase of the wound process and often causes an allergic reaction - a skin rash.

There are positive reviews of the local treatment of diabetic ulcers with Ximedon (RF patent No. 2063752, IPC A61K 31/505, 9/06, 1996). This is a new pyrimidine derivative comprising xymedon and boric acid. This drug inhibits the growth of blue-green pus bacillus, but representatives of anaerobic non-clostridial microflora of the bacteroid class belong to the main role in the spread of infection through the tendons and synovial sheaths. In addition, the bacterial landscape of the wound in a patient with SDS is very wide, therefore, Xymedon can only be used as a drug for the comprehensive treatment of SDS. It has some toxicity; in patients with diabetes mellitus, glucose tolerance should be more often determined.

All this speaks in favor of the search for new drugs for the prevention, treatment of diabetes and its complications, including those based on natural compounds. Advantages of the latter: a wide range of pharmacological effects with low toxicity, rare side effects and minimal contraindications. A complex combination of phyto compounds with accompanying substances can have a beneficial effect on the structure of the skin and the body as a whole: accelerate the regeneration of epidermal cells, penetrate deep into the subcutaneous tissue, while regulating the salt-water balance and tone, activate enzyme systems, normalize hormonal and general metabolism. The combination in the phytocomposition of certain medicinal plant materials suggests the presence of various types of activity and to obtain medicinal and therapeutic and prophylactic agents with desired properties (RF patent No. 2323275, IPC A61K 36 / 00.35 / 64, A61P 17/02, 2009; 2326683, A61K 36/38, 35/64, 2007; 2287995, A61K A61K 36/72 ,, 36/38, F61P 17/02, 2006; 2311175, A61K 36/72).

For several years, the authors have been researching and developing gels with phytocompositions for correcting and preventing the consequences of diabetes mellitus, namely “diabetic foot” syndrome (SDS) (Ogay MA, Stepanova EF. Development and research of gels with phytocompositions for prevention consequences of diabetes mellitus. Izv. Universities. North-Caucasian region, special issue, 2007, pp. 99-100; Ogay MA, Stepanova EF, Larionov LP, Petrov A.Yu. Pharmacological research and technology phytogels for the correction of the effects of diabetes. Voronezh State University, series: Chemistry. Biology. Pharmacy. No. 2, 2009, pp. 171-173; Ogai M.A., Stepanova E.F. Larionov L.P., Petrov A.Yu. Pharmacotechnological studies of external drugs - phytogels based on medicinal plant materials to prevent the consequences of diabetes. Kursk scientific and practical bulletin "Man and his health", No. 1, 2010, p.123-126 (prototype)). In these works, the wound healing activity of gels of various compositions, based on medicinal plant materials, was studied, which included, along with tinctures of other medicinal plants, tinctures of St. John's wort, sea buckthorn oil, and propolis tincture. As a result of these studies, the wound healing activity of the proposed gels in animals with experimental pathology was verified and it was concluded that it is possible to study the latter in the future for the prevention and treatment of some consequences of diabetes mellitus, including “diabetic foot”. However, the optimal qualitative and quantitative ratios of the components of medicines, including tinctures of St. John's wort, sea buckthorn and propolis, as well as the base, have not been investigated and proposed.

The objective of the invention is to develop a new soft dosage form - phytogel for the prevention and treatment of SDS.

The technical result consists in effective tissue regeneration due to the optimal qualitative and quantitative ratio of the components of the drug prepared according to the indicated technology at all three stages of the wound healing process (exudation, proliferation, epithelization).

The technical result is achieved by the fact that the wound healing agent for the prevention and treatment of wounds in patients with diabetes mellitus (phytogel) contains alcohol-water extract from perforated St. John's wort, which is prepared on a 40% alcohol solution at a rate of 1: 5, sea buckthorn oil, propolis tincture and as the base alloy PEO-400 and PEO-1500 (1: 1), with the following components, wt.%:

Alcohol-based extract from St. John's wort perforated 5.0-5.5 sea buckthorn oil 3.0-3.5 propolis tincture 2.0-2.5 the basis rest

The dosage form was tested at the Pyatigorsk State Pharmaceutical Academy at the Department of Drug Technology.

Figure 1 shows table 1, reflecting the composition of the proposed funds; figure 2 is a table 2 containing indicators of glucose and lipid spectrum in alloxan diabetes in rats; figure 3 - table 3 containing the average linear size of the wound in rats; figure 4 is a picture of a histological section in the control group; figure 5 is a picture of a histological section in the group treated with the drug levomekol; figure 6 is a picture of a histological section in the group treated with the proposed tool.

In experimental studies, the wound healing effect of the developed composition on linear skin wound models against the background of experimental alloxan diabetes was first revealed.

St. John's wort contains many diverse, biologically active substances: hypericin, flavonoid glycosides, tannins, essential oils, ascorbic acid, vitamin PP, carotene. Such a diverse composition explains the many-sided pharmacological properties of St. John's wort, tincture of St. John's wort herb is prepared on a 40% alcohol solution at a rate of 1: 5.

Sea buckthorn oil (Reg. No 000245 / 02-2003, FC Altayvitaminy) is the richest source of vitamin A, and also has a high content of vitamin E. It has an epithelizing effect, promotes wound healing, resorption of scars, and has analgesic, antiseptic, antioxidant and capillary-strengthening properties. It is used for burns, frostbite, as well as for the treatment of various skin diseases (RF patent №2325175; Mashkovsky MD Medicines. M: Medicine, 1974, v.11, p.8).

Propolis contains amino acids, organic acids, antibiotic and tannins, balms, essential oils, carbohydrates, mineral salts, vitamins, trace elements and other components that determine its diverse pharmacological properties. It has a bactericidal, antiviral, fungicidal, anti-inflammatory, analgesic, dermatological effect. Propolis promotes wound healing, is used for burns and frostbite (Sinyakov A.F. Bee pharmacy. M.: Physical education and sports, 1995, p.100-101, 104-145, RF patent 2028808, 1995). We used the finished product tincture of propolis (FS 42-3736-99, OAO Tatkhimpharmpreparaty).

The base used is water-soluble and contains a hydrophilic non-aqueous solvent PEO-400 and a water-soluble polymer PEO-1500, related to polyethylene oxides, which are physiologically indifferent compounds. When mixing the PEO-400 solvent with the PEO-1500 polymer, the latter acts as a thickener, which, depending on the amount in the total mass, provides the required consistency of the created dosage form. Polyethylene oxides provide the interstitial action of the dosage form, which provides suppression of deep microflora. PEO-400 absorbs more water and has a long period of osmotic action, and PEO-1500, which is also an osmotically active substance, absorbs purulent exudate due to the binding of water molecules to hydroxyl and ether groups. This basis is recommended for ointment dosage forms used in the local treatment of wounds in the 1st phase of the wound healing process (RF patent No. 2367469, IPC A61K 38/43, 9/06, 2008).

Despite the fact that individual components and their combinations are found in well-known therapeutic ointments for this purpose, the proposed combination and their ratio were not found. The claimed combination of components, taken in experimentally selected, optimal amount, provides the claimed technical result. Other ratios of the components taken do not allow to achieve the necessary therapeutic effect.

Sea buckthorn oil and propolis tincture on 80% ethyl alcohol were introduced in the form of finished products.

Alcohol-water extraction of perforated Hypericum herb, sea buckthorn oil and propolis tincture in an amount of 10 wt.% (5: 3: 2 ratio) were introduced into 90 wt.% Of the rest of the phytogel composition - a base, which was an alloy of polyethylene oxide 400 and 1500 taken in a ratio of 1: 1.

A wound healing agent was prepared as follows. In the reactor with a steam jacket, the refractory component PEO-1500 was melted, then PEO-400 was introduced and mixed. In the reactor-mixer, with a stirrer rotating at no more than 30 rpm, a molten base was fed into which propolis extracts and tincture of sea buckthorn oil were added in parts in the stated proportions. The homogenization of the gel was carried out using a stirrer until a mass was uniform throughout the volume for 30-40 minutes.

The finished phytogel was subjected to analysis for homogeneity, appearance, organoleptic properties, microbiological purity, qualitative and quantitative content of biologically active substances.

Example

In the experiment, white outbred rats of both sexes weighing 150-175.0 g were used. Animals were obtained from the nursery, underwent two-week quarantine and kept in standard vivarium conditions. In each experimental group there were six animals (n = 6). A model of diabetes mellitus was created by intraperitoneal administration of alloxan hydrate at a dose of 170 mg / kg. Diabetes developed on the 4th day after administration of the drug.

On the 4th day after intraperitoneal administration of alloxan hydrate, 1 drop of blood was taken from the tail vein and blood glucose, total cholesterol, triglycerides, high density lipoproteins (HDL) were measured (see Table 2). All of these indicators were measured using a CardioChek portable biochemical blood analyzer using appropriate test strips. Then calculated the content of low density lipoproteins (LDL) mmol / l according to the formula 1:

LDL = Total cholesterol-HDL-Triglycerides / 5 (1)

The change in the above indicators was recorded upward (except for HDL, the level of which decreased from 1.03 ± 0.01 to 0.52 ± 0.04).

Linear skin wounds were applied under anesthesia - chloral hydrate (300 mg / kg). The skin of the back was cut to its own fascia. The cut length averaged 25 mm. Then, at an equal distance from the edges of the wound 1 suture was applied, bringing together the edges of the wound. For the convenience of subsequent measurement of the size of the wounds, the sutures were imposed so that the epithelium of the lateral edges of the wound did not come into contact, and in this case the epithelization occurred from the final edges of the wound.

Evaluation of the wound healing effect was carried out according to the nature of the clinical course (the presence of suppuration, the time of complete rejection of the scab, the time and dynamics of the complete fusion of the edges of the wound).

The studied objects were applied to the wounds of animals - the first group of animals was not treated (control), the second group of animals was daily applied to the wound surface with levomekol (comparison drug), the third group of animals - phytogel. All preparations were applied throughout the entire period of the experiment until the wounds healed completely.

The effect of the developed phytogel on the healing time of a linear skin wound in rats with experimental alloxan diabetes (see table 3 in figure 3).

By the fifteenth day of the experiment, the studied phytogel significantly exceeds the comparison drug levomekol by 53.08%, the differences are significant (p ₁ <0.05 *). By the twentieth day of the experiment, the studied phytogel 1 was 39.90% more active than the reference drug.

Complete healing of linear skin wounds was observed in two experimental groups by the twenty-fifth day of the experiment, with the exception of the control group, where M averaged 1.30 mm even by the 25th day of the experiment.

For histological examination, sections of the formed scars of three groups of experimental animals with alloxan diabetes and a linear skin wound are presented. The first group - did not receive treatment (control); the second group is treated with a comparison drug (levomecol); the third group is treated with phytogel.

When a histological examination of the slice in the control group, wound healing occurred by secondary intention (see figure 4). The wound defect is filled with granulation tissue, in which cellular elements of connective tissue predominate. The resulting vessels, inside covered with endothelium, are viewed. Incomplete epithelization is observed from the edges of the wound. Under the fused epithelium, an incomplete granulation cavity is visible, partially filled with the remains of necrotic tissue. Layers of horizontally located fibroblasts and collagen fibers are not formed, which indicates a sluggish wound healing process.

Upon histological examination in the group treated with the comparison drug levomekol (see figure 5), epithelization of the wound surface is completely completed. The scab on the surface of the scar is absent. There is no superficial leukocyte-necrotic layer. The formed scar is wide; cellular elements of connective tissue prevail in it. A small amount of the vascular component of the granulation tissue, which is in the involution stage, is visible. Germination from below of collagen fibers is weakly expressed.

On histological sections of the third group treated with phytogel, fusion of the epithelium occurred (see Fig.6). Granulation tissue itself consists mainly of cellular elements of connective tissue.

As a result of a histological study of sections of the formed scars after a linear skin wound on the background of experimental alloxan diabetes, it was shown that the developed phytogel according to its indicators is the degree of epithelization and keratinization of the epidermis, the ratio of cellular and fibrous elements of connective tissue, the severity and involution of the vascular component of granulation tissue is not active inferior to the comparison drug levomekol.

Advantages of using the proposed composition: the developed phytogel promotes the healing of linear skin wounds on the background of experimental alloxan diabetes mellitus at all three stages of the wound healing process (exudation, proliferation, epithelization); histological characteristics of sections of a linear skin wound characterize the maturity of the formed scar.

Claims (1)

Wound healing agent for the prevention and treatment of wounds in patients with diabetes mellitus, characterized in that it contains an alcohol-water extract from perforated St. John's wort, which is prepared on a 40% alcohol solution at a rate of 1: 5, sea buckthorn oil, propolis tincture and as a base alloy PEO-400 and PEO-1500 (1: 1), with the following components, wt.%:
alcohol-water extract of hypericum perforated 5.0-5.5 sea buckthorn oil 3.0-3.5 propolis tincture 2.0-2.5 the basis rest

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