BG66767B1 - Biologically active composition - Google Patents

Abstract

The invention relates to a biologically active compositionwhich can be used for treatment of chronic wounds as well as forcosmetic purposes. The preparation solves the problem ofaccelerating the healing process of wounds of different origin andcondition. The biologically active composition according to theinvention contains a relatively low concentration of glycerin-stabilized alkaline proteinase, collagen hydrolyzate, magnesium andzinc ions, vitamins, antiseptics. The preparation thus produced has awide range of action and a number of positive effects, which makesit multifunctional. The proposed combination of seeminglyopposing agents allows the recovery of previously known chronicnon-healing wounds in patients with diabetes or suffering fromvascular diseases (arterial insufficiency), and meanwhile can be usedin a variety of skin defects such as stinging, inflammation in burninjuries or insects, hitting injury hematoma, for lichen or scabies ofpsoriasis, etc.

Description

Field of technology

The invention relates to a biologically active composition based on proteolytic enzymes, which is used in medicine, the pharmaceutical industry and cosmetics.

BACKGROUND OF THE INVENTION

The efforts of a number of researchers are focused on the development of such biologically active compositions, the action of which can reduce the healing time of wounds of different origins at the application of individual therapeutic doses. Such compositions can help to remove old cell debris and generate new tissue and therefore increase the rate of tissue healing. In addition, chronic wounds show a different treatment profile than normal acute wounds, as they usually remain inflamed for a long period of time.

Wound treatments such as iodine tincture, 3% hydrogen peroxide solution, dilute potassium permanganate solution, iodasept and other oxidants are known. Tessa designed to decontaminate the wound [1] and thus allow the body to build its own tissue. The main problem with these preparations is that they have a limited application - they oxidize (burn) not only the microorganisms in the wound, but also healthy tissue of the patient [1]. Their use can lead to redness, swelling and are only suitable for people with preserved protective abilities and good blood circulation, such as young patients. Daily practice shows that said disinfectants in patients with circulatory problems or diabetes usually lead to necrosis, deepening and widening of the wound and even amputation of a limb.

Compositions are known, the application of which are methods relying on the mechanical withdrawal of the exudate from the wound by means of a highly swollen patented "soft silicone material with an antiseptic included in the dressing" [2].

These dressings reduce the possibility of a large number of microorganisms developing in the wound because they constantly pull them out of the wound area. This of course supports the treatment, but it turns out that these methods do not lead to healing of chronic non-healing wounds. It is known to use ready-made patches containing antiseptics, for example chlorhexidine acetate [3], which are applied directly to the wound. However, the contact between the patch and the relief of the wound is not always good, the necrotized tissue and fibrin deposits are almost unaffected and the results in chronic non-healing wounds also do not lead to wound closure. It is usually difficult to determine a therapeutically effective dose to obtain the desired biological result. The latest trends in formulations with biological action are those that use enzymes combined with antiseptics with a broad spectrum of action [4,5]. A composition [6] has also been proposed according to which a polyethylene oxide gel containing collagenase from Clostridium histolyticum (a type of microorganism) and gentamicin provides bloodless enzymatic cleansing of the wound, stimulates the development of healthy granulations and does not interfere with the epithelialization process. Enzymes break down necrotic tissue, pus and fibrin deposits and thus allow the body to begin to build its own tissue. The antiseptic provides sterility in the wound area. In most cases, this is enough and the wounds heal, but especially when the blood circulation is poor or the patient is also diabetic, the patient needs additional support and acceleration of healing. The prevailing view is that a satisfactory result can only be achieved if ways are found to stimulate healthy cell proliferation, granulation and angiogenesis in the wound area. In the American patent [7] the question of the so-called "Wound Healing Peptides WHPs". They are obtained by the degradation of native collagen by human or bacterial collagenase. These peptides have various very positive properties - to accelerate the proliferative properties of healthy epithelial cells (granulation), enhance their properties to navigate and move to a particular chemical agent (chemotactic properties), accelerate capillary morphogenesis and more. Ways for their isolation are also described, for example, by temporary chemical modification in view of the specific one

Descriptions of issued patents for inventions № 12.1 / 17.12.2018 their absorption on specific carriers, elution in purified form and their conversion back to native form. For this purpose, preparations such as enzyme-degraded collagen are used [8].

There is a problem with slow-healing wounds, in which the so-called fibrin plaque. It is a clot formed by fibrinogen present in the blood and lymph, which, when in contact with activated platelets, as are abundant in such wounds, becomes an insoluble spatial network of fibrin. This is a characteristic tissue of the body. It naturally degrades slowly and needs to be degraded again with a proteolytic preparation, which in many cases requires treatment by a doctor - a specialist in the field. In addition, the slow process of degradation of the fibrin deposit further slows down the overall wound healing process.

To accelerate the granulation of the wound, various substances have been proposed that are not known to have found appropriate application in practice: pancreatic enzyme inhibitors [9], polypeptides and nucleic acids that stimulate the viability of keratinocytes and endothelial cells [7], co-enzyme Q-10 [10] or their contents remain a secret, such as a gel containing an enzyme - from Antarctic plankton with almost universal action [11].

Technical essence of the invention

The invention is a biologically active preparation based on proteolytic enzymes, characterized in that 100 g of preparation contain: 0.1-10 g of enzymatically hydrolyzed sterilized collagen, 0.01-0.04 g of magnesium oxide, 0.01-0.04 g zinc oxide, 0.9-18 mg vitamin A (retinol), 25-300 pg vitamin D (cholecalciferol), 0.2-50 mg vitamin E (alpha-tocopherol), 0.05-0.2 g chlorhexidine digluconate ( hibitan) and 10-500 STA proteolytic units [12] purified stabilized alkaline proteinase.

The biologically active composition according to the invention contains a relatively low concentration of glycerol gel stabilized alkaline proteinases, collagen hydrolyzate, magnesium and zinc ions, vitamins A, D, E and gibitan. The preparation thus created shows a wide range of action and a number of positive effects, which makes the preparation multifunctional.

The invention solves the problem of accelerating the healing process of wounds of various origins and conditions, and there is a manifestation of synergism - as unusual in the same preparation are combined opposite in their action on the body substances. On the one hand, these are enzymes that selectively break down the necrotized parts of the wound, and on the other such substances that stimulate, nourish the building of healthy tissue in the injured area - collagen hydrolyzate, magnesium and zinc ions, vitamins A, D, E, with guaranteed sterility [13]. The unexpected effect is that the unconventional ingredients for such cases do not inhibit the enzyme or reduce its storage activity, but lead to a synergistic effect on wound healing. This makes it possible to treat chronic wounds of diabetics, the elderly or those with problems with blood supply to the wound with one preparation. The new combination does not alter the storage stability of the enzyme. For one year at a temperature below 17 ° C the proteolytic activity of the new preparation did not change by more than 5%.

Judging by the excellent results (rapid healing of chronic wounds) obtained in the application of the composition according to the invention, in the treatment of wounds, quite unexpectedly in the enzymatic hydrolysis of denatured collagen by a very broad specificity protease, short peptides are obtained, which have similar effects to the high molecular weight peptides (Wound Healing Peptides -WHPs) described in U.S. Pat. No. 7 [7], which are obtained there by degrading native collagen using a very specific enzyme such as collagenases.

The advantages of the new preparation are the following. The proposed combination of seemingly opposite substances allows the recovery of hitherto known as non-healing chronic wounds, even in patients with diabetes or suffering from poor blood supply (arterial insufficiency), while it can be used for various skin defects such as stings, burns or inflammation. insects, various hematomas from stroke, lichens or scabs from psoriasis, etc.

Collagen hydrolyzate contains free amino acids and short peptides that can be

Descriptions of issued patents for inventions № 12.1 / 17.12.2018 embedded in the extracellular matrix and thus make the skin healthier, denser and more stable, without the need for these amino acids to be delivered by blood, which accelerates wound healing and reduces the likelihood of wrinkles.

The preparation supplies vitamins A, D and E, as well as magnesium and zinc ions, all involved in collagen metabolism, which makes the skin lifelong.

Alkaline proteinase dissolves the fibrin deposit that often appears in the wound and is characteristic of the body and thus accelerates its healing.

The preparation can also be used for mild inflammations caused by excessive sun exposure, touching a hot object, insect bites, pinching, cutting, scabs or lichens, etc.

The composition of the preparation provides a hydrophilic environment during the construction of the new tissue, and this leads to the natural environment for the construction of the tissue. In this way, the developed preparation accelerates healing, avoids the adhesion of the dressing, and reduces the likelihood of scarring.

The drug, penetrating in depth, shows an unexpected effect, as it dissolves even the hematomas (bruises from clotted blood), as well as fibrin deposits in the capillaries, the latter become more permeable and blood circulation improves (there is a controlled redness around the wound), i.e., which also accelerates wound healing.

The preparation keeps the environment sterile and thus enables even organisms with weak resistance forces, in which almost all processes are performed slowly, to rebuild their skin defect.

The use of the preparation according to the invention allows the application of skin autotransplantation by the same patient and thus healing of even extensive incurable chronic wounds can be achieved.

The use of the preparation according to the invention allows wound healing even in diabetics with insulin, psoriasis and elderly people with severe circulatory disorders.

The same preparation is used to clean the wound as well as to stimulate its healing.

Examples of the invention

Example 1 for the preparation of a 5% gel

Place 5 g of deproteinized starch in 100 g of aqueous-glycerol solution (50:50 rpm), mix well and the temperature of the mixture is gradually raised until a homogeneous mass is reached. After cooling to room temperature, 0.1 g of hydrolyzed sterilized collagen, 0.01 g of magnesium oxide, 0.01 g of zinc oxide, 0.9 mg of vitamin A (retinol), 25 pg of vitamin D (cholecalciferol) are added to 100 g of gel. ), 2 mg vitamin E (alpha-tocopherol), 0.2 g chlorhexidine digluconate and 10 STA proteolytic units [12] purified stabilized alkaline proteinase. The pH is monitored 5.0. The resulting preparation is packed in tubes of 10, 50 or 100 ml and stored at a temperature below 17 ° C.

Example 2 for the preparation of a 10% gel

In 100 g of aqueous-glycerol solution (50:60 v / t) place 10 g of deproteinized starch, mix well and the temperature of the mixture is gradually raised until a homogeneous mass is reached. After cooling, 2 g of hydrolyzed sterilized collagen, 0.02 g of magnesium oxide, 0.02 g of zinc oxide, 1.8 mg of vitamin A (retinol), 50 pg of vitamin D (cholecalciferol), 4 mg of vitamin 2 are added to 100 g of gel. E (alpha-tocopherol), 0.2 g chlorhexidine digluconate and 50 CTA proteolytic units purified and stabilized alkaline proteinase [12]. pH 7.0. The resulting preparation is packed in tubes of 10, 50 or 100 ml and stored at a temperature below 17 ° C. Shelf life two years.

Example 3 for the preparation of a 15% gel

Put 15 g of deproteinized starch in 100 g of aqueous-glycerol solution (50:80 rpm), mix well and the temperature of the mixture is gradually raised until a homogeneous mass is reached. After cooling, 5 g of hydrolysed sterilized collagen, 0.04 g of magnesium oxide are added to 100 g of gel,

Descriptions of issued patents for inventions № 12.1 / 17.12.2018

0.04 g zinc oxide, 3 mg vitamin A (retinol), 100 pg vitamin D (cholecalciferol), 6 mg vitamin E (alpha-tocopherol), 0.2 g chlorhexidine digluconate and 100 STA proteolytic units [12] purified and stabilized alkaline proteinase. pH 5.0. The resulting preparation is packed in tubes of 10, 50 or 100 ml and stored at a temperature below 17 ° C.

Example 4 for obtaining 15% wire

Place 15 g of deproteinized starch in 100 g of aqueous glycerol solution (10:90 rpm), mix well and the temperature of the mixture is gradually raised until a homogeneous mass is reached. After cooling, 5 g of hydrolyzed sterilized collagen, 0.04 g of magnesium oxide, 0.04 g of zinc oxide, 6 mg of vitamin A (retinol), 200 pg of vitamin D (cholecalciferol), 10 mg of vitamin E (100 g) are added to 100 g of gel. alpha-tocopherol), 0.5 g chlorhexidine digluconate and 200 STA proteolytic units [12] purified and stabilized alkaline proteinase. pH 9.0. The resulting preparation is packed in tubes of 10, 50 or 100 ml and stored at a temperature below 17 ° C.

Example 5 for the preparation of an 18% gel

Place 18 g of deproteinized starch in 100 g of glycerin, mix well and the temperature of the mixture is gradually raised until a homogeneous mass is reached. After cooling, 10 g of hydrolyzed sterilized collagen, 0.04 g of magnesium oxide, 0.04 g of zinc oxide, 18 mg of vitamin A (retinol), 300 pg of vitamin D (cholecalciferol), 50 mg of vitamin E (100 g) are added to 100 g of gel. alpha-tocopherol), 0.5 g chlorhexidine digluconate and 200 STA proteolytic units [12] purified and stabilized alkaline proteinase. pH 7.0. The resulting preparation is packed in tubes of 10, 50 or 100 ml and stored at a temperature below 17 ° C.

Example 6 for the preparation of a 2.5% polyacrylamide gel

Proceed as described in Example 4, but instead of starch, 2.4 g of acrylamide, 0.07 g of methylene bisacrylamide, the remaining components as in Example 1 are added, and finally 0.04 g of tetramethylethylenediamine and 0.05 g of ammonium persulphate are added. Allow to gel at room temperature, stir and pack the resulting preparation in tubes of 10, 50 or 100 ml. Store at room temperature below 17 ° C. Shelf life 2 years.

Example 7 for the preparation of a 2% polyacrylamide gel

Proceed as described in Example 4, but instead of starch, add 1.9 g of acrylamide, 0.08 g of methylenebisacrylamide, the remaining components as in Example 1, and finally add 0.03 g of tetramethylethylenediamine and 0.04 g of ammonium persulphate. Allow to gel at room temperature, stir and pack the resulting preparation in tubes of 10, 50 or 100 ml. Store at room temperature below 17 ° C. Shelf life 2 years.

Example 8 for the preparation of a 1.5% polyacrylamide gel

Proceed as described in Example 3, but instead of starch, 1.4 g of acrylamide, 0.07 g of methylene bisacrylamide, the remaining components as in Example 1 are added and finally 0.03 g of tetramethylenediamine and 0.04 g of ammonium persulphate are added. Allow to gel at room temperature, stir and pack the resulting preparation in tubes of 10, 50 or 100 ml. Store at room temperature below 17 ° C. Shelf life 2 years.

Example 9. Testing the effectiveness of application of the preparation

The skin defect is mechanically cleaned with a cotton swab soaked in rivanol or saline solution, or preferably with a boiled solution of sodium bicarbonate (1/2 teaspoon of baking soda in 200 ml of boiling water). The damaged area is covered with 1-2 mm of the preparation, preferably directly or such a layer is applied on gauze and it adheres to the skin every 24 hours. In normally healthy organisms and when the wound is superficial, usually after 4 to 6 dressings the area of damaged skin is cleared of pus and fibrin deposits (whitish areas), and pink or red healthy tissue appears at the bottom of the wound and the edges of the wound become well formed, slightly reddened due to increased blood circulation and begin to retract to the center of the wound. The wound is then treated in the same way until complete healing. Due to the fact that the wound is cleansed, that the emerging fibrin plaque is constantly removed, that the recovery process takes place in a humid natural environment, and that the inflammatory process ends quickly, the likelihood of scarring is low even in

Descriptions of issued patents for inventions № 12.1 / 17.12.2018 extensive wounds. The wound healing process is significantly slower than that of wound cleansing and depends very much on the degree of blood supply to the area where the wound is located.

Claims (3)

Patent claims Biologically active composition in the form of a gel containing proteolytic enzymes, characterized in that 100 g of it contain 0.1-10 g of hydrolyzed sterilized collagen, 0.01-0.04 g of magnesium oxide, 0.01- 0.04 g zinc oxide, 0.9-18 mg vitamin A (retinol), 25-300 pg vitamin D (cholecalciferol), 0.2-50 mg vitamin E (alpha-tocopherol), 0.05-0.2 g chlorhexidine digluconate and 10-500 CTA proteolytic units purified stabilized alkaline proteinase. Biologically active preparation according to Claim 1, characterized in that the gel is based on deproteinized starch in aqueous-glycerol medium. Biologically active preparation according to Claim 1, characterized in that the gel is based on polyacrylamide.