BG3731U1 - Gel composition for the treatment of varicose and diabetic wounds - Google Patents

Abstract

The composition, according to the utility model, will find application for the treatment of varicose and diabetic wounds. According to the utility model, the gel for wound healing contains c / wt. % / bioactive composition, which is a mixture of yeast hydrolysates from Saccharomyces cerevisiae, allantoin, gelatin hydrolyzate, calendula extract (Calendula officinalis), plantain leaf extract (Plantago major), the bioactive composition being in an amount of 25 to 30, aqueous extract of mucus of garden snails Helix aspersa in an amount of 25 to 40, copper sulphate in an amount of 0.001 to 0.005 ingredients in an amount of 12 to 14, preservative in an amount of 1 to 2 and deionized water up to 100. The gel for the treatment of varicose and diabetic wounds provides extremely high antibacterial, antifungal and regenerating effect. The application of the gel does not cause side effects. The treatment of the affected areas with the gel, according to the utility model, leads to complete healing of the wounds.

Description

Field of application

The composition according to the utility model will find application for the treatment of varicose and diabetic wounds.

BACKGROUND OF THE INVENTION

One of the most common causes of severe wounds is diabetes. The disease causes different types of wounds and damage to the limbs, which requires a different approach to treatment. In practice, combined treatment with various drugs and equipment is used to treat these wounds.

Decreased neutrophil chemotaxis and phagocytosis predispose patients with diabetes to infections. Insufficient control of diabetes increases the likelihood of pathogenic microorganisms such as Staphylococcus, Streptococcus and Candida albicans causing skin infections. Dermatophyte infections occur in both patients and healthy individuals, but life-threatening skin infections such as necrotizing fasciitis, malignant otitis externa, and mucormycosis (unspecified zygomycosis) are more common in diabetics and require special care.

The use of snail mucus as an active ingredient in therapeutic agents for the treatment of burns and skin diseases, as well as in cosmetic products is known from patents US 2009/026349 and WO 2009/002982 A2.

WO 2012097466 A1 discloses a pharmaceutical composition of a product comprising snail slime from Helix aspersa muller (Cryptophalus aspersus) (from 5% to 50%) and pharmaceutically acceptable additives and / or carriers for the preparation of a gel or cream with a viscosity of 50 Pas up to 1000 Pas. This composition in the form of a cream or gel is impregnated from gauze in the form of a patch, bandage or sock for application to ulcers or other similar skin conditions. The composition may contain extracts of calendula (Calendula officinalis), propolis and vegetable oils. The utility model relates to the composition, method for its preparation and its use as a drug or device for the prevention or treatment of ulcers or wounds with poor prognosis caused by metabolic pathologies such as diabetes, gout, Lesch Nyhan syndrome or other skin conditions , including psoriasis, keratosis, onychomycosis and dermatomycosis, among other diseases.

A drug is known under the trademark IRUXOL, which is a proteolytic enzyme for topical application. Each gram ointment of the preparation contains collagenases 0.6 IU and proteinases 0.12 IU. The action of the preparation is associated with the removal of necrotic tissue in dermal and subdermal wounds, pressure ulcers, burns, frostbite, gangrene; preparation for skin transplantation. The ointment is applied in a thin layer on the wound surface 1-2 times a day after preliminary cleansing. In order to avoid the risk of irritation of the surrounding skin, the application of the preparation requires additional application of zinc paste around the edges of the wound. The course of treatment is for a period of 1-2 weeks.

Technical essence of the utility model

The task of the utility model is to create a gel for the treatment of varicose and diabetic wounds, which provides a high regenerative, antibacterial and antifungal effect, and contains bioactive components of natural origin.

The problem is solved with a gel composition for the treatment of varicose and diabetic wounds, containing c / wt. % / bioactive composition, which is a mixture of yeast hydrolyzate from Saccharomyces cerevisiae, allantoin, gelatin hydrolyzate, calendula extract (Calendula officinalis), plantain leaf extract (Plantago major), the bioactive composition is in an amount of 25 to 30 , aqueous extract of the mucus of snails Helix aspersa in an amount of 25 to 40, copper sulfate in an amount of 0.001 to 0.005, ingredients in an amount of 12 to 14, preservative in an amount of 1.0 to 2.0 and deionized water up to 100.

The bioactive composition is a 100% mixture with the following qualitative and quantitative composition: hydrolyzate of yeast Saccharomyces cerevisiae and allantoin 40, gelatin hydrolyzate from 15 to 20, calendula extract (Calendula officinalis) from 20 to 30 and extract of plantain leaves (Plantago major ) of 15

5389 Descriptions of utility model registration certificates № 06.2 / 30.06.2020 to 20. The amount of allantoin is from 0.5 to 1. The bioactive composition contains DNA and RNA hydrolyzate from the yeast Saccharomyces cerevisiae, which are a mixture of 2'-deoxyadenosine, 2'-deoxyguanosine 2'-deoxycytidine, thymine deoxyriboside, adenosine and uridine, which are in a ratio of 1: 1: 1: 1: 1: 1.

According to the utility model, calendula extract (Calendula officinalis) and plantain leaf extract (Plantago major) are water-alcohol extracts. The aqueous extract of snail mucus from Helix aspersa has a protein concentration of 0.3 to 0.5 mg / ml.

According to the utility model, the ingredients included in the gel are carbopol and triethanolamine, and the preservative is 1,2-hexanediol.

According to the utility model, the gel for the treatment of varicose and diabetic wounds has a proven high biological activity:

• Rich in collagen, elastin, peptides and glycans, garden snail extract stimulates cell regeneration and promotes rapid healing of ulcers or wounds. The effect is enhanced by the addition of collagen hydrolyzate (collagen peptides) and hydrolyzate from the yeast Saccharomyces cerevisiae, which accelerate the regeneration of healthy cells and participate in wound healing processes.

• The snail mucus extract contains heparin - a bound glucose-amino-glycan that helps break down a blood clot and improves blood circulation.

• The action of peptides in mucus, with a proven antibacterial effect, is enhanced by the added extract of plantain (Plantago major) and calendula (Calendula officinalis), which are known for their pharmacological properties and wound healing effect.

• Allantoin helps to heal wounds, smooth the skin and cell regeneration, as well as a moisture-binding effect similar to urea. Its antimicrobial and anti-inflammatory action helps to treat many skin diseases, including the treatment of varicose veins. However, the human body cannot synthesize allantoin and therefore it is necessary to obtain additional.

• The addition of copper ions to the gel is beneficial in diabetes because they improve the action of insulin, which regulates the amount of sugar in the blood. Copper ions are involved in the formation of the structure of connective tissue proteins - collagen and elastin and other proteins important for the body.

The gel according to the present utility model is obtained as:

1. The starting water-soluble extract of the snail slime of Helix aspersa is mixed with the collagen peptide solution and an aqueous solution containing copper sulphate (source of copper ions) with constant stirring until complete homogenization.

2. To the solution thus obtained under item 1 is added a hydrolyzate of the yeast Saccharomyces cerevisiae and allantoin containing the appropriate combination of DNA and RNA, with continuous stirring until the solution is completely homogenized.

3. To the obtained solution under item 2 are added sequentially, with stirring the extract of calendula (Calendula officinalis) and broadleaf plantain (Plantago major).

4. The solution thus obtained under item 3 is added in a thin stream with stirring at slow speeds to the pre-obtained gel from the ingredients (carbopol and triethanolamine), the corresponding amount of preservative and deionized water up to 100%.

The gel for severe, diabetic and varicose wounds, according to the utility model, was tested to determine its effect. In order to establish the antibacterial and antifungal effect, in vivo studies of wounds caused by diabetes have been performed.

Experiments were performed with collected gauze material (provided by a clinical facility) on a diabetic wound before and after treatment with the developed gel.

Figure 1 shows a diabetic treated wound with the gel, according to the utility model and the dressing used for microbial tests. Gauze soaked in gel for severe diabetic and varicose wounds is used to treat diabetic wounds. After its removal from the wound, it is stored in a sterile container to check the antibacterial and antifungal properties of the product used.

5390 Descriptions to utility registration certificates № 06.2 / 30.06.2020

An extract was extracted from the gauze, which was studied for the presence of spores and mycelium of pathological fungal species. The antifungal activity of the gel for severe diabetic and varicose wounds was studied by applying two methods: 1) plaque culture method and 2) agar-diffusion method.

The results of the study show that even after 7 days of cultivation, no change in the color of resazurin was detected, which indicates a lack of spore development and growth of mold fungi (Fig. 2). Figure 2 shows the staining of the development of mold spores after: A) 24-hour cultivation; B) 7-day cultivation.

When the agar diffusion method is applied, the gel is added to the glucose-potato agar medium plates according to the utility model, which is spread over the entire surface of the agar. Gauze material from a treated diabetic wound is applied to the surface of the agar. The results obtained are shown in Figures 3 and 4.

It was found that at 24 hours after inoculation of agar medium with gauze extract from a diabetic wound soaked in the gel for severe varicose and diabetic wounds, there was no development of mold (Fig. 3): (A) upper surface; (B) lower surface.

It was found that even after 7 days of cultivation, the development of mold fungi (Fig. 4) on agar culture medium containing gel for severe wounds was not reported.

Figure 4 shows the lack of growth of mold spores after inoculation of agar medium with gauze extract from a diabetic wound soaked in the gel after 7 days of cultivation. (A) upper surface; (B) lower surface.

The performed microscopic analyzes show the presence of a bacterial infection before treatment with the gel, according to the utility model, which significantly decreases or disappears after treatment (Fig. 5). Similar studies were performed on gauze extracts taken from 4 different wounds. In the initial stage, before treating the wounds with the gel, a bacterial infection was established, which responded after treating the wound with the gel. Analyzes show that after treating the wounds with the gel, according to the utility model, no bacterial infection is observed. Figure 5 shows microscopic analyzes of the wound, before and after gel treatment, which show the presence of bacterial infection before wound treatment.

The composition of the bodies according to the utility model provides extremely high antibacterial, antifungal and regenerating effect. The application of the gel does not cause side effects. The treatment of the damaged areas with the gel, according to the beneficial effect, leads to complete healing of the wounds.

Exemplary implementation of the utility model

Example 1. All specified quantities are in wt. %.

The composition of the gel for the treatment of varicose and diabetic wounds, according to the utility model, contains:

1. Aqueous extract of the mucus of garden snails Helix aspersa - 40%, with a protein concentration of 3 mg / ml (determined by the method of Bradford);

2. Bioactive composition - 25%, containing a combination of the following components:

Communication: Quantity '1 A mixture of 4 D11K hydroxides and 2 PCC hydrolysates of cerevitfoe and allantoin: 2 ^m- deoxyadensions (DCC nucleoid) - 20 μM 2'-deoxyguanosine {D1IK nucleoside} - 20 mN 2 -di ⁺ Ezucleosmd) - 20 gpM 1'imine dszchzhsrnriooznd (DNA nucleoside) - 20 gpM Addnosine (RNA nucleoside! - 20 gpM Urniain (RNA nucleoside} - 20 mM Allantoin - 1% 40% 20% Gelatin hydrolyzate (collage peptides) 20% 3 Broad-leaved plantain extract {Pkmtagp major} 20%. 4 Marigold extract (Catendukt atfit-inalis} 20%

5391 Descriptions to utility registration certificates № 06.2 / 30.06.2020

3. Copper ions in the form of copper sulfate (CuSO ₄ ) - 0.001% (10 mg);

4. Ingredients total amount 12%, which are carbopol - 6% and triethanolamine - 6%;

5. Preservative 1,2-hexanediol - 1%

6. Deionized water up to 100%.

Method of preparing a solution of a bioactive composition:

Prepare a 20 μM solution of the corresponding DNA and RNA hydrolysis from the yeast Saccharomyces cerevisiae and 1% allantoin in 100 ml buffer containing 0.14 M sodium chloride and 0.010 M Tris-HCl, pH 7.5. The solution was heated on a water bath to 70 ° C for 5 minutes and vortexed until the components were completely dissolved.

Prepare a 20% solution of lyophilized gelatin hydrolyzate (collagen peptides) in 50 ml buffer containing 0.14 M sodium chloride and 0.010 M Tris-HCl, pH 7.5. The solution is vortexed until completely dissolved without heating. The gelatin hydrolyzate (collagen peptides) used was obtained by additional protolytic hydrolysis (to obtain shorter chain bioactive peptides) of Rousselot® ProTake P -P (Hydrolyzed gelatin), after using an enzyme cocktail containing three different proteases: trypsin alkalase and flavozyme.

Plantain extract (Plantago major) 50 ml (or 20%)

Preparation of plantain extract (Plantago major): Extraction of the biologically active substances from 100 g of dried plantain leaves was carried out by maceration in the dark with an aqueous-alcoholic solution containing 40% ethanol for 24 hours at room temperature. temperature, the ratio of solvent to raw material used being 2: 1. The extraction process is completed in an ultrasonic bath at room temperature for 30 minutes, in order to more completely recover the active substances. After the following operations: decantation, pressing, centrifugation, filtration, concentration on a vacuum rotary evaporator, the resulting extract is lyophilized. An aqueous-alcoholic extract containing 15% ethanol was prepared, 1 ml of which contained 400 mg of plantain (Plantago major).

Marigold extract (Calendula officinalis) 50 ml (or 20%)

Preparation of calendula extract (Calendula officinalis): Crushed, dried flower baskets of calendula (Calendula officinalis) were extracted by maceration with 45% propylene glycol (or aqueous-alcoholic solution containing 45% ethanol), in a ratio of solvent to raw material 4: 1 . The process is completed by ultrasonic extraction at room temperature for 30 to 45 minutes. After a series of operations, such as: decantation, pressing, centrifugation, filtration, concentration on a vacuum rotary evaporator, the resulting extract is lyophilized. An extract containing 15% ethanol and 35% glycerol was prepared, 1 ml of which contained an extract corresponding to 200 mg of calendula (Calendula officinalis).

An aqueous solution of copper sulphate is prepared by dissolving 10 mg of CuSO ₄ in 100 ml of deionized water.

The method for preparing the gel includes the following steps:

1. The starting water-soluble extract of the snail slime of Helix aspersa is mixed with the collagen peptide solution and an aqueous solution containing copper sulphate (source of copper ions) with constant stirring until complete homogenization.

2. To the solution thus obtained under item 1 is added the corresponding mixture of DNA and RNA hydrolyzate from yeast Saccharomyces cerevisiae and allantoin, with continuous stirring until complete homogenization of the solution at room temperature (25 ° C).

3. To the solution obtained under item 2, the extract of calendula (Calendula officinalis) and the extract of plantain (Plantago major) are added sequentially, with constant stirring.

4. The solution thus obtained under item 3 is added in a thin stream with continuous stirring at slow speed to the pre-obtained gel of the ingredients, which are carbopol and triethanolamine, the corresponding amount of preservative 1,2-hexanediol and deionized water up to 100%.

Example 2. All specified quantities are in wt. %.

The composition of the gel for diabetic and varicose wounds, according to the utility model contains:

5392 Descriptions to utility registration certificates № 06.2 / 30.06.2020

1. Aqueous extract of the mucus of garden snails Helix aspersa - 25% with a protein concentration of 0.5 mg / ml (determined by the method of Bradford);

2. Bioactive composition - 30%, combination of the following water-soluble components:

Commodity: Col n HLCIBiT ] 1 A solution of a mixture of 4 DNA kndroliit, n 2 RCHK hydroplishga of SavefatrorHyces cerevisiae and alangoin: 2'Adschon ^ nadenoshn (DNA n ukl «gtd) -40 ιηΜ 2'-deoxyluanosine (DNA nucleoside) - 40 mM 2 '-desitksn! entcdi n (D1ΙΚ nucleoside) - 40 Ώ1Μ Thyme desokeririboznd (DNA nucleoside) - 40 mM Adepshst {RNA nucleoside) * 40 mM Uridia (RNA nucleoside) - 40 mM Allantonp - 0.5% 40% 2 20% XtjjaLMHiju kndroli: sht (collagen ieptn.di) 15% 3 Pfntrtagn ttrajfir} 15% Marigold err extract (Caieiitluiu ujfieitrttiis) 30%

3. Copper ions in the form of copper sulfate (CuSO ₄ ) - 0.005%;

4. Ingredients total amount 14%, which are carbopol - 7% and triethanolamine - 7%;

5. Preservative 1,2-hexanediol - 2%;

6. Deionized water up to 100%.

Method of preparing a solution of a bioactive composition

1. Prepare a 20 mM solution of the corresponding DNA and RNA hydrolysis from the yeast Saccharomyces cerevisiae and 0.5% allantoin in 120 ml of deionized water. The solution was heated on a water bath to 80 ° C for 10 minutes and vortexed until the components were completely dissolved.

2. Prepare a 20% solution of lyophilized gelatin hydrolyzate (collagen peptides) in 45 ml of deionized water. The solution was vortexed until complete dissolution without heating at room temperature (25 ° C). Ready-made gelatin hydrolyzate of Rousselot® ProTake ™ -P (Hydrolyzed gelatin) was used.

3. Plantain extract (Plantago major) 45 ml (or 15%); INCI name: Plantago major leaf extract, CAS № 84929-43-1; EINECS / ELINCS №: 284-526-8; Chem / IUPAC.

4. Marigold extract (Calendula officinalis) 90 ml (or 30%), INCI name: Calendula officinalis Flower Extract; CAS № 84776-23-8; EINECS № 283-949-5; Chcm / IUPAC Name: Calendula officinalis.

5. Prepare an aqueous solution of copper sulphate by dissolving 5 0 mg in 100 ml of deionized water, homogenised on a vortex at room temperature (25 ° C).

The method for preparing the gel includes the following steps:

1. The starting water-soluble extract of the snail slime of Helix aspersa is mixed with the collagen peptide solution and an aqueous solution containing copper sulphate (source of copper ions) with constant stirring until complete homogenization.

2. To the solution thus obtained under item 1, a combination of DNA and RNA hydrolysis of the yeast Saccharomyces cerevisiae and allantoin cooled to 25 ° C is added, with continuous stirring until complete homogenization of the solution at room temperature (25 ° C).

3. To the solution obtained under item 2, the extract of calendula (Calendula officinalis) and the extract of plantain (Plantago major) at room temperature (25 ° C) are added sequentially, with constant stirring.

4. The solution thus obtained according to item 3 is added in a thin stream with stirring at slow speed, room temperature (25 ° C), to the previously obtained gel from the ingredients carbopol and triethanolamine, to the preservative 1,2-hexanediol and deionized water up to 100 %.

Claims (7)

5393 Descriptions to utility registration certificates № 06.2 / 30.06.2020 Claims A gel composition for the treatment of varicose and diabetic wounds, containing snail mucus Helix aspersa and herbal extracts, characterized in that it contains c / wt. % / bioactive composition, which is a mixture of hydrolysis of yeast from Saccharomyces cerevisiae, allantoin, gelatin hydrolyzate, calendula extract (Calendula officinalis), extract of plantain leaves (Plantago major), the bioactive composition is in an amount of 25 to 30 , aqueous extract of snail mucus Helix aspersa in an amount of 25 to 40, copper sulfate in an amount of 0.001 to 0.005, ingredients in an amount of 12 to 14, preservative in an amount of 1 to 2 and deionized water up to 100. Gel composition according to claim 1, characterized in that the bioactive composition contains water-soluble components and is a 100% mixture with the following qualitative and quantitative composition of the hydrolysis of yeast Saccharomyces cerevisiae and allantoin 40, gelatin hydrolyzate 15 to 20, calendula extract (Calendula officinalis) from 20 to 30 and leaf plantain extract (Plantago major) from 15 to 20. Gel composition according to claim 1, characterized in that the amount of allantoin is from 0.5 to 1. Gel composition according to claim 1, characterized in that the bioactive composition comprises the DNA and RNA hydrolysis of the yeast Saccharomyces cerevisiae, which are a mixture of 2'-deoxyadenosine, 2'-deoxyguanosine, 2'-deoxycytidine, thymine deoxyribosine, adeno uridine and are in a ratio of 1: 1: 1: 1: 1: 1. Gel composition according to claim 1, characterized in that the calendula extract (Calendula officinalis) and the leaf extract of plantain (Plantago major) are aqueous-alcoholic extracts. Gel composition according to claim 1, characterized in that the aqueous extract of Helix aspersa snail mucus has a protein concentration of 0.3-0.5 mg / ml. Gel composition according to claim 1, characterized in that the ingredients are carbopol and triethanolamine, and the preservative is 1,2-hexanediol.