RU2699663C1 - Agent in the form of gel with erythropoietin, having reparative and antioxidant activity - Google Patents

Abstract

FIELD: medicine; pharmaceutics.

SUBSTANCE: invention relates to medicine and pharmaceutical industry and represents a method for preparing an agent for local treatment of wounds in the form of gel with liposomal form of erythropoietin, consisting in preparation of mass by dissolving in 100 ml of water 0.05 g of Nipazol with subsequent addition of polyvinylpyrrolidone 20 % – 20.0 g in a mixture with sodium carboxymethyl cellulose 5 % – 5.0 g, the mass is mixed, left to swell for 20 minutes, then heated in a water bath until complete dissolution with further introduction of liposomal form of erythropoietin.

EFFECT: what is developed is a method for preparing the agent with high effectiveness of wound-healing stimuli in case of thermal injury.

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Description

The invention relates to medicine, namely to pharmacy, and can be used to treat wounds with thermal injury. The product obtained by dissolving 0.05 g of Nipazole in 100 ml of water, followed by the addition of polyvinylpyrrolidone 20% - 20.0 g in a mixture with 5% - 5.0 g sodium carboxymethyl cellulose, the mass is stirred, allowed to swell for 20 minutes, then heated in a water bath until completely dissolved with the further introduction of the liposomal form of erythropoietin. The technical result is a soft dosage form (gel). The gel is evenly distributed over the surface of the wound, creates a cooling and analgesic effect, absorbs exudate. The invention relates to medicine, in particular to pharmacy, for the development of medicinal gels containing erythropoietin. The inventive composition allows you to create a drug for the treatment of wounds in thermal injury with violations of the surface layers of the skin - up to III degree for local treatment. The use of this invention allows to stimulate wound healing during thermal injury due to the pleiotropic effect of erythropoietin.

Analogs of the present invention are transdermal dosage form with erythropoietin [Method for producing a drug with local erythropoietin: US Pat. 2611401 Ros. Federation, IPC A61K 47/10, A61K 38/18, A61K 47/12, A61K 47/38, A61K 47/30 / M.V. Osikov, E.V. Simonyan, O.T. Saedgalina; patent holder South Ural state. honey. University (RU). - No. 2015146627; declared 10/28/2015; publ. 02.21.2017. - 4 pp.] And a gel with erythropoietin [Local application and preparations of erythropoietin for healing skin wounds: US Pat. 2465003 ROS. Federation, IPC A61K 38/18, A61K 9/10, A61K 47/38, A61P 17/02 / Bader Augustinus; patent holder Heroys Kulzer GMBH (DE). - No.2010130890 / 15; declared 12/20/08; publ. 10/27/12, Bull. No. 30. - 36 c.].

Various compounds can be used as active substances in the liposomal gel for local therapy: antibiotics, immunomodulators, antihistamines, antioxidants, vitamins. Endogenous regulators of homeostasis, in particular erythropoietin, are also of interest. Its pleiotropic effects have been the subject of study by many researchers in recent years. Erythropoietin and its biotechnological derivatives are highly suitable for systemic use for specific regeneration of affected tissue in case of damage to the skin, mucous membrane, open and superficial skin wounds, including burn injuries, and contribute to the acceleration of healing. The prior art method for producing recombinant human erythropoietin with cultivation in growth and nutrient media to obtain injectable injection solutions [Method for producing a substance of recombinant human erythropoietin and nanocapsulated form of recombinant human erythropoietin using a substance obtained by the specified method: US Pat. 2518329 Ros. Federation, IPC C07K 14/505 (2006.01), C12N 15/16 (2006.01), B82B 1/00 (2006.01), A61K 9/51 (2006.01) / E.A. Nechaev, N.A. Varaksin, T.G. Ryabicheva [et al.]; patent holder scientific Center for Virology and Biotechnology "Vector" (RU). - No. 2012153074/10; declared 12/07/2012; publ. 06/10/2014, Bull. No. 16 .; Erythropoietin conjugated to polyethylene glycol: US Pat. 2433134 ROS. Federation, IPC C07K 14/505 (2006.01) / S.V. Sheremetyev, V.V. Zverev, S.A. Korovkin [et al.]; patent holder FORT Limited Liability Company (RU). - No.2010140709 / 10; declared 10/06/2010; publ. 11/10/2011, Bull. No. 31.]. The discovered antioxidant properties of erythropoietin can also contribute to limiting the secondary alteration zone during thermal injury [Osikov, M.V. Transdermal hydrocolloid coatings - a promising area for thermal injury / M.V. Osikov, E.V. Simonyan, G.P. Grigoryeva, O.T. Saedgalina, N.N. Nozhkina // Fundamental medicine. - 2014. - No. 10. - S. 1632-1639.]

The prototype of the invention are erythropoietin gels for treating wounds. In particular, the invention relates to special viscous or gelatin preparations based on polysaccharides, in particular cellulose derivatives, which contain erythropoietin and which are capable of stabilizing the latter and its slow and uniform release in the wound [Ointment for the treatment of burns, skin diseases and wound healing: pat . 2325175 ROS. Federation, IPC A61K 36/00 (2006.01), A61K 9/06 (2006.01), A61K 35/64 (2006.01), A61P 17/02 (2006.01) / E.K. Samitin (RU), D. Samitin (LV). - No. 2006120510/15; declared 06/13/2006; publ. 05/27/2008 .; Ointment for the treatment of burns, folliculitis, furunculosis, vasculitis and wound healing: US Pat. 2526152 Ros. Federation, IPC A61K 9/06, A61K 35/64, A61K 35/02, A61K 36/55, A61K 35/12, A61P 17/02 / patent holder M.E. Dzodzikova (RU). - No. 20111151228/15; declared 12/14/2011; publ. 08/20/2014 Bull. No. 23 .; Ointment for the treatment of burns, purulent and infected wounds of various etiologies and a method for the treatment of burns, purulent and infected wounds: US Pat. 2256441 ROS. Federation, IPC A61K 9/06, A61K 35/64, A61K 47/44 A61P 17/02 / E.O. Potapenkova, M.V. Maksimova (RU). - No. 2003134532/15; declared 11/25/2013; publ. 07/20/2005 .; Biological dressing for the treatment of burns: US Pat. 2314129 Ros. Federation, IPC A61F 13/02, A61L 15/32 / A.S. Ermolov, S.V. Smirnov, V.B. Khvatov [et al.]; patent holder of the Research Institute of Ambulance named after N.V. Sklifosovsky Department of Health of Moscow (RU). - No. 2006122784/15; declared 06/27/2006; publ. 01/10/2018 .; Dressing for closing and treating burns: US Pat. 2275179 ROS. Federation, IPC A61F 13/00 (2006.01), A61L 15/22 (2006.01), A61L 15/44 (2006.01) / A.A. Alekseev, A.E. Bobrovnikov, T. S. Vasilieva [et al.] (RU). - No. 2004122457/15; declared 07/22/2004; publ. 04/27/2006.]. This dosage form has a prolonged regenerative effect.

Disclosure of the invention.

The objective of the invention is the creation of a new drug with a complex effect on the various stages of the wound healing process.

The technical result to which the claimed invention is directed is to obtain a means with high efficiency stimulating wound healing in thermal injury due to the regenerating and antioxidant effects of erythropoietin. The effectiveness of the drug is ensured by the selected concentration of erythropoietin.

The problem is also solved by the fact that the means for treating wounds during thermal injury includes the liposomal form of erythropoietin described above, which ensures the stabilization of the erythropoietin molecule and contributes to its maximum prolonged release.

The problem is also solved by the fact that the means for treating wounds during thermal injury includes 20% of polyvinylpyrrolidone, which increases the bioavailability of erythropoietin, has a weak bactericidal effect, and increases transdermal absorption.

The problem is solved in that the liposomal gel with erythropoietin for the treatment of thermal injury, containing erythropoietin as the active substance, and 20% polyvinylpyrrolidone and 5% carboxymethyl cellulose Sodium salt as an excipient, additionally contains Nipazole preservative, in the following ratio of gel components, g, per 100 ml of the finished gel form (Fig. 1)

The optimal concentration of active and auxiliary substances in the product is determined experimentally.

The use of a liposomal soft dosage form containing erythropoietin is used for the first time.

To solve the above problem, the technological characteristics of the dosage form were studied: viscosity, adsorption activity, biopharmaceutical availability.

The technology is carried out under aseptic conditions as follows: 0.05 g of Nipazole is dissolved in 100 ml of water, followed by the addition of polyvinylpyrrolidone 20% (20.0 g) in a mixture with 5% sodium carboxymethyl cellulose (5.0 g), the mass is stirred, allowed to swell for 20 minutes, after which it is heated in a water bath until completely dissolved, followed by the introduction of a liposomal form of erythropoietin into a medium cooled to room temperature.

The proposed method differs from the existing ones in that, in addition to the biopolymer having reparative and barrier functions, an antioxidant and reparative drug is included. As a result, potentiation of the therapeutic effect, reduction of microbial contamination is achieved. The gel form has a cooling and analgesic potential. Absorb exudate on itself, thereby promoting early granulation. Plasticity provides an atraumatic effect. An example of a specific implementation.

Example 1

The composition of the gel mass for the preparation of 100 ml of gel: 0.05 g of Nipazole; polyvinylpyrrolidone 20% - 20 g; Carboxymethyl cellulose sodium salt 5% - 5.0 g, water 100 ml.

A mixture of gelling agents is prepared. 0.05% of Nipazole is pre-dissolved in a beaker in the entire volume of water, gelling agents are added in a mixture of polyvinylpyrrolidone 20% and 5% sodium carboxymethyl cellulose and left to swell for 30 minutes. The resulting mass is heated in a water bath until the components are completely dissolved. Cool at room temperature. Erythropoietin is introduced with constant stirring and left for 1 hour at room temperature to remove air. The resulting gel is evaluated by appearance, adsorption activity and biopharmaceutical availability.

Example 2

The composition of the gel mass for the preparation of 100 ml of gel: 0.05 g of Nipazole; Carboxymethyl cellulose sodium salt 15% (15.0 g), water 100 ml.

0.05% of Nipazole is pre-dissolved in a beaker in the entire volume of water, a gelling agent is added. Sodium salt of carboxymethyl cellulose 15% and left for 30 minutes to swell. The resulting mass is heated in a water bath until the components are completely dissolved. Cool at room temperature. Erythropoietin is introduced with constant stirring and left for 1 hour at room temperature to remove air. The resulting gel is evaluated by appearance, adsorption activity and biopharmaceutical availability.

Example 3

The composition of the gel mass for the preparation of 100 ml of gel: 0.05 g of Nipazole; polyvinylpyrrolidone 20% (20.0 g), water 100 ml.

0.05% of Nipazole is pre-dissolved in a beaker in the entire volume of water, a 20% PVP gelling agent is added and left to swell for 30 minutes. The resulting mass is heated in a water bath until the components are completely dissolved. Cool at room temperature. Erythropoietin is introduced with constant stirring and left for 1 hour at room temperature to remove air. The resulting gel is evaluated by appearance, adsorption activity and biopharmaceutical availability.

Example 4

The composition of the gel mass for the preparation of 100 ml of gel: 0.05 g of Nipazole; Gelatin 5% (5.0 g), water 100 ml.

0.05% Nipazole is pre-dissolved in a beaker in the entire volume of water, 5% Gelatin gelling agent is added and left to swell for 30 minutes. The resulting mass is heated in a water bath until the components are completely dissolved. Cool at room temperature. Erythropoietin is introduced with constant stirring and left for 1 hour at room temperature to remove air. The resulting gel is evaluated by appearance, adsorption activity and biopharmaceutical availability.

Example 5

The composition of the gel mass for the preparation of 100 ml of gel: 0.05 g of Nipazole; Hydroxypropyl methylcellulose 5% (5.0 g), water 100 ml.

In a beaker, 0.05% Nipazole is pre-dissolved in the entire volume of water, a 5% hydroxypropylmethyl cellulose gelling agent is added and the mixture is allowed to swell for 30 minutes. The resulting mass is heated in a water bath until the components are completely dissolved. Cool at room temperature. Erythropoietin is introduced with constant stirring and left for 1 hour at room temperature to remove air. The resulting gel is evaluated by appearance, adsorption activity and biopharmaceutical availability.

Received dosage forms were evaluated visually, as well as for compliance with technological parameters. In addition, adsorption capacity and biopharmaceutical parameters (erythropoietin release) are being studied.

The gel adsorption activity, which satisfies the technological parameters, is evaluated by its ability to adsorb dye methylene blue. To do this, 50 μl, 100 μl, 150 μl, 200 μl, 250 μl, 500 μl of erythropoietin are taken alternately, adjusted to the mark in a 50 ml volumetric flask with a 0.1% methylene blue solution. The flask is placed on a shaker and shaken for 1 hour at a speed of at least 120 '. The resulting solution is filtered. 2.5 ml of the filtrate was placed in a 100 ml volumetric flask, adjusted to the mark with water and the optical density of the solution was measured at 667 nm. In parallel, a blank experiment is conducted with a methylene blue solution under the same conditions. The adsorption capacity of 1 g of the drug should be at least 0.03 g of methylene blue. The results of the determination of adsorption activity (Fig. 2).

Evaluation of the degree of release of erythropoietin was evaluated by HPLC in a dissolution test. The results are presented in the figure (Fig. 3).

From the above examples, it is seen that the composition described in Example No. 1 is optimal in terms of technological parameters, adsorption activity, and ability to gradually release erythropoietin.

A method of obtaining a liposomal gel with erythropoietin for the treatment of thermal injury, during the implementation of the method, a gel is prepared (the mass is mixed, left to cool completely at room temperature, poured into a mold.

Thus, the invention makes it possible to organize the production of a stable highly effective means for treating wounds in thermal injury.

A brief description of the drawings.

Figure 1 - The ratio of the components of the gel, g, per 100 ml of the finished gel form.

Figure 2 - The results of determining the adsorption activity of the gels.

Figure 3 - Results of the assessment of the degree of release of erythropoietin.

Figure 4 - A method of obtaining a liposomal gel with erythropoietin for the treatment of thermal injury.

Claims (1)

A method of obtaining a means for local treatment of wounds in the form of a gel with a liposomal form of erythropoietin, when implementing the method, the mass is prepared by dissolving 0.05 g of Nipazole in 100 ml of water, followed by the addition of polyvinylpyrrolidone 20% - 20.0 g in a mixture with 5% sodium carboxymethyl cellulose - 5.0 g, the mass is stirred, left to swell for 20 minutes, after which it is heated in a water bath until completely dissolved, followed by the introduction of the liposomal form of erythropoietin.

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