RU2811124C1 - Ophthalmic medicinal films based on biostimulator from cattle brain tissue with addition of silver nanoparticles - Google Patents

Abstract

FIELD: biotechnology; medicine; ophthalmology.

SUBSTANCE: invention can be used to treat corneal injuries. A method has been proposed for producing an ophthalmic medicinal film based on a biostimulator from cattle brain tissue.

EFFECT: invention provides increased effectiveness of treatment of erosive and ulcerative pathologies of the cornea, acceleration of healing processes, minimization of exposure and contact with the pathological focus.

1 cl, 1 ex

Description

Field of technology to which the invention relates

The invention relates to medicine, in particular to ophthalmology, and can be used for the treatment of ulcerative injuries of the cornea.

State of the art

Pathologies of the anterior segment of the eyeball account for the majority of all eye diseases in animals, which is anatomically determined [Fedota, N.V. Analysis of the prevalence of diseases of the organ of vision in dogs and cats in Stavropol / N.V. Fedota, A.N. Shulunov , A. N. Kvochko // Trends in the development of veterinary surgery: materials of the International scientific and practical conference dedicated to the 95th anniversary of the Department of General, Private and Operative Surgery, Vitebsk State Academy of Veterinary Medicine, Vitebsk, November 03–04, 2021. – Vitebsk: Educational Institution “Vitebsk Order of the Badge of Honor” State Academy of Veterinary Medicine,” 2021. – P. 136-137. – EDN XEZWTW]. Treatment is most often based on minimally invasive techniques using eye drops and ointments of different composition and spectrum of action. Methods of administration into the conjunctival sac and onto the surface of the cornea are different: instillation, instillation, laying. The dosage forms used in these methods have a number of both advantages and disadvantages. The administration of liquid dosage forms has low labor costs, ease of use, and often minimal contact with the animal, which reduces its anxiety. However, the high frequency of exposure and the duration of the treatment period, low efficiency encourages researchers to find new forms for use in the anterior segment of the eye [Bilalov, E. N. Evaluation of the tolerability of the eye medicinal film "GlazAvir" in an experiment / E. N. Bilalov, F. A. Bakhritdinova, K. M. Imomalieva // Russian National Ophthalmological Forum. – 2018. – T. 2. – P. 442-446. – EDN VVGNCG; Aznabaev, M. T. Ophthalmic medicinal films in the prevention of infectious and inflammatory complications / M. T. Aznabaev, G. A. Azamatova, G. Ya. Gaisina // Saratov Journal of Medical Scientific Research. – 2018. – T. 14. – No. 4. – P. 933-938. – EDN VGZPJV].

A modern solution is the use of ophthalmic medicinal films for the treatment of corneal pathologies. This is a relatively new dosage form that has proven itself in the pharmaceutical market as a reliable medicine. Ophthalmic medicinal films provide prolonged delivery of the active substance to the affected area, which reduces the number of manipulations for drug administration.

The closest in technical essence and achieved effect and accepted by the authors as a prototype are ophthalmic medicinal films with various active ingredients, such as levofloxacin, 6-methyl-3-(thietan-3-yl)uracil, streptodecase and emoxin, papain and urea. The basis of the ophthalmic medicinal film is a film-forming substance. A number of researchers use polyvinyl alcohol, polyvinylpyrrolidone, methylcellulose, sodium carboxymethylcellulose, gelatin [Patent No. 2404779 C1 Russian Federation, IPC A61K 31/715, A61K 9/08, A61P 27/02. Ophthalmic medicinal film: No. 2009140514/15: application. 02.11.2009: publ. 11/27/2010 / G. A. Azamatova, M. T. Aznabaev, Yu. B. Monakov [etc.] ; applicant State Institution "Ufa Research Institute of Eye Diseases" of the Academy of Sciences of the Republic of Bashkortostan (UfNII GB AN RB). – EDN TBBNDY; Patent No. 2740923 C1 Russian Federation, IPC A61K 9/00, A61K 31/506, A61K 47/32. Ophthalmic medicinal film with 6-methyl-3-(thietan-3-yl)uracil, which has a wound-healing effect: No. 2020119998: application. 06/09/2020: publ. 01/21/2021 / A. F. Gabdrakhmanova, S. A. Kurbanov, S. A. Meshcheryakova, F. Kh. Kildiyarov; applicant Federal State Budgetary Educational Institution of Higher Education "Bashkir State Medical University" of the Ministry of Health of the Russian Federation. – EDN AFPPYV; Patent No. 2056819 C1 Russian Federation, IPC A61F 9/00. Ophthalmic medicinal film and method of its manufacture: No. 92001988/14: application. 10/22/1992: publ. 03/27/1996 / A. D. Romashchenko. – EDN BYYALP; Technology of production and study of acute toxicity of ophthalmic medicinal films with papain and urea / I. I. Romanovskaya, S. S. Dekina, R. I. Chalanova, E. P. Sotnikova // Chemical-Pharmaceutical Journal. – 2012. – T. 46. – No. 3. – P. 37-39. – EDN TMRBBJ].

The main disadvantage of existing ophthalmic medicinal films is the inclusion of an antibacterial component in their composition, which is especially important in the current conditions of increased antibiotic resistance of pathogenic microorganisms. Also, the imperfection of films comes down to high cost and unidirectional action [Review of drugs for the treatment of conjunctivoceratitis in dogs and cats / M. M. Gorokhova, A. N. Kvochko, A. N. Shulunova, N. V. Fedota // Trends development of veterinary surgery: materials of the International Scientific and Practical Conference dedicated to the 95th anniversary of the Department of General, Private and Operative Surgery, Vitebsk, November 03–04, 2021. – Vitebsk: Educational Institution “Vitebsk Order of the Badge of Honor” State Academy of Veterinary Medicine,” 2021. – P. 35-37. – EDN TYSNHD].

As a result of previously conducted tests of various film-forming substances, it was found that ophthalmic medicinal films of gelatin with the addition of glycerin, acting as a plasticizer, are optimal for a number of technical characteristics (availability, ease of use, physical properties) [Laboratory tests of film-forming substances for the production of ophthalmic medicinal films / M. M. Gorokhova, A. N. Shulunova, N. V. Fedota, A. N. Kvochko // International Bulletin of Veterinary Medicine. – 2022. – No. 2. – P. 53-58. – DOI 10.52419/issn2072-2419.2022.2.53. – EDN NMLKGQ.].

Disclosure of the Invention

The objective of the invention is to develop ophthalmic medicinal films based on a biostimulator from cattle brain tissue with the addition of silver nanoparticles.

The technical result that can be obtained using the present invention is to increase the effectiveness of treatment of erosive and ulcerative pathologies of the cornea, accelerate healing processes, minimize exposure and contact with the pathological focus.

The technical result is achieved using ophthalmic medicinal films based on a biostimulant from cattle brain tissue with the addition of silver nanoparticles, which includes taking 0.1 g of biostimulant into a test tube and adding 0.3 ml of a 0.9% sodium chloride solution with a micropipette, followed by stirring until homogeneous. consistency and adding 1 g of gelatin and adding 7 ml of distilled water, after which the test tube is left for 10 minutes at room temperature, and then the test tube is placed in a solid-state thermostat (Biosan BioTDB-100) at a temperature of 70°C, where the gelatin is brought to complete dissolution, stirring every 5 minutes for 35 minutes, then add 1 ml of glycerol to the film-forming solution with a micropipette and mix thoroughly, after the gelatin is completely dissolved, the solution of the film-forming substance is cooled to a temperature of +39°C and as it cools, 1 ml is added to the test tube with the prepared biostimulator solution using a micropipette film-forming solution, mix and place in a thermostat, after which drops of 0.5 ml are made with a micropipette onto a plastic tape and left at room temperature for 4 hours.

A biostimulant made from cattle brain tissue with the addition of silver nanoparticles was selected as the active ingredient. The biostimulator is represented by a complex of low molecular weight, water-soluble polypeptide biologically activated fractions. The components of the stimulant are able to overcome the blood-brain barrier and have an antioxidant and tissue-specific effect. Thanks to its unique properties, a biostimulator from brain tissue has wound-healing, proliferative and stimulating properties [Fedota, N.V. Hematological parameters and mathematical modeling of biorhythms of live weight gain in sheep under the influence of biostimulants: specialty 03.03.01 “Physiology”: dissertation for an academic degree candidate of veterinary sciences / N. V. Fedota. – Stavropol, 1998. – 133 p.].

Carrying out the invention

Ophthalmic medicinal films based on a biostimulant from bovine brain tissue with the addition of silver nanoparticles were prepared as follows.

At the first stage, the active substance is prepared for adding a film-forming component to the solution. To do this, in compliance with all aseptic rules, place 0.1 g of biostimulant into a test tube and add 0.3 ml of a solution of 0.9% sodium chloride solution using a micropipette and carefully mix until a homogeneous consistency is achieved.

Next, under aseptic laboratory conditions, 1 g of gelatin is poured into a test tube and 7 ml of distilled water is added using a micropipette (Thermo Scientific). After this, the test tube is left for 10 minutes at room temperature for the gelatin to soak and swell. Then place the test tube in a solid-state thermostat (Biosan BioTDB-100) at a temperature of 70°C, where the gelatin is brought to complete dissolution, stirring every 5 minutes. This will take 35 minutes. Next, add 1 ml of glycerol to the film-forming solution using a micropipette and mix thoroughly.

After the gelatin is completely dissolved, the solution of the film-forming substance is cooled to a temperature of +39°C to avoid coagulation of the protein components of the active substance. As the micropipette cools, 1 ml of the film-forming solution is added to the test tube with the prepared biostimulator solution, mixed and placed in a thermostat. Due to rapid thickening, a thin film is formed on the surface of the gelatin solution, making it difficult to pipet the film former. Therefore, the test tube with gelatin must be constantly kept in a thermostat at the desired temperature. Using a 0.5 ml micropipette, drop drops onto a plastic tape and leave at room temperature for 4 hours.

An example of a specific implementation: in the conditions of the scientific-diagnostic and medical veterinary center, as well as the Department of Physiology, Surgery and Obstetrics of the Federal State Budgetary Educational Institution of Higher Education "Stavropol State Agrarian University", an experimental production of ophthalmic medicinal films based on a biostimulator from cattle brain tissue with the addition of nanoparticles was carried out silver

At the first stage, the active substance was prepared for adding a film-forming component to the solution. To do this, in compliance with all aseptic rules, 0.1 g of biostimulant was placed in a test tube and 0.3 ml of 0.9% sodium chloride solution was added using a micropipette and carefully mixed until a homogeneous consistency was achieved.

Next, under aseptic laboratory conditions, 1 g of gelatin was poured into a test tube and 7 ml of distilled water was added using a micropipette (Thermo Scientific). After this, the test tube was left for 10 minutes at room temperature for the gelatin to soak and swell. Then the tube was placed in a solid-state thermostat (Biosan BioTDB-100) at a temperature of 70°C, where the gelatin was brought to complete dissolution, stirring every 5 minutes. This will take 35 minutes. Next, 1 ml of glycerol was added to the film-forming solution using a micropipette and mixed thoroughly.

After complete dissolution of the gelatin, the solution of the film-forming substance was cooled to a temperature of +39°C to avoid coagulation of the protein components of the active substance. As the micropipette cooled, 1 ml of the film-forming solution was added to the test tube with the prepared biostimulator solution, mixed and placed in a thermostat. Due to rapid thickening, a thin film is formed on the surface of the gelatin solution, making it difficult to pipet the film former. Therefore, the test tube with gelatin must be constantly kept in a thermostat at the desired temperature. Using a micropipette, drops of 0.5 ml were made onto a plastic tape and left at room temperature for 4 hours.

Testing for toxicity was carried out by placing ocular medicinal films based on a biostimulant from cattle brain tissue with the addition of silver nanoparticles on the cornea of laboratory animals, for which five giant rabbits and one mongrel dog were selected. Five minutes after the start of the experiment, the films began to lose their shape and after 20 minutes they appeared in the third eyelid area. It is worth noting that in all animals taking part in the experiment, these dosage forms were completely dissolved within 45 minutes; no allergic reactions, irritation or signs of inflammation were detected.

The proposed ophthalmic medicinal films did not spread, had and retained the correct shape, were elastic, translucent, without tears or cracks.

Compared to the prototype and other known technical solutions, the proposed invention has a number of advantages:

- Bidirectional action;

- High efficiency;

- No risk of allergic reactions and tolerance.

Formula

Ophthalmic medicinal films based on a biostimulant from cattle brain tissue with the addition of silver nanoparticles, including taking 0.1 g of a biostimulant into a test tube and adding 0.3 ml of a 0.9% sodium chloride solution with a micropipette, followed by mixing until a homogeneous consistency and adding 1 g gelatin and adding 7 ml of distilled water, after which the test tube is left for 10 minutes at room temperature, and then the test tube is placed in a solid-state thermostat (Biosan BioTDB-100) at a temperature of 70°C, where the gelatin is brought to complete dissolution, stirring every 5 minutes at for 35 minutes, then 1 ml of glycerin is added to the film-forming solution with a micropipette and mixed thoroughly, after the gelatin is completely dissolved, the solution of the film-forming substance is cooled to a temperature of +39°C and as it cools, 1 ml of the film-forming solution is added to the test tube with the prepared biostimulator solution using a micropipette, mixed and placed in a thermostat, after which drops of 0.5 ml are made with a micropipette onto a plastic tape and left at room temperature for 4 hours.

Essay

The invention relates to medicine, in particular to ophthalmology, and can be used to treat corneal injuries.

The technical result that can be obtained using the present invention is to increase the effectiveness of treatment of erosive and ulcerative pathologies of the cornea, accelerate healing processes, minimize exposure and contact with the pathological focus.

The technical result is achieved using ophthalmic medicinal films based on a biostimulant from cattle brain tissue with the addition of silver nanoparticles, which includes taking 0.1 g of biostimulant into a test tube and adding 0.3 ml of a 0.9% sodium chloride solution with a micropipette, followed by stirring until homogeneous. consistency and adding 1 g of gelatin and adding 7 ml of distilled water, after which the test tube is left for 10 minutes at room temperature, and then the test tube is placed in a solid-state thermostat (Biosan BioTDB-100) at a temperature of 70°C, where the gelatin is brought to complete dissolution, stirring every 5 minutes for 35 minutes, then add 1 ml of glycerol to the film-forming solution with a micropipette and mix thoroughly, after the gelatin is completely dissolved, the solution of the film-forming substance is cooled to a temperature of +39°C and as it cools, 1 ml is added to the test tube with the prepared biostimulator solution using a micropipette film-forming solution, mix and place in a thermostat, after which drops of 0.5 ml are made with a micropipette onto a plastic tape and left at room temperature for 4 hours.

Claims (1)

A method for producing an ophthalmic medicinal film based on a biostimulant from cattle brain tissue, including taking 0.1 g of a biostimulant into a test tube and adding 0.3 ml of a 0.9% sodium chloride solution with a micropipette, followed by stirring until a homogeneous consistency and adding 1 g of gelatin and adding 7 ml of distilled water, after which the test tube is left for 10 minutes at room temperature, and then the test tube is placed in a solid-state thermostat (Biosan BioTDB-100) at a temperature of 70°C, where the gelatin is brought to complete dissolution, stirring every 5 minutes for 35 minutes, then 1 ml of glycerin is added to the film-forming solution with a micropipette and mixed thoroughly, after the gelatin is completely dissolved, the film-forming solution is cooled to a temperature of +39°C and as it cools, 1 ml of the film-forming solution is added to the test tube with the prepared biostimulator solution by micropipette, mixed and placed in thermostat, after which drops of 0.5 ml are made with a micropipette onto a plastic tape and left at room temperature for 4 hours.