RU2404768C1 - Eye drops - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: eye drops for prophylactics and treatment of cataract, which contain such active reactants as biogenic compounds carnosine, taurine and glutathione. As carrier, eye drops contain low molecular microdispersed dextran ground for 9-11 minutes in vibration disk mill, and also contain benzalkonium chloride and purified water at certain ratios of components.

EFFECT: invention provides for prophylactics and treatment of cataract.

2 tbl, 4 dwg

Description

The invention relates to medicine, namely to ophthalmology, and is directed to the prevention and treatment of cataracts.

Cataract is a non-infectious disease of the eye, expressed in persistent opacification of the lens substance or lens capsule [1]. Possible causes of cataracts are metabolic disorders in the lens, heredity, trauma, ultraviolet radiation, diseases of the inner membranes of the eye [2]. The pathogenesis of cataracts is not well understood. Most scientists associate the development of cataracts with the destruction of the proteins that make up the lens. In this regard, the osmotic pressure rises in the lens, which in the initial stage leads to an increased flow of water into the lens and its swelling. Later, protein cleavage products diffuse through the bag of the lens into the intraocular fluid, the amount of water in the lens decreases, and it becomes dense. In the hydrolysis of β-crystallin (the main lens protein), a relative increase in the amount of water-insoluble and, therefore, opaque proteins in the lens occurs. Clinically proven the effect of oxidative stress on the development of cataracts [3]. A significant decrease in the level of endogenous antioxidants in the tissues of the eye (in particular, glutathione) leads to the activation of free radical reactions leading to the formation of reactive oxygen species (ROS), the oxidative modification of lipids and proteins of the eye tissues. In this case, lens proteins are most affected, the attack of which from the ROS side leads to their glycation, the formation of Schiff bases and cross-linking [4]. Hydrolysis can also be preceded by the oxidation of sulfhydryl groups of soluble proteins with a partial transition of cysteine to cystine. The development of cataracts is also associated with impaired carbohydrate metabolism and tissue respiration in the lens and impaired permeability of its capsule.

There is an opinion that conservative treatment of cataracts with the use of antioxidants helps to slow down the development of this disease [5].

Currently, there are a number of anti-cataract preparations containing amino acids, vitamins and other active ingredients of various origins [6]. All preparations known to the authors are presented in the form of eye drops.

The main requirements that eye drops must meet are:

- sterility;

- lack of mechanical impurities;

- comfort (isotonicity, optimal pH value);

- chemical stability;

- prolongation of action [7].

Known eye drops "Taufon" containing taurine as the active substance [8], used for cataracts of various nature.

Drops are known for the treatment of cataract "Vitiodurool" [8], which include glutathione.

Known eye drops for the treatment of cataracts [9], which include the carnosine dipeptide (β-alanyl-L-histitine) in the range of 1-5%, concentrated hydrochloric acid and water, while the pH of the medium is 6.5-7.2 . A disadvantage of the known solution is that the pH is somewhat shifted to the acidic region. Although it is known that the physiological region of the pH of the tear fluid is 7.4. A painless area of tolerance covers a pH of 7 to 9, and the deviation to the acid side is more sensitive to the eye [10]. In addition, the disadvantage of this invention is the unreasonably high content of carnosine (5%), while in a rabbit experiment, instillation of a carnosine solution with a concentration above 2% showed pupil constriction in 20% of cases [11].

Known pharmaceutical composition for the prevention of development and treatment of the initial stage of age-related cataracts (options) [12], containing as an active principle a mixture of two short-chain peptides D-pantetin and L-carnosine. The disadvantage of the invention is the high content of active substances (in the example used 5% solution of drugs), which can cause unwanted reactions from the tissues of the eye (edema, hyperemia, etc.). In addition, the composition of the described pharmaceutical composition lacks agents that provide the required pH level.

Known eye drops containing the antioxidant glutathione [13], in addition, lipoic acid, a chelating agent, an oxygenating agent and a substance that improves the permeability of the mucous membrane of the eye. Additional antioxidants in the form of taurine, vitamin E, vitamin C can be introduced into the composition of the drops. The composition is intended to increase the elasticity of the lens and its accommodation, as well as reduce eye discomfort caused by dryness, injuries, burns.

A common drawback of all the known compositions mentioned above is the absence of a prolongator or a mixture thereof, while prolongation of eye drops is one of the requirements for eye drops. It is known that the conjunctival sac can take only 1/5 drops of a drug substance: most of the solution is washed out within 30 seconds. To prevent leaching and prolong contact of the active substance with the cornea, increase the viscosity of aqueous solutions of eye drops [14].

The closest analogue to the proposed pharmaceutical composition are eye drops of reparative and anti-cataract action, containing taurine, carnosine, dextran, benzalkonium chloride and water [15]. The composition contains: taurine 2.0-6.5 g, carnosine - 2.0-7.0 g, dextran 2.5-10.3 g, benzalkonium chloride 0.025-0.200 g, purified water - up to 100 ml.

However, this composition does not provide sufficient effectiveness, because it contains one compound having antioxidant activity. In addition, it is known that dextran, being a polymer with a linear molecular structure, is poorly wetted and dissolves for a long time.

The objective of the invention is to create a more effective than the known, composition, providing prolonged contact of the active substances with the mucous membrane of the eye and is suitable to prevent and / or slow down the process of clouding of the lens of the eye, that is, for the prevention and treatment of cataracts.

The problem is solved in that the proposed pharmaceutical composition for the prevention and treatment of cataracts contains dextran, benzalkonium chloride, water and biogenic compounds carnosine and taurine as active active ingredients, and as a prolongator low molecular weight microdispersed dextran, crushed for 9-11 minutes in vibration disk mill, and additionally contains the active component of glutathione in the following ratio of components:

Taurine - 1.5-1.9 g Carnosine - 1.5-1.9 g Glutathione - not less than 0.01 g Dextran low molecular weight microdispersed - 10.5-15.0 g Benzalkonium chloride - 0.005-0.02 g Purified water - up to 100 ml

The increase in the effectiveness of the anti-cataract properties of the proposed pharmaceutical composition in comparison with the known preparations is based on the combined action of the active principle components of the created composition and the prolongation of this action due to the introduction of low molecular weight microdispersed dextran, crushed in a vibration disk mill for 9-11 minutes.

The advisability of introducing three active components into the composition of the composition is determined by the ambiguity and versatility of the pathological process in the lens. Disruption of metabolism, oxidative stress, chemical transformations of its proteins require a comprehensive approach to the prevention and treatment of cataracts. Carnosine, glutathione and taurine, although they are substances of an amino acid nature, have a general positive effect on the tissues of the eye, different in their mechanism of action. It is the combination of these substances in one composition that makes it possible to achieve a positive effect that is unexpected in comparison with analogues.

It is known that in a healthy eye, a high level of endogenous antioxidants, the most important of which glutathione and its dependent enzymes are considered, prevents the development of oxidation [16]. Glutathione is contained in the lens in an amount ten times greater than its content in any other tissues, which indicates its significant role in the biochemistry of the lens. One of the most important functions of glutathione is to maintain the transparency of the lens, due to:

- prevent damage to cells under the influence of oxidative stress,

- providing transport to the lens of the amino acids necessary for the synthesis of crystallins,

- activation of the synthesis of other proteins [17].

Under certain conditions, the level of glutathione in the lens decreases, which leads to pathological processes in it, therefore it is necessary to additionally introduce glutathione in the form of eye drops, making up for its deficiency. In addition, it is known that carnosine, having its own anti-cataract activity, is also able to increase the content of endogenous glutathione in the lens. The anticataractal effect of carnosine lies in its ability to inhibit protein glycation and cross-linking, thereby ensuring the transparency of the lens [4].

The presence of carnosine, which is an active biological pH stabilizer, in the composition of eye drops will make it possible not to introduce additional buffering agents into the composition. In this case, the pH of the claimed composition will be 7.0-7.5.

The above facts determined the introduction of carnosine and glutathione into the composition.

Taurine, having a reparative and metabolic effect, favorably affects the processes of protein metabolism in the lens, and thereby preventing clouding of the lens [18].

To establish the role of each of the components in the antioxidant defense of the body, the antioxidant activity of the components of the eye drops in vitro was determined.

To determine the antioxidant activity of 0.05 N, a solution of potassium permanganate in a 0.024 M solution of sulfuric acid was titrated with solutions of ascorbic acid, carnosine, glutathione, taurine until discoloration [19]. As an indirect indicator of antioxidant activity (AOA, mg / ml), the amount of substance in milligrams was used, which went to the bleaching of 1 ml of potassium permanganate solution. For a better presentation of the results, the value of antioxidant ability (AOS) was proposed, numerically expressed in the inverse value of AOA.

To determine the level of antioxidant activity of the studied substances, a solution of ascorbic acid, a well-known model antioxidant, was used.

Table 1 No. p / p Substances

,
мг/мл

,
mg / ml Σ (

-AOA _i ) ² S _AOA ΔAOA, mg / ml ε,% AOS one. Ascorbic
acid 0.280 2.0 · 10 ^-9 2.58 · 10 ^-4 6.32 · 10 ^-4 0.23 3571 2. Glutathione 0.350 2.0 · 10 ^-7 2.58 · 10 ^-3 6.32 · 10 ^-3 1.80 2857 3. Carnosine 51,700 2.1 · 10 ^-4 8.37 · 10 ^-2 0.21 0.39 19 four. Taurine - * - - - - 0

As can be seen from the data in Table 1, only glutathione has a comparable ascorbic acid activity, carnosine showed very low activity, taurine did not discolor the potassium permanganate solution. The results obtained make it possible to position glutathione as the leading active substance of the composition. In addition, it is known that carnosine, exhibiting a rather low antioxidant activity in vitro, when included in in vivo processes, increases it manyfold [4]. Therefore, carnosine in the claimed composition is introduced in a sufficiently large amount compared to the content of glutathione.

The use of a prolongator and thereby increasing the viscosity of eye drops is one of the most important requirements for eye drops. Low molecular weight dextran (M 40,000) was chosen as a prolongator. It is known that dextran ((C ₆ H ₁₀ O ₅ )] _n ), as a glucose polymer, is a high molecular weight substance with a linear molecular structure. The dissolution of compounds with a linear structure proceeds in two stages: swelling and dissolution [20]. Dextran because of this is poorly wetted and slowly dissolved. To improve this process in the claimed invention, the dextran was crushed using a vibrating disk mill for 9-11 minutes, followed by its dissolution in water.

The optimal dispersion time was selected as follows: grinding was performed using an RS-200 vibrating disk mill (RETSCH, Germany; Center for Nanostructured Materials and Technologies of BelSU) for 5, 10, 15 minutes. The size of the obtained particles was determined using an electron microscope. Dissolution of the substance in purified water was carried out using a magnetic stirrer.

The results of the study of the dependence of the solubility of low molecular weight dextran in water on the time of its grinding are presented in table 2 and figure 1, 2, 3.

The dissolution of dextran, crushed for 5 minutes, takes 50-60 minutes, which does not differ from the solubility of uncrushed dextran. The results of electron microscopy confirmed the absence of changes in the spatial structure of dextran (figure 1). The figure is dominated by large spherical particles.

Dissolution of dextran, crushed for 10 minutes, takes 10 minutes. The results of electron microscopy are presented in figure 2. Low molecular weight Dextran, crushed for 10 minutes, represents individual small spherical particles with cracks and kinks into which water penetrates during the manufacture of eye drops and, providing a wedging effect, ensures rapid dissolution.

Dissolution of dextran, ground for 15 minutes, takes 30 minutes. The results of electron microscopy are presented in figure 3. The low molecular weight dextran crushed for 15 min is small particles combined into sparingly soluble conglomerates.

table 2 Grinding time, min Description of Dextran Particles Dextran Particle Sizes The time of dissolution of dextran, min No chopping Large spherical particles from 7.60 to 80.96 microns 60 5 Large spherical particles from 7.60 to 80.96 microns 50-60 10 Small spherical and shapeless particles, single conglomerates mostly from 27.70 to 30.96 microns 10 fifteen Small shapeless particles combined in conglomerates particle size - up to 20.00 microns;
conglomerate size - up to 208.61 microns thirty

The addition of a low molecular weight microdispersed solution to the dextran solution made it possible to increase the viscosity of eye drops from 2 MPa · s to 10 MPa · s, and at the same time, reduce the preparation time of the solution by six times.

It is known that a preservative must be introduced into the composition of eye drops for packaging volumes exceeding the daily requirement in order to prevent the propagation of extraneous microflora in the open packaging during use. Benzalkonium chloride was chosen as a preservative.

Eye drops are prepared as follows. Benzalkonium chloride is dissolved in 50% of the volume of purified water, microdispersed dextran is added, mixed until the components are dissolved. After that, carnosine, taurine and glutathione are introduced into the resulting solution; adjust the volume to the required purified water. After complete dissolution of all components, the solution of eye drops is filtered and sterilized by the method of sterilizing filtration, then, under aseptic conditions, the bottles and bottles of eye drops are bottled and capped.

Thus, the introduction of three active components in the composition will prevent the destruction of the proteins that make up the lens, because the introduction of carnosine triggers the synthesis mechanism of endogenous glutathione and, in combination with the additional administration of glutathione from the outside, in general, significantly increases its level in the lens, therefore, the activity of free radical reactions leading to the formation of reactive oxygen species (ROS), the oxidative modification of lipids and proteins of the eye tissue decreases . In addition, an increased content of glutathione activates the synthesis of lens proteins. In addition, carnosine has its own anti-cataract activity, and the introduction of taurine improves metabolism and the regeneration process in the lens.

Comprehensive toxicity studies of developed eye drops have been carried out. It is concluded that the studied composition of eye drops refers to low-toxic drugs. A comparative study of the developed drops and the Taufon preparation manufactured by LENS-Pharm (Russia) confirmed their compliance with the Toxicity indicator.

The anti-cataract activity of the developed eye drops of the proposed composition (composition No. 1), composition No. 2 (not stated in the description of the invention, does not contain glutathione) was studied in comparison with the reference drug - Taufon produced by "LENS-Pharm" (Russia). It has been proven that eye drops of both formulations (No. 1 and No. 2) have a pronounced protective effect when visually assessing the transparency of the lens, not inferior to the reference drug - Taufon eye drops manufactured by LENS-Pharm (Russia).

When simulating selenitic cataract in the control group of animals not receiving treatment, a clearly visible pronounced clouding of the lens was observed in all rats. When applying eye drops of formulations No. 1, No. 2 and the comparison drug - eye drops of Taufon (from the first day after the administration of sodium selenite) in a dose of 1 drop in the form of instillation into the conjunctival cavity of each eye, a pronounced therapeutic effect was observed during the entire observation period. Moreover, in all groups receiving the drug, the visual distribution of the degree of damage to the lens did not significantly differ; only one rat out of 10 in the group treated with eye drops of composition No. 1 showed almost complete absence of lens opacification. Figure 4. cataract damage of the lens of rats is presented: A - control group; B - group treated with developed eye drops of compounds No. 1 and No. 2; C - group treated with Taufon eye drops; D is an isolated case of the absence of clouding of the lens in the group treated with eye drops of composition No. 1.

A morphometric study revealed greater anti-cataractal efficacy of eye drops of composition No. 1 compared with Taufon eye drops produced by LENS-Pharm (Russia) and composition No. 2.

References

1. Eye diseases. / Ed. V.G. Kopaeva. - M .: Medicine, 2002, p.249.

2. Bagirov N.A. Modern problems of cataractogenesis. / N.A. Bagirov. // Ophthalmol. Zh., 2000. - No. 6. - S.98-102.

3. Boldyrev A.A. Carnosine: an endogenous physiological corrector of the activity of the antioxidant system of the body [Text]. / A.A. Boldyrev, S.L. Stvolinsky, T.N. Fedorova. // Usp. fiziol. sciences. - 2007. - No. 38 (3). - S. 57-71.

4. Boldyrev A.A. Carnosine. - M.: Publishing House of Moscow State University. M.V. Lomonosov. - 1998.

5. Chylack L.T. Jr, Brown N.P., Bron A. et al. The Roche European American Cataract Trial (REACT): a randomized clinical trial to investigate the efficacy of an oral antioxidant micronutrient mixture to slow progression of age-related cataract. // Ophthalmic Epidemiol. - 2002. - No. 9. - P. 49-80.

6. B.N. Ames, M.K.Shigenaga, T.M. Hagen. Oxidants, antioxidants and the degenerative diseases of aging. // Proc. Natl. Acad. Sci. USA - 1993 .-- Vol.90. - P.7915-7922.

7. Zhilyakova E.T., Stepanova E.F., Novikova M.Yu. Harmonization of the requirements for eye drops in the light of the modern development of pharmaceutical production. // Materials of the international scientific-practical conference "Biologically active compounds of natural origin: herbal medicine, pharmaceutical marketing, pharmaceutical technology, pharmacology, botany." Belgorod, 2008. P.62-66.

8. Mashkovsky M.D. Medicines: In 2 tons. T.2. - M.: Publishing House New Wave LLC, 2004. - P.126.

9. Pat. RU No. 2071316, IPC A61K 9/00, A61K 31/415. Eye drops for cataracts. / Boldyrev A.A. and etc.; Applicant NPK LLP Bisl company, patent holders: Boldyrev AA, Ragimov AA, Formazyuk V.E., NPK LLP Bisl company. - No. 93039867/14; declared 08/05/1993; publ. 01/10/1997.

10. Morozov V.I., Yakovlev A.A. Pharmacotherapy of Eye Diseases: A Guide. M .: Medicine, 1989, p. 240.

11. Pat. RU 2201213, IPC A61K 9/08, A61K 38/05, A61F 9/00. Eye drops "Carnosine" / Bagrov S.N. and etc.; Applicant and patent holder of LLC Scientific and Experimental Production Eye Microsurgery. - No. 2001118086/14; declared 07/04/2001; publ. 03/27/2003.

12. Pat. RU 2352352, IPC A61K 38/04, A61K 38/05, A61P 27/12. Pharmaceutical composition for the prevention and treatment of the initial stage of age-related cataract (options). / Soustov L.V. and etc.; Applicant and patent holder Institute of Applied Physics RAS. - No. 2007144339/15; declared 12/03/2007; publ. 04/20/2009.

13. Pat. CA 2200501, IPC A61K 31/195. Glutathione antioxidant eye drops. / Hofmann R.F., Bottoni D.J .; - declared. 03/20/1997; publ. 09/18/1998.

14. Technology and standardization of drugs. Collection of scientific papers. - T.2. - Kharkov: IG "RIREG". - 2000, p. 292-293.

15. Application RU No. 2007148492, IPC A61K 31/00. Eye drops. / Zhilyakova E.T. and etc.; applicants GOU VPO "BelSU", LLC "Asclepius". - No. 2007148492/15, declared 12/21/2007; publ. 06/27/2009.

16. Kulinsky V.I. Active forms of oxygen and oxidative modification of macromolecules: benefits. Harm and protection. // Soros educational journal. - 1999. - No. 1. - S.1-7.

17. Parris M. Kidd. Glutathione: Systemic Protectant Against Oxidative and Free Radical Damage. // Altemative Medicine Review. - 1997. - Vol. 2, No. 3. - C.155-176.

18. Pat. SU 1617377, IPC G01N 33/52. A method for assessing the damaging effect of substances in contact with the cornea. / Babizhaev M.A. and etc.; applicant and patentee MNIIIGB them. Helmholtz. - No. 4383190 / 30-14; declared 02.24.88; publ. 12/30/90, Bull.№48.

19. Pat. RU 2170930, IPC G01N 33/50, G01N 33/52. A method for determining antioxidant activity. / Maksimova T.V. and etc.; applicant and patent holder Mosk. Medical Academy. I.M.Sechenova. - No.2000111126 / 14; declared 05.05.00; publ. 07/20/01.

20. Technology of dosage forms: Textbook in 2 volumes. T.1 / Ed. L.A. Ivanova. - M .: Medicine, 1991. - P.228.

Claims (1)

Eye drops for the prevention and treatment of cataracts containing dextran, benzalkonium chloride, water and biologically active compounds carnosine and taurine as the active ingredients, characterized in that as a prolongator they contain low molecular weight microdispersed dextran, crushed for 9-11 minutes in a vibrating disk mill, and additionally contain the active component of glutathione in the following ratio of components:
Taurine 1.5-1.9 g Carnosine 1.5-1.9 g Glutathione not less than 0.01 g Dextran low molecular weight microdispersed 10.5-15.0 g Benzalkonium chloride 0.005-0.02 g Purified water up to 100 ml

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