RU2533276C1 - Eye drops based on composition of pharmaceutically acceptable additive acid salts and methylethyl pyridonol and taurine containing composition of group b vitamins - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: invention represents eye drops containing a pharmaceutically acceptable additive acid salt and methylethylpyridinole, taurine and a composition of group B vitamins.

EFFECT: creating the combined preparation for treating traumatic and dystrophic eye injuries causing no irritant action and possessing improved pharmacological properties.

5 cl, 6 ex, 2 tbl

Description

FIELD OF THE INVENTION

The invention relates to ophthalmic preparations, and more particularly to eye drops, which can be used for therapeutic purposes with conjunctival or retinal hemorrhage, as well as with vitreous hemorrhage, with injuries, thermal and chemical burns in the eye area. The proposed drug has a beneficial effect in the degeneration of the macula and / or choroid, glaucoma, myopia and diabetic retinopathy.

State of the art

Emoxipin (3-hydroxy-6-methyl-2-ethylpyridine hydrochloride) is known as a biologically active substance with anti-aggregation, anti-hypoxic, angioprotective and pronounced cardioprotective effects. Emoxipin reduces the permeability of the vascular wall, improves microcirculation, in particular, by reducing the viscosity and blood coagulation, enhances the process of fibrinolysis, increases the resistance of the brain to hypoxia and ischemia, normalizes tissue metabolism, including in case of stroke and myocardial infarction, unstable angina.

In the context of ophthalmic applications, the beneficial properties of emoxipin contribute, in particular, to the resorption of post-traumatic intraocular hemorrhages and improve blood supply to the eye tissue during thrombosis of the central retinal vein and its branches. In addition, due to its retinoprotective effect, emoxipin helps protect the retina of the eye when exposed to high-intensity light used in laser coagulation.

Known ophthalmic preparations in the form of eye drops, representing a 1% aqueous solution of emoxipin with pharmaceutically acceptable additives (buffering agents, preservative, antioxidant, viscosity modifier). An example of such a drug is Emoxipin. Eye drops 1% "(FSUE Moscow Endocrine Plant, Russia). Such drops are indicated, in particular, for the treatment of hemorrhage in the anterior chamber of the eye and sclera (especially in elderly patients), inflammation and burns of the cornea, as well as when wearing contact lenses (protection of the cornea) and complications of myopia.

Taurine (2-aminoethanesulfonic acid) plays an important role in lipid metabolism, helps normalize the function of cell membranes, optimizes energy and metabolic processes, and preserves the electrolyte composition of the cytoplasm due to the accumulation of potassium and calcium ions. In the brain, it acts as a neurotransmitter that inhibits synaptic transmission and has anticonvulsant activity. It causes the normalization of ocular tissue metabolism in diseases of a dystrophic nature.

Known ophthalmic preparations in the form of eye drops, representing a 4% aqueous solution of taurine containing methylhydroxybenzoate as a preservative. An example of such a drug is Taufon. Eye drops 4% ”(FSUE Moscow Endocrine Plant, Russia). Subconjunctival instillation of such drops is indicated for degenerative lesions of the retina, including hereditary, corneal dystrophy, senile, diabetic, traumatic and radiation cataracts, as well as for corneal injuries as a stimulator of reparative processes.

Vitamin ophthalmic preparations in the form of drops are known for improving nutrition and energy metabolism in the tissues of the eye, contributing to the restoration of the cornea during wound injuries and burns, facilitating the treatment of blepharitis, keratitis, allergic conjunctivitis by the introduction of appropriate drugs. Vitamin drops are also indicated for cataracts. An example of such a preparation is Eyelight Vita Yellow eye drops (Hua Giang Pharmaceutical JSC, Vietnam), composition

Vitamin B1 5 mg Vitamin B2 0.2 mg Vitamin PP 40 mg Excipients (boric acid, borate sodium, nipagin, water for injection in sufficient quantities) up to 10 ml

The disadvantage of the considered drugs is a narrow range of indications for use.

The patent RU 2493841 (published September 27, 2013) discloses a synergistic preparation for the treatment of cardiovascular insufficiency, diabetes mellitus types I and II, diseases of the hepatobiliary system, characterized in that it contains 2-75% of a combination of active ingredients and pharmaceutically acceptable carriers, excipients and excipients, wherein the composition of the active components consists of a pharmaceutically acceptable salt of 2-ethyl-6-methyl-3-pyridin-3-ol and taurine in a weight ratio of from 9: 1 to 1: 9. Preferably, the preparation is presented in the form of a solution for injection containing 1.5% w / v. methylethylpyridinol succinate and 3.5% w / v taurine, as well as in the form of gelatin capsules and tablets. The description of the invention to the specified patent does not contain information that allows a specialist in this field to consider such a solution for injection as a potential ophthalmic agent.

In connection with the considered reasons, there is a need to expand the arsenal of drugs for the treatment of dystrophic and traumatic eye lesions, which, in particular, have a wider range of indications and greater efficiency.

Disclosure of invention

The aim of the invention is the creation of a combined drug for the treatment of traumatic and dystrophic lesions of the eye, which is not irritating and has improved pharmacological properties.

As a result of the studies, the inventors have found that the disadvantages of the prior art can be overcome by creating eye drops based on a pharmaceutically acceptable acid and methyl ethyl pyridinol addition salt containing a composition of vitamins of group B, which is selected from the group consisting of vitamin B1, vitamin B2, vitamin PP and their mixtures.

The main requirements for solutions intended for instillation into the conjunctival cavity are sterility, transparency, pH and osmolality values close to those of the tear fluid of a healthy body.

Sterility is necessary to ensure long-term storage of the drug before use. It also guarantees the absence of pathogenic microorganisms that release toxic waste products that can multiply on the conjunctiva after administration of the drug. In accordance with the invention, sterility in production is ensured by sterilization and aseptic packaging of containers containing eye drops, and during storage by the introduction of a preservative (sodium benzoate).

Transparency is a natural requirement for optical media. Indicators of osmolality (260-330 mOsm / kg) and pH (5.5-6.2), close to natural, create a comfortable environment for conjunctival cells, which eliminates their damage and unpleasant sensations of pain or burning in the eye. Theoretical and experimental methods for estimating component concentrations in preparations of a similar type are obvious to a person skilled in the art, the most famous of which is a theoretical calculation taking into account isotonic equivalents, cryometry under supercooling with a salt / ice mixture, and electron spectroscopy in the UV region. The introduction of the components of the buffer mixture (sodium hydrogen phosphate dodecahydrate, potassium dihydrogen phosphate) in appropriate amounts allows the osmolality and pH to be maintained within their natural boundaries.

Thus, the aim of the invention can be achieved by eye drops containing a composition of active substances, representing a mixture of a pharmaceutically acceptable acid and salt additive of methyl ethyl pyridinol and taurine and a composition of promoters selected from the group consisting of vitamin B1, vitamin B2, vitamin PP and their mixtures, these drops have the composition (in g / 100 ml):

Composition of active substances 1.00-5.00 The composition of the vitamins of group B 0.01-0.05 Stabilizer 0.15-0.35 Preservative 0.10-0.30 Buffer agent 1.00-1.25 Viscosity regulator 0.60-0.7 0 Water for injections up to 100 ml

In a first preferred embodiment of the invention, the active ingredient composition consists of a pharmaceutically acceptable salt of 2-ethyl-6-methyl-3-pyridin-3-ol selected from the group consisting of hydrochloride and succinate and taurine in a weight ratio of from 9: 1 to 1 : 9, the composition of the vitamins of group B is selected from the group consisting of vitamin B1, vitamin B2, vitamin PP and mixtures thereof, the stabilizer is selected from sodium sulfite and sodium metabisulfite, the buffering agent is selected from a mixture of sodium or potassium hydrogen phosphate and dihydrogen phosphate, the viscosity regulator is selected from g a group consisting of methyl cellulose and hydroxypropyl methyl cellulose, and the preservative is selected from the group consisting of sodium benzoate and nipagin.

In a second preferred embodiment of the invention, the composition of vitamins of group B consists of vitamin B2.

In a third preferred embodiment of the invention, the composition of vitamins of group B consists of vitamins B1 and B2.

In a fourth preferred embodiment of the invention, the composition of vitamins of group B consists of vitamins B1, B2 and PP.

Drops in accordance with the invention do not exhibit a locally irritating effect and do not cause systemic allergic reactions, which can be established in an adequate animal model.

Drops in accordance with the invention are suitable for use for 2 years, which was confirmed by comparing the physico-chemical and microbiological parameters obtained for the freshly packaged preparation and the preparation after storage. Quality indicators subject to mandatory control (pH, surface tension, electrical resistivity and refractive index) for all preparations during storage did not statistically significantly (p <0.05) differ from the initial values.

Drops in accordance with the invention have antihypoxic, anti-dystrophic and re-epithelializing effects, which can be shown in adequate in vitro and in vivo models.

The achievement of the technical result of the invention will be further shown in the preferred examples of its implementation.

The implementation of the invention

Examples 1-3. Eye drops

A. Preparation of a solution of the composition of active substances

In accordance with sanitary standards, a glass apparatus is prepared having a working volume of 0.8 l and equipped with a stirrer, thermometer and bubbler. 550-570 ml of water for injection with a temperature of 20-25 ° C is loaded into the apparatus, nitrogen is supplied through a bubbler and 6.0 g of methylethylpyridinol hydrochloride are added with stirring, they are completely dissolved, after which 54.0 g of taurine are dissolved. The volume of the solution was adjusted to 600 ml and purged with nitrogen. The mass ratio of methylethylpyridinol hydrochloride and taurine is 1: 9.

B. Preparation of eye drops

Under aseptic conditions, 670-700 ml of water for injection is placed in a glass apparatus having a working volume of 1.0 l, equipped with a stirrer and a thermometer, and 4.875 g of water-soluble methylcellulose, 1.500 g of sodium benzoate, 4.050 g of potassium dihydrogen phosphate, 4.875 g are successively dissolved in it. sodium dihydrogen phosphate dodecahydrate and 1,500 g of sodium sulfite. The resulting solution was divided into three parts of 250 ml, placing each part under aseptic conditions in a separate flask.

In three sterile glass devices having a working volume of 0.8 l and equipped with a stirrer and a thermometer, three compositions of promoting substances (B vitamins) are prepared by mixing the following weighed components:

Example Mass of Vitamin, mg Mass ratio B1 B2 PP one 0 25.0 0 - 2 22.5 2,5 0 B1 / B2 9: 1 3 4,5 0.5 20,0 B1 / B2 / PP 9: 1: 40

and dissolving them with stirring in 250 ml of the previously obtained solution. 200 ml of a solution of the composition of active substances prepared according to Method A above is added to each apparatus with stirring, and the volume of each solution is adjusted to 500 ml.

Transparent solutions are subjected to sequential sterilizing filtration through membrane filters with a cut-off capacity of 1.0 and 0.2 μm and in 10 ml portions are aseptically packaged in glass bottles, which are sealed with rubber caps and wrapped in aluminum caps, after which they are supplied with labels indicating the information necessary for identification and application, and placed in cardboard boxes.

The prepared eye drops have the following composition (g / 100 ml):

Composition of active substances 4.00 The composition of the promoting substances 0.01 Sodium sulfite 0.25 Sodium Benzoate 0.20 Sodium dihydrogen phosphate dodecahydrate 0.65 Potassium dihydrogen phosphate 0.54 Water Soluble Methyl Cellulose 0.65 Water for injections up to 100 ml

Example 4. Evaluation of the allergenicity of drops in vivo

The study was carried out on 9 rabbits of the breed "chinchilla" weighing 2.1-2.4 kg, kept in a 10-day quarantine under natural light and free access to feed and water in standard cages, one individual in each cage.

Within 2 weeks, the studied drops were instilled into the right eye of each rabbit (a group of three rabbits for the study of each drug) in the morning and evening. A sterile isotonic sodium chloride solution was instilled into the left (control) eye. Observation of animals during daylight hours was carried out throughout the entire study period, evaluating the following indicators: appearance (condition of the body, teeth and coat, type of eyes, limbs and ears), motor functions (activity, muscle tone, spatial orientation, temperament ), respiration, salivation, frequency of excretion and type of urine and excrement.

Within 2 weeks of the study, the appearance of the rabbits is unchanged: loss of hair and / or teeth, changes in indicators of motor and excretory functions were not noted. The excrement is decorated, the breathing is even, uncomplicated. Instillation of the drug does not cause animals anxiety and within the next hour does not cause edema, hyperemia and / or lacrimation.

At the end of the study, a visual examination did not reveal differences in the right and left eyes according to the state of the eyelids, cornea, iris and lens. Observations indicate the absence of locally irritating effect of any of the preparations provided in accordance with preferred embodiments of the invention.

Example 5. Evaluation of the antihypoxic effect of drops on a chemiluminescence (CL) model of a suspension of phospholipid liposomes induced by iron (II) ions

A. Obtaining a suspension of liposomes

To extract phospholipids from the total fraction of egg yolk phospholipids, one volume of egg yolk is homogenized for 30 minutes in 20 volumes of a chloroform-methanol mixture (2: 1 by volume), the homogenate is filtered through a defatted paper filter to separate the aggregated protein. The resulting lipid extract was washed with 0.74% aqueous KCl (1/3 of the total volume). After settling for 12 hours at 0-4 C, the upper water-methanol phase is carefully removed, the lower phase is poured into a round bottom flask and evaporated on a rotary evaporator. The dry lipid film is washed off with 3-4 ml of hexane and a 20-fold amount of chilled acetone is added, while the phospholipids precipitate in the form of white flakes. The precipitation of phospholipids with acetone is carried out 5 times. The resulting phospholipids are dissolved in a chloroform-methanol mixture. The concentration of phospholipids was determined gravimetrically.

To prepare multilayer phospholipid liposomes, the required amount of a chloroform-methanol solution of phospholipids is placed in a round bottom flask and the solvent is evaporated on a vacuum rotary evaporator. The dry lipid film was washed from the flask walls with 20 mM Tris-HCl buffer at pH 7.4. In order to form liposomes that are more uniform in size, the resulting liposome suspension is subjected to cryolytic treatment by triple freezing-thawing. Before use, the suspension of liposomes is incubated for at least 1 hour at 37 ° C.

B. Chemiluminescent measurements

Chemiluminescent measurements are carried out using an HLM-3 chemiluminometer (Bikap, Russia). Data is recorded and processed using the PCSGU250 USB laboratory and the software bundled with it (Velleman, Belgium).

The reaction medium with a total volume of 5 ml contains 0.1 ml of a suspension of liposomes with a concentration of phospholipid of 0.4 mg / ml in 20 mm Tris-HCl buffer with a pH of 7.4. An alcoholic solution of C-525 is introduced into the system to a final concentration of 1 μM. The initiation of lipid peroxidation was carried out by introducing a solution of FeSO ₄ to a final concentration of 7 μM, after which the kinetics of changes in the chemiluminescence intensity was recorded for 10-30 minutes.

For measurements, a phosphate buffer (50 mM KH ₂ PO ₄ , 100 μM EDTA, pH 7.4) is successively added to a chemiluminometer cell to a final volume of 5 ml, 0.3 μM HB, 10 μM LM and a certain volume of the test object. Initiation of the oxidation of LM is carried out by the introduction of 52 μM hydrogen peroxide. Test substances were added before the introduction of H ₂ O ₂ .

The kinetic model parameters characterizing the effectiveness of drugs as antioxidant and retinoprotective agents are the relative values of the amplitude t / t ₀ and the induction (latent) period h / h ₀ CL, calculated relative to the control. The lower the values of these values, the more active the drug is.

The data presented in table 1,

Table 1 A drug t / t ₀ h / h ₀ The control 1.00 1.00 Emoxipin 0.42 0.57 Example 1 0.28 0.37 Example 2 0.33 0.48 Example 3 0.36 0.51

show that the drops in accordance with the invention have more pronounced antioxidant properties compared with the well-known drug "Emoksipin. Eye drops 1% ”(standard deviation not more than 8%).

Example 6. Evaluation of re-epithelizing effect on a model of corneal injury

The experiments are carried out on 7 rabbits of the breed "Chinchilla", weighing 2-2.5 kg After epibulbar anesthesia with trepan 6.0 mm in diameter, an anterior incision is made of the anterior layers of the corneal stroma, after which this layer, under the level of the incision edge, is removed along with the epithelium.

The drug is instilled 3 times a day in the right eye. The left eye serves as a control - physiological saline is instilled into it.

Re-epithelialization and regeneration of corneal stroma tissue is evaluated by the dynamics of changes in the external manifestations of the inflammatory reaction, fibrin film on the defect of the cornea, stromal edema, and by the time the defect is filled with tissue and complete epithelization and are expressed in hours relative to the control eye (a minus sign indicates acceleration of processes). The results presented in table 2,

table 2 A drug The average time of registration of the state relative to the control eye, h BUT B AT G D Emoxipin -36 -27 -89 -33 -fifteen Example 1 -43 -34 -96 -40 -twenty Example 2 -39 -31 -92 -36 -17 Example 3 -37 -29 -91 -34 -16 A - the disappearance of signs of inflammation; B - the disappearance of the fibrin film; In - the disappearance of edema of the stroma; G - filling the defect with tanya; D - complete re-epithelization.

indicate a higher activity of the drops in accordance with the invention compared with the well-known drug "Emoxipin. Eye drops 1%. "

Claims (5)

1. Eye drops, characterized in that they contain a composition of active substances, representing a mixture of a pharmaceutically acceptable additive salt of acid and methylethylpyridinol and taurine, a composition of promoters selected from the group consisting of vitamin B1, vitamin B2, vitamin PP and mixtures thereof, stabilizer, preservative, buffering agent and viscosity regulator in the following amounts (g / 100 ml):
Composition of active substances 1.00-5.00 The composition of the vitamins of group B 0.01-0.05 Stabilizer 0.15-0.35 Preservative 0.10-0.30 Buffer agent 1.00-1.25 Viscosity regulator 0.60-0.70 Water for injections up to 100 ml 2. Eye drops according to claim 1, characterized in that the composition of the active components consists of a pharmaceutically acceptable salt of 2-ethyl-6-methyl-3-pyridin-3-ol, selected from the group consisting of hydrochloride and succinate, and taurine in weight ratio from 9: 1 to 1: 9, the composition of vitamins of group B is selected from the group consisting of vitamin B1, vitamin B2, vitamin PP and mixtures thereof, the stabilizer is selected from sodium sulfite and sodium metabisulfite, the buffering agent is selected from a mixture of hydrogen phosphate and dihydrogen phosphate sodium or potassium, the viscosity regulator is selected from the group oyaschey of methylcellulose and hydroxypropyl methylcellulose, and a preservative selected from the group consisting of sodium benzoate and nipagin. 3. Eye drops according to claim 1 or 2, characterized in that the composition of vitamins of group B consists of vitamin B2. 4. Drops according to claim 1 or 2, characterized in that the composition of vitamins of group B consists of vitamins B1 and B2. 5. Drops according to claim 1 or 2, characterized in that the composition of vitamins of group B consists of vitamins B1, B2 and PP.