RU2513997C1 - Combined ophthalmic preparation presented in form of eye drops and containing polyhexamethylene guanidine and taurine - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: preparation comprises branched polyhexamethylene guanidine in the form of hydrochloride, taurine, a promoting ingredient, additives and water. The promoting ingredient is specified in succinic acid or its pharmaceutically acceptable salt, while the additives are specified in a group consisting of physiologically acceptable alkaline or acidic agents and a salt tonic agent specified in physiologically acceptable sodium or potassium salts or mixtures thereof.

EFFECT: more effective therapeutic effectiveness for pathological conditions of the eyes, and a potential comorbid bacterial infection.

3 cl, 3 tbl, 4 ex

Description

FIELD OF THE INVENTION

The invention relates to pharmacology and ophthalmology. More specifically, it provides a preparation in the form of eye drops for treating pathological conditions of the eyes, such as ophthalmic hypertension and cataract, and also acting as a remedy for corneal injuries, especially complicated by bacterial infections. The drug in accordance with the invention can find application as an additional tool in preparing patients for surgical intervention for cataracts and as means against complications of bacterial origin in the postoperative period.

State of the art

In the broadest sense, the term "cataract" defines any change in optical uniformity or a decrease in the transparency of the lens of the eye. Many factors are involved in the etiology of cataracts, such as heredity, aging, diabetes, smoking, nutrition, the cumulative effect of X-ray and UV radiation, and endocrine and / or enzymatic imbalance in the body.

These factors negatively affect the processes of electrolyte metabolism, which, in particular, leads to disruption of cell osmoregulation and adversely affects the permeability of the membranes of the cornea and retina cells. In the publication Kopylov G.V., Vislabokov I.I., Mantsev V.V. // Biology. - 1989. - Ser. 3. T. 3. No. 1. S.62-66, it was shown that taurine normalizes electrolyte metabolism. Also in the literature there is evidence of the normalization of protein, carbohydrate and hormonal metabolism with the introduction of taurine. These factors contributed to the introduction of taurine in ophthalmic medical practice since the second half of the 1970s.

Ophthalmic hypertension is not a disease as such, but it is a pathological condition of increased intraocular pressure, which is normally in the range of 10-21 mm Hg. Persistent or frequently occurring ophthalmic hypertension can not only cause glaucoma, but also lead to other eye damage leading to loss of vision, such as retinal vein occlusion and non-glaucomatous destruction of the optic fiber layer [Minckler D.S. "Histology of optic nerve damage in ocular hypertension and early glaucoma." Surv. Ophthalmol. Vol. 33 (April 1989). P.401-411].

There is evidence that the instillation of taurine solutions positively affects the blood-ophthalmic barrier and significantly reduce intraocular pressure (IOP). In the article Bunin A.Ya., Yartseva E.I., Kolesnikova Yu.A., Ermakova V.N., Yakovleva A.A., Bernikova T.A., Komleva N.A. “The effect of taurine on intraocular pressure and the blood-ophthalmic barrier (experimental study)” [Vopr. oft. No. 1 (1978). P.22-24] on a model of prostaglandin hypertension in rabbits, it was shown that instillation of a 4% taurine solution delays the increase in IOP in time and statistically significantly decreases by 2-5 mm Hg. IOP of the experimental eye throughout the experiment. Similar results were obtained on the model of salt hypertension of the eyes in rhesus monkeys. Thus, taurine has a hypotensive effect on ophthalmotonus, which may be due to inhibition of secretion of aqueous humor.

Mechanical injuries of the cornea, especially those received in the absence of proper medical care (for example, in natural disasters, military operations, expeditions, etc.), are usually accompanied by bacterial infections, which can have irreversible consequences for the eye until it is lost.

Preparing a patient for ophthalmic surgery, in particular regarding cataracts, requires the eradication of bacterial infections that can cause postoperative complications, especially in the case of elderly patients or patients with diabetes mellitus (Suto C., Morinaga M., Yagi T., Tsuji C ., Toshida N. "Conjunctival sac bacterial flora isolated prior to cataract surgery". Infect. Drug Resist. No.5 (2012). P.37-41). First of all, the degree of infection with typical pathogens of conjunctivitis and blepharitis should be minimized. Therefore, there is a need for ophthalmic preparations with a wide profile of antibacterial activity, which also have hypotensive and reparative effects.

Various ophthalmic preparations containing taurine are known in the art. EP0778021 (publ. June 11, 1997) discloses eye drops to accelerate the regeneration of the cornea after damage, containing 0.5-3.0% taurine, sodium chloride, potassium chloride and sodium bicarbonate having a pH of from 5.5 to 8.0 and osmotic pressure from 250 to 450 mOsm.

The disadvantage of such taurine preparations is their susceptibility to contamination by pathogenic microorganisms during storage of opened packages, which makes their use ineffective and unsafe.

In the patent RU 2127099 (publ. 10.03.1999) disclosed eye drops containing (in wt.%):

sodium succinate hexahydrate 1.91 sodium citrate 5.5-aq 0.40 sorbitol 0.80 sodium chloride 0.31 L-glutamic acid 0.015 riboflavin mononucleotide phosphate 0.02 benzalkonium chloride 0.014 glucose 0.30 taurine 0.4 distilled water for injection rest

The use of drops can improve energy metabolism in the cells of the anterior and posterior corneal epithelium, as well as in keratoblasts in any process that disrupts this metabolism.

In the patent RU 2404768 (publ. 10.03.1999) disclosed eye drops for the prevention and treatment of cataracts, containing:

taurine 1.5-1.9 g carnosine 1.5-1.9 g glutathione not less than 0.01 g low molecular weight dextran microdispersed 10.5-15.0 g benzalkonium chloride 0.005-0.02 g purified water up to 100 ml

In patent RU 2295331 (publ. 20.03.2007) eye drops of reparative and anti-glaucoma action are disclosed, containing:

taurine 4 g cyanocobalamin 0.025 g dextran 5.0 g benzalkonium chloride 0.01 g purified water up to 100 ml

The use of benzalkonium chloride as a preservative can reduce the degree of contamination of the drug with microorganisms after starting use from an open package, however, preservatives of this type have a strong local irritant effect and are allergens, which is their drawback. Therefore, there is a need for new effective and safe treatments for cataracts.

Disclosure of invention

As a result of extensive research, the authors of the present invention found that the disadvantages of the prior art can be overcome by creating a combined ophthalmic preparation in the form of eye drops containing polyhexamethylene guanidine and taurine and further comprising sodium succinate.

Thus, the present invention, aimed at expanding the arsenal of drugs, provides an ophthalmic preparation in the form of drops containing a combination of active components, a promoting component, excipients and water, in which the combination of active components consists of a branched polyhexamethylene guanidine of the formula (I)

n ₁ , n ₂ , and n ₃ are 1-3, az is 0.15-1.10 with a molecular weight distribution of M _W / M _n from 5.4 to 9.3 with a weight average molecular weight of M _W ranging from about 3800 to 6300 and a number average molecular weight M _n in the range of about 600 to 1100 present in the form of the hydrochloride and taurine;

the promoting component is selected from succinic acid or its pharmaceutically acceptable salt, with the following content of components (in wt.%):

polyhexamethylene guanidine hydrochloride 0.03-0.5 taurine 1.0-5.0 promoter component 0.5-2.0

excipients selected from the group consisting of physiologically tolerated alkaline or acid agents present in amounts required to maintain the pH of the drug in the range from 6.0 to 7.8, and a salt tonic agent selected from physiologically tolerated sodium or potassium salts and their mixtures present in an amount that provides the tonicity of the drug, comparable to the tonicity of the natural tear fluid of a healthy person.

Preferably, in the ophthalmic preparation, the physiologically tolerated acid agent is hydrochloric acid, present in an amount required to provide a pH of 6.0 ± 0.1 of the preparation.

Also preferably in the ophthalmic preparation, the physiologically tolerated alkaline agent is a mixture of potassium dihydrogen phosphate and sodium can hydrophosphate, present in amounts required to maintain the pH of the preparation in the range of 7.2 to 7.7.

One of the technical results of the present invention is to accelerate the restoration of corneal tissue during traumatic injury complicated by an attached bacterial infection. Another technical result is an improvement in the course of cataracts. Another technical result is the possibility of reducing the drug load of allergenic antibiotics in preparing the patient for ophthalmic surgery, for example, about cataracts.

A comparative study of the action of the drug "Taufon, eye drops 4%" and the drug in accordance with the invention on a model of traumatic cataract in rabbits revealed the advantage of the proposed drug over the known one, which is expressed in a smaller area and intensity of turbidity of the damaged area of the lens. During the study, no side effects were found, as well as signs of intolerance and allergic effect of the proposed drug.

Thus, the authors confirm the achievement of the claimed technical result of the invention.

The implementation of the invention

The invention will now be illustrated by examples of its implementation.

Example 1. The preparation of eye drops that do not contain a buffer mixture

Under aseptic conditions, pyrogen-free distilled water taken in a volume of 80-85% of the volume of the finished product is loaded into a weighed sterile container. While stirring, 0.05 g of polyhexamethylene guanidine hydrochloride (PHMG-GC), 1.00 g of taurine, 2.00 g of sodium succinate and 0.70 g of sodium chloride are dissolved in water, after which the pH of the solution is adjusted to 6.0 ± 0. 1 by adding a 0.1 M hydrochloric acid solution. The mass of the resulting mixture was adjusted to 100.00 g by the addition of pyrogen-free distilled water.

Sterilization of the solution is carried out at 120 ° C for 10-12 minutes, after which the finished preparation is packaged in 5 ml portions in sterile bottles of high pressure polyethylene with droppers, provided with screw caps and placed in boxes indicating the information necessary for identification and use.

Example 2. Preparation of eye drops with a buffer mixture

Under aseptic conditions, pyrogen-free distilled water taken in a volume of 85-90% of the volume of the finished product, which is heated to 60 ° C, is loaded into a weighed sterile container. With stirring, 0.03 g of potassium dihydrogen phosphate, 0.27 g of sodium monohydrogen phosphate and 0.50 g of sodium succinate are dissolved in hot water. The resulting solution was cooled to 25 ° C and 0.50 g of PHMG-GC, 5.00 g of taurine and 0.1 g of sodium chloride were added to it with stirring. The mixture is stirred until the components are completely dissolved.

Sterilized filtration is carried out through a 0.2 micron filter, 2 ml portions are aseptically packed in high-pressure polyethylene bottles with droppers, provided with screw caps and placed in boxes with information necessary for identification and use.

Example 3. The study of therapeutic effects on a model of traumatic cataract in rabbits

The study was carried out on 24 rabbits of the breed "White Giant" with an average weight of 4.75 kg 4-5 months of age with a model of traumatic cataract, divided into groups of 6 individuals (n in table 2).

After drug myosis caused by twofold instillations in the conjunctival cavity of a 1% solution of pilocarpine hydrochloride with an interval of 15 minutes, laser coagulation of the pigment rim of the pupil is performed with localization of coagulates 1.5-2.0 mm outward from the optical center of the lens in the meridian 8-10 causing traumatic cataract. 15–40 coagulates are applied to the iris of each eye. Laser radiation parameters: pulse energy 1.6-2.2 W, pulse duration 0.6 sec.

In accordance with the objectives of the study, all animals are divided into 4 groups (table 1).

Table 1 Group The drug and dosage regimen one The drug in accordance with example 1; 1 mg / kg rabbit weight 1 time per day subconjunctival for 10 days 2 The preparation in accordance with example 2; 1 mg / kg rabbit weight 1 time per day subconjunctival for 10 days 3 “Taurine, eye drops 4%”, 1 mg / kg rabbit weight 1 time per day subconjunctively for 10 days four Water for injection in 0.3 ml rabbit 1 time per day subconjunctival for 10 days

In the first week, lens biomicroscopy and the study of its transparency are carried out daily, and then weekly in transmitted light using a SchL-2B slit lamp and a head-mounted binocular microscope (HBO-2) using ophthalmoscopic lenses of various optical powers.

In addition, in all rabbits, immediately after cataract modeling, as well as after 1 and 2 months of therapy with the compared drugs, photographic recording of the lens opacity is performed using the Retcam-2 apparatus (manufactured by Massive Research, USA). The results of dynamic observation of rabbits are recorded in a special registration card.

30 minutes after the laser exposure, the pigment border of the pupil in the areas of application of laser coagulates acquires a milky white color. On the second day of the experiment, the pupil gains its previous size and exposes the part of the lens exposed to laser radiation. It represents a site of intense opacification of the capsule and the anterior cortical layers of the horseshoe-shaped crystalline lens (projection of the pigmented border of the narrowed pupil at the time of laser exposure). In this case, a thin area of turbidity, repeating the contour of the edge of the pupil, is surrounded by a halo of subcapsular vacuoles and areas of developing turbidity that do not extend more than 1.5 mm from the main "contour" turbidity.

Starting from the third or fourth day, the clouding of the lens areas under consideration reaches maximum intensity, depth and area and stabilizes. Traumatic cataract acquires a tendency to reverse (extremely insignificant) development starting from the 30th day of the experiment. However, even after 60 days of treatment with the studied drugs, in no case was a complete regression of lens opacity observed.

The area of the lens opacification is determined using a micrometric grid superimposed on a full-sized image of the lens, made using the apparatus "Retcam-2", according to the formula

S = N + n / 2,

where S is the area of turbidity, mm ² ; N is the number of millimeter squares fully “occupied” by turbidity, n is the number of millimeter squares partially “occupied” by turbidity.

The severity of clouding of the lens is evaluated on a four-point scale: "0" - the full transparency of the lens; "1" - gentle cloudy cloudiness; “2” - pronounced turbidity, behind which a weakened reflex from the fundus is still guessed; "3" - intense dense opacification of the lens in the absence of a reflex from the fundus in its projection. Due to the fact that the clouding of the lens is not uniform, the average clouding coefficient is calculated by multiplying the severity of clouding in each of its sections by its area, followed by summing and dividing by the total clouding area.

The results of dynamic observation of lens opacities in rabbits in the groups shown in table 1 are presented in table 2.

table 2 Group n Observation stage 30 days 60 days Area, mm ² Severity Area, mm ² Severity one 6 4.8 ± 0.6 2.3 ± 0.3 4.1 ± 0.6 2.1 ± 0.3 2 6 4.1 ± 0.5 2.0 ± 0.4 3.7 ± 0.5 1.9 ± 0.2 3 6 6.1 ± 0.6 2.9 ± 0.2 5.2 ± 0.6 2.3 ± 0.3 four 6 6.4 ± 0.7 3.1 ± 0.3 6.1 ± 0.5 2.7 ± 0.3

The drug in accordance with examples 1 and 2 of the invention, according to the results of the examination on the 30th day of the experiment, was statistically significantly more effective than the drug "Taufon, eye drops 4%". In control group 4, the area, depth and intensity of turbidity remained more stable than in animals treated with therapy.

According to the results of further observation of rabbits, carried out from the 30th to the 60th day of the experiment, a more noticeable stabilization and slowdown of the development of opacities in the lenses (area, depth and severity of opacities) was established, especially in group 2 treated with the drug in in accordance with examples 1 and 2 of the invention.

Example 4. Determination of the spectrum of bactericidal efficacy of drugs in vitro and the minimum bacteriostatic (inhibitory growth) concentration (MIC)

The spectrum of bactericidal efficacy of drugs in an in vitro experiment is characterized by the suspension method in accordance with guidance P 4.2.2643-10 “Methods of laboratory research and testing of disinfectants to assess their effectiveness and safety”. A 0.05% solution of polyhexamethylene guanidine hydrochloride of the formula (I) (G-PHMG) and preparations prepared in accordance with Examples 1 and 2 of the invention are examined.

The minimum bacteriostatic (BMD) concentration is determined by the micromethod of serial two-fold dilutions in Müller-Hinton medium (Oxoid) in accordance with the normative document "Guidelines for determining the sensitivity of microorganisms to antibacterial drugs" (MUK 4.2 1890-04).

Preparations with a concentration of the active component of 1000 μg / ml, prepared on pyrogen-free distilled water, diluted 128 μg / ml of culture medium. Studies are performed in 96-well plates for immunological studies. Prepare a series of two-fold dilutions of the test drug in medium II Muller-Hinton (Oxoid) in a volume of 50 μl. For microorganisms that do not grow well on ordinary nutrient media, 5% defibrinated lamb blood is added to the above medium.

An inoculum of bacterial cultures from isolated colonies is prepared as a suspension according to the McFarland turbidity standard (0.5) in physiological saline. The suspension is diluted in medium II Muller-Hinton to a concentration of 10 ⁸ CFU / ml The prepared suspension is added to the wells of the plates (50 μl / well) and dilutions are prepared in the concentration range from 128 to 0.0003 μg / ml. Inoculated plates are incubated for 18 hours at 37 ° C. The lowest concentration of the study drug, in which there is no visible growth of microorganisms, is an estimate of the IPC.

To determine the MPC of Candida albicans from an isolated colony, a suspension is prepared in Saburo culture medium according to the MacFarland turbidity standard (0.5), which corresponds to 10 ⁶ CFU / ml for fungal cultures. A series of double dilutions of the preparations is prepared in Saburo's medium in a volume of 100 ml. After plating the yeast cultures, the plates are incubated for 48 hours at 30 ° C.

The results of the action on the IPC series of consecutive double dilutions are presented in table 3.

Table 3 Microorganism (Strain) G-PHMG A drug Example 1 Example 2 Eschenchia coli (ATCC 27952) 2 four 8 Proteus mirabilis (5) one one four Staphylococcus aureus (ATCC 29213, methicillin sensitive) 2 four 8 Staphylococcus aureus (ATCC 43300, resistant to methicillin) 2 four four Candida albicans (ATCC 24433) four 8 16 Streptococcus oralis (1105007) one 2 four Streptococcus pyogenes (151023) 2 8 8

The data obtained indicate that the proposed drug has a higher activity compared with a solution of PHMG-GC.

Claims (3)

1. An ophthalmic preparation in the form of drops containing a combination of active components, a promoting component, excipients and water, characterized in that the combination of active components consists of a branched polyhexamethylene guanidine of the formula (I)

n ₁ , n ₂ and n ₃ are 1-3, az is 0.15-1.10 with a molecular weight distribution of M _W / M _n from 5.4 to 9.3 with a weight average molecular weight of M _W ranging from approximately 3800 to 6300 and a number average molecular weight M _n in the range of about 600 to 1100 present in the form of hydrochloride and taurine;
the promoting component is selected from succinic acid or its pharmaceutically acceptable salt, with the following content of components (in wt.%):
polyhexamethylene guanidine hydrochloride 0.03-0.5 taurine 1.0-5.0 promoter component 0.5-2.0
excipients selected from the group consisting of physiologically tolerated alkaline or acid agents present in amounts required to maintain the pH of the drug in the range from 6.0 to 7.8, and a salt tonic agent selected from physiologically tolerated sodium or potassium salts and their mixtures present in an amount that provides the tonicity of the drug, comparable to the tonicity of the natural tear fluid of a healthy person. 2. The ophthalmic preparation according to claim 1, characterized in that the physiologically tolerated acid agent is hydrochloric acid, present in the amount required to provide a pH of 6.0 ± 0.1. 3. The ophthalmic preparation according to claim 1, characterized in that the physiologically tolerated alkaline agent is a mixture of potassium dihydrogen phosphate and sodium can hydrophosphate, present in amounts required to maintain the pH of the drug in the range from 7.2 to 7.7.