Ophthalmic compositions containing mucoadhesive polysaccharides stimulating corneal epithelium recovery

FIELD: medicine, pharmaceutics.

SUBSTANCE: group of inventions relates to medicine, particularly to ophthalmology. An ophthalmic composition for treating keratoconjunctival damages and inflammations consists of an aqueous solution containing arabinogalactan 1 to 10 wt %, one or more preserving agents specified in a group consisting of sodium merthiolate, thimerosal, phenylmercuric nitrate or phenylmercuric acetate, phenylethyl alcohol, methyl-, ethyl-, propyl parabene, chlorhexidine acetate or gluconate or chlorobutanol, and containing no benzalkonium chloride. The ophthalmic composition is applied as a lacrimal substitute recommended for people wearing contact lenses.

EFFECT: group of inventions provides treating corneal erosions caused by wearing contact lenses.

16 cl, 5 tbl, 7 dwg

 

The present invention relates to ophthalmic compositions containing mucoadhesive polysaccharides can stimulate the recovery of the corneal epithelium. More specifically, the invention relates to ophthalmic solutions containing arabinogalactan, having a protective effect on the corneal epithelium, which is especially recommended for use as artificial tears to stimulate healing of lesions of the cornea and is especially applicable for people who wear contact lenses.

As you know, the cornea is an anterior fibrous sheath of the eyeball, the cornea is only her sixth part, and the rest represents the sclera; these two structures are in contact and form an indivisible whole. In fact, the cornea is less curved than the sclera, so it gives the impression that the cornea is forward. Due to its specific characteristics (transparency and absence of vessels), as well as due to the shape, which makes it a fine concave-convex lens, the cornea is an essential element of the refractive system of the eye.

The front surface of the cornea is convex and elliptical, its horizontal diameter is slightly larger than the vertical, and therefore, the cornea has a different curvature in these two directions. This difference is due to the Oh physiological astigmatism. The posterior corneal surface, opposite, concave, has the same diameter and radius in all directions. The thickness of the cornea varies from approximately 0.5 mm in the Central part to about 0.7 mm at the periphery.

Histologically, the cornea consists of five layers, which are arranged in the following order (from outside to inside): the epithelium, the membrane of Bowman, stroma, the membrane of Descemet and endothelium.

The cornea performs its optical function due to its high transparency and flatness of the surface in contact with air. The last feature is caused by the presence of tear film that covers the epithelium, which is uneven due to the presence of network MicroSCADA in the outer layer. Tear film makes the epithelium is smooth, uniform and gives him a high optical properties.

Tear film that covers the conjunctiva of the eyeball and conjunctiva of the eyelids, consists of three superposed on each other layers, which are arranged in the following order (from outside to inside): the lipid layer, water layer and mucous layer. The mucin produced by the goblet cells of the conjunctival epithelium, making the entire surface of the epithelium smooth, which creates the opportunity for a homogeneous distribution of the water component of the slimy film on the layer; the lipid layer, which secrete meibomiae gland and the glands of Zeis, who meet the function of preventing the evaporation of the tear film.

As already mentioned, transparency is a key feature of the cornea. Transparency is possible due to the absolute absence of vessels in this tissue, the structural characteristics of stroma and certain specific physiological processes that regulate the circulation of water and swelling of the cornea and prevent the wetting of the cornea, causing the level of hydration to the normal value of about 78%.

Other physiological characteristics of the cornea is its reflectivity (reflection light from the surface), which is associated with the integrity of the epithelium, and permeability - are an integral function of the water cycle and/or penetration of foreign substances, such as pharmaceuticals.

Significant sensitivity to various stimuli related, ultimately, with abundant innervation of the membrane, which decreases in old age and in the presence of some inflammatory and dystrophic-degenerative changes.

The most frequently detected changes of the cornea can be an initial symptom of inflammatory processes, dystrophic or degenerative nature or explicit manifestation of the disease. Changes mechanical-traumatic nature include: erosion of the cornea, caused by surface abrasion of the corneal epithelium or foreign bodies, for example, metallicheskie, glass or plastic particles, wood and crop residues; rupture or perforation caused by objects penetrating the eye with a special power that are embedded in some depth; recurrent erosion of the cornea, which consist of spontaneous episodes of deterioration or peeling of the layers of the corneal epithelium; burns caused by weak acids or weak bases, high temperature or ultraviolet radiation.

The destruction of the corneal epithelium in any case increases the risk of invasion of pathogens, with potentially disastrous results. Conjunctiva is actually one of the body tissues colonized by microorganisms from the very first moments of extrauterine life, and it should be noted that among the usual resident microorganisms, such as Streptococcus (St. viridans), staphylococci (S. epidermidis, S. aureus), hemophilic Bacillus, Propionibacterium acnes, there are some, possessing all the characteristics that are inherent in true pathogens.

The ecosystem of the conjunctiva is a system that strives to be balanced and biologically active and remains so until the tissue, microbial or environmental factors that disrupt this balance, thereby making the environment of the conjunctiva favorable territory for micro is organizmov or for the owner. Tissue factors depend on the histological structure of the tissue of the conjunctiva and secrets, the components of the tear film: these factors can act as associated causes in the pathogenesis of various infections of the cornea, caused by bacteria, viruses and fungi. They can colonize the tissue of the cornea, especially in the presence of lesions of the cornea and to give rise to keratitis, including very severe, disrupting the function of the eye, such keratitis can cause ulcers of the cornea. Bacterial keratitis is characterized by acute pain, ulceration of the epithelium and sometimes corneal stroma and the presence of detachable conjunctiva. Etiological reasons for this defeat eyes are S. aureus, St. pneumoniae, Ps. aeruginosa, Citrobacter, Klebsiella, Enterobacter. In addition, Mycobacterium chelonei and Mycobacterium fortuitum can cause chronic ulcers of the cornea. As for the etiology of fungal infections, fungal keratitis cause, mainly saprophytic filamentous fungi and yeast, colonizing areas of damaged epithelium of the cornea. Finally, keratitis caused by Acanthamoeba spp., is a rare form of chronic keratitis caused by this parasite, which could even threaten vision and is characterized by annular multifocal suppuration corneal stroma, followed by ulcers.

All changes of the cornea (traumatic, inflammatory or is egenerative type), which cause tissue destruction or loss of substance, quickly heal through physiological mechanisms that partly differ from mechanisms other tissues due to the lack of corneal vessels. The healing process which is affected by the characteristics of the lesion, and which, undoubtedly, is more problematic in the presence of purulent ulcers, always starts at the epithelial level, not only in the case of surface damage limited to this layer, but also in cases involving the parenchyma. Thus, the epithelium is involved in the healing of lesions damage stroma, as illustrated in the formation of epithelial cap", is necessary in order to regulate the development of subsequent events collagen regeneration.

Simple wound epithelium or abrasions heal quickly due to the high mitotic activity of cellular elements in this area. In fact, at the time of injury is temporary interruption of the physiological mechanism shedding of superficial cells, as well as the movement and migration of intact adjacent cells to the damaged area. An interrupt stops when devoid of epithelium region is fully covered, and regeneration of epithelial elements provides restoration of normal structural characteristics.

If the damage is about what ranchero epithelium, his complete healing enables the recovery of high reflecting properties and transparency of the membrane.

Specific and frequent cause of corneal erosion are contact lenses. Causes erosions may include a variety of physiological factors, toxic or mechanical nature, and erosion of the cornea, caused by contact lenses, can result, in particular, their application, difficulties of their introduction and removal, problems in the relationship of the lens and the cornea is damaged lenses or foreign bodies trapped under the lens. These erosion can occur when you use and hard and soft lenses, but they are more common in people who use hard lenses. The study included 500 users of soft contact lenses, showed that one third of them were quite serious eye problems, and these problems could lead to infections, and that almost 50% of them had symptoms of lung lesions at least one eye (R.J. Derick et al., CLAO J., 1989 Oct-Dec; 15(4):268-70).

Thus, erosion of the cornea - a problem that can occur in anyone, but contact lenses, will certainly cause abrasion of the cornea, and this increases the risk: users of contact lenses suffer from lesions of the cornea is approximately three times more often than those who do not use lenses.

The use of contact lenses can outproduce a number of other problems, which occur due to the influence of the lens on the normal flow of oxygen to the corneal epithelium or due to pathological conditions, such as conjunctivitis with hyperplasia of papillae or vascularization of the cornea, or more frequent dry eye syndrome". The latter, also known as "dry eye, or keratoconjunctivitis sicca, is a disorder caused by reduction of the quantity and quality of tears. Typical symptoms of pathology are irritation and burning sensation in the eyes, feeling of sand or foreign bodies, photophobia, pain, and "the fog before my eyes." In the end, you may experience the plagues that threaten vision.

For the treatment of "dry eye" or weakening of associated symptoms suggested many formulations of artificial tears for periodic instillation on the conjunctiva (or code of the conjunctiva), to ensure the replacement of the lacrimal fluid and relieve the feeling of dryness in the eyes. To increase the retention on the surface of the cornea, as well as to provide good portability, these drugs, usually made viscous by the addition of high molecular weight medium - usually water-soluble polymers, synthetic, semi-synthetic or natural origin. Based on the fact that it takes a long time to save on the surface of the cornea, artificial tear should the te properties close as possible to the dispersion properties of mucin, that is, it should behave the extent possible as malomyotismon substance, so preference was given to compositions based on macromolecular compounds of natural origin, such as cellulose derivatives (in particular, esters of cellulose, such as carboxymethylcellulose, and alcohol derivatives of cellulose ethers, such as hypromellose), glycosaminoglycans (in particular, hyaluronic acid is a polysaccharide found in many tissues and fluids of humans and animals, which is widely used in ocular preparations), polysaccharides having suitable rheological properties (e.g.the polysaccharide extracted from tamarind gum, TSP).

In addition, to provide the necessary lubricating properties of the surface of the cornea, artificial tears always have a certain viscosity (even if the preferred solutions of this viscosity is sharply reduced when the product is subjected to shear stress, such that observed in the blink), which can cause blurred vision, and leave residues on the cornea or on the edge of the eyelids.

Given the above problems associated with the use of contact lenses, it is extremely important that the potential product, use the nd as an additive to the tear fluid, had - in addition to the good tolerability and lack of irritant effects on the eyes - a low viscosity, so as not to cause fuzziness of view and does not leave residue on the lens and the edges of the eyelids. Equally important for such a product the ability to ensure the integrity of the corneal epithelium and prevent any negative interactions and/or reactions with the material of the contact lens.

According to the present invention it was found that ocular solutions containing specific natural polysaccharide polymers, arabinogalactan, which are compounds with almost negligible viscosity, have sufficient mucoadhesives to avoid rapid drying at the surface of the cornea, and in addition to the good tolerability have a significant ability to stimulate the epithelium. In fact, it was demonstrated that within the research associated with the present invention, these compositions are introduced into the eye with the damaged epithelium, accelerate healing. In the proposed composition associated with the use of contact lenses, which are able to stimulate the healing of corneal erosion that may occur, preventing the deterioration of epithelial damage and any complications. As already mentioned, another important characteristic of the product that makes it innovative and perfect is for this purpose, is that with appropriate mucoadhesive properties, it does not change the viscosity of aqueous solutions and, therefore, does not affect vision as subjects wearing contact lenses, and those that do not use them. At the same time mucoadhesive properties allow the product to set different connection with the mucous membrane surface of the conjunctiva and cornea, and these links allow the product longer be stored in preocular region, and thereby allow us to manifest the epithelium and hydrating activity that characterizes the product.

Eye solutions for use as artificial tears containing as polymer for lubrication of the cornea arabinogalactan already described in U.S. patent No. 4039662 (Hecht et al., transferred Alcon Laboratories Inc.). In this case, however, regenerating epithelium properties of the polysaccharide has not been given much importance, they were not used and, most importantly, its usability was due to the need for the presence in the composition of benzalkonium chloride. According to the description of this document, in fact, the introduction of benzalkonium chloride is a substance that has been used in ophthalmic compositions as biocide - is a critical element for the proposed drug, because it is believed that this compound is combined with the policies of the reed or forming complexes with the polysaccharide, is the reason for the stability of the product in the adsorbed state on the surface of the cornea, that is, it performs the function of the stabilizer precorneal film. In accordance with this observation, in the cited document proposes eye solutions, based on the arabinogalactan and containing as a binding ingredient benzalkonium chloride.

On the other hand, according to the research carried out within the framework of the present invention, the epithelium and the protective properties of the surface of the cornea, which are arabinogalactan and mucoadhesive properties, giving them a long time to save precorneal the film, even in the absence of any significant viscosity, are not associated with the simultaneous presence of benzalkonium chloride. Therefore, from the regenerating epithelium compositions containing arabinogalactan of the present invention, benzalkonium chloride excluded because, if the composition must include preservatives, can be used many other products suitable for this purpose.

As you know, arabinogalactane represent a class of long, densely branched polysaccharides with a molecular weight of from 10,000 to 120,000 daltons, and these polysaccharides have a Central structure consisting of a chain constructed of units galactopyranose In nature they are found in a variety of microbial systems, in particular mycobacteria, which they are in complex with peptidoglycan and mycotic acid, forming a sheath cells. Rich sources of arabinogalactans (mainly in glycoprotein form) are many edible and inedible plants. Many plants with confirmed immunostimulating properties, such as Echinacea purpurea, Baptisia tintoria and Thuja occidentals, contain significant amounts of arabinogalactans. Woody tissue of plants of the genus Larex especially rich arabinogalactane, especially Larex occidentalis, but also Larex dahurica (originating from Central Asia), for example, Larex dicidua (European) and Larex leptolepis (Japanese). In fact, larch wood is the most common industrial source of arabinogalactan from this source this polysaccharide extract, to be used not only for industrial purposes, for example, in the cosmetic industry, but also and primarily in the food industry as diet and food ingredient, rich in fiber, beverages, as well as immunomodulatory agent.

Thus, the present invention relates to ophthalmic compositions for use as artificial tears with keratoplasties and healing epithelium activity, containing from 1 to 10 wt.% arabinogalactan in an aqueous solution, characterized in that it is contains benzalkonium chloride. In particular, the most suitable concentration of arabinogalactan for the intended application of the present invention are the concentrations of components from 3 to 5 wt.% polysaccharide in aqueous solution.

According to preferred variants of implementation of the present invention arabinogalactan used in the proposed compositions is arabinogalactan derived from larch and related to the pharmaceutically acceptable category. In particular, arabinogalactan, preferably used in the preparations according to the present invention (arabinogalactan CAS# 9036-66-2), is the commercial name FiberAid®AG and is produced by LAREX®(Cohasset, MS, USA) for U.S. patent No. 5756098. This arabinogalactan is in the form of a fine white powder (molecular weight = 45 kilodaltons), dispersible, but not completely soluble in cold water.

All arabinogalactan isolated from larch, are polysaccharides that do not contain nitrogen. One third of the molecules are built from the main circuit related to the type of (1→3)-β-D-galactopyranose, while the rest consist of side groups attached by (1→6) to each unit of galactose, the size of which varies from monosaccharides to oligosaccharides. Distribution of side groups are heterogeneous. Often the side group is the Wallpaper disaccharide β-D-Galp-(1→6)-β-D-Galp or β-L-Arap-(1→3)-α-L-Araf. Less often monomer β-D-Galp or the monomer α-L-Araf. Units of galactose and arabinose are in a molar ratio of approximately 6:1.

Morphological studies of arabinogalactan showed that this polymer has a high conformational freedom and can take many different forms, but the main chain usually has a rigid structure, triple helix, while the side groups form a flexible branches with plenty of outdoor hydroxyl groups (R. Chandrasekaran, S. Janaswamy, 2002, Carbohydrate Research). This is the reason underlying mucoadhesive characteristics of the polymer, which makes it able to form hydrogen bonds with molecules of mucin eyes.

In some specific embodiments, the implementation of the present invention, the ophthalmic composition of the addition of arabinogalactan also contains one or more funds, regulatory toychest, which gives the solution of the desired osmolarity. Because the proposed ophthalmic solution may be isotonic or slightly hypotonic with respect to the tear fluid regulating toychest funds will be present in the composition in such amount to provide the osmolarity of 150 to 300 mOsm/L. Preferably referred to one or more funds, regulatory toychest selected from the group consisting of mannitol, sodium chloride, chlorine is IDA potassium, dextrose, boric acid and sorbitol.

As arabinogalactan also becomes stable in buffered aqueous solutions with different pH values, other ingredients that can be added, like known in the field of pharmacology, represent one or more ophthalmologist acceptable acid or base, as proofreaders pH, and/or one or more buffers. In particular, suitable buffers can be selected from the group consisting of phosphate buffer, borate buffer, citrate buffer, bicarbonate buffer, tributary (trihydroxystilbene). In addition, the compositions according to the invention can with advantage be used in other buffer systems.

If the product is not Packed in units containing the same dose, the composition may also contain preservatives and antimicrobials, except benzalkonium chloride. Possible preservatives, compatible with the product, in particular, are thimerosal sodium or thimerosal, a mercury-phenyl nitrate or mercury-phenyl acetate, phenethyl alcohol, methyl-, ethyl - and propylparaben, chlorhexidine acetate or gluconate and chlorbutanol.

Finally, to compositions containing arabinogalactan according to the invention, if necessary, can be added even complexing agents such as edetate or EDTA.

Another aspect of this is about the invention relates to the use of eye solution containing from 1 to 10 wt.% and preferably from 3 to 5 wt.% arabinogalactan in an aqueous solution for the production of ophthalmic compositions with keratoplasties and healing epithelium activity. As already noted, of the composition referred to ophthalmic compositions may be excluded benzalkonium chloride, without creating any problems.

Specifically ophthalmic composition obtained by the use of arabinogalactan according to the present invention, an artificial tear that shows people using contact lenses. In some specific embodiments, the implementation ophthalmic composition is an artificial tear, is indicated for the treatment keratoconjunctivitis damage and inflammation and, in the latter case, it may specifically be an ophthalmic composition, recommended for the treatment of corneal erosion caused by the use of contact lenses.

Below by means of non-limiting examples are presented some examples of compositions containing arabinogalactan, applicable as artificial tears with keratoplasties and healing epithelium activity according to the invention.

Composition 1
arabinogalactan2 wt.%
mannitolq.s. up to 300 mOsm/kg
phosphate buffer with a pH of 7.4q.s. to 100 wt.%
Composition 2
arabinogalactan3 wt.%
mannitolq.s. up to 300 mOsm/kg
deionized waterq.s. to 100 wt.%
Composition 3
arabinogalactan6 wt.%
NaClq.s. up to 300 mOsm/kg
deionized waterq.s. to 100 wt.%
Composition 4
arabinogalactan2 wt.%
mannitolq.s. up to 300 mOsm/kg
mercury-phenyl-nitrate0.002 wt.%
phosphate buffer with a pH of 5.9q.s to 100 wt.%
Composition 5
arabinogalactan5 wt.%
NaClq.s. up to 300 mOsm/kg
thimerosal sodium0,008 wt.%
deionized waterq.s. to 100 wt.%

For the preparation of the above compositions accurately weighed polymer was dispersible in deionized water or phosphate buffer and kept stirring until complete dissolution. To the thus obtained solution was added a suitable amount of other excipients (mannitol, NaCl and/or preservative), continuing to mix until dissolved.

The compositions thus obtained, can be sterilized by filtration through a membrane with a pore diameter of 0.22 μm.

Specific features of the present invention and its advantages will become more apparent in relation to one of the specific variants of its implementation, are described below only for illustrative purposes, together with the results of the experiments performed on them; also the data for comparison with the prior art. Some experimental results are also illustrated by the attached drawings, where:

Fig. 1 shows treason is their viscosity compositions, containing arabinogalactan in an aqueous solution of the present invention, as changing the concentration of the polysaccharide;

Fig. 2 shows the time trend of preservation on the surface of the cornea of a solution containing arabinogalactan according to the invention, moreover, the time saving is measured by determining the fluorescent derivative of arabinogalactan;

Fig. 3 shows the test results of the Schirmer I have rabbits that were injected with atropine sulfate (AS)to experimentally cause dryness of eyes; one eye mentioned rabbits have also introduced a composition containing arabinogalactan according to the invention (AG-Sol), as treatment;

Fig. 4 shows the histogram of the test results in damage to the corneal epithelium (visual examination using a slit lamp after the introduction of fluorescein) on the same rabbits, as in Fig. 3;

Fig. 5 graphically represents the results of the test on the healing of lesions of the cornea, experimentally induced in rabbits, which were then treated with the composition according to the invention (AG) or, for comparison, the saccharide gum tamarind (TSP) or hyaluronic acid (HA);

Fig. 6 and 6a graphically represent, with varying detail, the results of the test on the healing of lesions of the cornea, experimentally induced as in the tests of Fig. 5, where the composition of the present invention (AG-Sol) is ravniaetsia with a similar solution, also contains benzalkonium chloride (AG-Bz); and

Fig. 7 shows test results of cytotoxicity from the point of view of the viability of the cells after contact with the solution containing arabinogalactan according to the invention (AG-Sol), or with a similar solution containing benzalkonium chloride (AG-Bz).

Test viscosity

Solutions arabinogalactane (FiberAid® AG LAREX®, Cohasset, PCs Minnesota, USA), hereinafter referred to as AG, in various concentrations(0,2; 0,5; 0,8; 1; 2; 3; 4; 5; 6; 7; 8; 9 and 10 wt.%) tested for viscosity using a rotational viscometer Rheostress RS 150 (Haake) with coaxial cylinders (Z40 and Z41) at a constant temperature of 25 °C. the viscosity of the solution is shown graphically in Fig. 1 of the attached drawings.

Rogramme was obtained for values of the velocity gradient from 0 to 200 with-1on the charts evaluated the correlation between shear (τ) and velocity gradient (γ) by means of mathematical processing using software Rheowin. Thus discovered that solutions of the product AG have Newtonian rheological behavior, and the thixotropy is not present.

Newtonian behavior of solutions AG is due to the fact that they are not viscous is a desirable feature in the case of solutions that are injected into the eyes of a person wearing contact lenses; as a result, the liquid penetrates into the space between the lens and rogovichi, what s causing fuzziness of view.

Research mucoadhesive

1. The study of mucoadhesive on a solid matrix

Mucoadhesive properties of arabinogalactan according to the invention (FiberAid® AG, Larex®) was evaluated by measuring the force required to separate the two mucous surface, between which is placed the sample. The mucous surface consisted of a water dispersion of 25% gastric mucin pigs (TCI, Tokyo), adsorbed on filter paper (Saettone et al., 1989, Int. J. Pharm., 51:203-212).

The equipment used consisted of micrometer scale, a movable platform and a computerized system capable of recording the force required to separate two surfaces (the analyzed sample and mucous layer)as a function of the elongation (software TP 5008, TiePie Engineering, Leeuwarden, the Netherlands).

The work of adhesion was measured for solid matrices, consisting of the study of polysaccharide polymer, with a diameter of 13 mm and a thickness of 1 mm, which was obtained by pressing the appropriate number of polymeric material by the hydraulic press (Perkin-Elmer) at a pressure of 3000 kg/cm2.

The results were compared with the work of adhesion measured using solid matrices of the same size, consisting of a polysaccharide gum tamarind (TSP), known from the literature as mucoadhesive obtained in the same manner. The results obtained is shown in the following table.

TABLE 1
The work of adhesion for the studied polymers (mean ± error of the mean, n = 20)
AGTSPThe mucin/mucin
The work of adhesion (erg/cm2)282,77355,87118,00
Standard error23,6041,739,00

2. Rheological evaluation of the strength of adhesion of the mucin-polymer

To explore mucoadhesive properties of arabinogalactan (AG) in more detail, we applied the method Saettone et al. (Saettone et al., 1994, Journal of Ocular Pharmacology, 10:83-92) and Hassan and Gallo (Hassan, Gallo, 1990, Pharm. Res., 7:491-495), which consists of measuring changes in viscosity, resulting in dispersion of mucin after addition of the polymer. In this method the component viscosity caused by bioadhesive (ηb), is calculated from the equation ηb= ηt- ηm- ηpwhere ηt, ηmand ηp- viscosity coefficients of the system, mucin and polymer, respectively. The value of ηbthen normalizability according to ur next is the ranking Δη/η=η bp.

Then measured the change in the viscosity due to the interaction of mucin-polymer in aqueous solutions containing: (i) 15% mucin (ηm); (ii) AG and other polymers in the respective effective concentrations ηpthat was used as the standard; (iii) a mixture of mucin-polymer in the same concentrations that were specified previously (ηt).

Used the following songs:

1. An aqueous solution of gastric mucin pigs (MGS; TCI, Tokyo), 15% wt./wt.;

2. An aqueous solution of AG, 5% wt./wt. (FiberAid® AG, LAREX®);

3. An aqueous solution of the TSP, of 0.5% wt./wt. (Farmigea, Pisa);

4. An aqueous solution of hyaluronic acid, 0.2% wt./wt. (Chemofin, Milan);

5. Aqueous dispersion of mucin, 15% wt./wt., and AG, 5% wt./wt.;

6. Aqueous dispersion of mucin, 15% wt./wt., and TSP of 0.5% wt./wt.;

7. Aqueous dispersion of mucin, 15% wt./wt., and hyaluronic acid, 0.2% wt./wt.

Viscosimetrically the measurements were performed using a rotational viscometer Rheo-stress RS 150 (Haake) with coaxial measuring cylinders (Z40 and Z41) at a constant temperature of 32°C. Rogramme received when the value of the velocity gradient from 0 to 500-1.

Solutions of the polymers showed Newtonian behavior, and the obtained curves are used to evaluate the linear correlation between shear (τ) and velocity gradients (indicated by γ or D) by means of mathematical processing and, performed using the software Rheowin.

Hyaluronic acid, mucin, and the variance of the mucin-polymer showed a pseudoplastic-type behavior. Mathematical correlation of the obtained graphs defined using software Rheowin, was as follows: τ=aDb. Viscosity in Newtonian compositions and apparent viscosity of a pseudoplastic-type compounds was calculated by choosing a certain value D, and then received τ from the respective equations (D=200-1).

Viscosity values for all compositions are given below in table 2, and the value of the parameter Δη/η for polymers used are shown in table 3.

TABLE 3
Δη/η investigated compositions
TrackGS (% wt./wt.)AG (% wt./wt.)TSP (% wt./wt.)Hyaluronic acid (% wt./wt.)Δη/η
515,05,0--17,21
6 15,0-0,5-14,55
715,0--0,23,02

The results show that an aqueous solution of 5% wt./wt. AG has a much lower viscosity (1,38 MPa∙s)than the polymer taken as standard (9,16 MPa∙s and 24,40 MPa∙s for TSP and HA, respectively), although the normalized value Δη/η AG has the same value as the value of the TSP. This suggests that AG is able to various kinds of interaction with mucus covering the surface of the conjunctiva and cornea. Therefore, despite the fact that AG - non-viscous polymer which does not cause a lack of vision and does not interfere with the use of contact lenses, AG sets of interaction, allowing long product stored in preocular region, and thereby provides long-lasting protection and hydration of the corneal surface.

Studies of the interaction of contact lenses with polymer

To assess the degree of interaction of contact lenses with the test polymer was obtained fluorescent derivative of arabinogalactan (FITC-AG), as described below.

Accurately weighed quantity of arabinogalactan is a (FiberAid® AG, Larex®), equal to 1 g, was dissolved in 10 ml of dimethyl sulfoxide containing a few drops of pyridine. To this solution was added 0.1 g of fluorescein isothiocyanate (Sigma-Aldrich, Germany), and then 20 mg of dibutyltindilaurate (Sigma-Aldrich, Germany). Thus obtained mixture was heated for 2 hours to 95°C. After precipitation and washing with ethanol to remove unreacted products, FITC-AG was filtered and dried at 80°C (A.N. de Belder & K. Granath, 1973, Carbohydrate Research, 30:375-378).

For the study was prepared by following the solutions in deionized water:

1. 5% wt./wt. FITC-AG;

2. Fluorescein sodium (SF) (Sigma-Aldrich, St. Louis, USA) 0,0223% wt./wt.

The SF concentration was chosen such that it had the same fluorescence, and 5% wt./wt. a solution of FITC-AG. For studies used soft contact lenses daily wear Focus Dailies®, Ciba Vision, Germany).

Contact lenses were immersed in 1 ml of artificial tears without proteins, to which was added 50 μl of 5% wt./wt. solution of FITC-AG or 0,0223% wt./wt. SF and was left alone for 30 minutes. Then the lens was washed by immersing them in 100 ml of artificial tears for 15 minutes with slow stirring with a magnetic stirrer. The treated lenses were examined with a wood's lamp with a wavelength of 365 nm and compared with untreated contact lenses as standard.

None of the lenses was not found, ujena visible traces of fluorescence - this proves that the polymer AG is not retained on the surface of the lens when used conditions.

Artificial tear had the following composition, expressed in mg/100 ml of deionized water: MgCl2- 4,75; CaCl27,97; KHCO3- 260,00; NaCl - 754,00 (Burgalassi et al., 1999).

Test ferning

The innermost layer of the tear fluid consists of mucous glycoproteins, which in normal conditions are secreted goblet cells of the conjunctiva. One of the most important physical features of this mucus is the ability to form after drying at room temperature the crystals, resembling a fern leaf.

Various aspects of the crystallization of mucus, tears can serve as useful indicators of conditions of stability of the tear film, and these aspects are divided into four types:

Type I: figure "fern" homogeneous between an individual "fern leaves no gaps. "Fern leaves" large, tightly spaced branches.

Type II: "fern leaves" still a lot, but some of the leaves are smaller and not so widely branched. Between the "leaves" are notable gaps.

Type III: partial ferning: "fern leaves" small and weakly branched; between the "leaves" there are significant gaps.

Type IV: the ferning is missing, there are threads Il the conglomerates, representing degenerative slimy material, mixed with desquamated cells.

The first two types of ferning typical for normal eyes with the good condition of the mucous layer and the tear film. Type III appears to be a transitional form and indicates the difficulty of maintaining the integrity and function of mucus. Type IV indicates marked changes of the components of tears.

All eye solutions, which crystallized in the form of fern leaves, comparable with types I and II are structurally similar to the mucous glycoproteins produced by goblet cells of the conjunctiva.

Thus, the test ferning was performed to evaluate the potential ability of AG to crystallize with characteristics similar to those of mucus present on the surface of the eye. The test consisted of drying at room temperature (25±1 °C) for 24 h on a glass slide previously obtained mixture, which consisted of 10 μl of 2.5% wt./wt. solution AG and 2 μl of artificial tears.

a 2.5%solution of AG was obtained by dispersion under stirring for a suitable amount of polymer (FiberAid® AG, Larex®) in deionized water. The residue after drying was studied using polarizing microscope Reichert-Jung Microstar with 10-fold magnification.

In the crystallization solution of AG was formed like PA is erotico branched structure, very similar to the structures obtained from the lacrimal fluid of humans. The result confirmed by tests on rheological interaction, supports the hypothesis that the investigated polymer compatible with glycoprotein structures of eye mucus. Therefore, we can assume that AG can replace natural mucous component in lack of the latter due to pathological reasons.

Biological tests

1. Time to save on the surface of the cornea

This study used a fluorescent derivative of arabinogalactan (FITC-AG), obtained as described above in the section on interaction studies contact lenses with the polymer.

Quantification of FITC-AG in biological samples was performed by fluorometric analysis. The equipment consisted of a fluorescence detector Shimadzu RF-551 with the appropriate software. Detection was at a wavelength of excitation 490 nm and the wavelength of emission 514 nm.

The test was performed on new Zealand rabbits, albino animals with a weight of 2-2,5 kg In the lower conjunctival SAC of rabbits were injected with 50 µl of the aqueous composition 5% wt./wt. FITC-AG. At suitable time intervals after instillation(1, 3, 5, 10, 20, 30, 45 and 60 min) were sampled tears (1,0 μl) at the edge of the lower conjunctival IU the spacecraft through microcapillary (Microcaps, Drummond Scientific, PCs, new Jersey, USA), avoiding any contact with the corneal epithelium. Sample tears transferred in Eppendorf tubes and then investigated by fluorometric analysis after diluting 50 ál of water.

In the accompanying Fig. 2 shows the percentage attenuation of the fluorescence FITC-AG in the tear fluid over time; 100% accepted fluorescence of the product after 1 min, the Fluorescence was always high after 10 min after injection, decreased after 20 min and was still measurable after 60 min, which indicates the long-term preservation of AG in precorneal area.

2. Induction and treatment of dry eye syndrome"

This study used a solution of arabinogalactan (FiberAid® AG, Larex®) in water (hereinafter referred to as AG-Sol with concentration of AG 5% wt./wt., which was made by adding as a result of isotopically appropriate number mannitol (to 4.41% wt./wt.) and had a pH 6,46.

To obtain the composition of the AG-Sol precisely weighed polymer was dispersible in deionized water (Milli-Q, Millipore), and thus obtained solution was heated up to 80 °C for 30 minutes under stirring with a magnetic stirrer. After cooling, was added mannitol.

Thus obtained solution was filtered in a fume hood laminar flow of air through a sterile filter (Sartorius Minisart) with a pore size of 0.22 μm was Packed in a glass LAF is ons.

To assess the effect of arabinogalactan on lacrimation, in cases of damage to the epithelium of the cornea, caused by the syndrome of "dry eye", performed the experiment in vivo.

The tests were performed using new Zealand rabbits, albino with a weight of 2-2,5 kg

In both eyes of the animals 3 times a day (at 9:00, 13:00 and 17:00), 5 days in a row, put one drop of 1.0% aqueous solution of atropine sulphate (AS), to reduce the formation of tears and cause "dry eye" (S. Burgalassi et al., 1999, Ophthalm. Res., 31, 229-235). After 5 min after each injection AS right in the eyes of the animals were instillirovti 50 μl of the composition AG-Sol.

Animals were subjected to the test Schirmer I on day 0 (before treatment) and at 2-nd, 3-rd, 4-th and 5-th day after the start of treatment. The test involved measuring the amount Sekretareva tears once a day. The determination was performed before each infusion (AS and AG-Sol) by means of strips of filter paper (Alfa Intes), which was placed in the lower fornix of the conjunctiva of each eye of the rabbit. The degree of lifting of tears along the strip of filter paper was measured in mm the Values obtained for the eyes treated only with atropine sulphate was compared with that obtained for the eyes treated with a solution of arabinogalactan according to the invention.

The initial value of the secretion of tears was measured in a group of control animals that did not receive any pharmacological treatment.

p> The results obtained are shown graphically in Fig. 3 of the accompanying drawings. The figure shows that the solution AG-Sol protects against the development of dry eye.

In addition, in the 3rd, 4th and 5th days after the start of treatment, the degree of damage to the epithelium of the cornea, caused by dry eye, was estimated using examination with a slit lamp equipped with a blue color filter, after staining of the cornea 10 ál of water with 1% wt./wt. solution fluorescein. In case of damage of the epithelium on the surface of the cornea was observed fluorescent region.

The results are shown below in table 4, as the percentage of eyes with the fluorescence, of the total number of treated eyes. The same results are depicted in the histogram in the accompanying Fig. 4, which graphically compares the percentage of eyes with fluorescence after treatment with a solution of AG-Sol, in relation to the percentage of eyes with fluorescence, control animals that received only AS.

TABLE 4
The results obtained by examination with a slit lamp
No.Day 3Day 4 Day 5
DX (without treatment)SXDX (without treatment)SXDX (without treatment)SX
1-+----
2-----+
3---+--
4-+-+++
5---+--
6-+/td> -+--
7---+-+
8+---++
9--++-+
10-----+
11-+---+
12--+---
amount142627
the presence of fluorescence, %8,3333,3316,6750,0016,67with 58.33

According to the results shown in table 4 and in Fig. 4, one can notice that in untreated animals the percentage of eyes with fluorescence, increased day by day, and by the 5th day has reached almost 60%, while in the animals treated with a solution of AG-Sol, the percentage of eyes with fluorescence, between the 4th and 5th day of treatment remained constant, and it is much lower - about 17%.

3. Evaluation of ability to recover epithelium

The test, described below, allows us to estimate the time of repairing the damage of the cornea of the rabbit after the introduction of the following songs:

1. The aqueous solution AG-Sol (5% AG, to 4.41% mannitol);

2. Polysaccharide TS (TSP), a 0.04% aqueous solution;

3. Hyaluronic acid (HA), 0,00144% aqueous solution.

It should be noted that the purpose of the products according to the invention is the creation of deputies of the lacrimal fluid, not as viscous as artificial since the and of the prior art, based on polysaccharides so that they are most applicable for people using contact lenses, so the concentration of standard solutions 2) and 3)containing the TSP and HA, was selected to create solutions with the same rheological behavior (i.e. Newtonian) and the same viscosity as the solution AG-Sol.

The test was performed on new Zealand rabbits, albino animals with a weight of 2-2,5 kg Rabbits were anestesiologi intramuscular injection (0.15 ml/kg Zoletil 100®(Laboratories Virdac, France), and the eye kept open using blueparrott, and anestesiologi its surface, installarea 10 μl of oxybuprocaine hydrochloride (Novesina®, MiPharm, Italy).

To cause the defeat of the cornea, paper microbiological disc diameter of 6 mm, impregnated with 10 µl of n-heptanol, was placed in the Central region of the cornea for 60 seconds. After you remove the disk surface of the eye thoroughly washed with 1 ml of physiological solution (Burgalassi et al., 2000, Eur. J Ophthalmol., 10, 71-76).

At time 0 (immediately after removal of the impregnated paper and washing) and through 3, 7, 18, 24, 27, 29, 31, 34, 41, 44, 48 and 51 hours after creating the defeat of the eye surface were stained with 10 µl of water with 1% wt./wt. solution fluorescein sodium, and the diameter of the lesion was measured using a special micrometer.

Treated animals received 100 μl investigated what's the solution in the affected eye 5 times per day (at 9:00, 11:00, 13:00, 15:00 and 17:00).

In one group of animals with the lesion of the cornea is allowed to heal on its own; this group was used as control.

In Fig. 5 of the accompanying drawings graphically depicts the trend corneal healing in groups of animals depending on time. Data are presented as the percent reduction of the area affected, where 100% is assumed size at time 0. The same information is shown numerically in table 5 below.

0
TABLE 5
Time, hControl (area, %)Standard errorAG-Sol (area, %)Standard errorTSP (area,%)Standard errorHA (area,%)Standard error
01000100010001000
3100010010001000
787,19,1472,3582,91879,1673,07691,0714,593
18554,144,44061,0335,07559,2937,591
2431,624,3529,2363,06531,8592,11741,863of 3.64
2726,91of 1.5718,8391,83323,6271,63730,952,183
2925,2054,545 15,8381,21115,5082,27320,1213,362
3116,29of 4.387113,6572,68814,3842,101
3411,724,283,812at 1,0477,0981,7827,2131,249
417,122,098of 1.0620,9812,5681,3365,0832,663
445,22,30,6950,6950,8170,6532,8471,663
483,7 3,70,00100,170,170,1110,099
5100000000

On the above table and Fig. 5 you can see that starting from 27 hours after administration of the compositions, only the composition of the present invention, the AG-Sol, shows the rate of recovery, significantly different from control values. In fact, AG-Sol demonstrates statistically significant differences through 27, 29, 31, 34 and 41 hours after the start of treatment, and for other compositions the difference is statistically significant only for the TSP after 29 hours.

4. Evaluation of the healing epithelium ability in the presence of benzalkonium chloride

In the test evaluated the healing time of lesions of the cornea of the rabbit after the introduction of the following songs:

1). The aqueous solution AG-Sol (5% AG, to 4.41% mannitol).

2). The aqueous solution AG-Bz (5% AG, 4,00% mannitol, 0.01% benzalkonium chloride).

The test was performed on new Zealand rabbits, albino animals with a weight of 2-2,5 kg Krolik who was anestesiologi intramuscular injection (0.15 ml/kg Zoletil 100 ®(Laboratories Virdac, France), and the eye kept open using blueparrott, and anestesiologi surface, installarea 10 μl of oxybuprocaine hydrochloride (Novesina®, MiPharm, Italy).

To cause the defeat of the cornea, paper microbiological disc diameter of 6 mm, impregnated with 10 µl of n-heptanol, was placed in the Central region of the cornea for 60 seconds. After you remove the disk surface of the eye thoroughly washed with 1 ml of physiological solution (Burgalassi et al., 2000, Eur. J Ophthalmol., 10, 71-76).

At time 0 (immediately after removal of the impregnated paper and washing) and through 3, 7, 18, 24, 27, 29, 31, 34, 41, 44, 48 and 51 hours after creating the defeat of the eye surface were stained with 10 µl of water with 1% wt./wt. solution fluorescein sodium, and the diameter of the lesion was measured using a special micrometer.

Treated animals were injected in the damaged eye 100 ál of the investigated solutions 5 times a day (in 9:00, 11:00, 13:00, 15:00 and 17:00).

In one group of animals with the lesion of the cornea is allowed to heal on its own; this group was used as control.

In Fig. 6 and 6a, with varying degrees of accuracy graphically shows the trend of the healing of lesions of the cornea in the groups of animals over time. Data are presented as the percent reduction of the area affected, where 100% is assumed size at time 0.

You may notice that on ichie of benzalkonium chloride modifies the rate of recovery of AG, lowering her complete healing in animals treated with the composition of AG-Bz occurred after 51 hours, while in the animals treated with AG-Sol, healing occurred after 44 hours. That is why the use of both products composition, aimed at protecting the cornea, it is impractical.

Histological evaluation of the treated corneas

To estimate, did the recovery process of the epithelium in addition to providing complete healing of experimentally induced lesions also to the correct stratification of epithelial cells with restoration of the natural structure of the cornea, performed the histological examination.

The biopsy of cornea, taken 24 hours after the creation of lesion of cornea after complete healing (when fluorescein was no longer detected in the epithelium) and 1 week after healing, were fixed in a 10% solution of paraformaldehyde in 0.1 M phosphate buffer with a pH of 7.4. Then the samples were subjected to several washes with buffer for 12 hours, obezvozhivani in battery solutions of ethanol of increasing concentration and placed at 4°C in a special resin for light microscopy (JB-4, Embedding kit, Polysciences Inc.). Drenched in resin fabric cut by microtome and stained by Nissly, and then were subjected to microscopy.

24 hours after the creation of the defeat became obvious signs of physiological the systematic reduction mechanism in the form of displacement and migration of adjacent integral epithelial cells in the damaged area with the formation of a monolayer. The epithelium was restored natural thickness (55-60 μm in the Central area of the cornea) and layered structure only after complete healing, when physiological mechanism desquamation of cells was restored to full coverage minus the area and the regeneration of epithelial elements provided recovery of normal structural characteristics. The condition remained unchanged after a week of healing - this indicates that the regeneration process is complete.

The treatment produced after the creation of lesion of cornea, made the same products reported in the previous section 3, devoted to biological tests, has led to a reduction in healing time of lesions in different degrees depending on the product. This led to the assumption that the different impact of the used polymers on the mechanism of reparation. Therefore, it became necessary to investigate the recovery phase of the epithelium to identify any differences in the morphological aspect of the fabric.

With this purpose, the method of assessing the recovery of the epithelium (section 3, biological tests) was repeated, creating the defeat and making treatment of the polymer compositions described above. Animals were scored before complete healing of lesions of the cornea, that is, when the equipment used could not measure the defeat, what about the delay fluorescein on the surface of the eye was still determined visually.

The biopsy of cornea receiving such treatment, were subjected to histological examination (procedure was the same as described above), and in each case was measured thickness recovered epithelium.

The measurement is performed each time at the same distance from the edge of the resumption of growth, and focus on the differences in the thickness of the epithelium, converted after various influences. In fact, the results were as follows:

- untreated control (self-healing) - 6 ám;

the sample treated with AG-Sol (5% AG, to 4.41% mannitol) - 18 microns;

the sample treated with 0.5% solution of polysaccharide TS - 18 microns;

the sample treated with 0.2% solution of hyaluronic acid - 12 microns.

These results show that each of the observed cases, the epithelium, is ready to recreate the defective area, had no physiological thickness, although in samples treated with the compositions of the AG-Sol and polysaccharide TS, the thickness of the epithelium was much higher than its thickness in the samples that were subjected to self-healing.

Monitoring of relevant images (not shown) also revealed morphological differences between samples of the cornea. In fact, there are significant differences in the layout of the new epithelium in the four studied cases.

The sample obtained from the control eye, demonstriruya very well-organized structure, the stratification is weak or not evident.

- Epithelium treated with a composition of AG-Sol showed a great similarity with the natural epithelium in terms of tissue structures.

The sample treated with polysaccharide TS contained poorly stratified newly formed tissue, despite the fact that the thickness of the epithelium was the same as the thickness of the epithelium in the sample treated with AG-Sol; in fact often found a few large cells, much larger than normal.

Finally, the HA treatment resulted in discrete layering of the new epithelium, even if the final obtained thickness was less.

These results make us conclude that the higher the rate of healing of lesions of the cornea after treatment with the composition comprising AG, caused by stimulation of regeneration of epithelial elements, which is caused by this product. Higher ability of epithelial cells to replication leads to early layering fabric, which thus allows you to control a greater number of cells to cover the defective area.

Studies of cytotoxicity

Evaluated the cytotoxicity of solutions containing AG separately or together with benzalkonium chloride (Bz). The original polymer solutions were prepared (1. 5% wt./wt. AG; 2. 5% wt./wt. AG + is 0.01% wt./wt. Bz) directly in modified And lsom environment Dulbecco (DMEM, Sigma Chemical Co., St. Louis, Missouri, USA). These solutions were then diluted 1:2, 1:5, 1:10, 1:100 and 1:1000, still using DMEM.

Used a cell line consisting of epithelial cells of the cornea of the rabbit (transformed RCE SV40, N. 95081046, ECACC, .g. b). Line RCE was created by infection of primary cultures of epithelial cells of the cornea of the rabbit recombinant retrovirus SV-40. Cells had a typical "cobblestone" morphology of the epithelium and the ability to layering, as well as the development of desmosomes and microvilli.

Nutrient medium for the cells was modified by Epsom environment Dulbecco (DMEM, Sigma Chemical Co., St. Louis, Missouri, USA) with the addition of a nutrient mixture Ham F12 (1:1), L-glutamine (1% vol./about., 2 mm), penicillin (100 IU/ml), streptomycin (0.1 mg/ml), amphotericin B (0.25 microgram/ml), inactivated by high temperatures serum bovine embryos (15% vol./about.) (Gibco, UK), insulin (5 μg/ml), epidermal growth factor (10 ng/ml) (Sigma Chemical Co., St. Louis, Missouri, USA). Cells were grown at 37°C in a saturated water vapor atmosphere containing 5% CO2.

Assessment of the degree of toxicity of the solutions tested on the RCE cells, was performed by colorimetric method using a commercially available reagent for assessing cell proliferation WST-1 (Roche Diagnostics® S.p.A., Monza). The test is based on the splitting due to the activity of mitochondria salt tetrazole the WST-1 to soluble colored compounds (formazan). Because this transformation can only be made by living cells, the number of obtained formazan directly proportional to the number of living cells.

The RCE cells were placed on 96-well plates (Coming Costar® Italia, Milan) 5·103cells per well. After 24 hours at 90% confluence culture medium was removed and replaced with test solution (100 μl solution). After 60 minutes of contact (at 37 °C in humidified atmosphere containing 5% CO2) reaction medium was removed, and cells were twice washed in DMEM F12; then to each well was added 100 μl of fresh medium and 10 μl of reagent WST-1. Cells were incubated for another 2 hours at 37 °C in humidified atmosphere containing 5% CO2), and the tablet was carefully shaken for 9 seconds; spectral absorption capacity of the environment was measured at 450 nm using a suitable spectrophotometer (Microtiter reader 550®Bio-Rad Laboratories, Hercules, California). The optimal time of incubation with the reagent WST-1 was identified in a number of preliminary experiments.

The spectrophotometer readings were determined by comparison with the "idle" well, containing only a mixture of medium and WST-1 (100 and 10 μl, respectively) without cells. The results were expressed as the percentage of absorbance of treated wells (Abstr) relative to untreated wells (control, Abswith), which contained cells treated only environment without the pharmaceutical product according to the following formula:

The results are depicted graphically in Fig. 7, in which cell viability is expressed as percent with respect to the logarithm of the concentration of AG, when the cells were treated with solutions containing only AG, or a mixture of AG + Bz. All tested concentrations of AG showed no cytotoxicity. Adding Bz, especially in higher concentrations caused an increase in toxicity. Thus, the presence of benzalkonium chloride leads to significant cytotoxicity and, in addition, as shown in experiments in vivo, reduces regenerative epithelium activity of AG.

The present invention has been disclosed with respect to certain specific options, but you need to understand that the specialist in the art may make changes or modifications without departing from the scope of the attached claims.

1. Ophthalmic composition for treating keratoconjunctivitis damage and inflammation that contains:
- an aqueous solution comprising from 1 to 10 wt.% arabinogalactan,
one or more preservatives selected from the group consisting of thimerosal, sodium, thimerosal, the mercury-phenyl nitrate or mercury-phenyl acetate, phenethyl alcohol, m is Teal, -, ethyl-, propyl paraben, chlorhexidine acetate or gluconate or chlorbutanol and does not include benzalkonium chloride.

2. Ophthalmic composition according to claim 1, containing from 3 to 5 wt.% arabinogalactan in aqueous solution.

3. Ophthalmic composition according to claim 1 or 2, where the said arabinogalactan is arabinogalactan larch related to the pharmaceutically acceptable category.

4. Ophthalmic composition according to claim 1, additionally containing one or more funds, regulatory toychest.

5. Ophthalmic composition according to claim 4 where the above-mentioned one or more funds, regulatory toychest, are present in aqueous solution in such an amount to make the solution osmolarity of 150 to 300 mOsm/L.

6. Ophthalmic composition according to claim 5 where the above-mentioned one or more funds, regulatory toychest, selected from the group consisting of mannitol, sodium chloride, potassium chloride, dextrose, boric acid and sorbitol.

7. Ophthalmic composition according to claim 1, additionally containing one or more pharmaceutically acceptable acid or base as a pH corrector.

8. Ophthalmic composition according to claim 1, additionally containing one or more buffers.

9. Ophthalmic composition of claim 8 where the above-mentioned buffers selected from the group consisting of phosphate buffer, borate buffer, citrate buffer, bicarbonate buffer, Tris buffer (trihydroxystilbene Metan).

10. Ophthalmic composition according to claim 1, additionally containing one or more complex-forming means.

11. Ophthalmic composition of claim 10 where the above-mentioned complexing agent is EDTA.

12. The use of ophthalmic composition for the treatment keratoconjunctivitis damage and inflammation, containing an aqueous solution comprising from 1 to 10 wt.% arabinogalactan, one or more preservatives selected from the group consisting of thimerosal, sodium, thimerosal, the mercury-phenyl nitrate or mercury-phenyl acetate, phenethyl alcohol, methyl-, ethyl-, propyl paraben, chlorhexidine acetate or gluconate or chlorbutanol and does not include benzalkonium chloride.

13. The application of section 12 where the above-mentioned ophthalmic composition contains from 3 to 5 wt.% arabinogalactan.

14. Use on 12 or 13 where the above-mentioned ophthalmic composition is an artificial tear, recommended for people using contact lenses.

15. The application of section 12 where the above-mentioned ophthalmic composition is an artificial tears for the treatment of kerato-conjunctival lesions and inflammations.

16. The application of clause 15 where the above-mentioned artificial tear is intended for the treatment of corneal erosion caused by the use of contact lenses.



 

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13 cl, 3 tbl, 3 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: group of inventions relates to veterinary. Self-emulsifying veterinary antihelminth composition, contains: a) from approximately 15 to approximately 25% wt/vol. benzimidazole antihelminth medication; and (b) water-immiscible system of solvents, which contains lactone solvent in quantity from approximately 10% to approximately 40% wt/vol, ether oil in quantity from approximately 10 to approximately 35 wt/vol. and surface-active substance in quantity from approximately 30% to approximately 60% wt/vol. Self-emulsifying veterinary antihelminth composition, contains: a) from approximately 15% to approximately 25% wt/vol. triclabendazole; and b) water-immiscible system of solvents, which contains γ-hexalactone, 1,8-cineole and glycerides of polyethyleneglycol caprylic/capric acid. Methods of treating warm-blooded animals from parasites, which represent helminthes, include introduction one of claimed self-emulsifying veterinary antihelminth compositions to said animal. Method of preparing self-emulsifying veterinary antihelminth composition includes mixing (1) benzimidazole antihelminth medication with (2) water-immiscible system of solvents, which contains lactone solvent, ether oil and surface-active substance.

EFFECT: group of inventions ensures increased efficiency of animal treatment.

33 cl, 8 tbl, 8 ex, 4 dwg

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to medicine and pharmaceutical industry, particularly to a galenic form for transmucosal-buccal administration of at least one active substance of the family of triptanes. The form contains at least: the above active substance in the form of a base and in the form of a salt, and an aqueous-alcoholic solution of the concentration of at least 15 degrees. Said active substance is found in the state of stable and complete dissolution in the aqueous-alcoholic solution to enable rapid absorption of said active substance through the oral and/or pharyngeal mucosa.

EFFECT: invention also refers to a method for preparing said galenic composition, as well as to using it.

16 cl, 3 tbl, 9 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: claimed is liquid composition of elsamitrucin for treatment of tumorour diseases or conditions, including: a) at least one stable salt of elsamitrucin, which represents product of free elsamitrucin base and organic acid and can exist as stable solid substance and does not include pseudo-salts or salts, obtained in situ, which exist only in solution, and b) pharmaceutically acceptable carrier, and claimed liquid composition does not contain antioxidants or methanol, ethanol, chloroform, n-butanol or t-butanol as admixtures, and claimed stable salt of elsamitrucin preserves at least 90% of its anti-tumor activity within 18 months in liquid form at appropriate storage temperature.

EFFECT: it is shown, that compositions, namely disclosed in claimed invention water solutions of true elsamitrucin salts, can be preserved and introduced without reduction before application.

16 cl, 7 dwg, 7 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention relates to medicine and concerns irrigation solution, which is applied for endoscopic examination and surgical operations in urology and gynecology. Irrigation solution contains water solution of glycine and antiseptic, where as antiseptic introduced is nipagin (methylparaben) with the following ratio of component composition, wt %: glycine 1.0-3.0%; nipagin (Methylparaben) 0.01-0.3%; purified water.

EFFECT: invention ensures reduction of risk of postoperative purulent-septic complications, sanifying influence on urinary bladder during operation, both bactericidal and mechanical, extension of field of irrigation solution application in washing systems in early postoperative period, and therapeutic-diagnostic manipulations in urology and gynecology.

1 cl

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention relates to field of medicine and veterinary. Stable pharmaceutical composition for injections, contains water solution of 2-ethyl-6-methyl-3-oxypyridine succinate and auxiliary substance, and as auxiliary substance carbonic acid is applied, composition is obtained by mixing with the following component ratio (wt %): 2-ethyl-6-methyl-3-oxypyridine succinate 1-10; carbonic acid 0.01-0.5; water for injection to 100. Method of composition obtaining by i.1 is characterised by the fact, that filling and sealing ampoules on syringe filling machine is carried out in steam of carbonic acid, and sterilising filtration of 2-ethyl-6-methyl-3-oxypyridine succinate solution before filling is carried out under pressure of carbon dioxide.

EFFECT: pharmaceutical composition does not possess allergenic properties and is highly stable during entire normative storage term.

5 cl, 4 tbl, 8 ex

Disinfectant // 2490008

FIELD: medicine.

SUBSTANCE: disinfectant refers to veterinary medicine, particularly to disinfectants used for air sanitation and surface disinfection in livestock houses in the presence of animals, including poultry. The agent has an antiseptic effect, which allows using it in acute, chronic and sub-clinical mastitis, vaginitis, endometritis and other inflammatory processes of varying aetiology. An active ingredient is presented by a complex of dimethyl sulphoxide and crystalline iodine, succinic acid, fumaric and citric acids which provides better drug absorption and prolonged action. It is prepared in the form of mother liquor diluted in water immediately before use.

EFFECT: invention enables prolonging the shelf life when in use.

7 tbl

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to pharmacy, medicine and veterinary science, particularly new drugs for inflammatory and allergic diseases. The assigned problem preparing a non-toxic, easy-to-use and storage-stable drug in the form of eye drops, intranasal drops and spray and causing anti-inflammatory and antiallergic action is solved by creating a composition containing a pigment-mineral complex of sea-urchin shell (0.05-2.0%) containing spinochromes B, D, dimer polyhydroxynaphthoquinone and a mineral ingredient (calcium, magnesium, phosphor, sodium, potassium) in the form of water-soluble salts, a co-solvent (0.01-10%), a preserving agent (0.01-0.2%), an antioxidant (0.01-0.2%), an acidity regulator (to pH 6.0-8.0) and water (to 100%).

EFFECT: what is described is the experimentally specified method for preparing the agent according to the invention eliminating the oxidation of the pigment-mineral complex as early as the stage of preparing.

3 cl, 3 tbl, 1 dwg, 6 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: group of inventions refers to a liquid oral solution for treating angiotensin II mediated disorders and conditions, and to a method for preparing it. The liquid oral solution contains valsartan in the concentration of 5 mg/ml or less, a wetting agent, a preserving agent, a flavouring agent, a buffer system and water with pH of the solution making 4.5-5.9. The wetting agent is poloxamer 188 described by the structure HO(CH2CH2O)a(CH(CH3)CH2OH)b(CH2CH2O)cH, wherein a is equal to 75, b is equal to 30, and c is equal to 75, with average molecular weight 8350. The buffer system contains alkali citrates and citric acid, alkali acetates and acetic acid, alkali succinates and succinic acid, or any mixtures thereof. The method for preparing the above solution involves mixing the ingredients added then with valsartan when heated.

EFFECT: group of inventions provides preparing the liquid solution of valsartan with an extended storage period.

8 cl, 12 tbl, 1 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to chemical-pharmaceutical industry and represents a composite enterosorbent of a silicone polymer specified in a group containing methyl monosilane acid xerogel or methyl monosilane acid hydrogel, differing by the fact that contains at least one ingredient specified in a group: lactulose, inulin, lignin, fructooligosaccharide, alginic acid in the form of pharmaceutically acceptable salts, chitosan, pectin, gum resin, beta-glucan in the amount of 0.1 to 10 portions per 1 weight portion of monosilane acid hydrogel or xerogel.

EFFECT: invention provides creating an agent to provide normalising the intestinal microflora and relieving the manifesting intoxication.

3 cl, 6 ex

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