Ophthalmic solution for uv-a rays protection of internal structures of eyeball or for treating keratoconus by transepithelial crosslinking

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to the pharmaceutical industry and represents an ophthalmic solution containing riboflavin and vitamin E for the UV-A rays protection of the internal structures of an eyeball and for treating keratoconus by transepithelial crosslinking.

EFFECT: invention provides extending the range of products for the UV-A rays protection of the internal structures of the eyeball and for treating keratoconus by transepithelial crosslinking.

9 cl, 10 dwg

 

The technical FIELD TO WHICH the INVENTION RELATES.

The present invention generally relates to compositions for the treatment and methods of treatment of keratoconus and, more specifically, to a new ophthalmic solution, applicable for the protection of the internal structures of the eyeball from UV-a rays or treatment with corneal cross-linking.

The LEVEL of TECHNOLOGY

In the book [18] provides an overview describing the problems and methods of introduction and assimilation of ophthalmic solutions.

Corneal (collagen cross-linking (C3) using Riboflavin (vitamin B2), briefly called Riboflavin - C3, is an innovative method of treatment of patients with keratoconus and ectasia of the cornea and consists in the introduction of Riboflavin and irradiated with ultraviolet light (UV-a) to strengthen corneal tissue[1], [2].

The treatment method of cross-linking is relatively simple: Riboflavin pour in drop by drop in the eye (irrigate the eye) and the cornea is irradiated with an appropriate dose of UV-a for five minutes; then repeat the procedure six times in a row to the total time of UV-irradiation was 30 minutes.

The most important clinical indicator, which should be taken into account when determining the suitability for cross-linking, is the thickness of the cornea, which should not be less than 400 microns.

The aim is that conservative treatment of keratoconus is to delay or hopefully, eliminating the need for transplantation (transplantation) of the cornea and improve the visual performance of patients, improving their quality of life[6], [7].

The method of cross-linking was used for the treatment of keratoconus, pathology, characterized by progressive thinning of the cornea due to abnormal sagging of the parenchyma of cornea due to the low adhesion of the collagen fibers of which it is composed. When using UV-a radiation and Riboflavin between adjacent collagen molecules of the cornea formed new relationships and processed the cornea becomes thicker and stronger [3]. The cornea contains several layers of collagen fibres, the thickness of parenchyma; cross connection, the so-called "matching", which connect between different layers of collagen, which are crucial for the rigidity of the cornea. The purpose of the cross-linking of corneal tissue is to increase the degree of hardness due to the formation of a larger number of these cross-linking.

Local (topical) application (application) of Riboflavin on decapitalizing the cornea with a permeability of about 200 microns and UV-a-irradiation of Riboflavin molecules determines the chemical imbalance of the Riboflavin molecules with the subsequent formation of free radicals. Molecule of Riboflavin become unstable and stabiliser who are by linking two of the collagen fibrils. Between the collagen fibrils are formed a number of biochemical "bridges" (i.e., crosslinking), which leads to an overall strengthening of the corneal tissue [3].

In fact, the treatment is carried out after removal of the outer layer of the cornea (i.e., the corneal epithelium, corneal epithelium). This way of implementation of the cross-linking (C3-R) keratoconus and ectasia of the cornea assumes that the removal of the corneal epithelium to facilitate the penetration of 0.1% of a standard solution of Riboflavin-dextran (e.g., solution, manufactured by SOOFT ITALIA S. r.l. under the trademark RICROLIN™) underneath the stroma, and the standards of this treatment to normalize these conditions. According to supporters of this method, removal of the epithelial layer is necessary to ensure (guarantee) the best adsorption of Riboflavin solution inside the corneal stroma and thereby the maximum efficacy of the treatment.

Unfortunately, removal of the corneal epithelium can cause itching and burning in the eyes the next day after treatment and in the following days and temporary blurred image; these symptoms are well known and are saved to restore the corneal epithelium and are usually treated in the days after C3-R [4-7] eye drops containing non-steroidal anti-inflammatory drugs (NSPs, NSAID) eye drops based substitutes natural SL is C and analgesics and the use of therapeutic contact lenses.

Some authors argue that it is possible to carry out treatment C3-R according to the standard procedure without removal of the corneal epithelium and that such treatment would be effective and safe, as shown by the obtained clinical results. According to this method, the treatment should be carried out without the removal of the corneal epithelium (decapitalization). The goal is to avoid the characteristic of the first method of pain in patients caused by the removal of the epithelium, to carry out treatment in the outpatient setting and, in particular, to avoid the risk of postoperative infections characteristic of the treatment, which involves the removal of the epithelium with subsequent exposure of the lower layers of the cornea. Proponents of this mode of carrying out the treatment is offered to enter the Riboflavin in the eye (to irrigate the eye for a longer period of time to ensure the best adsorption of Riboflavin into the stroma before UV-a-irradiation [8].

As to whether to delete or not the epithelium for the treatment of keratoconus and ectasia of the cornea by cross-linking, in the literature there are contradictory opinions.

C3-R treatment was studied and carried out after removal of the corneal epithelium to facilitate the penetration of Riboflavin into the corneal stroma. According to the authors of the information in the literature there is no information about the research on ODA the division, gets whether the Riboflavin and the extent to which penetrates the corneal stroma [9] with or without excision of the corneal epithelium.

The implementation of the cross-linking without prior decapitalization criticized by many authors, who claim that in this way, Riboflavin does not pass through the epithelium and that while not shown, it effectively penetrate or not penetrate, and if it penetrates, how, standard 0.1% solution of Riboflavin - dextran in the stroma of the cornea without removing the epithelium, and whether effective UV-a irradiation in transepithelial the way and after removal of the corneal epithelium.

After attempts to find an effective substance for the treatment of keratoconus by cross-linking without removal of the corneal epithelium Dr. Sporl proposed [17] to use benzalkonium chloride to increase the permeability of the epithelium, and Dr. Pinelli proposed to use a mixture of surfactants with Riboflavin.

In the Italian patent application No. MI2007A002162 [16] revealed a new solution for the treatment of keratoconus method transepithelial cross-linking, containing Riboflavin and benzalkonium chloride.

Experiments conducted by the applicants on the cornea of a person, the results of which are shown below in this description, lead to the conclusion that the second method, implemented using a standard solution is whether the composition, Dr. Pinelli [16], it is not possible to overcome the difficulties caused by the removal of the corneal epithelium, as it may break down, this increases the risk of infection and changes in the mechanisms of reparation, which are opened (exposed) lower layers.

For the implementation of the cross-linking of the cornea, it would be desirable to have a composition comprising Riboflavin, which could penetrate the corneal epithelium in a relatively short time and which had not struck the corneal epithelium, which would avoid postoperative symptoms.

The INVENTION

Applicants conducted a thorough study to determine how much Riboflavin penetrates alone or in mixture with other products ("enhancers permeability) through the cornea of a person with a prior removal or without prior removal of the corneal epithelium, as well as the efficacy and safety of subsequent treatment using UV-a radiation.

Identified several candidate substances selected from the group consisting of essential and conditionally essential (such as arginine, cysteine, glycine, glutamine, histidine, Proline, series and tyrosine) amino acids, coenzyme Q, vitamin E, L-Proline, glycine, lysine hydrochloride, L-leucine, L-arginine and compounds, p is designated to stimulate the production of metalloproteinases MMR, examined in more detail later in this description, which can be effectively used as carriers ("permeability enhancers") in ophthalmic solutions, suitable for introduction Riboflavin, in particular, with the standard solution of Riboflavin-dextran through the epithelium of the cornea. Thus obtained ophthalmic solutions, which can be sold in the form of eye drops, or in the form of a gel or in the form of aqueous solutions or emulsions, or applied to, a therapeutic contact lens, can be used for the treatment of keratoconus method transepithelial cross-linking, thereby maintaining the corneal epithelium.

Ophthalmic solutions may ultimately contain excipients, such as acetic acid, or the above-mentioned substances before mixing with Riboflavin can be treated with acetic acid or other excipients.

In addition, the invention provides the use of at least one substance selected from the group consisting of essential and conditionally essential amino acids, coenzyme Q, vitamin E, L-Proline, glycine, lysine hydrochloride, L-leucine, L-arginine and compounds designed to stimulate the production of metalloproteinases MMR, discussed below in more detail herein, for the preparation of ophtalmologic the RCM solution, contains Riboflavin, designed to protect the internal structures of the eyeball from UV-a radiation or for the treatment of keratoconus method transepithelial cross-linking and using appropriate ophthalmic solution containing Riboflavin, as a carrier ("enhancer permeability") in compositions suitable for administration of Riboflavin through the corneal epithelium.

Further, the present invention provides a method of making ophthalmic solution, which consists in adding to a solution of Riboflavin at least one of the identified above media.

Each offer as a carrier substances can be added alone or in combination with other proposed media to a solution containing Riboflavin in a concentration selected within the range of concentrations indicated in the following description of typical variants of the invention.

Scope of the invention defined in the attached claims.

BRIEF DESCRIPTION of FIGURES

In Fig.1 presents a visual, fluorometrically, colorimetric assessment scale adopted for evaluating the passage of a 0.1% solution of Riboflavin through the cornea after application transepithelial way.

In Fig.2A presents fluoroscopically the image slice of the cornea that Tr is capitellum way put fourth test (test) composition (irrigate with it) after 15 minutes.

In Fig.2b presents fluoroscopically the image slice of the cornea that transepithelial way put fourth test (test) composition after 30 minutes.

In Fig.2c presents fluoroscopically the image slice of the cornea treated by the method of transepithelial cross-linking with the use of the fourth new solution, which after processing can be detected intense fluorescence due to the passage of the fourth composition and relevant stiffness of the fabric.

In Fig.3A shows the degree of curvature of the cornea treated transepithelial cross-linking with the use of a standard solution.

In Fig.3b shows the degree of curvature of the cornea treated transepithelial cross-linking with the use of the fourth new test solution.

In Fig.4 shows an image taken with a scanning electron microscope, on which you can see the leafs in the slice of the cornea affected by keratoconus.

In Fig.5 presents were taken with a scanning electron microscope magnified image of the cornea of Fig.4.

In Fig.6 shows the image obtained using a scanning microscope, on which you can see the leafs in the slice of the cornea affected by keratoconus, after transepithelial cross-linking carried out with the application of the fourth new the th test composition.

In Fig.7 shows the image obtained using a scanning electron microscope, showing the morphology of microvilli and superficial layers of the epithelium in healthy (normal) the cornea.

In Fig.8 shows an image taken with a scanning electron microscope, showing the cornea treated with the standard dose of UV-a-irradiation after transepithelial applying standard 0.1% solution of Riboflavin-dextran.

In Fig.9 shows the image taken with a scanning electron microscope, showing the cornea treated with the standard dose of UV-a-irradiation after transepithelial applying the fourth new test composition.

In Fig.10 shows an image taken with a scanning electron microscope, showing the cornea treated with the standard dose of UV-a-irradiation after transepithelial application of saline.

The IMPLEMENTATION of the INVENTION

All testing is carried out on the human cornea donors, obtained from the Azienda Ospedaliera Napoli 1 - Banca called OCCHI ("Eye Bank") - Regione Campania - Ospedale dei Pellegrini, after consent, as provided for in the Protocol of explantation and in accordance with the resolution of the ethics Committee - No. 0009304/2009 - Decision No.1269.

See penetration (penetration) of the tested compositions through the whole of the human cornea, i.e., be the removal of the epithelium, moreover, the cornea has a thickness of between 500 and 600 microns, the composition represent the standard solution Riboflavin-dextran, a composition prepared from Riboflavin-dextran plus benzalkonium chloride, proposed in [16] and [17], and new test (test) compositions obtained by blending Riboflavin at least one substance selected from the group comprising vitamin E, coenzyme Q, L-Proline, glycine, lysine hydrochloride, L-leucine, in certain concentrations.

The concentration of the substances used to create the (implementation) of new test compositions comprise the following intervals:

vitamin E: the range of concentrations is from 0.0001 mg % ml to 2000 mg % ml According to a more preferred variant of the invention intervals concentrations range from 0.01 mg % to 1500 mg % ml According to a further preferred variant of the invention intervals concentrations range from 10 mg % ml to 1000 mg % ml According to a further preferred variant of the invention a concentration equal to about 500 mg % ml;

vitamin Q: the concentration range is from 0.0001 mg % ml to 2000 mg % ml According to a more preferred variant of the invention intervals concentrations range from 0.01 mg % to 1500 mg % ml According to a further preferred variant of the invention, the intervals of concentrations ranging from 1 mg % ml to 1000 mg % ml With the public even more preferred variant of the invention the concentration is equal to, approximately, 100 mg % ml;

L-Proline: the concentration range is from 0.0001 mg % ml to 2000 mg % ml According to a more preferred variant of the invention intervals concentrations range from 0.001 mg % up to 100 mg % ml According to a further preferred variant of the invention intervals concentrations range from 0.005 mg % ml to 10 mg % ml According to a further preferred variant of the invention intervals concentrations range from 0.01 mg % ml to 1 mg % ml According to a further preferred variant of the invention the concentration is about 0.1 mg % ml;

glycine: the concentration range is from 0.0001 mg % ml to 2000 mg % ml According to a more preferred variant of the invention intervals concentrations range from 0.001 mg % up to 100 mg % ml According to a further preferred variant of the invention intervals concentrations range from 0.005 mg % ml to 10 mg % ml According to a further preferred variant of the invention intervals concentrations range from 0.01 mg % ml to 1 mg % ml According to a further preferred variant of the invention the concentration is about 0.1 mg % ml;

lysine hydrochloride: the concentration range is from 0.0001 mg % ml to 2000 mg % ml According to a more preferred variant of the invention intervals concentrations range from 0.001 mg % up to 100 mg % ml According to a further preferred variant of the invention intervals concentrations range from 0.005 mg % ml to 10 mg % ml According to a further preferred variant of the invention intervals concentrations range from 0.01 mg % ml to 1 mg % ml According to a further preferred variant of the invention the concentration is about 0.05 mg % ml;

L-leucine: concentration range is from 0.0001 mg % ml to 2000 mg % ml According to a more preferred variant of the invention intervals concentrations range from 0.001 mg % up to 100 mg % ml According to a further preferred variant of the invention intervals concentrations range from 0.005 mg % ml to 10 mg % ml According to a further preferred variant of the invention intervals concentrations range from 0.01 mg % ml to 1 mg % ml According to a further preferred variant of the invention, the concentration equal to approximately 0.08 mg % ml

New solutions are applicable for the treatment of keratoconus method transepithelial cross-linking or to protect the eyeball from UV-a radiation, is produced by mixing one or more of the above substances at specified intervals concentrations, with a solution containing Riboflavin, for example with a solution of Riboflavin-dextran at a concentration selected from a range of concentrations from 0.0001% to 0.5%. According to a more preferred variant of the invention, the intervals of the concentration of the Riboflavin-dextran comprise from 0.001% to 0.4%. According to another more predpochtitel the final variant of the invention intervals concentrations range from 0.005% to 0.3%. According to a further preferred variant of the invention, the intervals of concentrations are from approximately 0.01% to 0.2%. According to a further preferred variant of the invention the concentration is about 0.1%.

The test results show that each of the substances identified groups applicable to facilitate the penetration of Riboflavin and, in particular, a standard solution of Riboflavin-dextran through the epithelium of the cornea and to protect the cornea from UV-a radiation.

The cornea used (taken into account) in the test rejected the Bank of the cornea due to unfitness for transplantation, are stored in the respective solutions, and to test it again checked by optical microscopy and by studying endothelial cells.

Use only the cornea with high transparency, thickness in the range from 500 to 600 microns, with good endothelial mosaic, in accordance with what is proposed in [10-15].

The cornea have such a way as to cover the cylindrical chamber containing 0.4 ml of a given (predetermined) solution of sodium hyaluronate plus xanthan gum. On the surface of the cornea impose waterproof sealing metal ring of the same diameter as the cylindrical box. Then on the corneal surface nano is Yat subjects of the composition, containing a fluorescent substance (Riboflavin). Measuring the fluorescence of the solution inside the box at various points, you can determine how much new solutions penetrates the cornea and what time is spent on it.

The effectiveness of these substances as carriers in compositions for transepithelial introduction Riboflavin and compositions obtained by mixing Riboflavin at least one of these substances are described in the following examples, tests, only to illustrate and not to limit.

For brevity, only describes the test results obtained during the processing of the cornea following songs:

1) standard 0.1% solution of Riboflavin-dextran;

2) standard 0.1% solution of Riboflavin-dextran+0.01% benzalkonium chloride in accordance with [16];

3) the first new test composition containing 0.1% Riboflavin-dextran+vitamin E TPGS (D-alpha Tocopheryl polyethylene glycol 1000 succinate) at a concentration of 500 mg % ml;

4) the second new test composition containing 0.1% Riboflavin-dextran+vitamin Q 100 mg % ml;

5) the third new test composition containing 0.1% Riboflavin acid+L-Proline, 0.1 mg %+glycine 0.1 mg %, lysine hydrochloride 0.05 mg %+L-leucine 0.08 mg %;

6) of the fourth new test composition containing 0.1% Riboflavin-dextran+vitamin is (D-alpha Tocopheryl polyethylene glycol 1000 succinate) 500 mg % ml+vitamin Q 100 mg % ml+L-Proline, 0.1 mg %+glycine 0.1 mg %+lysine hydrochloride 0.05 mg %+L - leucine 0.08 mg %.

Each of these six compositions applied to the surface of the cornea, selected and placed, as described above, and after 15 minutes and after 30 minutes assess the soaking corneal stroma together with the presence of fluorescent substances in a solution of sodium hyaluronate+xanthan gum 0.4 ml, placed inside a container for the treated cornea.

Determination of the penetration of Riboflavin into the corneal stroma spend, making the slices stroma and the subsequent evaluation under a fluorescent microscope.

The presence of Riboflavin in solution of sodium hyaluronate+xanthan gum 0.4 ml, which demonstrates the penetration through the cornea, evaluated qualitatively using visual and fluorescent scale, as shown in Fig.1, and quantitatively by colorimetric scale. Two figures about each sample colors denote respectively the number of parts of a standard solution of Riboflavin, which gives the color shown, and the number of parts of a solution of xanthan gum and sodium hyaluronate. Standard scale determine, preparing a dilution solution of Riboflavin - dextran 0.1% xanthan gum+sodium hyaluronate in the following proportions (units/ml): 50/0, 40/10, 30/20, 20/30, 10/40, 0/50. Prepare a visual scale and fluorimetric scale matching specific values in units/ml, and each dilution assessed the t in scale from 10 to 0. On a colorimetric scale is expected to be minimal percentage of yellow, equal to 20% in the absence of Riboflavin, it corresponds to the colorimetric range of substances, selected as the diluent.

Evaluation on the visual scale is carried out in standard conditions of illumination by direct comparison of the experimental sample with a standard sample and method of digital photography. Fluorimetric estimation is performed using a fluorescence scanning microscope equipped with a digital camera, in a dark room. A relative measure for the visual and colorimetric scale conducts third researcher on the basis of the average values obtained by two methods.

Colorimetric assessment is made by placing the material at the end of the experiment, the inside of the cylindrical chamber (and therefore under the cornea) in a transparent container, and with the help of computer analysis, scanning at high resolution predefined (ready) solutions and estimating the percentage of yellow color using Photoshop™ 7.0 and monochrome filter. This way you can compare detective the percentage of yellow color in the experimental sample with a precisely defined concentration, expressed in units/ml of standard 0.1% solution Riboflavin is a, as shown in Fig.1.

In Fig.2A presents fluoroscopically the image slice of the cornea treated by the method of transepithelial cross-linking, after the application of the fourth new solution (irrigation fourth new solution) for 15 minutes. In Fig.2b presents fluoroscopically the image slice of the cornea treated by the method of transepithelial cross-linking, after the application of the fourth new solution (irrigation fourth new solution) for 30 minutes. In Fig.2C presents fluoroscopically the image slice of the cornea treated by the method of transepithelial cross-linking the fourth new solution. This last figure, you can see that Riboflavin has penetrated across the cornea and after cross-linking the tissue became more rigid.

The tests show that:

a) after 15 minutes transepithelial application (irrigation) standard 0.1% solution of Riboflavin-dextran corneal stroma partially saturated, and fluorescent solution is not found in the substance inside the container, and colorimetric range is imposed on the range (compatible with spectrum) of a solution of sodium hyaluronate+xanthan gum 0.4 ml (a score of 0, as in Fig.1 and the percentage of yellow color on above 20%);

b) after 30 minutes transepithelial application (irrigation) the zoom is 0.1% solution of Riboflavin-dextran corneal stroma, apparently, fully impregnated with a fluorescent solution; it is possible to estimate the fluorescence in solution sodium hyaluronate+xanthan gum 0.4 ml in the container 2 to 3 points (Fig.1), and the percentage of yellow color, above a certain computer method is in the range of 75% to 80%;

C) after 15 minutes transepithelial application (irrigation) of the first new test solution (test, test solution) 0.1% Riboflavin-dextran+vitamin E TPGS (D-alpha-Tocopheryl polyethylene glycol 1000 succinate) at a concentration of 500 mg % ml, the cornea is completely soaked and the inside of the container has a fluorescent solution (score 2-3 in Fig.1, the percentage of yellow color is 72-76%);

g) after 30 minutes transepithelial application (irrigation) of the first new test solution all layers of the cornea completely soaked, and the inside of the container has a high concentration of Riboflavin, which demonstrates the good permeability of corneal tissue for the product upon contact with a surface (3 - 4 in Fig.1, the percentage of yellow color is 79 - 84%);

d) the composition proposed in [16], and new test solutions of the second and third containing, respectively: benzalkonium chloride 0.01%; vitamin Q 100 mg % ml; L-Proline, 0.1 mg %, glycine 0.1 mg %, lysine hydrochloride 0.05 mg % and L-leucine 0.08 mg %, demonstrate, in the same condition is the conditions, better penetration (penetration) of Riboflavin in quantitative terms, and from the point of view of speed of penetration than one standard solution (Riboflavin - dextran 0.1%) (score 3-4 in Fig.1 through 15 minutes and score 4-6 after 30 minutes, with the percentage of yellow color 70-79% after 15 minutes and 78-86% after 30 minutes);

e) the fourth new test composition gives the results, even surpassing the results of any other test compositions. Different concentrations of coloring substances detected using fluorescence and computer analysis of fluorescent substances inside the container after 15 minutes and after 30 minutes of application (irrigation) of the product on the surface of the epithelium, are extremely high (score 5 - 6 in Fig.1 after 15 minutes, the percentage of yellow color is 88 - 91%; score 6-7 after 30 minutes, the contents of the yellow above 90%"); a higher concentration of fluorescent substances in solution under the cornea, after transepithelial application (irrigation) fourth new test solution (test solution) becomes particularly evident after 15 minutes, especially compared with the results obtained by applying the standard solution, which is not even found in the solution inside the container through the same time. This can explain the, if we assume that the permeability enhancers, in a joint application (mixing), show a synergistic effect, facilitating the passage of Riboflavin through the corneal epithelium.

Illustrated results show that at least the following substances:

- vitamin E

where R1 denotes CH3or N; R2 denotes CH3or N; R3 represents CH3, can bring only as an example, vitamin E TPGS (D - alpha-Tocopheryl polyethylene glycol 1000 succinate);

- coenzyme Q

in the oxidized form,

in the form of Polukhina,

in restored form,

regardless of the number of isoprenoid units of coenzyme Q; only as an example of coenzyme Q10;

- L - Proline

;

- glycine

;

- lysine

or lysine hydrochloride;

- L-leucine

;

alone or in combination with each other, eventually in combination with excipients, such as acetic acid, and the concentration selected in the above intervals, promote the penetration of Riboflavin through the corneal epithelium over to Odie time intervals, than the time intervals required for a standard solution of Riboflavin-dextran, and in quantities sufficient for subsequent cross-linking.

The combination of all of these compounds with Riboflavin exhibits an unexpected synergistic effect, giving the best results from the point of view of concentration of product that passes through the tissue of the cornea, and from the point of view of the speed with which he penetrates these tissues.

Applicants check the in vitro efficacy of transepithelial cross-linking on human cornea, causing the fourth new test composition on the human cornea and irradiating UV-rays when the flow capacity of 3 mW/cm in accordance with a standard Protocol. The cornea for testing is prepared as in the previous experiment, locking it on the appropriate support and dealing (irrigation) standard 0.1% solution of Riboflavin-dextran and fourth new test composition on the surface of the epithelium (i.e., transepithelial) of the cornea within thirty minutes. Then spend a standard UV-a-irradiation within thirty minutes, irradiation is carried out stepwise (Paladino), 5 minutes each stage, with the previous re-applying each solution on the surface of the cornea. At the end of the experiment, the degree of stiffness of the cornea is evaluated as follows: each cornea is oderzhivayut for the tail section length of 2 mm forceps to the cornea, located in a horizontal position, and measure the angle formed by the opposite end of the cornea relative to horizontal.

In Fig.3A shows the cornea after transepithelial cross-linking, by using a standard 0.1% solution of Riboflavin-dextran, which is bent down about 40°, and Fig.3b shows another cornea after cross-linking, by using a fourth new test composition and without prior removal of the corneal epithelium. When comparing the two figures it is obvious that transepithelial cross-linking with the application of the fourth new test composition strengthens the cornea, which is bent downwards only 25°, as required.

For additional confirmation of the effectiveness of transepithelial cross-linking with the application of the fourth new test compositions of this treatment is carried out in vitro on end parts of the human cornea from patients with keratoconus who underwent perforating keratoplasty; in these cases, instead of the destruction of the cornea, explantions the patient, it was used in the laboratory, drawing attention to the fact that in this case, avoid diathermy during the operation, so remain in the surface layers of the cornea (the transplantation of full-thickness). Limbs (edges) of the cornea FIC is irout on a suitable support. Standard 0.1% solution of Riboflavin-dextran is applied to the first cornea, and the fourth new test solution is applied to the second cornea, wetting the surface of the cornea solutions for thirty minutes without removing the epithelium. Then carry out a standard 340 nm UV-a-irradiation only the cornea, the processed fourth test solution with a flow capacity of 3 mW/cm for thirty minutes, divided into stages for 5 minutes each with the previous re-applying the composition to the surface of the cornea. The other end section of keratoconus, fixed on a suitable substrate, is treated only by application to the cornea of a standard solution of 0.1% Riboflavin - dextran without further UV-a-irradiation. At the end of the experiment conducted to study the parenchyma of cornea under the scanning electron microscope.

In Fig.4 shows an image of the leafs slice of the cornea of a patient with untreated (raw) keratoconus under a scanning microscope. In Fig.5 provides an enlarged image of a fragment of Fig.4. These two figures show the weakening of the leafs at the end of the cornea that cause standard 0.1% solution of Riboflavin-dextran, without UV-a-irradiation.

In Fig.6 shows the image under a scanning microscope, which shows how are the leafs slice of the cornea affected keratocan the catfish, after transepithelial cross-linking with the use of the new fourth of the test solution. The tail section of keratoconus (conical cornea), treated the fourth new solution applied transepithelial, and UV-a-irradiation according to the standard Protocol, within thirty minutes, shows a very tight (thick) distributed and compact the leafs of the cornea, which show that they formed a new biochemical cross connection.

Good results were obtained also with the new test compositions of the first, second and third. The results obtained with the application of new test compositions also refute the objections of some authors claiming a hypothetical weakening effect cross-linking in exercising its transepithelial method as epithelium inhibits UV-a rays. The assumption of decreased effectiveness of transepithelial cross-linking is incorrect, as demonstrated by histological samples obtained in the case when the Riboflavin as a carrier (permeability enhancer) add at least one proposed in this description of the substances and irradiation is carried out in accordance with standard Protocol.

In addition, using a scanning electron microscope (h) conduct a comparative study of transepithelial the aqueous cross-linking on the morphological integrity or lack of integrity of the epithelial layers of the cornea and microvilli on the surface of epithelial cells. This study carried out to assess tolerability transepithelial cross-linking with the application of the fourth new test composition on the epithelium of the cornea compared with the standard 0.1% solution of Riboflavin-dextran and with a mixture of standard composition+benzalkonium chloride 0.01%, proposed in [16].

This test is administered because epithelial cells are the first organic structure irradiated by UV-A, and may be damaged due to the absorption of these rays. Apparently, none of the existing literature is not considered this aspect, and in the literature there are no relevant data.

The characteristics considered as the most reliable for assessing the viability of epithelial cells, obtained in the course of cytological study of "impressions", "print" and, in particular, the electron microscope of the microvilli of the cells of the surface layer of the corneal epithelium. The presence of an outer membrane bends (microvilli) whole cell surface elements containing high concentrations of the mucines and enough glycocalyx, promotes optimal binding of the free mucin, which consists of a deep layer pedagogicznej tear film. On the contrary, due to disease (pathological) loss of microvilli defines the difficulty and what GeSHi layer of the tear film to the surface of the eyes and irritation phenomena epithelium, caused by dysfunction of pedagogicznej tear film and, as a consequence, inflammation.

The morphology of microvilli examined under a scanning electron microscope after transepithelial treatment in vitro UV-a rays in accordance with standard doses. The human cornea thickness from 500 to 600 microns incubated for thirty minutes with a balanced salt solution, standard 0.1% solution of Riboflavin-dextran, with a standard solution+benzalkonium chloride 0.01%, and the fourth new test composition, respectively.

Pre-study the morphology of microvilli and the surface layer of the corneal epithelium, is not subjected to any processing, to demonstrate how to look epithelial cells and their microforming not subjected to any photochemical processing.

In Fig.7 given the image under a scanning microscope, which shows the morphology of microvilli and the surface layer of the epithelium in normal (healthy) the cornea, and Fig.8 presents made scanning microscope image of the cornea (corneal samples), exposed to UV-a-irradiation after application of the standard 0.1% solution of Riboflavin-dextran in accordance with a standard Protocol. When compared with Fig.7 shows that all the epithelial layers of the lost and the membrane is of Omena naked.

A similar situation is observed in the samples of the cornea is treated with a composition containing standard solution+benzalkonium chloride 0.01% as proposed in [16].

In Fig.9 presents made by using electron microscope image of the cornea is treated with UV irradiation in the standard dose after processing the fourth new test composition. When compared with Fig.7 and 8 you can see the preservation of the epithelial layers of cell nuclei and slit-like compounds. In addition, there is a significant decrease in the density of microvilli, while the remaining microforming are morphologically intact, deeper located cytoplasmic cell membrane is not exposed to danger. Similar results were obtained with the first, second and third new test compositions.

In Fig.10 shows obtained with a scanning electron microscope image of the cornea irradiated with UV-a at standard dose, using, for comparison, only saline solution. Epithelial layers are broken, many cells have lost their nuclei, and almost all of slit-like compounds and microforming lost.

The obtained results can be summarized as follows:

1) samples of the cornea affected by UV-a-irradiation with pre-incubation with standard 0.1% solution of Riboflavin-d is xtran or composition, containing standard solution+benzalkonium chloride 0.01% according to the standard Protocol, undergo a total loss of all epithelial layers and Romanova membrane becomes naked (Fig.8). In fact, when using only a standard solution, which ultimately add benzalkonium chloride, the epithelium is destroyed under the action of UV-a radiation. This suggests that cross-linking carried out without removal of the epithelium and using either the standard solution or composition Riboflavin-dextran+benzalkonium chloride, will not allow patients after treatment to avoid irritation caused by surgical removal of the epithelium;

2) samples of the cornea treated with the standard dose UV-a-irradiation after incubation with compositions containing Riboflavin-dextran and at least a carrier selected from the above media, and, in particular, with the fourth new test composition retain their epithelial layers, the cell nucleus and slot connection. There is a noticeable decrease of the density of microvilli, while the remaining microforming are morphologically intact. Deeper located cytoplasmic cell membrane of the cornea is a whole;

3) in the samples of the cornea treated with the standard dose UV-a-irradiation after incubation with SB is linzirowanii salt solution (BSS), there is destruction of the epithelial layer, the loss of numerous cells, the endoplasmic engines, and almost all slit-like compounds and microvilli.

Using a scanning microscope also received many image samples of the cornea treated with new first, second and third compositions, but in this specification, they are not shown because they look almost identical images of the cornea treated with a fourth new test composition.

These results make us conclude that at least vitamin E, vitamin Q or coenzyme Q, tested amino acids such as L-Proline, glycine, lysine hydrochloride and L-leucine helps to protect the epithelium of the cornea and to facilitate the penetration of Riboflavin through the epithelium of the cornea. Of the new solutions proposed in this application, the fourth composition exhibits the best performance from the point of view of time of penetration, and from the point of view of preservation of the corneal epithelium.

While it is not clear why the proposed substances contribute to the achievement of such remarkable results and why substances used in the fourth test composition, have a synergetic effect. Without limiting the invention by any theory, the applicants suggest that less visible lesions of the corneal epithelium, the cat is found that the field of processing of the fourth new test composition, can be caused cytoreductive effect of vitamin E such as vitamin E-TPGS, and/or by the presence in solution of at least one essential or part of essential amino acids. Perhaps the vitamin E effect on the glutathione oxidase and peroxydisulfate, which are enzymes involved in repair of the epithelium, or essential or conditionally essential amino acids have cytoreductive action and, probably, also will contribute to effect cross-linking. In addition, a more complete impregnation of the fabric of the fourth new solution determines, probably, not only the strengthening effect of cross-linking, but also the best the layers of the corneal epithelium during the dangerous UV-rays.

With new solutions Riboflavin-dextran and at least one carrier (permeability enhancer), selected among the substances proposed in this description, it is possible to carry out cross-linking, for example, for the treatment of keratoconus, without prior removal of the corneal epithelium. The fact that the corneal epithelium is not removed, allows to avoid:

1) illness caused by irradiation, which typically occur in the first days after cross-linking in accordance with the standard Protocol;

2) the necessity of applying therapeutic contact lenses after treatment and, above all,

3) risk poslove which include infections of the cornea due to the removal of the corneal epithelium.

In addition, the refusal of surgical intervention can be treated on an outpatient basis, eliminating the need for operating and the operating microscope.

New solutions can also be entered in the form of eye drops or gel or apply to therapeutic contact lenses prior to exposure to sunlight, especially in summer, to enhance the effect of natural cross-linking due to the irradiation of Riboflavin sunlight.

Furthermore, new solutions could protect the internal structures of the eyeball from UV-a rays and so they can be used, for example, to protect against macular degeneration and/or prevention of cataracts in people with increased risk her education due to the fact that they are many hours are on the sun.

Benzalkonium chloride in a concentration of from 0.0001%-0.02%, preferably, at a concentration of about 0.01%, can be added only for as long as possible to keep the new solutions are not in single dose containers. If the new solutions will be produced and sold in single dose containers for single use, adding benzalkonium chloride is not required. In addition, the composition can be added preservatives, antimicrobial agents, antifungal agents, excipients (e.g., acetic acid) and, in General, any wind is estvo, used in ophthalmology for the preparation of a stable and sterile ophthalmic solutions and/or for better absorption.

As indicated above, applicants believe that the good performance of the composition obtained by adding to the Riboflavin at least one substance selected from vitamin Q, L-Proline, glycine, lysine hydrochloride, L-leucine, partly due to the fact that the latter substances are essential or conditionally essential amino acids, which are cytoreductive function, increasing levels of metalloproteinases MMR. This leads to the conclusion that it is possible to obtain results similar to the results obtained using the new test tracks, the first, second, third and fourth, by adding to a solution of Riboflavin, for example to a standard solution of Riboflavin-dextran at least one substance used to increase metalloproteinases MMR, presumably, at a concentration of from 0.00001% to 0.5%.

Among these substances used to increase (improve) the metalloproteinase MMR that deserve mention genistein(5,7-dihydroxy-3-(4-hydroxyphenyl)chromen-4-one), phytoestrogens, cytokines and the so-called non-steroidal anti-inflammatory drugs (drugs) (NSPs, NSAID). Among NCPs used to made the Oia ophthalmic solution Riboflavin, worth mentioning: acetylsalicylic acid(2-acetoxybenzoic) acid, flufenamic acid(2-{[3-(trifluoromethyl)phenyl]amino}benzoic acid), meclofenamic acid (2-[(2,6-dichloro-3-were)amino]benzoic acid), mefenamovaya acid(2-(2,3-dimetilfenil)aminobenzoic acid), niflumova acid(2-{[3-(trifluoromethyl)phenyl]amino}nicotinic acid), telenova acid(2-[(3-chloro-2-were)amino]benzoic acid), benorilate(4-(acetylamino)phenyl 2-(atomic charges)benzoate), carprofen((RS)-2-(6-chloro-9H - carbazole-2-yl)propanoic acid), celecoxib(4-[5-(4-were)-3-(trifluoromethyl)pyrazole-1-yl]benzosulfimide), tenoxicam(piroxicam of cinnamate or [9-methyl-10,10-dioxo-8-(pyridine-2-ylcarbonyl)-10$1^{6}-thia-9-azabicyclo[4.4.0]DECA-1, 3, 5, 7-tetraen-7-yl](E)-3-phenylprop-2-ENOAT), diflunisal(2',4'-debtor-4-hydroxybiphenyl-3-carboxylic acid), diclofenac(2-[2-[(2,6-dichlorophenyl)amino]phenyl]acetic acid), droxicam(2H,5H - 1,3-oxazino(5,6-C)(1,2)benzothiazin-2,4(3H)-dione, 5-methyl-3-(2 - pyridinyl)-, 6,6-dioxide), etodolac((RS)-2-(1,8-diethyl-4,9-dihydro-3H-pyrano[3,4-b]indol-1-yl)acetic acid), etoricoxib(5-chloro-6'-methyl-3-[4-(methylsulphonyl)phenyl]-2,3'-bipyridine), fenoprofen(2-(3-phenoxyphenyl)propanoic acid), flurbiprofen ((RS)-2-(2-forbiden-4-yl)propanoic acid), ibufenac (4-isobutylbenzene acid), ibuprofen((RS)-2-(4-(2- methylpropyl " phenyl) propanoic the acid), indometacin(2-{1-[(4-chlorophenyl) carbonyl]-5-methoxy-2-methyl-1H-indol-3-yl}acetic acid), Ketoprofen((RS)-2-(3-benzoylphenyl)propanoic acid), Ketorolac ((±)-5-benzoyl-2,3-dihydro - 1H_-pyrrolizine-1-carboxylic acid, 2-amino-2-(hydroxymethyl)-1,3-propandiol), lornoxicam((3E)-6-chloro-3-[hydroxy(pyridin-2-ylamino)methylene]-2-methyl-2,3-dihydro-4H-thieno[2,3-e][1,2]thiazin-4-one 1,1-dioxide), lumiracoxib ({2-[(2-chloro-6-forfinal)amino]-5-were}acetic acid), meloxicam (4-hydroxy-2-methyl-N-(5-methyl-2-thiazolyl)-2H-1,2-benzothiazin-3-carboxamide-1,1-dioxide), Metamizole (sodium [(2,3-dihydro-1,5-dimethyl-3-oxo-2-phenyl-1H-pyrazole-4-yl) methylamino] methanesulfonate), naproxen ((+)-(S)-2-(6-methoxynaphthalene-2-yl)propanoic acid), nimesulide(N-(4-nitro-2-phenoxyphenyl)methanesulfonamide), oxaprozin(3-(4,5-diphenyl-1,3-oxazol-2-yl)propanoic acid), parecoxib(N-{[4-(5-methyl-3-phenylisoxazol-4-yl)phenyl]sulfonyl}propanamide), piroxicam((8E)-8-[hydroxy-(pyridin-2-ylamino)methylidene]-9-methyl-10,10-dioxo-10λ6-thia-9-azabicyclo[4.4.0]DECA-1,3,5-trien-7-one), rofecoksib(4-(4 - methylsulfinylphenyl)-3-phenyl-5H-furan-2-one), sulindac({(1Z)-5-fluoro-2-methyl-1-[4-(methylsulfinyl)benzylidene]-1H-inden-3-yl}acetic acid), sudoxicam(4-hydroxy-2-methyl-N(2)-thiazolyl-2H-1,2-benzothiazin-3-carboxamide 1,1-dioxi is), tenoxicam((3E)-3-[hydroxy(pyridin-2-ylamino)methylene]-2-methyl-2,3-dihydro-4H-thieno[2,3-e][1,2]thiazin-4-one 1,1-dioxide), valdecoxib(4-(5-methyl-3-phenylisoxazol-4-yl) benzosulfimide).

In addition, the applicant believes that it is possible to obtain results similar to the results obtained with the first, second, third and fourth new test composition by adding to a solution of Riboflavin, for example, to a solution of Riboflavin-dextran at least one essential or conditionally essential amino acid.

Among potentially applicable substances most promising seems to be L-arginine. It can be mixed with a solution of Riboflavin, for example with a standard solution of Riboflavin-dextran, followed by the addition of one or more of the above substances to produce new solutions. Although haven't checked all of these substances, applicants seems logical to assume that all essential or conditionally essential amino acids can be used effectively to facilitate the penetration of Riboflavin through the epithelium of the cornea, thereby protecting the cornea from UV-a-irradiation from sunlight absorbed by Riboflavin. In particular, it seems reasonable to consider L-arginine as suitable media, like suggested above substances, as it is logical to assume that L-arginine, the amino acid predestiny is nitric oxide, can deliver nitric oxide (NO) on the periphery. The use of nitric oxide (NO) for treatment of patients with keratoconus using topical application to the cornea epithelium substances in which NO dissolves, prevents poor solubility NO.

It was experimentally verified as L-arginine and other essential amino acids protect microforming epithelium from UV-a-irradiation during cross-linking. As it turned out that other essential or conditionally essential amino acids, particularly L-arginine, applicable for the preparation of ophthalmic solutions, which contribute to the implementation of transepithelial cross-linking, at the same time preserving the epithelium of the cornea, the applicants consider it logical that they should be added to the solution of Riboflavin, for example, a standard solution of Riboflavin-dextran, in an approximate concentration of from 0.00001% to 0.5%.

According to a more preferred variant of the invention, the concentration of L-arginine or other essential or conditionally essential amino acid is from 0.001% to 0.4%. According to a further preferred variant of the invention, the concentration range is from 0.005% to 0.3%. According to a further preferred variant of the invention, the concentration range is from 0.01% to 0.2%. According to a further preferred variant of the invention the concentration is ia L-arginine or other essential or conditionally essential amino acid is about 0.1%.

Alternatively, you can add L-arginine and/or any other essential or conditionally essential amino acid in any of the proposed new solutions.

Dosage new ophthalmic solutions depends on the revealed pathology and its severity. I believe that the dose can range from one drop in the eye a day to a maximum of one drop per hour.

The claimed invention is an integral part of this description. Full disclosure of the priority of Italian patent applications VA2009A000052 and VA2010A000044 on the name of the same applicants entered into this description by reference.

LINKS

1. Seiler T., E. Spoerl, Huhle M, Kamouna A. Conservative therapy of by leading to numerous refractive enhancement of collagen cross-links. Invest Ophthalmol. Vis. Sci. 1996; 37: S1017.

2. Seiler T., Quurke AW. Iatrogenic keratectasia after LASIK in a case of forme fruste, leading to numerous refractive. J Cat Refract Surg. 1998; 24: 1007-1009.

3. Spoerl e, Huhle m, Seiler T. "Induction of cross-links in corneal tissue", Exp Eye Res. 1998; 66: 97-103.

4. Mazzotta C, Traversi C, Baiocchi s, Sergio P., Caporossi T., Caporossi A. Conservative treatment of leading to numerous refractive by riboflavin-uva-induced cross-linking of corneal collagen: a qualitative investigation" Eur J Ophthalmol. 2006; 16: 530-5.

5. Caporossi A., Baiocchi, S., Mazzotta C, Traversi C, Caporossi T. "Parasurgical therapy for leading to numerous refractive by riboflavin-ultraviolet type A rays induced cross-linking of corneal collagen: preliminary refractive results in an Italian study" J Cataract Refract Surg. 2006;32: 837 - 45.

6. Spoerl e, Seiler T. "Techniques for stiffening the cornea" J Refract Surg. 1999; 15: 711-713.

7. G. Hagele, Boxer Wachler BS. "Corneal Collagen Crosslinking with Riboflavin (C3-R) for corneal stabilization" Presented at the International Congress of Corneal Cross inking (CCL). December 9 - 10,2005. Zurich, Switzerland.

8. Pinelli, R. "C3-Riboflavin for the treatment of leading to numerous refractive J Cataract &Refractive Surgery Today Europe. 2006; 1: 49-50.

9. Pinelli R. "Eyeword", 2007; 5: 34-40.

10. Linda J. Muller, Elisabeth Pels, Gijs F. J. M. Vrensen: "The specific architecture of the anterior stroma accounts for maintenance of corneal curvature". Br J Ophthalmol 2001; 85:437 - 443 (April).

11. Mau T. Tranl, Robert N. Lausch 2 and John E. Oakes: "Substance P Differentially Stimulates IL-8 Synthesis in Human Corneal Epithelial Cells" Investigative Ophthalmology and Visual Science. 2000; 41: 3871- 3877.

12. L. J. Muller, L. Pels and GF. Vrensen: "Novel aspects of the ultrastructural organization of human corneal keratocytes" Investigative Ophthalmology & Visual Science, Vol 36, 2557-2567.

13. G. Perrella, P. Brusini, R. Spelat, P. Hossain, A. Hopkinson, H. S. Dua: "Expression of haematopoietic stem cell markers, CD133 and CD34 on human corneal keratocytes" British Journal of Ophthalmology 2007; 91: 94- 99.

14. Tadashi Senoo, and Nancy C. Joyce: "Cell Cycle Kinetics in Corneal Endothelium from Old and Young Donors" Investigative Ophthalmology and Visual Science. 2000; 41: 660-667.

15. L. J. Mtiller, L. Pels and GF. Vrensen: "Ultrastructural organization of human corneal nerves" Investigative Ophthalmology & Visual Science, Vol 37,476 - 488.

16. Italian patent application No. MI2007A002162, 14 Nov. 2007, R. Pinelli, "Collirio per il trattamento del cheratocono con tecnica cross - linking trans - epiteliale".

17. R. Pinelli, A. J. Kannellopoulos, B. S. B. Wachler, E. Spoerl, A. Ertan, S. L. Trokel, "C3 - Riboflavin treatments: Where did we come from? Where are we now?", Cataract &Refractive Surgery Today Europe, Summer 2007.

18. Ashim K. Mitra, "Ophthalmic Drug Delivery Systems", Second Edition, Revised And Expanded, Marcell Dekker Inc., NY, 2003.

1. Ophthalmic solution containing Riboflavin and vitamin E, to protect the internal structures of the eyeball from UV-a rays or for the treatment of keratoconus method transepithelial cross-linking.

2. Ophthalmolo the practical solution on p. 1, which contains Riboflavin, vitamin E, and optionally at least one compound selected from the group consisting of coenzyme Q, L-Proline, glycine, lysine hydrochloride, L-leucine and L-arginine.

3. Ophthalmic solution under item 1, which contains a solution of Riboflavin in a concentration selected from a range of concentrations from 0.0001% to 0.5%, and Riboflavin provide in the form of a solution of Riboflavin-dextran, optionally containing dextran at a concentration of 20 wt. -%, and vitamin E in a concentration selected from a range of concentrations from 0.0001 mg % up to 2000 mg %.

4. Ophthalmic solution on p. 3, which further comprises at least one of the following connections:
- coenzyme Q concentration selected from a range of concentrations from 0.0001 mg % up to 2000 mg %;
- L-Proline in a concentration selected from a range of concentrations from 0.0001 mg % up to 2000 mg %;
- glycine in a concentration selected from a range of concentrations from 0.0001 mg % up to 2000 mg %;
- lysine hydrochloride at a concentration selected from a range of concentrations from 0.0001 mg % up to 2000 mg %;
- L-leucine at a concentration selected from a range of concentrations from 0.0001 mg % up to 2000 mg %;
- L-arginine in a concentration selected from a range of concentrations from 0.00001% to 0.5%.

5. Ophthalmic solution on p. 3, which contains a solution of Riboflavin-dextran concentration is about 0.1%, vitamin E at a concentration of about 500 mg %; and at least one of the following connections:
- coenzyme Q in a concentration of about 100 mg %;
- L-Proline in a concentration of about 0.1 mg %;
- glycine in a concentration of about 0.1 mg %;
- lysine hydrochloride in a concentration of about 0.05 mg %;
- L-leucine at a concentration of about 0.08 mg %;
- L-arginine in a concentration of about 0.1%.

6. Ophthalmic solution on p. 3, which contains a solution of Riboflavin-dextran 0.1%, vitamin E TPGS (D-alpha-Tocopheryl polyethylene glycol 1000 succinate) 500 mg %, coenzyme Q 100 mg %, L-Proline, 0.1 mg %, glycine 0.1 mg %, lysine hydrochloride 0.05 mg % and L-leucine 0.08 mg %.

7. Ophthalmic solution according to any one of paragraphs. 1-6 which is prepared in the form of eye drops or ophthalmic gel or in a form suitable for application to therapeutic contact lenses.

8. The use of Riboflavin for making ophthalmic solution containing vitamin E, to protect the internal structures of the eyeball from UV-a rays or for the treatment of keratoconus method transepithelial cross-linking.

9. Enhancer permeability through the epithelium of the cornea ophthalmic solution containing Riboflavin, to protect the internal structures of the eyeball from UV-a rays or for the treatment of keratoconus method transepithelial cross-linking, comprising vitamin E.



 

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SUBSTANCE: invention refers to medicine, specifically to pharmaceutical compositions possessing prolonged antiarrhythmic actions and applicable to correct the cardiac arrhythmia, including that of the ischemic origin. What is presented is a pharmaceutical composition with the prolonged antiarrhythmic acitivity containing bis[2-(diethylamino)]-N-(2,6-dimethylphenyl)acetamide L-glutaminate, L-glutamic acid, 2-aminoethane sulpho-acid and excipients.

EFFECT: prolonged antiarrhythmic action and stability of the dosage form (tablets, capsules for oral application or solution for injections).

3 cl, 4 tbl

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to manufacturing pharmaceutical drugs. A drug represents a solution of N,N'-diaminodiphenylsulphone in a mixture of solvents: ethanol and water and at least one of the solvents specified in a group including propylene glycol, benzyl alcohol and glycofurol. Preferentially, the solution contains ethanol 25%, propylene glycol 58%, benzyl alcohol 1%, glycofurol 5% and water 11%.

EFFECT: what is prepared is the dosage form of N,N'-diaminodiphenylsulphone used for cerebral infarction, epilepsy, a traumatic marrow injury, a craniocerebral injury, leprosy, pneumocystis carinii infections, and any condition requiring fast and full absorption of the formulation.

13 cl, 6 dwg, 1 ex

FIELD: medicine.

SUBSTANCE: invention relates to veterinary, in particular to obstetrics, gynecology and reproduction biotechnology, and can be used for treating animals in case of clinical mastitis. Medication for treating animals with clinical mastitis includes: trivit - 10.0 ml, cefotaximum - 750 mg, prednisolone - 10.0 mg, nystatin - 325.0 ml. Medication is introduced to ill animals intracycternally in dose 10.0 ml with interval 12 hours after milking.

EFFECT: invention provides increased efficiency of treating clinical mastitis by complex impact on all sides of pathological process.

4 tbl, 3 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to medicine, more specifically to new dosage forms of cephem compounds effective for treating bacterial infections.

EFFECT: dosage forms are stable; they show the improved solubility and are especially applicable for parenteral administration.

9 cl, 29 tbl, 8 ex

FIELD: chemistry.

SUBSTANCE: invention represents method of obtaining alimemazine tartrate solution for introduction by injection, characterised by the fact that sodium sulfate and ascorbic acid are dissolved with constant mixing in water for injections, which has temperature 20-25°C and was preliminarily bubbled with carbon dioxide for 15-25 minutes, after complete dissolution alimemazine tartrate is introduced and mixed for 10-20 minutes, filtered by method of sterilising membrane filtration, obtained solution is packed into dimming glass ampoule with application of gas protection by carbon dioxide with further sterilisation of solution at temperature 100-105°C for 30 minutes.

EFFECT: method improvement.

2 cl, 3 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to a concentrated acidic component for producing a hemodialysis solution. The acidic component contains the following ingredients in an amount of producing 1 litre of the solution in water purified for hemodialysis: 204.7-215.0 g of sodium chloride NaCl, 6.2-9.0 g of calcium chloride CaCl2*2H2O, 3.56-7.12 g of magnesium chloride MgCl2*6H2O, 5.22-10.44 g of potassium chloride KCl, 0.021-6.28 g of acetic acid and 0.02-6.2 g of succinic acid. Also, the invention refers to the hemodialysis solution containing the above concentrated acidic component, water purified for hemodialysis, and bicarbonate component. The invention also refers to a method for producing the hemodialysis solution with the method involving supplying the concentrated acidic component into a hemodialysis apparatus, diluting with water purified for hemodialysis, and mixing with bicarbonate component. What is also declared is a kit for producing the concentrated acidic component containing sodium chloride, calcium chloride, magnesium chloride, potassium chloride, succinic acid in the form of dry agents and acetic acid in the form of a liquid agent.

EFFECT: invention provides higher effectiveness by a biochemical compatibility with blood plasma of the hemodialysis solution.

36 cl, 18 tbl, 16 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: group of inventions refers to medicine, namely ophthalmology, and aims at treating cataract. A drug preparation for treating cataract based on the active substance sodium dihydroazapentacene polysulphonate (azapentacene) is presented in the form of ophthalmic drops. Stabilising is ensured by using a borate buffer and potassium chloride, a borate buffer and sodium chloride, a phosphate buffer, a quaternary ammonium derivative, Nipaging and a preserving agent of mercury derivatives. The ingredients are used in the declared amounts.

EFFECT: using the group of inventions provides the enhanced antimicrobial properties and the reduced toxicity of the ophthalmic drops, thereby increasing the clinical effectiveness in cataract.

6 cl, 1 ex

FIELD: medicine.

SUBSTANCE: invention refers to a new pharmaceutical composition in the form of spray for oral administration, containing an aqueous solution of sildenafil citrate as an active substance in the amount of 3-30 wt %. The composition contains stabilising agents in the form of pharmaceutically acceptable calcium salts taken in an amount required to reduce pH to 3.0 to 6.0. The calcium salts are specified in a group consisting of calcium lactate pentahydrate, calcium lactate trihydrate, calcium gluconate, calcium malate, calcium glycolate, calcium chloride hexahydrate and calcium chloride dihydrate. What is also described is a method for preparing the composition by introducing the pharmaceutically acceptable calcium salts as the stabilising agents into the aqueous solution of sildenafil citrate.

EFFECT: pharmaceutical composition in the form of oral spray is free from organic solvents and applicably for sexual dysfunction.

13 cl, 8 ex

FIELD: chemistry.

SUBSTANCE: invention represents pharmaceutical composition for correction and therapy of manifestations of amyloid intoxication in patients with brain pathologies, which are characterised by the fact that it contains melatonin 3-10 mg and memantine 5-300 mg.

EFFECT: effective treatment of patients, including cases of moderate cognitive disorders.

4 cl, 2 ex, 6 tbl, 7 dwg

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to pharmaceutical industry and represents a topical composition containing a combination of natural salt or pure sodium chloride and glucose mixed in ratio 1:1-30 (wt/wt) as an active ingredient in an amount effective for treating bacterial vaginosis caused by Cardnerella vaginalis together with a pharmaceutically acceptable carrier.

EFFECT: invention provides the higher pharmacological activity.

4 cl, 4 ex, 5 tbl

FIELD: food industry.

SUBSTANCE: biologically active food additive strengthening the organism adaptive power and body defences and having anti-inflammatory and antioxidant activity contains vegetal origin components represented by a complex extract of devil's-club root, Rhaponticum carthamoides root, Hedysarum neglectum Ledeb root, celery roots and leaves, rhodiola rosea root, Japanese angelica tree roots, boschniakia rossica roots, Hungarian sainfoin herb, magnolia-vine fruits; additionally the additive contains chitosan, trepang fermentative hydrolysate, ascorbic acid, taurine, glutathione, nicotinamide, vitamin B1, vitamin B2, vitamin B6, vitamin B12, folic acid, anhydrous calcium chloride, magnesium chloride, zinc chloride, bee honey at preset ingredients ratio.

EFFECT: biologically active food additive promotes effective strengthening of the organism adaptive power and body defences and human aging retardation.

4 tbl, 6 ex

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