GB2493568A

GB2493568A - Ophthalmologic composition comprising a viscoelastic polysaccharide and a dye

Info

Publication number: GB2493568A
Application number: GB1114317.9A
Authority: GB
Inventors: Ja Rgen Nachbaur; Gillian Rodden; Andre Wolfstein; Claude Moissonnier; Pascal Bernard; Gildas Lorec; Nicole Bielefeldt; Brian Rathert; Lidia Nachbaur; Mario Gerlach
Original assignee: Carl Zeiss Meditec AG
Current assignee: Carl Zeiss Meditec AG
Priority date: 2011-08-09
Filing date: 2011-08-09
Publication date: 2013-02-13
Anticipated expiration: 2031-08-09
Also published as: GB201114317D0; GB2493568B; WO2013020917A1

Abstract

Ophthalmologic composition, comprising an aqueous solution of at least one viscoelasÂtic polysaccharide, wherein the at least one polysacchaÂride is covalently bound to at least one dye. Preferably, the dye is a modified and/or unmodified aminoanthracenedione and/or a modified and/or unmodified nitrophenyldiazenylbenzeneamide. A method for producing an ophthalmoÂlogic composition, in which at least one dye is covaÂlently bound to at least one water-soluble, viscoelastic polysaccharide is also disclosed. The composition may be used in ophthalmology, in particular for use in cataract surgery.

Description

Ophthalmologic composition comprising an aqueous solution of at least one viscoelastic polysaccharide

Field of the invention

S The invention relates to an ophthalmologic composition, comprising an aqueous solution of at least one viscoelas-tic polysaccharide. The invention further relates to a method for producing an ophthalmologic composition.

Prior art

Cataract is a common disease especially in elderly peo-ac ple, where the crystalline lens gradually becomes less transparent. This opacity of the natural lens leads to the loss of visual acuity. To restore vision, cataract surgery has to be performed. In a surgical step called phacoemulsification, the natural lens is destroyed and * 15 the pieces are aspirated out of the eye. In the next step, an artificial lens is injected into the eye and visual acuity is re-established.

* : Immediately before phacoemulsification, the anterior **.*.* * -chamber is usually filled with an ophthalmologic composi-tion, comprising a transparent viscoelastic solution. The viscoelastic ophthalmologic composition is used as a sur-gical aid to protect intraocular tissues (for example the corneal endothelium during phacoemulsification), as a space maintainer (for example to maintain the anterior chamber of the eye) and to facilitate intraocular maneu-vers, for example to make a controlled capsulorhexis.

Viscoelastic compositions are water based solutions con- S taming a polysaccharide like hyaluronic acid, hy- droxyropylmethylcellulose, chondroitin sulfate or mix- tures thereof that are routinely used in cataract sur-gery. The viscoelastic composition might differentiate in the molecular weight of the polysaccharide dissolved in the solution, in the concentration of the polysaccharide and in the viscosity of the solution. They are generally well tolerated following brief exposure to intraocular tissues.

Most currently available ophthalmologic compositions corn-prising viscoelastic polysaccharides are transparent. A disadvantage of the transparency is that it is difficult to determine or control the amount and locaticn of the ophthalmologic compositions in the eye. It is of impor- tance that at the end of the surgery all of the ophthal-roologic composition is removed from the eye to prevent a * trabecular meshwork obstruction and thus the elevation of * intraocular pressure which may lead to g]aucoma. Due to the transparency of the used polysaccharides, it is dif-ficult to judge if residual ophthalmologic composition is still left in the eye. Furthermore, poor visibility of the viscoelastic ophthalmologic composition within the ** eye also prohibits exact visualization of the amount in-jected.

EP 1 263 363 B1 suggests the use of a viscoelastic oph-thalmologic composition based on aqueous solutions of hyaluronic acid and hydroxypropylmethylcellulose. To avoid the above-named problems associated with transpar-ency the ophthalmologic solution also contains trypan blue to stain the viscoelastic solution.

From WO 01/66053 Al, a further ophthalmologic composition is known, which is composed of a mixture of a viscoelas-tic polysaccharide with a vital dye, that is with a dye, which can be applied to living cells without killing them.

However, therein, both the trypan blue and the vital dye have to possess a substantially higher solubility in the ophthalmologic composition than in the surrounding tissue in order not to stain the tissue. However, in particular upon longer dwell of the ophthalmologic composition in the eye, for example during surgery, it cannot be ex-cluded that a certain proportion of the dye diffuses into the surrounding tissue and stains it. Moreover, there ex-ist only comparatively few dyes, which can be applied to living cells without permanently damaging them.

It has been proposed to formulate a viscoelastic material containing fluorescein as a dye in the past (Zirm et al, British Journal of Ophthalmology 67, 1983, 259-263) These materials however did not have the desired advan- tages in ocular surgery. Dyes like fluorescein can dif- fuse out of the ophthalmologic composition, too, as a re- sult of which they also stain the surrounding tissue. An-other problem is that fluorescein has to be irradiated with UV-light to show fluorescence and to make the vis- coelastic solution visible. In order to minimize the dif-fusion of the dye from the ophthalmologic composition, US 4764360 Al uses a high molecular weight polymer dye with a molecular weight preferably exceeding 30,000 Dal- ton (u) to maintain the dye in the ophthalmologic solu-tion.

Detailed description of the invention

It is the object of the present invention to improve an ophthalmologic composition of the initially mentioned type such that it is simpler and safer in application. A further object of the invention is to provide a method for producing such an ophthalmologic composition.

According to the invention, the objects are solved by an ophthalmologic composition having the features of claim 1 as well as by a method for producing an ophthalmologic composition according to claim 11. Advantageous develop-ments of the invention are specified in the respective dependent claims, wherein advantageous developments of the ophthalmologic composition are to be regarded as ad-vantageous developments of the method and vice versa.

In an ophthalmologic composition according to the inven- * tion, comprising an aqueous solution of at least one vis-coelastic polysaccharide, it is provided that the at * 2 least one polysaccharide is covalently bound to at least one dye. Therein, within the scope of the invention, chemical compounds are to be understood by dye, which in- * dude at least one chromophore molecule structure, which I.....

* 25 absorbs light in the wavelength range visible to the hu-man between about 380 nm and about 800 nm and preferably does not exhibit any fluorescence or phosphorescence. By the covalent bond of the dye to the at least one viscoe-lastic polysaccharide, it is reliably excluded that the dye diffuses into adjoining tissue and undesirably stains it during use of the ophthalmologic composition. More- over, unlike fluorescent dyes such as fluorescein, rhoda-mine or the like, it is not required to irradiate the s ophthalmologic composition with DV light to visualize it.

This allows a substantially simpler handling of the oph-thalmologic composition. In addition, there is a priori no risk that the concerned tissue is damaged by the irra-diation with high-energy uv light. Further, a user -for example a surgeon -can even recognize minor traces with-out additional auxiliary means such as DV lamps or the like due to the integral coloration of the ophthalmologic composition, whereby the use of the ophthalmologic compo-sition according to the invention in addition becomes is substantially simpler and safer -for example within the scope of eye surgeries. In simplest configuration, the ophthalmologic composition is composed of an aqueous so-lution of a single viscoelastic polysaccharide, to which the dye is covalently bound. Alternatively, the ophthal- mologic composition can include plural different viscoe- lastic polysaccharides and/or further additives. Inde- * * pendently thereof, it can also be provided that two or more different dyes are covalently bound to a viscoelas-tic polysaccharide.

In an advantageous development of the invention, it is provided that the polysaccharide is cellulose, a cellu-* .Se.s * lose ether with methyl and/or ethyl and/or propyl groups,

S

in particular hydroxypropyl methylcellulose, hydroxyethyl methylcellulose and/or methylcellulose, a glycosaminogly-can, in particular hyaluronic acid, chondroitin sulphate, dermatan sulphate, heparin, heparan sulphate, keratan sulphate, alginic acid, polymannuronic acid, polygulu-ronic acid, polyglucuronic acid, amylose, amylopectin, callose, chitosan, polygalactomannan, dextran, xanthan and/or a mixture thereof. Hereby, in particular the vis-coelastic properties of the ophthalmologic composition can be adapted to the respective purpose of employment and use in optimum manner. Therein, basically, it can also be provided that the ophthalmologic composition in-cludes two or more polysacchartdes of the same type, which only differ with regard to the molecular proportion of covalently bound dye.

In a further advantageous development of the invention, it is provided that the at least one dye is bound di-rectly and/or via a spacer to the polysaccharide. Hereby, the dye can be particularly simply adapted to the present reactive groups of the concerned polysaccharide and be covalently bound to it. The use of a spacer is for exam-ple advantageous if the reactive group of the dye could not be bound to a corresponding reactive group of the polysaccharide or only within the scope of multi-stage reactions. Furthermore, the use of a spacer is advanta- geous if the absorption behavior of the dye would other-wise be affected in undesired manner by the covalent binding to the polysaccharide. Finally, by the use of a * * 25 spacer the viscoelastic properties of the polysaccharide can be influenced as needed.

In a further advantageous development of the invention, * it is provided that the at least one polysaccharide is * obtainable by a reaction of the polysaccharide with at least one reactive dye from the group of modified and/or unmodified aminoanthracenedjone and/or modified and/or unmodified nitrophenyldiazenylbenzenamine. The use of an aminoanthracenedione or of an aminoanthracenedione de-rivative and/or of a nitrophenyldiazenylbenzenamine or of a nitrophenyldiazenylbenzenamine derivative as a dye, S which is covalently bound to the polysaccharide, therein offers the advantage that the color of the viscoelastic polysaccharide and thereby of the ophthalmologic composi-tion is specifically adjustable nearly in the entire visible wavelength range. Furthermore, these two dyes or dye groups are characterized by a great fastness to wash-ing and light. Thus, the ophthalmologic composition is particularly simply and safely to handle and additionally has high storage stability.

In a further advantageous development of the invention, it is provided that the at least one polysaccharide is obtainable by reaction with at least one reactive dye of the general formula (I) ::iiii"iii'iiiiiiiiii'iii: * ...e.

wherein in the general formula (I) at least one of the substituents R1 to R9 is an amino group, and the remain-ing substituents R1 to R8, which are different from the amino group are selected from hydrogen, halogen, C1-C4- alkyl, 01-C4-alkoxy, C1-C4-alkoxy-C1-04-alkyl, C1-C4-alkyl- 01-C4-alkoxy, C1-C4-alkoxy-C1-04-alkoxy, aryloxy, acetamide, propionamide, butyramide, isobutyramide, sul- phonate, (aminophenyl) -N-alkylacetamide, (aminophenylsul-phonyl)alkyl sulphate, alkylbenzenesulphonamide, tn (alkyl)benzenarnine, hydroxy, (alkylsul-phonyl) benzenamine, and (alkenylsulphonyl) benzenamine, and/or wherein at least two adjacent substituents R, R.i with x=l to 3 and/or 5 to 7 form a 3-, 4-, 5-, 6-or 7-membered homocyclic or heterocyclic radical, wherein said cyclic radical can be unsaturated or aromatic. Therein, all of the stereoisomers, racemic mixtures and position isomers are to be considered as included. With the aid of a dye of the general formula (I), the color properties of the polysaccharide covalently bound to it can be varied within wide limits, wherein combinations of different dyes of the general formula (I) can basically also be used to achieve a specific coloration. By at least one of the radicals R1 to R3 being an amino group, the dye can additionally be coupled to a plurality of functional groups in chemically particularly simple manner. There- fore, different viscoelastic polysaccharides with corre-spondingly different functional groups can also quickly and simply be coupled to the dye of the general formula (I) without the requirement of costly multi-stage reac-tions, thereby resulting significant cost advantages. All * reactive dyes of the general formula (I) exhibit a high * 25 extinction coefficient in the visible range of the light *:c* spectrum.

In a further advantageous development of the invention, it is provided that the at least one polysaccharide is obtainable through reaction with at least one reactive BC dye of the general formula (II), R8 10 P5 (II) wherein in the general formula (II) at least one of the substituents R1 to R5 is an amino group and at least one of the substituents R5 to Rio is a nitro group, and wherein the remaining radicals R1 to Rio, which are dif- ferent from the amino group and the nitro group are se-lected from hydrogen, halogen, 01-C4--alkyl, C1-C4-alkoxy, C1-C4-al koxy-C--C4-alkyl, Ci-C4-alkyl-Ci--04-alkoxy, C-C4-alkoxy-C1-C4-alkoxy, aryloxy, acetamide, propionamide, butyramide, isobutyramide, suiphonate, (arninophenyl)-N-- alkylacetamide, (aminophenylsulphonyl)alkyl sulphate, al-kylbenzenesulphonamide, tn (alkyl) benzenamine, hydroxy, (alkylsulphonyl) benzenamine, and (alkenylsul-phonyl)benzenamine, and/or wherein at least two adjacent substituerits R, R+1 with y=l to 4 and/or 6 to 9 form the cyclic radical with: * R=hydrogen, halogen, amino, hydroxyl, nitro, C1-04--alkyl, * 20 C1-C4-alkoxy, Ci-Cq--alkoxy-Ci-C4-alkyl, C1-C4-alkyl--C1-C4-alkoxy, and/or Ci-C4-alkoxy-C1--cq-alkoxy. Therein, all of the sterecisomers, racemic mixtures and position isomers are to be considered as included. With the aid of a dye -10 -of the general formula (II), the color properties of the polysaccharide covalently bound to it can also be varied within wide limits, wherein combinations of different dyes of the general formula (II) can basically also be used to achieve a specific coloration. By at least one of the radicals R1 to R5 being an amino group and at least one of the radicals R5 to R10 being a nitro group, a par-ticularly high extinction coefficient in the visible range of the light spectrum results. With the aid of the at least one amino group, the dye can additionally be co-valently bound to a plurality of functional groups in chemically particularly simple manner. Therefore, differ-ent viscoelastic polysaccharides with correspondingly different functional groups can also quickly, simply be coupled to the dye of the general formula (II) without the requirement of costly multi-stage reactions, thereby resulting significant cost advantages.

E'urther advantages arise by the zero-shear viscosity of the aqueous solution being between 20,000 and 8,000,000 mPa.s, preferably between 50,000 and 500,000 mPa.s. Here-by, the ophthalmologic composition has a particularly good flow behavior and is correspondingly well to handle in typical applications. * .

In a further advantageous development of the invention it is provided that the molecular weight of the colored polysaccharide is between 500,000 u and 5,000,000 u, preferably between 800,000 u and 2,500,000 u. Hereby too, *..*.* * : a particularly good flow behavior and a correspondingly * * good manageability of the ophthalmologic composition is 3D achieved. In addition, plural dye molecules can be cova-lently coupled to a polysaccharide molecule without this -11 -resulting in substantial variations of the properties of the viscoelastic polysaccharide.

In a further advantageous development of the invention, it is provided that the concentration of the colored polysaccharide in the ophthalmologic composition is be-tween 0.5% and 5% (w/w), preferably between 1% and 3% (w/w). By a concentration between 0.5% and 5% (w/w), within the scope of the invention, in particular concen-trations of 0.5 %, 0.6 %, 0.7 %, 0.8 %, 0.9 %, 1.0 %, 1.1 %, 1.2 %, 1.3 %, 1.4 %, 1.5 %, 1.6 %, 1.7 %, 1.8 %, 1.9 %, 2.0 %, 2.1 %, 2.2 %, 2.3 %, 2.4 %, 2.5 %, 2.6 %, 2.7%, 2.8%, 2.9%, 3,0%, 3,1%, 3,2%, 3,3%, 3,4%, 3,5 %, 3,6 %, 3,7 %, 3,8 %, 3,9 %, 4.0 %, 4.1 %, 4.2 %, 4.3 %, 4.4 %, 4.5 %, 4.6 %, 4.7 %, 4.8 %, 4.9 %, and 5.0 % as well as corresponding intermediate values are to be understood. Therein, within the scope of the inven-tion, percentage information is to be understood as mass percent unless otherwise stated. By use of the polysac- charide in the indicated range of concentration, a par-ticularly good flow behavior and a correspondingly good manageability of the ophthalmologic composition is achieved for typical application cases.

S..... * *

In a further advantageous development of the invention, it is provided that the ophthalmologic composition is prepared for use in ophthalmology, in particular for use in a phaooemulsification method. Due to the integral, non-washable and simply adjustable coloration of the oph-S.....

* thalmologic composition, the viscoelastic solution can be **.

* introduced into the eye in a particularly simple and con-trolled manner without herein being the risk of washout of the dye or of undesired coloration of the surrounding -12 - tissue. The entire ophthalmologic composition can be re-moved from the eye without residue in correspondingly simple and safe manner for example after a cataract sur-gery.

S A further aspect of the invention relates to a method for producing an ophthalmologic composition, in which at least one dye is covalently bound to at least one water-soluble, viscoelastic polysaccharide. By the covalent binding of the at least one dye to the at least one vis-coelastic polysaccharide, it is reliably excluded that the dye diffuses into adjoining tissue and undesirably stains it during the use of the ophthalmologic composi- tion. Moreover, unlike fluorescent dyes such as fluo- rescein, rhodamine or the like, it is not required to ir-radiate the ophthalmologic composition with UV light to render it visible. This allows a substantially simpler handling of the ophthalmologic composition. In addition, there is a priori no risk that the concerned tissue is damaged by the irradiation with high-energy UV light.

Furthermore, a user -for example a surgeon -can even recognize minor traces without additional auxiliary means such as UV lamps or the like due to the integral cobra-tion of the ophthalmologic composition, whereby the use of the ophthalmologic composition according to the inven- * , 25 tion -for example within the scope of eye surgeries-ad-ditionally becomes substantially simpler and safer. In simplest configuration, the ophthalmologic composition is produced with a viscoelastic polysaccharide, to which a single dye type is covalently bound. Alternatively, it can be provided that plural different viscoelastic poly-saccharides and/or further additives are used to produce -13 -the ophthalmologic composition. Independently thereof, it can also be provided that two or more different dyes are covalently bound tD a viscoelastic polysaccharide, where-upon the colored viscoelastic polysaccharide -optionally with further additives -is used for producing the oph-thalmologic composition.

Furthermore, it can be provided that a dye with plural reactive groups is used. Hereby, the dye advantageously can be used as a cross-linker for cross-linking plural polysaccharides, whereby the viscoelastic properties of the polysaccharide can be specifically adjusted among other things. Alternatively or additionally, of course, non-colored cross-linkers such as for example divinyl suiphones or diepoxies can also be used for cross-linking for example alcohol or other functional groups of plural polysaccharides.

In a further advantageous development of the invention, it is provided that the at least one dye is used in a mo- lar ratio between 1:10 and 1:30 with regard to the poly-saccharide and/or in a concentration of between 0.0005% and 1%, preferably between 0.01 and 0.3%, based on the * weight of the ophthalmologic composition. Therein, within * the scope of the invention, by a molar ratio between 1:10 ****** * and 1:30, in particular molar ratios of 1:10, 1:11, 1:12, 1:13, 1:14, 1:15, 1:16, 1:17, 1:18, 1:19, 1:20, 1:21, 1:22, 1:23, 1:24, 1:23, 1:26, 1:27, 1:28, 1:29, and 1:30 as well as corresponding intermediate values are to be * : understood. Within the scope of the invention, by a con- * centration of between 0.0005% and 1%, in particular con-centrations of 0.0005 %, 0.001 %, 0.011 %, 0.021 %, 0.031 %, 0.041 %, 0.051 %, 0.061 %, 0.071 %, 0.081 %, -14 - 0.091 %, 0.101 %, 0.111 %, 0.121 %, 0.131 %, 0.141 %, 0.151 %, 0.161 %, 0.171 %, 0.181 %, 0.191 %, 0.201 %, 0.211 %, 0.221 %, 0.231 %, 0.241 %, 0.251 %, 0.261 %, 0.271 %, 0.281 %, 0.291 %, 0.301 %, 0.311 %, 0.321 %, 0.331 %, 0.341 %, 0.351 %, 0.361 %, 0.371 %, 0.381 %, 0.391 %, 0.401 %, 0.411 %, 0.421 %, 0.431 %, 0.441 %, 0.451 %, 0.461 %, 0.471 %, 0.481 %, 0.491 %, 0.501 %, 0.511 %, 0.521 %, 0.531 %, 0.541 %, 0.551 %, 0.561 %, 0.571 %, 0.581 %, 0.591 %, 0.601 %, 0.611 %, 0.621 %, 0.631 %, 0.641 %, 0.651 %, 0.661 %, 0.671 %, 0.681 %, 0.691 %, 0.701 %, 0.711 %, 0.721 %, 0.731 %, 0.741 %, 0.751 %, 0.761 %, 0.771 %, 0.781 %, 0.791 %, 0.801 %, 0.Bll %, 0.821 %, 0.831 %, 0.841 %, 0.851 %, 0.861 %, 0.871 %, 0.881 %, 0.891 %, 0.901 %, 0.911 %, 0.921 %, 0.931 %, 0.941 %, 0.951 %, 0.961 %, 0.971 %, 0.981 %, 0.991 %, and 1.000 % as well as corresponding intermedi-ate values are to be understood. By the use of the dye in the specified molar ratios and/or concentrations, the ophthalmologic composition has an optimum colorfulness for typical applications.

In a further advantageous development of the invention, it is provided that a dye with an amino group is used and covalently bound to the polysaccharide by way of an Ugi reaction. The Ugi reaction is a multi-component reaction * * 25 involving a ketone or aldehyde, an amine, an isocyanide and a carboxylic acid. The products of the Ugi reaction are bis-arnides. The Ugi reaction is an uncatalyzed reac-tion but is usually complete within minutes after adding :4 the isocyanide. The main advantages of using an Ugi reac- tion are the inherent high atom economy as only a mole- cule of water is lost and the high product yields. Ac- -15 -cordingly, the dye including the amino group can be bound quickly, simply and with high yields to polysacoharides comprising functional keto groups, aldehyde groups and/or carboxylic acid groups. An example for a suitable poly- S saccharide is hyaluronic acid. Hyaluronic acid is a poly- saccharide with repeating disaccharide units of glu- curonosyl---l,3-N-acety1glucosamine linked by 3-1,4-glycoside (glycosidic) bonds and has the general formula O321/

OH

In a further advantageous development of the invention, it is provided that for producing the ophthalmologic com-position the polysaccharide coupled to the dye and/or a physiologically acceptable salt thereof is dissolved and/or dispersed in an amount of between 0.05 percent by weight and 5 percent by weight possibly together with a * . buffer system and/or with further agents and/or excipi-S.....

* . ents in a protic solvent, in particular in water. Within the scope of the invention, by an amount cf between 0.05 percent by weight and 5 percent by weight, in particular amounts of 0.05 %, 0.15 %, 0.25 %, 0.35 %, 0.45 %, *r 0.55 %, 0.65 %, 0.75 %, 0.85 %, 0.95 %, 1.05 %, 1.15 %, *:" 1.25 %, 1.35 %, 1.45 %, 1.55 %, 1.65 %, 1.75 %, 1.85 %, 1.95 %, 2.05 %, 2.15 %, 2.25 %, 2.35 %, 2.45 %, 2.55 %, 2.65 %, 2.75 %, 2.85 %, 2.95 %, 3.05 %, 3.15 %, 3.25 %, -16 - 3.35 %, 3.45 %, 3.55 %, 3.65 %, 3.75 %, 3.85 %, 3.95 %, 4.05 °, 4.15 %, 4.25 %, 4-35 %, 4.45 %, 4.55 %, 4.65, 4.75 %, 4.85 %, 4.95 %, and 5.00 % as well as correspond-ing intermediate values are to be understood. Hereby, the property profile of the ophthalmologic composition can be adapted to different purposes of application in optimum manner.

In a further advantageous development of the invention, it is provided that in order to bond the at least one dye, the viscoelastic polysaccharide is functionalized and the at least one dye is covalently bound to the added functional group of the polysacchar±de. In other words, first, a functional group is introduced into the viscoe-lastic polysaccharide, which is subsequently reacted with the at least one dye, in order to covalently bind the at least one dye to the polysaccharide. In this manner, the at least one dye can be simply bound to different poly-saccharides with correspondingly specific functional groups largely independent of its precisely present reac- tive group. Alternatively or additionally, it can be pro-vided that the at least one dye is first functionalized and subsequently covalently bound to the viscoelastic polysaccharide via its newly created functional group by means of a desired type of reaction.

Further features of the invention appear from the claims as well as based on the following embodiments. The fea- tures and feature combinations mentioned above in the de- * scription as well as the features and feature combina-4t**** * tions mentioned below in the embodiments are usable not only in the respectively specified combination, but also -17 -in other combinations without departing from the scope of the invention.

Preferred embodiments of the invention The present invention generally provides a colored oph-thalmologic composition which can be completely removed after cataract surgery. The ophthalmologic composition comprises a viscoelastic solution based on a polysaccha-ride to which at least one dye is covalently bound. This means for example that a surgeon can visualize even very small traces of the viscoelastic ophthalmologic composi- tion. By way of covalently binding the dye to the poly- saccharide it is also ensured that the dye does not dif-fuse out of the solution and therefore does not stain the surrounding tissues.

The ophthalmologic composition according to the invention can for example be used as eye drops. The viscoelastic property of the solution of the color labeled polysaccha-ride therein provides for a stable and long lasting tear film. Alternatively, it can be provided that the ophthal-mologic composition according to the invention is used as caring solutions for contact lenses. Hereby, uncolored or * colored contact lenses can be better recognized and sim- * pler be removed from the contact lens container. * *.

In the ophthalmic surgery, the ophthalmologic composition according to the invention can be used for tilling up the vitreous body as well as for stabilizing the anterior chamber and for protection of the highly sensible endo-*.***.

* thelial cell layer of the cornea during surgery on the anterior eye portions, especially the surgery of the cat-aract. As the viscoelastic polysaccharide, to which the -18 -the dye is covalently bound, for example hyaluronic acid is suited.

Suitable dyes that can be covalently bound to hyaluronic acid or another polysaccharide are based on aminoanthra-cenedione derivatives of the general formula (I) wherein in the general formula (I) at least one of the substituents R1 to R8 is an amino group, and the remain-ing substituents R1 to R8, which are different from the amino group, are selected from; hydrogen, halogen, Ci-C4-alkyl, 01-04-alkoxy, C1-C4-alkoxy- C1-C4-alkyi, 01-C4-alkyl-01-04-alkoxy, Ci-C4-alkoxy-01-04-alkoxy, aryloxy, acetamide, propionamide, butyramide, isobutyramide, suiphonate, (aminophenyl) -N- is alkylacetamide, (aminophenylsulphonyl)alkyl sulphate, al-kylbenzenesulphonamide, tri(alkyl)benzenamine, hydroxy, (alkylsulphonyl)benzenamine, and (alkenylsul-phonyl)benzenamine, and/or wherein at least two adjacent substituents R, R+1 with x'=l to 3 and/or 5 to 7 form a 3-, 4-, 5-, 6-or 7- membered homocyclic or heterocyclic, unsaturated or aro-matic radical.

-19 -Alternatively or additionally a reactive dye from the group of ( (nitrophenyl)diazenyl)benzenamine derivatives having the general formula (II) (II) may be covalently bound to the polysaccharide, wherein in the general formula (II) at least one of the substituents R1 to R5 is an amino group and at least one of the sub- stituents R6 to Rio is a nitro group, and wherein the re-maining radicals Rj to R10, which are different from the amino group and the nitro group, are selected from: hydrogen, halogen, Ci-C4-alkyl, C1-04-alkoxy, 01-C4-alkoxy- C1-C4-alkyl, 01-04-alkyl-01-C4-alkoxy, 01-C4--alkoxy--01-04-alkoxy, aryloxy, acetamide, propionamide, butyramide, isobutyramide, sulphonate, (aminophenyl) -N- alkylacetamide, (arninophenylsulphonyl)alkyl sulphate, al-kylbenzenesulphonamide, tn (alkyl) benzenamine, hydroxy, (alkylsulphonyl)benzenamine, and (alkenylsul- * phonyl)benzeriamine, and/or * ti, * . 0 * wherein at least two adjacent substituents R, R,1 with 23 y=l to 4 and/or 6 to 9 form the cyclic radical * to R) with: -20 -Rhydrogen, halogen, amino, hydroxyl, nitro, C1-C4-alkyl, Cj-04-al koxy, C1-04-alkoxy-C1--C4-alkyl, Ci-C4-alkyl--C1-C4--alkoxy, and/or Ci-C4--alkoxy-ci-04-alkoxy.

Since both the dyes of the general formula (I) and the dyes of the general formula (II) have free amino groups, they can be bound to a polysaccharide having free keto, aldehyde and/or carboxyl groups without further derivati-zation or functionalization with the aid of the so-called [Jgi reaction. The Ugi reaction is a 4 component condensa- tion (U-4CC) , which allows the synthesis of a- aminoacylamide derivatives from aldehydes, amines, car-boxylic acids and isocyanides. Therein, the Ugi reaction proceeds according to the general reaction scheme: 0 N' + R P c 0 P H PXH + ÷ R'OH r In the specific case, thus, the dyes of the general for-mulas (I) und (II) function as the amino component and the hyaluronic acid functions as the carboxylic acid corn-ponent of the Ugi reaction. For example, acetaldehyde can * be used as the aldehyde. For example, cyclohexanenitrile is suitable as the isocyanide. Generally, low molecular alcohols have proven as the solvent in the Ugi reaction.

S

* Preferably, the reactants are prepared in high concentra-I*tst * tions with cooling and react very fast, that is within a few minutes at room temperature. Lewis acids known per se -2]. - can be employed in the conversion in particular of steri-cally demanding educts for catalysis.

In the case of hyaluronic acid, which has the general formula

OH

the dyes of the general formulas (I) and (II) thus can react with the free carboxyl groups of the hyaluronic ac-id with the aid of their amino groups in a reaction step and hereby be covalently bound to the hyaluronic acid.

The carboxylic acids employed within the scope of an tigi reaction can advantageously be diversely varied such that other polysaccharides with free carboxyl groups can also be used without previous functionalization within the scope of an ugI reaction. Polysaccharides without car- is boxyl groups can be subjected to a preceding derivatiza-tion or functionalization for later use in the tJgi reaction. Conversely, polysaccharides with free amino *:** groups can of course also be reacted with dyes having free carboxyl groups.

An advantage of the dyes of the general formulas (I) and (II) is in their basically very high extinction coeffi-0ie' * dent in the visible range of the light spectrum (about * U...

* * 400 nm to about 800 nm) . Furthermore, the color of the dyes of the general formulas (I) and (II) can be simply varied in a wide color range by use of corresponding sub- -22 - stituents and/or position isomers, whereby the ophthalmo- logic composition can be simply adapted to different pur-poses of use.

In the following table 1, various embodiments for dyes of the general formula (I) are specified.

Table 1: Embodiments for dyes of the general formula (I)

Example 1 0 NH2

jjj1S0aNa o I-IN nti Name and sodium l-amino-4-(4-(N-methyl- Synonyms acetamido)pheriylamino)-9, lO-dioxo-9, 10-dihydroanthracene-2-sulfonate; Acid Blue 41; GAS 2666-17-3

Example 2 0 NH2

SO N a

B I o * *0*** * *

* * Name and sodium 2-(3-(4-amino-9,lO-dicxo-3-sulfonato- *:*::* Synonyms 9, 10-dihydroanthracen-l- ylamino)phenylsulfonyl)ethyl sulphate; Reac-tive Blue 19 (Remazol Brilliant Blue R) ; GAS 2580-78-1 S.. * 0

-23 -

Example 3 0 NH2 cc

0 HNI, Name and N-(4-amino-3-methoxy-9, 10-dioxo-9, 10- Synonyms dihydroanthracen-l-yl) -4-methylbenzenesulfonamide; Disperse Red 86; CAS 81-68-5

Example 4 0 NH2

Name and 1-amtno-4-hydroxy-2-(2- Synonyms methoxyethoxy) anthracene-9, lO-dione; Dis-perse Red 59; CAS 17869-10-2

Example 5 0 NH2

crJcIS03N3 o HNxL * . Name and sodium l-amino-4-(mesitylamino)-9,10-dioxo-Synonyms 9, 10-dihydroanthracene-2-sulfonate; Acid Blue 129; GAS 6397-02-0

* Example 6 0 NH2

*** ..* * * **..* * S --0 OH -24 - Name and 1-amino-4-hydroxyanthracene-9,10-dione; Dis-Synonyms perse Red 15; CAS 116-85-8

Example 7 0 NH2 cot

Synonyms 1-amino-4-hydroxy-2-phenoxyanthracene-9, 10-dione; Disperse Red 60; CAS 17418-58-5

Example 8 0 NH2

SO3Na+ Name and sodium l-amino--9,.10-dioxo--4-(3- Synonyms (vinylsulfonyl)phenylamino)-9, 10- dihydroanthracene-2-sulfonate; UniBlue A So-dium Salt; CAS 14541-90-3 In the following table 2, various embodiments for dyes of the general formula (II) are specified. * .

Table 2: Embodiments for dyes of the general formula (II)

Example 1 -o

)N \\ NH2 * * Name and (E)-4-((4-nitrophenyl)diazenyl)benzenamine; Synonyms Disperse Orange 3; CAS 730-40-5 -25 -

Example 2 CI H2

Name and (E)-4-((2--chjoro-4- Synonyms nitroptienyl)diazenyl)benzenarnine; 4-[(2- chloro-4-nitrophenyl)azo]aniline; CAS 52735-98-S

Example 3

Name and (E)-2,5-dirnethyl-4-((4- Synonyms nitrophenyl)diazenyl)benzenamine; 4-[ (4-nitrophenyl) azoJ -2, 5-xylidine; GAS 6492-50-8

Example 4 -0

Name and (E)-2-methoxy-4-((4- Synonyms nitrophenyl)diazenyl)benzenamine; 4-[ (4-nitrophenyl)azo]-o-anisidine; GAS 101-52-0

* Example 5 -o -

* .* + P \ / * * rH Name and (E)-3-methoxy-4-((3-methyl-4- Synonyms nitrophenyl)diazeny]jbenzenam±ne; 4-Amino-2-methoxy-3 -methyl-4-nitroazobenzene; GAS -26 - 144829-57-2

Example 6 -o \

N //

o N-_(,_NH2 Name and (E) -2-methoxy-5-methyl-4-((4-Synonyms nitrophenyl) diazenyl) benzenamine; Disperse Red 31; CAS 2475-43-6

Example 7 -o

HN

Name and N-[5-arnino-2-[(p- Synonyms nitrophenyl)azo]phenyl]acetamide; CAS 26311-09-1

Example 8 -o

N 1/

* a * * ..

* . Name and (E)-2-methyi-4-((4- * . Synonyms nitrophenyl)diazenyl)benzenamine; 4-[(4-nitrophenyl)azo}-o-toluidine; CAS 84255-13-0 * a * a *a

Example 9 -o

N 1;

a..... * a a

* a.... a

-27 - Name and (E)-2,5-dimethoxy-4-((4- Synonyms nitrophenyl)diazenyl)benzenamine; 2,5-dimethoxy-4--(4-nitrophenylazo)aniline; CAS 6358-51-6

Example 10 -o -H2N d

U

Name and (E)-l-((4-nitrophenyl)diazenyl)naphthalene- Synonyms 2-amine; 1-(4' -Nitrophenylazo) -2-naphthylamine; Solvent Red 5; CAS 3025-77-2

Example 11

OH

Name and (E)-4-amino-3-((4-Synonyms nitrophenyl) diazenyl) naphthalene-l-sulphonic acid; Acid Red 74; CAS 6300-18-1 *:": However, it is to be emphasized that besides the Ugi re- : action, basically, other chemical reaction types can also be used for covalently binding the concerned dye to the concerned viscoelastic polysaccharide. For example, dyes with a free alcohol group can be bound to hyaluronic acid by an esterification reaction. *0s

Labeling of hyaluronic acid (HA) can for example also be obtained by the reaction of CNBr-activated HA at vicinal -28 - diols and a dye comprising a reactive amine group accord-ing to the following reaction scheme:

HA HA HA NH

LOH CNBr L0 NH2-R ____ ___ OH L0>NH LQH

HUH R

wherein HA denotes hyaluronic acid and NH2-R denotes the S reactive dye comprising an amine group. The reaction may also involve the hydroxyl groups in two hyaluronic acid molecules.

It is also possible to use a dye comprising an isothiocy-anate group (denoted by SCN-R) , which targets mostly the primary hydroxyl groups of HA or other viscoelastic poly-saccharides: S..... * . S

SCN-R + HA-CH2-OH NH * **

R

* The reaction is conducted at 37°C overnight. The result-S.....

* * 15 ing color labeled polysaccharide is stable for over a month.

-29 -A further type of reaction for covalently binding a dye to hyaluronic acid includes activation of a hyaluronic acid derivative with N-hydroxysuccinimide coupled di-phenylphosphoric acid. The resulting adduct is reacted with a dye comprising a reactive hydrazide group (denoted by [R]CO(NN)=O) according to the following general reac-tion scheme: N-O-F 0 RyNNH2 H HA-COO-N(nBu)4 r HA 0 HA Another method to chemically bond a reactive dye to hya-luronic acid (HA) or other polysaccharides comprising carboxylic groups is depicted in the following reaction scheme: NHR1 0 H2N HA-cOOH R1N=c=NR2 (EDC),JJJ, R HAfX + RlH)NHR2 The reaction employs a dye comprising a hydrazide group.

*:*$* The first step is to form an 0-acylurea of HA by reaction of HA with l-ethyl-3-(3-dimethylantinopropyl)carbodiimide (EDO) having the formula -30 - The resulting intermediate is then reacted with the reac-tive dye to form the color labeled HA.

Another possibility is the reaction between the carbox-ylic groups of HA and dyes comprising a carbodiimide group in the presence or absence of primary amines at pH 4.5 according to the general reaction scheme: $ 0 0 R1NCS Hg0 HACOOH II II R-NH2 -RJ_R...N..RI -R-N=C=N-R1 *

H H H

The reaction of carbodiimides proceeds rapidly at room temperature. The final group is an N-acylurea or 0-acylurea. The carbodiimides are synthesized in a 2-step reaction: the reaction of a dye comprising a primary amine and isothiocyanate to form a thiourea and the reac-tion of said thiourea and HgO to produce the reactive dye comprising a carbodiimide group. * * * *

*:c* HA esters can also be produced by a nucleophilic reaction of a reactive dye comprising a halide with a quaternary salt of hyaluronic acid: S.... * .

S

* * II HA-CQO NRf+ Br-R DMSO -31 - Another way to molecularly label the viscoelastic poly- saccharide is given by the cross-linking of HA with di-epoxies: OH -H' 0 OH

- HA

In acidic conditions, the ester of HA is formed by link- ing the epoxy to HA at the acidic position. In basic con-ditions, the ether is produced by linking the epoxy to HA at the primary alcohol position.

Another way to crosslink and therefore functionalize HA at the primary alcohol position is to use divinylsulfone as cross-linker: THA-CH2OH + _________ Alternatively, it can be provided herein that a dye with a free hydroxy group is used and bound to the hyaluronic acid with the aid of the cross-linker. * .. * * * **

Alternatively or additionally to hyaluronic acid, basi- cally, other viscoelastic polysaccharides such as for ex-ample hydroxypropylmethylcellulose, chondroitin sulfate or mixtures thereof can also be used.

-32 - An ophthalmologic composition comprising an aqueous solu-tion of a viscoelastic polysaccharide which is covalently bound to one or several dyes of the general formulas (I) and/or (II) may be used in a phacoemulsification method.

During the phacoemulsification, the anterior chamber of the eye is filled with the ophthalmologic composition comprising the colored viscoelastic solution. The viscoe-lastic ophthalmologic composition is used as a surgical aid to protect intraocular tissues, for example the cor-neal endothelium, during the phacoemulsification, as a space maintainer to maintain the anterior chamber of the eye, and to facilitate intraocular maneuvers, for example to make a controlled capsulorhexis. Because the dye is covaleritly bound to the polysaccharide, the dye does not diffuse out of the solution and therefore does not stain the surrounding tissues.

After the capsulorhexis the colored ophthalmologic compo- sition may be completely removed from the anterior cham-ber as the surgeon can visualize even very small traces of the viscoelastic ophthalmologic composition.

* . . *0 * * * * * ** * . S * *S

S * *.* * * *

* ** S..

S

Claims

<claim-text>-33 -Carl Zeiss Meditec AG Attorney's file: 42571/42571GB 09/08/2 011 Claims 1. Ophthalmologic composition, comprising an aqueous so-lution of at least one viscoelastic polysaccharide, characterized in that the at least one polysaccharide is covalently bound to at least one dye.</claim-text> <claim-text>2. The ophthalmologic composition according to claim 1, characterized in that the polysaccharide is cellulose, a cellulose ether with methyl and/or ethyl and/or propyl groups, in particular hydroxypropyl methylcellulose, hy-droxyethyl methylcellulose and/or methylcellulose, a glycosaminoglycari, in particular hyaluronic acid, chondroitin sulphate, dermatan sulphate, heparin, *t-.-.* * 20 heparan sulphate, keratan sulphate, alginic acid, po-lymannuronic acid, polyguluronic acid, polyglucuronic *:c-acid, amylose, amylopectin, callose, galactcmannan, dextran, xanthan and/or a mixture thereof. * * * 25</claim-text> <claim-text>3. The ophthalmologic composition according to claim 1 or 2, -34 -characterized in that the at least one dye is bound to the polysaccharide directly and/or via a spacer.</claim-text> <claim-text>4. The ophthalmologic composition according to any one of claims 1 to 3, characterized in that the at least one polysaccharide is obtainable by a reaction of the polysaccharide with at least one re- active dye from the group of modified and/or unmodi-fied aminoarithracenedione and/or of modified and/or unmodified nitrophenyldiazenylbenzenamine.</claim-text> <claim-text>5. The ophthalmologic composition according to claim 4, characterized in that the at least one polysaccharide is obtainable by re-action with at least one reactive dye of the general formula (I)I U I 0 (I)wherein in the general formula (I) at least one of * : the substituents R1 to P8 is an amino group, and the * * remaining substituents R1 to R8, which are different from the amino group: -35 - -are selected from: hydrogen, halogen, C1-C4-alkyl, C1-C1--alkoxy, C1-C4-alkoxy--C1-04--alkyl, Ci-C4-alkyl-Ci-C4-alkoxy, Ci-C4-alkoxy-Ci-C- alkoxy, aryloxy, acetamide, propionamide, bu- S tyramide, isobutyramide, sulphonate, (amino- phenyl) -N-alkylacetamide, (aminophenylsul- phonyl) alkyl sulphate, alkylbenzenesulphona- mide, tri(alkyl)benzenamine, hydroxy, (alkyl- sulphonyl)benzenamine, and (alkenylsul-phonyl)benzenarnine; and/or -wherein at least two adjacent substituents Rx, R,,1 with x=l to 3 and/or 5 to 7 form a 3-, 4-, 5-, 6-or 7-membered homocyclic or heterocyc-lic, unsaturated or aromatic radical.6. The ophthalmologic composition according to claim 4 or 5, characterized in that the at least one polysaccharide is obtainable by re-action with at least one reactive dye of the general * * formula (II) *****. * *</claim-text> <claim-text>1% (II) -36 -wherein in the general formula (II) at least one of the substituents R1 to R5 is an amino group and at least one of the substituents R5 to R10 is a nitro group, and wherein the remaining radicals R1 to R10, S which are different from the amino group and the nit-tro group: -are selected from hydrogen, halogen, 01-04-alkyl, C1-04-alkoxy, C1-C4-alkoxy-C1-04--alkyl C1-04-alkyl-C1-C4-alkoxy, 01-C4-alkoxy-C1--C4- alkoxy, aryloxy, acetamide, propionamide, bu- tyramide, isobutyramide, suiphonate, (amino- phenyl) -N--alkylacetamide, (aminophenylsul- phonyl)alkyl sulphate, alkylbenzenesulphona- nide, tri(alkyl)benzenamine, hydroxy, (alkyl- sulphonyl)benzenamine, and (alkenylsul-phonyl)benzenamine; and/or -wherein at least two adjacent substituents R, R+1 with y=l to 4 and/or 6 to 9 form the cy-clic radical * * 0 0.*000 * * with: R=hydrogen, halogen, amino, hydroxyl, nitro, C1-C4-alkyl, Ct-04-alkoxy, 01-04-alkoxy-01--04- alkyl, C1-C4-alkyl-01-04-alkoxy, and/or C1-C4- * 25 alkoxy-C1-04-alkoxy. 0.0.0 * 0</claim-text> <claim-text>-37 - 7. The ophthalmologic composition according to any one of claims 1 to 6, characterized in that the zero-shear viscosity of the aqueous solution is $ between 20,000 and 8,000,000 mPa.s, preferably be-tween 50,000 and 500,000 mPa.s.</claim-text> <claim-text>B. The ophthalmologic composition according to any one of claims 1 to 7, characterized in that the molecular weight of the colored polysaccharide is between 500,000 u and 5,000,000 u, preferably between 800,000 u and 2,500,000 u.</claim-text> <claim-text>9. The ophthalmologic composition according to any one of claims 1 to 8, characterized in that the concentration of the colored polysaccharide in the ophthalmoloqic composition is between 0.5% and 5% (w/w), preferably between 1% and 3% (w/w) S..... * Stie....* 10. The ophthalmologic composition according to any one of claims 1 to 9, characterized in that it is prepared for use in ophthalmology, in particu- * lar for use in a phacoemulsification method. S5..S</claim-text> <claim-text>S</claim-text> <claim-text>-38 - 11. A method for producing an ophthalmologic composition, in which at least one dye is covalently bound to at least one water-soluble, viscoelastic polysaccharide.</claim-text> <claim-text>12. The method according to claim 11, characterized in that the at least one dye is used in a molar ratio between 1:10 and 1:30 with regard to the polysaccharide and/or in a concentration of between 0.0005% and 1%, preferably between 0.01 and 0.3%, based on the weight of the ophthalmologic composition.</claim-text> <claim-text>13. The method according to claim 11 or 12, characterized in that a dye with an amino group is used and covalently bound to the polysaccharide by way of an Ugi reac-tion.</claim-text> <claim-text>14. The method according to any one of claims 11 to 13, characterized in that S.....for producing the ophthalmologic composition the polysaccharide coupled to the dye and/or a physio-logically acceptable salt thereof is dissolved and/or dispersed in an amount of between 0,05 percent by weight and 5 percent by weight possibly together with a buffer system and/or with further agents and/or ex-dfl t* cipients in a protic solvent, in particular in water.-39 - 15. The method according to any one of claims 11 to 14, characterized in that in order to bond the at least one dye, the viscoelas-tic polysaccharide is functionalized and the at least one dye is covalently bound to the added functional group of the polysaccharide.*Ifl.. * *Iale ISI * * * a. * * a * U.a..-.. * .*1111* * )</claim-text>