"Stable liquid composition of chondroitinsulfate and glucosamine"

FIELD OF THE INVENTION

The present invention concerns a stable liquid composition comprising a glycosaminoglycan and/or optionally N-acetylated glucosamine and a cyclodextrin for oral administration. More particularly, it is an object of the present invention to provide a formulation consisting of a solution containing, as active ingredients thereof, chondroitinsulfate and glucosamine or chondroitinsulfate alone or glucosamine alone, useful for the prevention or of the slowdown of arthrosis, with a higher stability in respect of formulation already existing on the market. BACKGROUND OF THE INVENTION

Glycosaminoglycans are a family of polysaccharides each formed by a mixture of chains wherein each chain is a compound consisting of a repetitive sequence of a uronic acid-hexosamine disaccaride, said uronic acid unit, glucuronic acid or iduronic acid, being linked in α 1— »4 or β l-»3 to said hexosamine unit, glucosamine or galactosamine, and said uronic acid and hexosamine units being sulfated at different extent.

Beside heparin and heparan sulfate, also dermatan sulfate, hyaluronic acid and chondroitinsulfate belong to the glycosaminoglycans' family. Physiologically, glycosaminoglycans are organized in proteoglycans formed by a protein core wherein the glycosaminoglycan is linked by a linkage region. These structures are in charge of the control of biochemical reactions by the uptake and the release of proteins and growth factors (J.F. Kennedy, C. A. White, Bioactive Carbohydrates, 1983, Ellis Horwood Ltd, 211-227). In particular, chondroitinsulfate is located in cartilages to give them elasticity and to control their resistance. In addition, it has an inhibiting effect on collagenase and elastase, which are cartilage depolymerizing enzymes. In a healthy subject, the cartilage destruction and reconstruction processes are in equilibrium but, when the cartilage destruction process is more rapid than the reconstruction one, a pathological condition occurs, for example arthrosis or arthritis.

In nature, chondroitinsulfate exists in two structural forms: chondroitin-4- sulfate (or

ChSA) and chondroitin-6-sulfate (or ChSC). It consists of a mixture of chains wherein each chain is a compound (molecule) showing a repetitive sequence of a glucuronic acid-galactosamine disaccharide wherein the glucuronic acid is linked to galactosamine by a β 1— »3 bond, said galactosamine being 4-0-sulfated (ChSA) or 6-0-sulfated (ChSC). hi the same molecule, clusters of ChSA and ChSC coexist. The proteoglycans containing chondroitinsulfate are formed by a hyaluronic acid skeleton onto which protein chains (protein core) bearing branches of condroitisulfate chains are grafted. A highly viscous macromolecule is thus formed whose viscosity is due to the very high molecular mass and also to the structural characteristics of its components.

Glucosamine is the monosaccharide starter of the biosynthesis of the glycosaminoglycans and, hence, its presence in a high amount shifts the equilibrium of the cartilage metabolism toward the biosynthesis. Unless otherwise specified, in the present description the term "chondroitinsulfate" designates chondroitinsulfate in sodium salt form or of another of its salts, the dosages given herein below being referred to sodium chondroitinsulfate. The term "glucosamine" designates the D-glucosamine (2-amino-2-deoxy-D-glucose), as such or in form of an acid addition salt thereof, the dosages given herein below being referred to glucosamine base. PRIQR ART

It is known that chondroitinsulfate is used in various forms as integrator in the prevention of arthrosis and in the treatment of different pathologic conditions of cartilage joints, such as inflammatory processes and osteoarthrosis, as described for example in EP 704216. Pharmaceutical or "nutraceutical" compositions containing chondroitinsulfate in association with glucosamine for their use in the protection and reparation of connective tissue are described in US 5,364,845 and US 5,587,363. Pharmaceutical forms for the oral administration of chondroitinsulfate, in tablets, capsules granules and solutions are already existing on the market. Among said forms, the solution is the most immediately available additive for the oral administration in the prevention of arthrosis. However, an aqueous solution of

chondroitinsulfate tends to turn yellow, even in the presence of preservatives. As a consequence, a pharmaceutical or "nutraceutical" composition in form of an aqueous solution of chondroitinsulfate becomes unpleasant during the time, even though the loss of activity is not appreciable. Thus, there is a need of avoiding the coloring of such solutions, which are in form of a gel and whose viscosity is due to the macromolecular nature of chondroitinsulfate, in order to improve their organoleptic character and to assure the maximal activity of all of their components at the moment of use. Glucosamine too, as free base or in form of one of its salts such as the hydrochloride or the sulfate, is not very stable in aqueous solution and tends to give a yellow coloration very probably due to oxidation products.

Li order to avoid the formation of such oxidation products and to keep the formation of moulds and bacteria under control, known preservatives such as sodium benzoate, parabens, ethylenediaminotetracetic acid (EDTA), as such or as sodium salt, or potassium sorbate are added to the oral liquid preparation of chondroitinsulfate and/or glucosamine.

Moreover, in order to avoid the oxidative degradation, antioxidants such as sodium metabisulfite or sodium sulfite are added to aqueous solutions containig chondroitinsulfate and/or glucosamine. However, all these substances do not allow an effective conservation of the solutions during the time and the inhibition of the formation of the yellow color. The above-mentioned EP 704216 document, for example, discloses a gel formulation containing 1.2 g of an organic or inorganic chondroitinsulfate salt in admixture with potassium sorbate and sodium benzoate as preservatives. Among the organic salts, the glucosamine salt is also mentioned. It has been observed that the above- mentioned preservatives in such a composition do not preserve the components from oxidation, all the more in the presence of glucosamine.

The use of cyclodextrins, for example α-cyclodextrin, β-cyclodextrin, γ-cyclodextrin, hydroxypropyl-β-cyclodextrin, hydroxypropyl-γ-cyclodextrin for solubilizing compounds which are not water soluble or for avoiding incompatibility between the various components of a solution is well known in the literature.

Document US 2005/0256083 discloses a pharmaceutical ophthalmic composition comprising a cyclodextrin, a guanidine cationic compound and sorbic acid. In addition, it cites a series of documents wherein the use of cyclodextrins is reviewed. Among these cited documents, in particular, JP 1016728 discloses an ophthalmic aqueous solution containing a drug, for example sodium hyaluronate, chondroitinsulfate or lysozyme chloride, an antiseptic cationic surface-active agent, such as benzalconium chloride or chlorhexidine hydrochloride, and a cyclodextrin or a derivative thereof. In this case, the cyclodextrin serves to inhibit clouding and precipitation. SUMMARY OF THE INVENTION

It has now been found that the addition of a cyclodextrin to a solution of chondroitinsulfate, of glucosamine, of antioxidants such as sodium metabisulfite and of preservatives such as sodium benzoate or potassium sorbate allows the concurrent presence of the two active principles, of the antioxidants and of the preservatives in a solution which does not tend to yellow and that remains intact for a longer time. hi particular, it has been found that cyclodextrins complex for example sodium benzoate (as described by SDMER E. et al. Thermochimica acta , 1987, vol. 116, pp. 249-256), which is a preferred preservative, and concurrently also favorably interact with glucosamine by including it in their skeleton (as also described by SAKAIRI N. et al. J. Chem. Soc. Chem. Commun, 1991, n. 5, pp. 289-290). DETAILED DESCRIPTION

It is an object of the present invention to provide a composition for oral administration in form of an aqueous solution which comprises

(a) chondroitinsulfate at a concentration of from 5% to 15%, or optionally N- acetylated glucosamine at a concentration of from 6% to 20%, or a mixture of said components, each at the above defined concentration;

(b) a cyclodextrin, at a concentration of from 4% to 10%;

(c) antioxidants at a total concentration of from 0.2 to 0.4%;

(d) preservatives at a total concentration of from 0.1% to 0.3%. In the present formulation, the chondroitinsulfate will be in form of an alkaline metal or alkaline-earth metal salt thereof such as a Na, K, Mg or Ca salt and glucosamine

will be in free amine form or as an acid addition salt thereof such as the sulfate or the hydrochloride salt. N-acetyl-D-glucosamine is used as such.

Beside the essential components (a)-(d) above, the composition of the present invention may also contain sweetening or flavoring agents destined to the optimization of the organoleptic characteristics of the oral formulation. Said composition may be a pharmaceutical or nutraceutical (as defined in US 5,364,845 and US 5,587,363) composition.

The sweetening agents may be natural, optional reduced sugar such as sucrose, dextrose, xylitol, mannitol or sorbitol, or synthetic product such as sodium saccharine or aspartame. Synthetic sweeteners may be present in a percentage of from 0.1 to 5% while the natural, optional reduced sugar may be present in a percentage of from 10% to 20%, preferably of from 15% to 20%. The flavoring agents will be chosen by the expert in the field in relation to the organoleptic properties of the composition. The flavoring agents may be selected from the group consisting of pharmaceutically or nutritionally acceptable flavors and tastes of synthetic and natural oils, the latter extracted from plants, leaves, flowers, fruits and their combinations, such as cinnamon, peppermint, anise and citron leaves, bitter almond, citrus fruits, in particular orange and/or lemon, linden and grapefruit oils. Also chocolate, vanilla or eucalyptus flavor and essences of fruit, in particular apple, pear, peach, strawberry, cherry, apricot, orange, lemon and grapes may be advantageously used. Preferred flavoring agents are those giving mint or fruit, such as grape, cherry or citrus, in particular orange and lemon, flavors or mixtures thereof. Flavoring agents, usually supported on a solid matrix, are generally present at a concentration of from 0.5 to 1.5%. The compositions of the present invention may also contain thickening agents, such as microcrystalline cellulose, hydroxypropylcellulose, carboxymethylcellulose or polyvinylpyrrolidone, in a total amount of from 0.1% to 0.2%. The liquid compositions of the present invention are aqueous solutions, preferably in dosage unit forms, each containing (a) chondroitinsulfate in an amount of from 400 mg to 1500 mg and/or optionally N- acetylated glucosamine in an amount of from 400 to 1500 mg;

(b) cyclodextrin, which may be natural, such as α-cyclodextrin, β-cyclodextrin and γ-cyclodextrin, or modified, such as hydroxypropyl-β-cyclodextrin, hydroxypropyl-α-cyclodextrin, hydroxypropyl-γ-cyclodextrin, methyl-β- cyclodextrin or sulfobutyl-β-cyclodextrin, in an amount of from 100 to 1000 mg; (c) antioxidants, such as sodium metabisulfite or sodium sulfite, in a total amount of from 3 to 35 mg; and

(d) preservatives, such as sodium benzoate, sorbic acid and its salts, in particular potassium sorbate, EDTA or a salt thereof, in particular sodium EDTA, p- hydroxybenzoic acid esters (parabens), such as methylparaben and ethylparaben, in a total amount of from 2 mg to 20 mg; in aqueous solution in admixture with common excipients.

The dosage unit preferably consists of an amount of from 4 to 30 g of preferably deionized water containing the components (a)-(d), sweetening and flavoring agents and, optionally, thickening agents. In said dosage units, the sweetening agents such as those mentioned above may be present in an amount of from 400 mg to 3 g, preferably from 600 mg to 3 g per dosage unit. The flavoring agents may be present in the compositions of the present invention in an amount of from 20 mg to 150 mg of flavoring agent per dosage unit. The optional thickening agents such as those mentioned above may be present in an amount of from 10 mg to 30 mg per dosage unit.

A particularly advantageous compositions comprises

(a) chondroitinsulfate and/or optionally N-acetylated glucosamine at a dosage of from 500 mg to 1200 mg and/or from 500 mg to 1500 mg, respectively;

(b) β-cyclodextrin, hydroxypropyl-β-cyclodextrin or sulfobutyl-β-cyclodextrin; (c) sodium metabisulfite as antioxidant;

(d) preservatives selected from the group consisting of sodium benzoate, potassium benzoate, potassium sorbate and sodium EDTA; in a unit dose consisting of an amount of from 4 g to 30 g of aqueous solution containing said components (a)-(d), sweetening agents and flavoring agents. The unit doses may be in form of welded sachets, little bottles in glass or plastics or any other container rendered available by the current technology, including

multidose containers.

The compositions of the present invention may be prepared by the known methods of the pharmaceutical technique.

It is another object of the present invention to provide a method for the preparation of said compositions, which comprises

(A) dissolving the total amount, or at least 50% thereof, of cyclodextrin in water by heating at 35-45°C under stirring to a clear solution, by subsequent cooling of the solution to room temperature and further addition of the preservatives, of the antioxidants and, if any, of the flavoring agents; (B) optionally, adding the optionally N-acetylated glucosamine under stirring to the solution obtained in (A) and kept below 20°C, to a complete solution; (C) adding a solution, previously prepared by dissolving the optional sweetening agent and chondroitinsulfate in water by heating at 55-65°C under stirring to a clear solution, to the solution obtained in step (A) or in step (B) and, after cooling to room temperature, adding the remaining amount of cyclodextrin, if any, thereto; and wherein, when step (B) is carried out, chondroitinsulfate may be absent in step

(C).

If another carrier, for example a thickening agent, is included in the composition, it will be preferably added in step (C).

In general, the total amount of cyclodextrin is used in step (A). It could be advantageous, in the case of the preparation of compositions in which both chondroitinsulfate and the optionally N-acetylated glucosamine are present in a small volume of composition (corresponding to 4-7 g), to introduce the cyclodextrin, in particular β-cyclodextrin, in two portions, i.e. about 60% of the calculated amount in step (A) and the remaining amount in step (C).

In the case of higher volumes (corresponding for example to 8-30 g of compositions), it could be advantageous to add a thickening agent such as those mentioned herein above. The formulations obtainable according to the present invention allow an easy administration even at high combination dosages of chondroitinsulfate and/or

optionally N-acetylated glucosamine in liquid form.

The following examples illustrate the invention without, however, limiting it.

Example 1

Solution for the oral administration of 500 mg of chondroitinsulfate and of 500 mg of glucosamine hydrochloride.

(A) In a blender with sleeve and a blade stirrer, 4375 g of hydroxypropyl-β- cyclodextrin and 12 kg of deionized water are added. The temperature is maintained at 40 ⁰ C and the mixture is vigorously stirred until complete solution. The temperature is brought to 25°C, then 68.75 g of sodium EDTA, 150 g of sodium metabisulfite and 500 g of commercial lemon flavoring agent are added. Stirring is continued at room temperature until a clear solution is obtained.

(B) The temperature of the solution obtained in step (A) is brought to 20 ⁰ C and 3125 g of glucosamine hydrochloride are showered thereinto. Stirring is continued until a clear solution (Solution B) is obtained. (C) In a second blender with sleeve and a blade stirrer, 9375 g of xylitol in 15 kg of water are poured, the mixture is heated at 60 ⁰ C under vigorous stirring to obtain a solution. Under the same stirring and at the same temperature (60 ⁰ C), 3125 g of sodium chondroitinsulfate are added. The clear solution is then cooled to room temperature, 62.5 g of microcrystalline cellulose (Avicel RC-591) are added and the mixture is stirred until complete dispersion to obtain a solution (Solution C). Solution C thus obtained is added to the previously prepared Solution B under moderate stirring and the resulting solution is passed through a steel sieve 1.5 mm mesh. Thus, about 50 kg of a solution are obtained, to be introduced in 6250 unit doses of the following composition Sodium chondroitinsulfate 500 mg

Glucosamine hydrochloride 500 mg(*)

Hydroxypropyl-β-cyclodextrin 700 mg

Sodium metabisulfite 24 mg

Sodium EDTA ll mg Xylitol C 1500 mg

Flavoring agent 80 mg

Microcrystalline cellulose 10 mg

Deionized water to 8000 mg

(*) corresponding to 416.66 mg of D-glucosamine base.

Example 2 Solution for the oral administration of 1200 mg of chondroitinsulfate and of 1500 mg of glucosamine hydrochloride.

The preparation is carried out as described in Example 1 by adding, in step (A), 3182 g of hydroxypropyl-β-cyclodextrin in 12 kg of deionized water, 64 g of sodium EDTA, 145 g of sodium metabisulfite and 455 g of commercial orange flavoring agent. In step (B), 6818 g of glucosamine hydrochloride are added and in step (C) there are added 9090 g of xylitol C, 5454 g of sodium chondroitinsulfate and 72.7 g of microcrystalline cellulose in 15 Kg of deionized water. The resulting solution is passed through a steel sieve 1.5 mm mesh. Thus, about 50 kg of a solution are obtained, to be introduced in 4545 unit doses of the following composition:

Sodium chondroitinsulfate 1200 mg

Glucosamine hydrochloride 1500 mg(*)

Hydroxypropyl-β-cyclodextrin 700 mg

Sodium metabisulfite 32 mg

Sodium EDTA 14 mg

Xylitol C 2000 mg

Flavoring agent 100 mg

Microcrystalline cellulose 16 mg

Deionized water to 11,000 mg

^• esponding to 1246.42 mg of D-glucosamine base.

Example 3

Solution for the oral administration of 600 mg of chondroitinsulfate The preparation is carried out as described in Example 1 by adding, in step (A), 4375 g of hydroxypropyl-β-cyclodextrin in 12 kg of deionized water, 37.5 g of sodium benzoate, 75 g of potassium sorbate, 150 g of sodium metabisulfite and 50 g of commercial lemon flavoring agent. Step (B) is not carried out because glucosamie is not provided in the formulation, hi step (C) the prepared solution contains 9375 g of

xylitol C, 3750 g of sodium chondroitinsulfate and 62.5 g of hydroxypropyl cellulose in 15 kg of deionized water. Thus, about 50 kg of a solution are obtained, to be introduced in 6250 unit doses of the following composition:

Sodium chondroitinsulfate 600 mg Hydroxypropyl-β-cyclodextrin 700 mg

Sodium metabisulfite 24 mg

Sodium benzoate 6 mg

Potassium sorbate 2 mg

Xylitol C 1500 mg Flavoring agent 8 mg

Hydroxypropyl cellulose lO mg

Deionized water to 8000 mg

Example 4

Solution for the oral administration of 1500 mg of glucosamine hydrochloride By operating as described in Example 1, in step (A) a solution of 4375 g of hydroxypropyl-β-cyclodextrin, 68.75 g of sodium EDTA, 150 g of sodium metabisulfite and 50 g of commercial peppermint flavoring agent in 12 kg of deionized water is prepared. In step (B) 9375 g of glucosamine hydrochloride are added to obtain Solution B. In step (C) a solution of 9375 g of xylitol C and 62.5 g of hydroxypropyl cellulose in 15 kg of deionized water (Soluzione C) is added to Solution B. Thus, about 50 kg of a solution are obtained, to be introduced in 6250 unit doses of the following composition:

Glucosamine hydrochloride 1500 mg(*) hydroxyprόpyl-β-cyclodextrin 700 mg Sodium metabisulfite 24 mg

Sodium EDTA 11 mg

Xylitol C 1500 mg

Flavoring agent 8 mg

Hydroxypropyl cellulose 10 mg Deionized water to 8000 mg

(*) corresponding to 1250 mg of D-glucosamine base.

Example 5

Solution for oral administration of 400 mg of chondroitinsulfate and of 500 mg of glucosamine hydrochloride

(A) In a blender with sleeve and a blade stirrer, 375 g of β-cyclodextrin and 24 kg of deionized water are added. The temperature is maintained at 40°C and the mixture is vigorously stirred until complete solution. The temperature is brought to 25 ⁰ C, then 110 g of potassium sorbate, 240 g of sodium metabisulfite and 800 g of commercial lemon flavoring agent are added.

(B) The temperature of the solution obtained in step (A) is brought to 2O ⁰ C and 5000 g of glucosmine hydrochloride are showered thereinto. Stirring is continued until a clear solution (Solution B) is obtained.

(C) m a second blender with sleeve and a blade stirrer, 7500 g of xylitol in 15 kg of water are poured, the mixture is heated at 60°C under vigorous stirring to obtain a solution. Under the same stirring and at the same temperature (60 ⁰ C), 4000 g of sodium chondroitinsulfate are added. The clear solution is then cooled to room temperature, 250.2 g of β-cyclodextrin are added and the mixture is stirred until complete dispersion to obtain a solution (Solution C). Solution C thus obtained is added to the previously prepared Solution B under moderate stirring and the resulting solution is passed through a steel sieve 1.5 mm mesh. Thus, about 50 kg of a solution are obtained, to be introduced in 10,000 unit doses of the following composition: Sodium chondroitinsulfate 400.0 mg

Glucosamine hydrochloride 500.0 mg(*) β-cyclodextrin 65.5 mg

Sodium metabisulfite 24.0 mg Potassium sorbate ll.O mg

Xylitol C 750.0 mg

Flavoring agent 80.0 mg

Deionized water to 5000.0 mg

(*) corresponding to 416.66 mg of D-glucosamine base. Example 6

Solution for oral administration of 400 mg of chondroitinsulfate and of 500 mg of

glucosamine hydrochloride

(A) In a blender with sleeve and a blade stirrer, 375 g of β-cyclodextrin and 24 kg of deionized water are added. The temperature is maintained at 40°C and the mixture is vigorously stirred until complete solution. The temperature is brought to 25°C, then 90 g of sodium benzoate, 20 gr of potassium sorbate, 240 g of sodium metabisulfite and 800 g of commercial lemon flavoring agent are added.

(B) The temperature of the solution obtained in step (A) is brought to 20°C and 5000 g of glucosmine hydrochloride are showered thereinto. Stirring is continued until a clear solution (Solution B) is obtained. (C) In a second blender with sleeve and a blade stirrer, 7500 g of xylitol in 15 kg of water are poured, the mixture is heated at 6O ⁰ C under vigorous stirring to obtain a solution. Under the same stirring and at the same temperature (60°C), 4000 g of sodium chondroitinsulfate are added. The clear solution is then cooled to room temperature, 250.2 g of β-cyclodextrin are added and the mixture is stirred until complete dispersion to obtain a solution (Solution C). Solution C thus obtained is added to the previously prepared Solution B under moderate stirring and the resulting solution is passed through a steel sieve 1.5 mm mesh. Thus, about 50 kg of a solution are obtained, to be introduced in 10,000 unit doses of the following composition:

Sodium chondroitinsulfate 400.0 mg Glucosamine hydrochloride 500.0 mg(*) β-cyclodextrin 65.5 mg

Sodium metabisulfite 24.0 mg

Na Benzoate 9.0 mg

Potassium sorbate 2.0 mg Xylitol C 750.0 mg

Flavoring agent 80.0 mg

Deionized water to 5000.0 mg

(*) corresponding to 416.66 mg of D-glucosamine base.

Example 7 Solution for oral administration of 400 mg of chondroitinsulfate and of 500 mg of N- acetylglucos amine

(A) In a blender with sleeve and a blade stirrer, 375 g of β-cyclodextrin and 24 kg of deionized water are added. The temperature is maintained at 40°C and the mixture is vigorously stirred until complete solution. The temperature is brought to 25°C, then 110 g of potassium sorbate, 240 g of sodium metabisulfϊte and 800 g of commercial lemon flavoring agent are added.

(B) The temperature of the solution obtained in step (A) is brought to 20°C and 5000 g of N-acetylglucosamine are showered thereinto. Stirring is continued until a clear solution (Solution B) is obtained.

(C) In a second blender with sleeve and a blade stirrer, 7500 g of xylitol in 15 kg of water are poured, the mixture is heated at 60°C under vigorous stirring to obtain a solution. Under the same stirring and at the same temperature (60°C), 4000 g of sodium chondroitinsulfate are added. The clear solution is then cooled to room temperature, 250.2 g of β-cyclodextrin are added and the mixture is stirred until complete dispersion to obtain a solution (Solution C). Solution C thus obtained is added to the previously prepared Solution B under moderate stirring and the resulting solution is passed through a steel sieve 1.5 mm mesh. Thus, about 50 kg of a solution are obtained, to be introduced in 10,000 unit doses of the following composition: Sodium chondroitinsulfate 400.0 mg

N-acetylglucosamine 500.0 mg β-cyclodextrin 65.5 mg

Sodium metabisulfϊte 24.0 mg

Potassium sorbate 11.0 mg

Xylitol C 750.0 mg

Flavoring agent 80.0 mg Deionized water to 5000.0 mg

Example 8 Solution for oral administration of 400 mg of chondroitinsulfate and of 500 mg of N- acetylglucosamine

(A) In a blender with sleeve and a blade stirrer, 375 g of β-cyclodextrin and 24 kg of deionized water are added. The temperature is maintained at 40°C and the mixture is vigorously stirred until complete solution. The temperature is brought to

25 °C, then 90 g of sodium benzoate, 20 gr of potassium sorbate, 240 g of sodium metabisulfite and 800 g of commercial lemon flavoring agent are added.

(B) The temperature of the solution obtained in step (A) is brought to 20°C and 5000 g of N-acetylglucosamine are showered thereinto. Stirring is continued until a clear solution (Solution B) is obtained.

(C) In a second blender with sleeve and a blade stirrer, 7500 g of xylite 1 in 15 kg of water are poured, the mixture is heated at 6O ⁰ C under vigorous stirring to obtain a solution. Under the same stirring and at the same temperature (60°C), 4000 g of sodium chondroitinsulfate are added. The clear solution is then cooled to room temperature, 250.2 g of β-cyclodextrin are added and the mixture is stirred until complete dispersion to obtain a solution (Solution C). Solution C thus obtained is added to the previously prepared Solution B under moderate stirring and the resulting solution is passed through a steel sieve 1.5 mm mesh. Thus, about 50 kg of a solution are obtained, to be introduced in 10,000 unit doses of the following composition:

Sodium chondroitinsulfate 400.0 mg

N-acetylglucosamine 500.0 mg β-cyclodextrin 65.5 mg

Sodium metabisulfite 24.0 mg

Na Benzoate 9.0 mg

Potassium sorbate 2.0 mg

Xylitol C 750.0 mg

Flavoring agent 80.0 mg

Deionized water to 5000.0 mg