KR20200031059A - Sulfated hyaluronic acid-based hydrogel and pharmaceutical composition comprising the same - Google Patents

Abstract

The present invention relates to sulfated hyaluronic acid-based hydrogel formed by conducting, with a crosslinking agent, a reaction of -COOH group of a crosslinking monomer comprising at least one selected from the group consisting of sulfated hyaluronic acid, derivatives thereof, and salts thereof. The sulfated hyaluronic acid-based hydrogel according to the present invention can be effectively used as an anti-cancer agent for preventing or treating arthritis, eye diseases, etc.

Description

Sulfurated hyaluronic acid-based hydrogels and pharmaceutical compositions comprising the same {SULFATED HYALURONIC ACID-BASED HYDROGEL AND PHARMACEUTICAL COMPOSITION COMPRISING THE SAME}

The present invention relates to a hydrogel based on sulfated hyaluronic acid and a pharmaceutical composition comprising the same.

Hyaluronic acid is a polysaccharide molecule with significant viscoelastic properties. Hyaluronic acid acts as a lubricant and shock absorber and is present in the joint cavity as a basic component of synovial fluid that protects chondrocytes against inflammatory cytokine activity (Asari et al., Arch Histol Cytol, 1995 , 58: 65-76; Brun et al., Osteoarthr Cartil, 2003, 11: 208-16; Stove et al., J Orthop Res, 2002, 20: 551-5). Hyaluronic acid has long been used as a “viscosupplement” or lubricant to treat degenerative osteoarthritis or the like in its own or derivative form.

Sulphated hyaluronic acid and its derivatives have known anti-inflammatory and anti-VEGF efficacy. However, in order to use sulfated hyaluronic acid as a drug having actual efficacy, it is necessary to increase the residual rate in the body and to have physical properties suitable for use. The known sulfated hyaluronic acid polymer solution is rapidly decomposed in the body, and it is difficult to control physical properties according to the use.

Accordingly, it is necessary to develop a sulfated hyaluronic acid-based material that exhibits anti-inflammatory and anti-VEGF efficacy, has resistance to decomposition, prolongs the efficacy in the body, and can control physical properties for each use.

International Publication WO2002-032407

The present invention is to provide a new hydrosulfate-based hydrogel based hydrogel for controlling the physical properties of the sulfated hyaluronic acid in the body for long-term efficacy and use.

In addition, the present invention is to provide an anti-inflammatory or anti-angiogenic pharmaceutical composition containing the sulfated hyaluronic acid-based hydrogel as an active ingredient.

The present invention provides a sulfated hyaluronic acid based hydrogel formed by reacting a -COOH group of a crosslinking monomer comprising at least one selected from the group consisting of sulfated hyaluronic acid, its derivatives and salts with a crosslinking agent.

In addition, the present invention provides an anti-inflammatory or anti-angiogenic pharmaceutical composition comprising the sulfated hyaluronic acid-based hydrogel as an active ingredient.

In addition, the present invention provides a pharmaceutical composition for the prevention or treatment of arthritis, prevention or treatment of eye diseases, anti-cancer, comprising the sulfated hyaluronic acid-based hydrogel as an active ingredient.

The hydrogel based on sulfated hyaluronic acid according to the present invention has an anti-inflammatory or anti-angiogenic effect, can inhibit decomposition in the body, prolong the anti-inflammatory or anti-angiogenic effect, and facilitate the physical properties of the gel to suit the application. Can be adjusted.

The hydrogel based sulfated hyaluronic acid according to the present invention can be effectively used as an anti-cancer agent for the prevention or treatment of arthritis and eye diseases.

1 is a 1 H NMR results for confirming the sulfated substitution of sulfated hyaluronic acid (SHA).
Figure 2 is a 1H NMR results for confirming the hydrogel based on the sulfated hyaluronic acid of the present invention.
3 is a graph of enzyme resistance evaluation of a hydrogel based on sulfated hyaluronic acid of the present invention.
Figure 4 is a graph of cytotoxicity evaluation of the hydrogel based on the sulfated hyaluronic acid of the present invention.
Figure 5 is a VEGF-treated sample of HUVEC cells and VEGF-treated samples of HUVEC cells (tube formation) and VEGF-treated samples of HUVEC cells produced by the treatment of sulfated hyaluronic acid-based hydrogel (SCO Gel) ( tube formation) This is an optical micrograph of the effect of inhibition.
Figure 6 is a graph of the anti-inflammatory efficacy of the sulfated hyaluronic acid-based hydrogel of the present invention.

Hereinafter, the present invention will be described in more detail to aid understanding of the present invention. At this time, the terms or words used in the present specification and claims should not be interpreted as being limited to ordinary or dictionary meanings, and the inventor appropriately defines the concept of terms in order to explain his or her invention in the best way. Based on the principle that it can be done, it should be interpreted as a meaning and a concept consistent with the technical idea of the present invention.

The present invention provides a sulfated hyaluronic acid based hydrogel formed by including at least one selected from the group consisting of sulfated hyaluronic acid, its derivatives and salts. The sulfated hyaluronic acid-based hydrogel of the present invention is formed by reacting a -COOH group of a crosslinking monomer containing at least one selected from the group consisting of sulfated hyaluronic acid, its derivatives, and salts with a crosslinking agent.

Conventional sulfated hyaluronic acid, its derivatives and salts are known to have anti-inflammatory and anti-VEGF efficacy, but the polymerized solution containing the sulfated hyaluronic acid, its derivatives and salts is rapidly decomposed in the body, and properties according to the use There was a difficult problem of control. Thus, the present invention provides a hydrogel based on a sulfated hyaluronic acid formed by including at least one selected from the group consisting of sulfated hyaluronic acid, its derivatives, and salts, thereby inhibiting decomposition in the body and inhibiting the body of sulfated hyaluronic acid in the body. Efficacy can be prolonged and the physical properties of the gel can be easily adjusted to suit the application. The hydrogel based on sulfated hyaluronic acid according to the present invention has an anti-inflammatory or anti-angiogenic effect, and can be effectively used as an anti-cancer agent for the prevention or treatment of arthritis and eye diseases.

The sulfated hyaluronic acid means that at least one of the alcoholic hydroxyl groups of hyaluronic acid is sulfated and substituted, and the sulfated hyaluronic acid derivative means other chemically modified sulfated hyaluronic acid or substituted sulfated hyaluronic acid. . For example, sulfated hyaluronic acid derivatives include sulfated hyaluronic acid-poloxamer derivatives, sulfated hyaluronic acid-polyethylene glycol derivatives, sulfated hyaluronic acid- (CH ₂ ) _n- CH ₃ derivatives, sulfated hyaluronic acid-benzyl esters It may be composed of one or more selected from derivatives, sulfated hyaluronic acid-chitosan derivatives, sulfated hyaluronic acid-PLGA derivatives, sulfated hyaluronic acid-gelatin or sulfated hyaluronic acid-collagen derivatives. The sulfated hyaluronic acid salt is selected from, for example, sodium sulfated hyaluronate, calcium sulfated hyaluronate, magnesium sulfated hyaluronate, zinc sulfated hyaluronate, cobalt sulfated hyaluronate or tetrabutylammonium sulfated hyaluronate It may be composed of one or more. Alternatively, the sulfated hyaluronic acid may be represented by, for example, the following formula.

[Formula]

In the above formula, R ₁ and R ₂ may each independently be SO ₃ H or H, and n is an integer of 1 or more.

At least one or more selected from the group consisting of the sulfated hyaluronic acid, its derivatives and salts may have a weight average molecular weight (Mw) of 800 to 5,000 kDa. More preferably, it may be 1,000 to 4,000 kDa, and more preferably 1,200 to 3,000 kDa. By using sulfated hyaluronic acid, its derivatives or salts within the weight average molecular weight range, reactivity with a crosslinking agent and ease of hydrogel production can be improved, and physical properties of the prepared hydrogel can be improved.

Hereinafter, unless otherwise indicated for convenience, sulfated hyaluronic acid refers to both sulfated hyaluronic acid and its derivatives and salts.

Conventionally, the sulfated hyaluronic acid is a primary -OH group, that is, the C6 site of hyaluronic acid is sulfated and substituted, so that the sulfated hyaluronic acid is used as a crosslinking monomer to crosslink with a crosslinking agent. It was difficult to form a hydrogel based on sulfated hyaluronic acid.

Accordingly, the present invention provides a new sulfated hyaluronic acid based hydrogel formed by reacting a -COOH group of a crosslinking monomer comprising at least one selected from the group consisting of sulfated hyaluronic acid, its derivatives and salts with a crosslinking agent.

At this time, the crosslinking monomer may be at least one selected from the group consisting of sulfated hyaluronic acid, derivatives and salts of which some or all of the alcoholic hydroxyl groups of hyaluronic acid are sulfated. Hyaluronic acid has a total of four -OH groups, including three secondary -OH groups, in addition to the primary -OH groups. The OH group may be sulfated first, and as the reaction proceeds further, the secondary -OH group may be sulfated and the total substitution rate may be 400% when all -OH of hyaluronic acid is sulfated substituted. . The crosslinking monomer comprising at least one or more selected from the group consisting of the sulfated hyaluronic acid, its derivatives and salts according to an embodiment of the present invention, may be 30 to 400% sulfated substituted in the alcoholic hydroxyl group of hyaluronic acid have. That is, in the case of a hydrogel based on sulfated hyaluronic acid formed by crosslinking using -COOH of sulfated hyaluronic acid as in the present invention, since -COOH group is used instead of -OH group during crosslinking, primary ( It is possible to use sulfated hyaluronic acid in which primary -OH groups are all sulfated, and furthermore, sulfated hyaluronic acid which is sulfated and substituted to both the primary -OH group and the secondary -OH group is used. You can also use In addition, it is also possible to use a sulfated hyaluronic acid in which the alcoholic hydroxyl group of hyaluronic acid is partially sulfated, and a part of the primary -OH group is sulfated. The sulfated hyaluronic acid according to an embodiment of the present invention maximizes efficacy and duration by using sulfated hyaluronic acid having a substitution rate suitable for each purpose in consideration of safety, stability, anti-inflammatory efficacy, anti-angiogenic efficacy, and physical properties. can do.

The hydrogel based on the sulfated hyaluronic acid of the present invention may use a diamine-based crosslinking agent as the crosslinking agent. The sulfated hyaluronic acid-based hydrogel of the present invention can form a hydrogel by reacting the -COOH group of the crosslinking monomer with the crosslinking agent by using a diamine-based crosslinking agent as the crosslinking agent. The diamine-based crosslinking agent is, for example, diaminobutane, diaminohexane, diaminooctane, adipic acid dihydrazide (ADH), polyethylene glycol diamine ( PEG-Diamine), cystamine, and the like, and most preferably diaminohexane. At this time, 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide (EDC), 1- [3- (dimethylamino) propyl] -3 as a catalyst -Ethyl carbodiimide methide (1- [3- (Dimethylamino) propyl] -3-ethylcarbodiimide methiodide), diisopropylcarbodiimide (DIC), N, N'-dicyclohexylcarbodiimide (N , N'-Dicyclohexylcarbodiimide (DCC), and the like, a carbodiimide-based catalyst may be used, and N-Hydroxysuccinimide (NHS) is used to assist EDC activity. Sulfo-N-Hydroxysuccinimide (sulfo-NHS), 1-hydroxybenzotriazole (HOBt), and the like can be used. These catalysts may serve to react -COOH groups of sulfated hyaluronic acid with -NH ₂ of a crosslinking agent.

As described above, the hydrogel based on sulfated hyaluronic acid according to the present invention has anti-inflammatory or anti-angiogenic effects. In addition, the hydrogel based on sulfated hyaluronic acid according to the present invention can prolong the efficacy of anti-inflammatory or anti-angiogenic effects by slowing the rate of degradation in the body. Accordingly, the hydrogel based on the sulfated hyaluronic acid according to the present invention can be effectively used as a prophylactic or therapeutic agent for arthritis, a prophylactic or therapeutic agent for eye diseases, anticancer agent, and the like.

In addition, the present invention provides an anti-inflammatory or anti-angiogenic pharmaceutical composition comprising the sulfated hyaluronic acid-based hydrogel as an active ingredient. The pharmaceutical composition according to the present invention can be used for preventing or treating arthritis, preventing or treating eye diseases, and anticancer.

The “pharmaceutical composition” may include other chemical components such as a hydrogel based on the sulfated hyaluronic acid of the present invention, a diluent, and a carrier. Accordingly, the pharmaceutical composition may include a pharmaceutically acceptable carrier, diluent, or excipient, or a combination thereof, as necessary. The pharmaceutical composition facilitates administration of the compound into an organism. There are various techniques for administering the compound, including, but not limited to, oral, injection, aerosol, parenteral, and topical administration.

In addition, the present invention provides a method for preventing or treating arthritis in mammals by administering a hydrogel based on sulfated hyaluronic acid as an active ingredient. At this time, the administration of the hydrogel based on sulfated hyaluronic acid may be preferably introduced as an injection preparation. Representative examples that can be treated with a hydrogel based on the sulfated hyaluronic acid of the present invention, for the treatment of osteoarthritis or rheumatoid arthritis, or gout or calcium pyrophosphate deposition disease (calcium pyrophosphate) dihydrate deposition disease), or may be used to reduce or prevent adhesions that may form after a surgical procedure. The hydrogels based on the sulfated hyaluronic acid of the present invention may be particularly useful for treating chronic or acute inflammation.

“Treatment” as used herein refers to stopping, delaying or alleviating the progression of the disease when used on an object exhibiting an onset symptom, and “preventing” indicates an onset symptom when used on an object that does not exhibit an onset symptom but is at such a high risk. It means stopping, delaying, or alleviating.

The following examples are presented, and are intended to be purely illustrative, to provide those skilled in the art with a complete disclosure and description of how the compounds, compositions, and methods provided herein are made and evaluated. Accordingly, the examples are not intended to limit the scope of what the inventors regard as their invention. There are many variations and combinations of reaction conditions, such as component concentration, desired solvent, solvent mixture, temperature, pressure, and other reaction parameters and conditions that may be used to optimize product properties such as purity, yield, and the like. . These are also considered to be within the scope of this application. Any combination of the elements described above in all possible variations is encompassed by the present invention unless otherwise indicated herein or otherwise clearly contradicted by context.

[ Preparation Example 1] -Synthesis of sulfated hyaluronic acid (sulfated HA)

HA-TBA derivatives incorporating tetrabutyl ammonium salt (TBA) into hyaluronic acid (HA) using ion exchange resin were added to 3 mg / mL of dimethylformamide (DMF), an organic solvent. Sulfur trioxide pyridine complex was dissolved at a concentration, and 15 mol of the HA-TBA derivative monomer was added, followed by reacting with nitrogen gas at 5 ° C. for 2 hours, and then sulfuric acid purified through ethanol precipitation. Hyaluronic acid derivatives were obtained.

Example 1

After dissolving the sulfated hyaluronic acid (SHA) prepared in Preparation Example 1 in water at 6 wt%, 100 moles of EDC, 100 moles of NHS, 100 moles of NHS, and diaminootane 20 relative to 100 moles of monomers of sulfated hyaluronic acid (SHA) A hydrogel was prepared by adding a molar portion and reacting at 40 ° C by overnight.

[ Experimental example  One: SHA  And Sulfated  Hyaluronic acid based Hydrogel  Confirm]

The sulfated hyaluronic acid (SHA) prepared in Preparation Example 1 was confirmed by 1H NMR analysis and elemental substitution to determine the degree of sulfate substitution, and the sulfated hyaluronic acid-based hydrogel prepared in Example 1 was subjected to 1H NMR analysis. It was confirmed, and the results are shown in FIGS. 1 and 2.

Referring to the NMR data of FIG. 1, the sulfated hyaluronic acid (SHA) prepared in Preparation Example 1 completely disappeared from the peak of 3.4 ppm in FIG. 1 data, and the primary -OH group of hyaluronic acid, that is, the C6 site It was confirmed that all were sulfated substituted sulfated hyaluronic acid (sulfated HA), and the sulfated hyaluronic acid-based hydrogel prepared in Example 1 confirmed that crosslinking agent peaks appeared between 1.0-1.6 ppm in FIG. 2 data. It was confirmed that a hydrogel based on sulfuric acid hyaluronic acid was prepared.

Example 2

A sulfonated hyaluronic acid-based hydrogel was prepared in the same manner as in Example 1, except that diaminohexane was used instead of diaminooctane as a diamine-based crosslinking agent.

Comparative Example 1

After dissolving the sulfated hyaluronic acid (SHA) prepared in Preparation Example 1 in NaOH at 20 wt%, 1,4-butanediol diglycidyl ether (BDDE) as a crosslinking agent is sulfated hyaluronic acid monomer With respect to 5 mol%, it was overnight at 30 ° C. However, in the case of Comparative Example 1 in which C6 sites were all sulfated-substituted sulfated hyaluronic acid (sulfated HA) as a crosslinking monomer, and BDDE was used as a crosslinking agent, a solution could not be formed even after the reaction was completed without forming a hydrogel. Analysis in the form of was impossible.

[Experimental Example 2: Measurement of viscoelasticity of hydrogel]

The hydrogels based on the sulfated hyaluronic acid prepared in Examples 1 to 2 were measured for viscoelasticity using an MCR 302 rheometer equipped with a 25mm steel plate at 25 ° C with 4% strain and 2.5 Hz. In addition, changes in viscoelasticity after autoclave sterilization of the hydrogels based on the sulfated hyaluronic acid prepared in Examples 1 to 2 at 121 ° C for 10 minutes were measured. Table 1 shows the results.

Before sterilization After sterilization G '(Pa) G '' (Pa) G '(Pa) G '' (Pa) Example 1 551 132 221 47 Example 2 679 197 240 63

Referring to Table 1, it can be confirmed that the hydrogels based on the sulfated hyaluronic acid prepared in Examples 1 to 2 can be manufactured to have viscoelasticity in a wide range from hard to soft properties depending on the application purpose, and Autoclave if necessary. It was confirmed through sterilization that it can have a desired viscoelastic range even after final sterilization.

[ Experimental example  3: Enzyme resistance evaluation]

The enzyme resistance of SHA-COOH (SCO) Gel of Example 2 is shown in FIG. 3 compared to hyaluronic acid-COOH hydrogel (HA-COOH Gel).

After mixing 1 g of each hydrogel with 10 ul of hyaluronic acid degrading enzyme Hyaluronidase 500 uint / g solution, the viscoelasticity was measured using a MCR 302 rheometer equipped with a 25mm steel plate at 37 ° C with 4% strain and 2.5 Hz. Did. The results are shown in FIG. 3.

Referring to FIG. 3, in the case of hyaluronic acid-COOH hydrogel (HA-COOH Gel) prepared under the same conditions, the decomposition of the gel over time is fast, whereas in the case of SHA-COOH Gel of Example 2, the properties of the degrading enzyme It was confirmed that the drop was significantly improved.

[ Experimental example  4: Cytotoxicity evaluation]

Using the SHA-COOH (SCO) Gel of Example 1, hyaluronic acid (HA), and sulfated hyaluronic acid (SHA), cytotoxicity of two types of cells, FLS and Chondrocyte, was evaluated.

Two types of cells, FLS and Chondrocyte, were seeded in 96 wells at 8.0x10 ³ and 3.4x10 ⁴ cells / well, and then overnighted to grow cells. Each sample was treated with 1 mg / ml. After the sample treatment, the viability of the cells after an additional 24 hour incubation was measured using a resazaurin assay. The results are shown in FIG. 4.

Referring to FIG. 4, all samples of SHA-COOH (SCO) Gel, hyaluronic acid hydrogel (HA), and sulfated hyaluronic acid (SHA) of Example 1 were compared to the control group in which no samples were treated FLS, Chondrocyte 2 It was confirmed that the viability of the type of cells was 80% or more, and there was no cytotoxicity.

[ Experimental example  5: HUVEC Antiangiogenic  Efficacy confirmation]

The tube formation formed when 3D culture was performed using Matrigel of the VEGF-treated and VEGF-untreated sample group of HUVEC cells is shown in FIG. 5.

In the VEGF-treated group / VEGF-untreated group, HUVEC cells were seeded in matrigel-coated 96 wells to confirm the degree of tube formation after 16h incubation through an optical microscope. In addition, in order to evaluate the efficacy of inhibiting the formation of tubes in each sample, HUVEC cells were seeded in 96 wells coated with matrigel, treated with a certain concentration of VEGF, and sulfated hyaluronic acid hydrogel of Example 1 (SCO) Gel) after treatment for 16 hours after incubation (incubation), the degree of blood vessel (tube) formation was observed through an optical microscope, and the results are shown in FIG. 5.

Referring to FIG. 5, it was confirmed that when the SCO Gel of Example 1 was treated, almost no blood vessels were formed.

[ Experimental example  6: Anti-inflammatory efficacy evaluation]

Using the sulfated hyaluronic acid-based hydrogel of Example 1 (SCO Gel), hyaluronic acid (HA), and sulfated hyaluronic acid (SHA), evaluation of anti-inflammatory efficacy of Chondrocyte and FLS 2 cells was performed.

Chondrocyte, FLS 2 types of cells are respectively 3.4x10 ^{4 in} 96 wells and After seeding with 8.0x10 ³ cells / well, TNF-alpha was treated with 10 ng / ml and 1 ng / ml to activate cells with overnight, and each sample was treated with 5 mg / ml and 1 mg / ml. . After 24 hours of incubation after sample treatment, the cytokine secretion of cells in the medium was measured using ELISA, and the results are shown in FIG. 6.

Referring to FIG. 6, IL-6 and IL-8 secretion of Chondrocyte cells was reduced to 60% or less compared to the control group in which the sulfated hyaluronic acid-based hydrogel of Example 1 (SCO Gel) was not treated with any sample. , It was confirmed that IL-6 and IL-8 secretion of FLS cells was reduced to 30% or less.

[ Experimental example  7: Anticoagulation reaction evaluation]

Using the sulfated hyaluronic acid-based hydrogel of Example 1 (SCO Gel), heparin (Heparin), and sulfated hyaluronic acid (SHA), the degree of inhibition of the activity of Factor IIa and Factor Xa using a chromogenic assay Was measured. Table 2 shows the results.

Anti-IIa activity [Unit / mg] Anti-Xa activity [IU / mg] Heparin 209 209 SHA 0.13 0.17 Example 1 0.05 0.12

Referring to Table 2, all samples of sulfated hyaluronic acid (SHA) had an activity of inhibiting Factor IIa and Factor Xa over 1200 times lower than heparin. In particular, it was confirmed that the activity of the hydrogel (SCO Gel) based on sulfated hyaluronic acid of Example 1 was lower than 1500 times that of heparin, so that an anticoagulation reaction hardly occurred.

Claims (12)

Sulfurized hyaluronic acid-based hydrogel formed by reacting a -COOH group of a crosslinking monomer comprising at least one selected from the group consisting of sulfated hyaluronic acid, its derivatives and salts with a crosslinking agent.
According to claim 1,
The crosslinking agent is a hydrogel based on sulfated hyaluronic acid, which is a diamine-based crosslinking agent.
According to claim 2,
The diamine-based crosslinking agent is diaminobutane, diaminohexane, diaminooctane, adipic acid dihydrazide (ADH), polyethylene glycol diamine (PEG-Diamine) And a hydrogel based on at least one sulfated hyaluronic acid selected from the group consisting of cystamine.
According to claim 1,
The crosslinking monomer comprising at least one selected from the group consisting of the sulfated hyaluronic acid, its derivatives and salts is a hydrogel based on sulfated hyaluronic acid in which some or all of the alcoholic hydroxyl groups of hyaluronic acid are sulfated substituted.
According to claim 4,
The cross-linking monomer comprising at least one selected from the group consisting of the sulfated hyaluronic acid, its derivatives and salts, 30 to 400% of the alcoholic hydroxyl group of hyaluronic acid is sulfated substituted hydrogel based hydrogel .
According to claim 1,
At least one selected from the group consisting of the sulfated hyaluronic acid, derivatives and salts thereof is a hydrogel based on sulfated hyaluronic acid having a weight average molecular weight (Mw) of 800 to 5,000 kDa.
According to claim 1,
The sulfated hyaluronic acid based hydrogel is a sulfated hyaluronic acid based hydrogel used for anti-inflammatory or anti-angiogenic.
A pharmaceutical composition for anti-inflammatory or anti-angiogenesis comprising the hydrogel based on sulfated hyaluronic acid according to claim 1 as an active ingredient.
A pharmaceutical composition for the prevention or treatment of arthritis, comprising the hydrogel based on sulfated hyaluronic acid according to claim 1 as an active ingredient.
A pharmaceutical composition for the prevention or treatment of eye diseases, comprising the hydrogel based on sulfated hyaluronic acid according to claim 1 as an active ingredient.
A pharmaceutical composition for anticancer comprising the hydrogel based on sulfated hyaluronic acid according to claim 1 as an active ingredient.
A method of treating arthritis in a mammal comprising the step of administering a hydrogel based on sulfated hyaluronic acid according to claim 1.

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