TW202135834A - Anti-inflammatory or anti-angiogenic pharmaceutical composition - Google Patents

Abstract

The present invention relates to an anti-inflammatory or anti-angiogenic pharmaceutical composition comprising at least one selected from the group consisting of sulfated hyaluronic acid having the mean sulfate-substitution ratio of 200 to 300% for alcoholic hydroxyl groups per the repeating unit of hyaluronic acid, derivatives, and salts thereof.

Description

Anti-inflammatory or anti-angiogenic pharmaceutical composition

The present invention relates to an anti-inflammatory or anti-angiogenic pharmaceutical composition. Cross-reference of related applications

This application claims priority based on Korean Patent Application No. 10-2019-0173213 filed on December 23, 2019, and the entire disclosure of this article is incorporated as a part of this patent specification.

Hyaluronic acid (HA) is an unsulfated glycosaminoglycan (glycosaminoglycan), which is a natural polysaccharide, among which D-glucuronic acid (GlcUA) and N-acetyl-D-glucosamine (GlcNAc) Repeating sequences are connected. Hyaluronic acid exists in various areas of animals, such as vitreous body, cartilage, synovial fluid, placenta and skin and is in the form of mucopolysaccharides.

Hyaluronic acid involves the extracellular matrix to absorb water or impart elasticity between cells, and is involved in cell signal transduction, wound healing and morphogenesis. In particular, hyaluronic acid inhibits the phagocytosis of macrophages and exhibits anti-inflammatory activity.

Because of this activity, hyaluronic acid has been used as a therapeutic agent for arthritis and as an adjuvant for wound healing and eye and middle ear surgery. In addition, hyaluronic acid has excellent biocompatibility, and its use in tissue engineering and regenerative medicine is gradually increasing.

At the same time, sulfated hyaluronic acid is known for its anti-inflammatory and anti-VEGF effects. However, the anti-inflammatory and anti-VEGF effects may vary greatly depending on the sulfate substitution rate of sulfated hyaluronic acid. Since sulfated hyaluronic acid is not a natural substance, it may be toxic according to the substitution rate. In addition, it is known that sulfated hyaluronic acid has anticoagulant activity like heparin. Depending on the intended use, the anticoagulant activity may appear in the form of unexpected side effects. In order to be a drug with practical effects, it is important that sulfated hyaluronic acid has excellent anti-inflammatory and anti-angiogenic effects, does not exhibit cytotoxicity and does not have blood anticoagulant activity to eliminate unexpected side effects. Therefore, there is a need to develop sulfated hyaluronic acid that exhibits anti-inflammatory and anti-VEGF effects, and does not exhibit cytotoxicity and blood anticoagulant activity.

[Prior Technical Document]

[Patent Document]

Korean Patent Publication No. 10-2017-0120991

technical problem

The present invention provides an anti-inflammatory or anti-angiogenic pharmaceutical composition comprising sulfated hyaluronic acid, its derivatives and salts with high potency and low toxicity. The sulfated hyaluronic acid, its derivatives and salts are excellent It has anti-inflammatory and anti-angiogenic effects, and does not show cytotoxicity and blood anticoagulant activity. Technical solutions

The present invention provides an anti-inflammatory pharmaceutical composition, the pharmaceutical composition comprising at least one selected from the group consisting of: sulfated hyaluronic acid, its derivatives and salts, each hyaluronic acid in the sulfated hyaluronic acid The acid repeating unit has an average sulfate substitution rate of 200% to 300% for the alcohol hydroxyl group.

The present invention also provides an anti-angiogenic pharmaceutical composition, which comprises at least one selected from the group consisting of: sulfated hyaluronic acid, derivatives and salts thereof, each hyaluronic acid in the sulfated hyaluronic acid The repeating unit has an average sulfate substitution rate of 200% to 300% for the alcoholic hydroxyl group.

The present invention also provides a pharmaceutical composition for the treatment or prevention of inflammatory or angiogenic eye diseases, the composition comprising at least one selected from the group consisting of: sulfated hyaluronic acid, its derivatives and salts Each repeating unit of hyaluronic acid in the sulfated hyaluronic acid has an average sulfate substitution rate of 200% to 300% for alcoholic hydroxyl groups.

The present invention provides a pharmaceutical composition for treating or preventing arthritis, the composition comprising at least one selected from the group consisting of: sulfated hyaluronic acid, its derivatives and salts, the sulfated hyaluronic acid Each repeating unit of hyaluronic acid has an average sulfate substitution rate of 200% to 300% for alcoholic hydroxyl groups.

The present invention also provides an anticancer agent, which comprises at least one selected from the group consisting of: sulfated hyaluronic acid, derivatives and salts thereof, each repeating unit of hyaluronic acid in the sulfated hyaluronic acid is Alcoholic hydroxyl groups have an average sulfate substitution rate of 200% to 300%. Beneficial effect

The present invention can provide an anti-inflammatory or anti-angiogenic pharmaceutical composition, the pharmaceutical composition comprising high-potency, low-toxicity sulfated hyaluronic acid, its derivatives and salts, the sulfated hyaluronic acid, its derivatives and Salts have excellent anti-inflammatory and anti-angiogenic effects, and do not show cytotoxicity and blood anticoagulant activity.

The anti-inflammatory or anti-angiogenic pharmaceutical composition according to the present invention can be effectively used as an agent for preventing or treating arthritis, eye diseases, etc. and an anticancer drug.

Hereinafter, the present invention will be described in more detail to assist the understanding of the present invention. Based on the concept that the inventor can properly define terms, interpret his own invention in the best way, the terms or words used in the specification and the scope of the patent application should not be construed as limited to general or dictionary meanings, and the term or The words should be interpreted as meanings and concepts consistent with the technical idea of the present invention.

The present invention provides an anti-inflammatory pharmaceutical composition, which comprises at least one selected from the group consisting of: sulfated hyaluronic acid, its derivatives and salts, each repeating hyaluronic acid in the sulfated hyaluronic acid The units have an average sulfate substitution rate of 200% to 300% for alcoholic hydroxyl groups. The present invention also provides an anti-angiogenic pharmaceutical composition, the composition comprising at least one selected from the group consisting of: sulfated hyaluronic acid, derivatives and salts thereof, each of which is transparent The repeating unit of the acid has an average sulfate substitution rate of 200% to 300% for the alcohol hydroxyl group.

It is known that conventional sulfated hyaluronic acid, its derivatives and salts have anti-inflammatory and anti-VEGF effects. However, the research on the anti-inflammatory and anti-VEGF efficacy and toxicity based on the sulfate substitution rate and molecular weight is still insufficient. They are also known to have blood anticoagulant activity like heparin. Therefore, there is a problem that unexpected side effects may occur during actual use. Therefore, in the present invention, each repeating unit of hyaluronic acid in sulfated hyaluronic acid, its derivatives and salts has an average sulfate substitution rate of 200% to 300%, so as to exhibit high efficiency and low toxicity for anti-inflammatory Sex and anti-angiogenesis effect, and will not show blood anticoagulant activity. These high-potency and low-toxic anti-inflammatory and anti-angiogenic pharmaceutical compositions can be effectively used as agents for preventing or treating arthritis, agents for preventing or treating eye diseases, and anticancer agents.

The sulfated hyaluronic acid means that at least one of the alcoholic hydroxyl groups of hyaluronic acid is substituted by sulfate, and the sulfated hyaluronic acid derivative means other chemically modified sulfated hyaluronic acid or substituted sulfated Hyaluronic acid. For example, the sulfated hyaluronic acid derivative can be composed of one or more of the following: sulfated hyaluronic acid-poloxamer derivative, sulfated hyaluronic acid-polyethylene glycol derivative, Sulfated hyaluronic acid-(CH ₂ ) _x -CH ₃ derivatives (where x is an integer from 1 to 20), sulfated hyaluronic acid-(CH ₂ ) _y -NH ₂ derivatives (where y is 1 to 20 Integer), sulfated hyaluronic acid-benzyl ester derivatives, sulfated hyaluronic acid-chitosan derivatives, sulfated hyaluronic acid-PLGA derivatives and sulfated hyaluronic acid-gelatin or sulfated hyaluronic acid Acid-collagen derivative. The sulfated hyaluronate may be composed of one or more selected from the following: sulfated sodium hyaluronate, sulfated potassium hyaluronate, sulfated calcium hyaluronate, sulfated magnesium hyaluronate, sulfated hyaluronate Zinc phosphate, sulfated cobalt hyaluronate, or sulfated tetrabutylammonium hyaluronate. Alternatively, the sulfated hyaluronic acid can be represented by, for example, the following formula:

[Chemical formula]

Wherein, R ₁ and R ₂ can each independently be SO ₃ H or H, and n is 1 or an integer greater than 1.

For convenience, unless otherwise indicated, the term "sulfated hyaluronic acid" hereinafter refers to all sulfated hyaluronic acid, its derivatives and salts.

Each repeating unit of hyaluronic acid has a total of four -OH groups, in addition to one primary -OH group, it also includes three secondary -OH groups. During the sulfate substitution reaction, the more reactive primary -OH group is first replaced by sulfate, and as the reaction proceeds further, the secondary -OH group may be replaced by sulfate. When all -OH groups of hyaluronic acid are replaced by sulfate, the total sulfate substitution rate can be 400%. Herein, the sulfated hyaluronic acid having a sulfate substitution rate of 100% or more is one in which the reactive primary -OH group is first substituted by sulfate and then the secondary -OH group is also substituted by sulfate. The present invention relates to sulfated hyaluronic acid having an average sulfate substitution rate of 200% to 300% per repeating unit, that is, the sulfated hyaluronic acid has an average of two of the four alcoholic hydroxyl groups per repeating unit of hyaluronic acid. Or three are replaced by sulfates. When the average sulfate substitution rate of the sulfated hyaluronic acid is less than 200%, the anti-inflammatory and anti-angiogenic effects are obviously insufficient. When the average sulfate substitution rate of the sulfated hyaluronic acid is greater than 300%, cytotoxicity occurs. Specifically, the sulfate substitution rate may be 210 to 300%, 210 to 280%, 210 to 260%, 220 to 300%, 220 to 280%, 220 to 260%, 240 to 300%, 240 to 280% , Or 240 to 260%.

The weight average molecular weight (Mw) of the sulfated hyaluronic acid, its derivatives or salts may be 500 to 3,000 kDa. More preferably, it may be 1,000 to 2,500 kDa, and even more preferably 1,400 to 2,200 kDa. Sulfated hyaluronic acid, its derivatives or salts within the above-mentioned weight average molecular weight range have excellent anti-angiogenic effects, hardly cause blood anticoagulation, and have a long residence time in the body, so they are suitable for actual medicines, and It is not too viscous, so it is suitable for production and use.

As mentioned above, the sulfated hyaluronic acid having an average sulfate substitution rate of 200% to 300% per repeat unit according to the present invention has excellent anti-inflammatory or anti-angiogenic effects. It also does not exhibit cytotoxicity and blood anticoagulant activity. Therefore, the pharmaceutical composition containing the sulfated hyaluronic acid according to the present invention can be effectively used as an agent for preventing or treating arthritis, as an agent for preventing or treating eye diseases, as an anticancer agent, and the like.

The "medical composition" may include the sulfated hyaluronic acid of the present invention and other chemical components, such as diluents, carriers, and the like. Therefore, the pharmaceutical composition may include pharmaceutically acceptable carriers, diluents, excipients, or combinations of them as needed. The pharmaceutical composition promotes the administration of the compound into the living body. There are many different methods for administering the compound, including, but not limited to oral, injection, aerosol, parenteral and topical administration.

The present invention can also provide a method for preventing or treating arthritis in mammals by administering a pharmaceutical composition containing the sulfated hyaluronic acid. As a representative example, the pharmaceutical composition containing the sulfated hyaluronic acid of the present invention can be used to treat osteoarthritis or rheumatoid arthritis, or other inflammatory arthritis, such as gout or calcium pyrophosphate dihydrate deposition disease , Or it can be used to reduce or prevent adhesions that may form after surgery.

The present invention can also provide a method for preventing or treating inflammatory eye diseases and angiogenic eye diseases by administering the pharmaceutical composition containing the sulfated hyaluronic acid of the present invention. As a representative example, the pharmaceutical composition containing the sulfated hyaluronic acid of the present invention can be used for inflammatory or angiogenic eye diseases, such as dry eye, uveitis, and macular degeneration And diabetic retinopathy.

The pharmaceutical composition containing the sulfated hyaluronic acid of the present invention may be particularly useful for treating chronic or acute inflammation.

As used herein, the term "treatment" means stopping, delaying or improving the progression of the disease in an individual showing symptoms of the disease. The term "prevention" means stopping, delaying or ameliorating the symptoms of an individual who is at risk of showing symptoms of the disease, even if he or she does not show the symptoms.

The following examples are presented to provide a complete disclosure and description of how to make and evaluate the compounds, compositions, and methods provided herein for those with ordinary knowledge in the technical field to which the invention belongs, and these examples are only exemplary. Therefore, these embodiments are in no way intended to limit the scope of the invention viewed by the inventors. There are many variations and combinations of reaction conditions, such as component concentrations, required solvents, solvent mixtures, temperature, pressure, and other reaction parameters and conditions that can be used to optimize product characteristics (such as purity, yield, etc.). These are also considered to be within the scope of this application. Unless otherwise indicated herein or clearly contradicted by the context, the present invention encompasses any combination of all possible variations of the above elements.

[Example 1]

Dissolve hyaluronic acid (HA)-tetrabutylammonium salt (TBA) derivative (where TBA is introduced into HA) in dimethylformamide (DMF) which is an organic solvent to make the concentration 3 mg/mL . 25 mol times the sulfur trioxide pyridine complex of the HA-TBA derivative monomer was added to it. After reacting for 1 hour at 5°C under nitrogen, ethanol precipitation for purification was performed to produce sulfated hyaluronic acid substituted with 200% sulfate.

[Example 2]

Example 2 was performed in the same manner as in Example 1, except: adding 30 mol times the sulfur trioxide pyridine complex of the HA-TBA derivative monomer; and after reacting at 5°C for 0.5 hours under nitrogen, Perform ethanol precipitation for purification to produce sulfated hyaluronic acid substituted with 210% sulfate.

[Example 3]

Example 3 was performed in the same manner as in Example 1, except: adding 20 mol times the sulfur trioxide pyridine complex of the HA-TBA derivative monomer; and after reacting at 5°C under nitrogen for 2 hours, Perform ethanol precipitation for purification to produce sulfated hyaluronic acid substituted with 240% sulfate.

[Example 4]

Example 4 was performed in the same manner as in Example 1, except that: 30 mol times the sulfur trioxide pyridine complex of the HA-TBA derivative monomer was added; and after reacting at 5°C under nitrogen for 3 hours, Perform ethanol precipitation for purification to produce sulfated hyaluronic acid substituted with 280% sulfate.

[Comparative Example 1]

Comparative Example 1 was performed in the same manner as in Example 1, except: adding 10.6 mol times the sulfur trioxide pyridine complex of HA-TBA derivative monomer; and after reacting at 5°C under nitrogen for 2 hours, proceed Ethanol precipitation for purification to produce sulfated hyaluronic acid substituted by 110% sulfate.

[Comparative Example 2]

Comparative Example 2 was performed in the same manner as in Example 1, except: adding 18 mol times the sulfur trioxide pyridine complex of the HA-TBA derivative monomer; and after reacting at 5°C under nitrogen for 2 hours, Perform ethanol precipitation for purification to produce sulfated hyaluronic acid substituted with 180% sulfate.

[Comparative Example 3]

Comparative Example 3 was performed in the same manner as in Example 1, except: adding 50 mol times the sulfur trioxide pyridine complex of the HA-TBA derivative monomer; and after reacting at 5°C under nitrogen for 3 hours, Perform ethanol precipitation for purification to produce sulfated hyaluronic acid substituted with 320% sulfate.

[Comparative Example 4]

Comparative Example 4 was performed in the same manner as in Example 1, except that: adding 100 mol times the sulfur trioxide pyridine complex of the HA-TBA derivative monomer; and after reacting at 5°C under nitrogen for 3 hours, Perform ethanol precipitation for purification to produce sulfated hyaluronic acid substituted with 370% sulfate.

[Comparative Example 5]

Comparative Example 5 was performed in the same manner as in Example 1, except: adding 16 mol times the sulfur trioxide pyridine complex of the HA-TBA derivative monomer; and after reacting at 5°C under nitrogen for 2 hours, Perform ethanol precipitation for purification to produce sulfated hyaluronic acid substituted with 150% sulfate.

[Experimental Example 1: Determination of Sulfate Substitution Rate]

The synthesis of the sulfated hyaluronic acid (SHA) prepared in Examples 1 to 4 and Comparative Examples 1 to 5 was confirmed by 1H NMR analysis. Measure the sulfate substitution rate by elemental analysis (EA). The molecular weight is determined by MALLS/RI analysis. The results are shown in Figure 1 (NMR analysis results) and Table 1 (EA and MALLS/RI analysis results).

Figure 1 shows the NMR analysis results of Example 1. Referring to Figure 1, as the sulfation progresses, the peak at 3.4 ppm disappears, and the peak appears between 4.1 and 4.3 ppm, indicating the introduction of sulfate groups.

Referring to Table 1, the sulfate substitution rate of the sulfated hyaluronic acid of Examples 1 to 4 falls within the range of 200% to 300%. It was found that the sulfate substitution rate of the sulfated hyaluronic acid of Comparative Examples 1 to 5 fell outside the above-mentioned range.

[Experimental Example 2: Evaluation of Anti-inflammatory Effect]

Use the sulfated hyaluronic acid (SHA) prepared in Examples 3 and 4 and Comparative Examples 1 to 4 in two types of cells (chondrocytes and fibroblast-like synovial cells (FLS)) for anti-inflammatory Effectiveness evaluation.

Two types of cells (chondrocytes and FLS) were ^{seeded into 24 wells at 5×10 4} and 2×10 ⁴ cells/well, and treated with 5 ng/ml and 2 ng/ml TNF-α The cells were activated overnight. Replace the medium, and then treat the cells with 1 mg/ml of each sample. After 24 hours of incubation after processing the sample, ELISA was used to measure the cytokine secretion of the cells in the culture medium. The results are shown in Figure 2.

Referring to Figure 2, it was found that in the two cell types of chondrocytes and FLS, the higher the sulfate substitution rate, the better the inhibitory effect on the secretion of IL-6 and IL-8 cytokine. Comparative Examples 1 and 2 (the replacement rate falls within the 100% band) showed insufficient anti-inflammatory effects, and the level was similar to that of HA. However, Examples 3 and 4 and Comparative Examples 3 and 4 (with a substitution rate of 200% or higher) inhibited the secretion of IL-6 and IL-8 in both cell types.

In addition to the sulfate substitution rate, in order to determine the efficacy based on the molecular weight, the samples of Examples 3 and 4 (in which the molecular weight was adjusted to a small value only by pyrolysis) were used to perform the above-mentioned cell experiment. The results are shown in Figure 3.

Referring to Figure 3, Examples 3 and 4 (with a sulfate substitution rate of 240 to 280%) also exhibit excellent anti-inflammatory effects at a molecular weight of 300 to 2000 kDa.

In addition to the secretion of IL-6 and IL-8, the secretion of MMP13, which is an inflammation-related factor, was measured by ELISA using the above cell culture samples. The results are shown in Figure 4.

Referring to Fig. 4, Example 1 (its sulfate substitution rate is 200%) exhibits an excellent inhibitory effect on MMP13 secretion. However, in Comparative Example 3 and Comparative Example 4 (whose sulfate substitution rate was 300% or higher), the secretion tended to increase.

[Experimental example 3: Evaluation of anti-angiogenesis efficacy]

The effectiveness of inhibiting the tube formation and proliferation of HUVEC cells was determined.

In order to determine the effectiveness of inhibiting the angiogenesis of HUVEC cells, Matrigel was used to conduct 3D culture of VEGF-treated HUVEC cells, and the use of the sulfated hyaluronic acid samples (Examples 2 and 3 and Comparative Examples 1 and 4) ) Whether angiogenesis (tube formation) is inhibited when processing the HUVEC cells treated with VEGF. In addition, HUVEC cells treated with VEGF were seeded in 96 wells coated with Matrigel, and HUVEC cells were treated with Avastin. Avastin is an anti-VEGF agent and used as the sulfated hyaluron. Acid comparison agent. After 6 hours of incubation, the formation of blood vessels (tubes) was measured through an optical microscope. The results are shown in Figure 5.

Referring to Fig. 5, in the Aisiting treatment group and the hyaluronic acid (HA) treatment group, the level of the blood vessel structure formed was similar to that of the control group. However, in Examples 2 and 3, and Comparative Example 4 (whose acid salt substitution rate is 200% or higher), it was found that the tube was hardly formed. However, in Comparative Example 1 (in which the sulfate substitution rate was less than 200%), a blood vessel structure was formed.

In order to determine the effect of inhibiting the proliferation of HUVEC cells, a sample group treated with or without VEGF, FGF and FBS was used as a control group, and different concentrations of sulfated hyaluronic acid were used (Examples 2 to 4 and Comparative Examples 1, 3 And 5) HUVEC cells were treated with Aisiting (which is an anti-VEGF agent), and whether cell proliferation was inhibited was determined.

HUVEC cells were seeded into 96 wells at 7,500 cells/well, and then treated with 100 ng/ml VEGF, followed by various concentrations of sulfated hyaluronic acid and Oncetin samples. Incubate the cells for 72 hours while checking the number of cells (cell population) in the Incucyte device. The results are shown in Figures 6 and 7.

Referring to Fig. 6, in the group treated with the samples of Examples 2 to 4, cell proliferation tended to be inhibited according to the concentration. At high concentrations, the proliferation of HUVEC cells was inhibited at the level of the negative control group not treated with VEGF, FGF and FBS. The anti-VEGF agent Aisiting inhibited cell proliferation at the level of the negative control group without VEGF even at a relatively low concentration. Although a high concentration is required, the sulfated hyaluronic acid of Examples 2 to 4 blocks various growth factors and therefore exhibits a better cell proliferation inhibitory effect than Aisiting. Unsulfated hyaluronic acid did not show the effect of inhibiting cell proliferation at high concentrations, but showed the effect of cell proliferation. In Comparative Example 1, even at high concentrations, the confluence of the cells was similar to that of the VEGF-treated group, and the effect of inhibiting the proliferation of HUVEC cells was insufficient. In Comparative Example 5, a higher concentration than those in Examples 2 to 4 is required to inhibit the proliferation of HUVEC cells, so the cell proliferation inhibitory effect is low. In Comparative Example 3, the cells did not proliferate even at low concentrations.

[Experimental Example 4: Evaluation of Cytotoxicity]

The sulfated hyaluronic acid (SHA) prepared in Examples 3 and 4 and Comparative Examples 1 to 4 were used to evaluate the cytotoxicity of two types of cells, HUVEC and chondrocytes, respectively.

Two types of cells, HUVEC and chondrocytes, were seeded in 96 wells at 7,500 cells/well, and treated with 1 mg/ml of each sample. The cells were incubated for 72 hours, and the cell population was examined using the Incucyte equipment. The results are shown in Figure 8.

Referring to Fig. 7, in the sample group treated with Example 3 or 4 or Comparative Example 1 or 2, the rate of cell proliferation may be affected, but no toxicity was observed. However, cytotoxicity was observed in the sample group treated with Comparative Example 3 or 4. Therefore, in order to develop sulfated hyaluronic acid as a therapeutic agent, the substitution rate needs to be 300% or less.

[Experimental Example 5: Evaluation of Blood Anticoagulation]

Using the sulfated hyaluronic acid in Examples 2 and 3 and Comparative Examples 1, 3 and 4, blood anticoagulation tests were performed. In particular, the sample of Example 2 was heat-treated to prepare sulfated hyaluronic acid (SHA) having molecular weights of 1,700 kDa, 1,000 kDa, 400 kDa, and 200 kDa, and the influence of molecular weight was also evaluated. Using each sulfated hyaluronic acid, the activity inhibition of factor IIa and factor Xa was measured by chromogenic analysis. The results are shown in Table 2.

Referring to Table 2 and Figure 8 above, the activity of the sulfated hyaluronic acid in inhibiting factor IIa and factor Xa is 500 times or more lower than that of heparin. The activity of Example 3 for inhibiting factor IIa and factor Xa was significantly lower than that of Comparative Example 3 or 4 (which has a high sulfate substitution rate), and it was found that Example 3 had almost no anticoagulant effect. Also, in the aspect of Example 2, it was found that when the molecular weight is 1,000 kDa or higher, the activity of inhibiting factor IIa and factor Xa is further reduced.

[Figure 1] is the 1H NMR result for identifying the sulfated substitution of the sulfated hyaluronic acid (SHA).

[Figure 2] The graphs shown illustrate the evaluation of the anti-inflammatory efficacy of the sulfated hyaluronic acid of Examples 3 and 4, and Comparative Examples 1 to 4.

[Figure 3] The graph shown illustrates the evaluation of the anti-inflammatory efficacy of the molecular weight of sulfated hyaluronic acid according to the present invention.

[Figure 4] The graph shown shows the secretion of MMP13 during treatment with the sulfated hyaluronic acid of Example 1 and Comparative Examples 1 and 3 to 4.

[Figure 5] is an optical micrograph used to observe the inhibitory effect of sulfated hyaluronic acid treatment on angiogenesis (tube formation) in HUVEC cell samples treated with VEGF.

[Figure 6] The graph shown illustrates the evaluation of the cell proliferation inhibitory effect when HUVEC cells are treated with the sulfated hyaluronic acid of Examples 2 to 4, and Comparative Examples 1, 3, and 5.

[Figure 7] The graphs shown illustrate the cytotoxicity evaluation of the sulfated hyaluronic acid of Examples 3 and 4, and Comparative Examples 1 to 4.

Claims (9)

An anti-inflammatory pharmaceutical composition comprising at least one selected from the group consisting of: sulfated hyaluronic acid, its derivatives and salts, each repeating unit of hyaluronic acid in the sulfated hyaluronic acid is Alcoholic hydroxyl groups have an average sulfate substitution rate of 200% to 300%. The anti-inflammatory pharmaceutical composition of claim 1, wherein the weight average molecular weight (Mw) of at least one selected from the group consisting of the sulfated hyaluronic acid, its derivatives and salts is 500 to 3,000 kDa. An anti-angiogenesis pharmaceutical composition comprising at least one selected from the group consisting of: sulfated hyaluronic acid, its derivatives and salts, each repeating unit of hyaluronic acid in the sulfated hyaluronic acid For alcoholic hydroxyl groups, it has an average sulfate substitution rate of 200% to 300%. The anti-angiogenic pharmaceutical composition of claim 3, wherein the weight average molecular weight (Mw) of at least one selected from the group consisting of the sulfated hyaluronic acid, its derivatives, and salts is 500 to 3,000 kDa. A medical composition for the treatment or prevention of arthritis, the composition comprising at least one selected from the group consisting of: sulfated hyaluronic acid, derivatives and salts thereof, each of the sulfated hyaluronic acid The repeating unit of hyaluronic acid has an average sulfate substitution rate of 200% to 300% for alcoholic hydroxyl groups. The medical composition of claim 5, which is used to treat or prevent arthritis, osteoarthritis, rheumatoid arthritis, gout, or calcium pyrophosphate dihydrate deposition disease. A medical composition for the treatment or prevention of inflammatory or angiogenic eye diseases, the composition comprising at least one selected from the group consisting of: sulfated hyaluronic acid and its salts, the sulfated hyaluronic acid Each repeating unit of hyaluronic acid in the acid has an average sulfate substitution rate of 200% to 300% for alcoholic hydroxyl groups. The medical composition of claim 7, which is used to treat or prevent dry eye, uveitis, macular degeneration, or diabetic retinopathy. An anticancer agent comprising at least one selected from the group consisting of: sulfated hyaluronic acid, derivatives and salts thereof, each repeating unit of hyaluronic acid in the sulfated hyaluronic acid has an alcoholic hydroxyl group The average sulfate replacement rate is 200% to 300%.

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