Sulfated Hyaluronic Acid

Abstract

The purpose of the present invention is to provide a novel use of sulfated hyaluronic acid. The present invention provides a composition for a subject who has been exposed to radiation or who is at a risk of being exposed to radiation, the composition being for reducing and / or preventing one or more adverse effects associated with the exposure to radiation. The composition is characterized by including a sulfated hyaluronic acid. The present invention also provides a composition that contains as an active ingredient a fibroblast growth factor for use at an affected site where bleeding needs to be arrested, and that further contains a sulfated hyaluronic acid to protect the fibroblast growth factor.

Description

[Technical field]

[0001] The present invention relates to a novel use of sulfated hyaluronic acid. [Background technology]

[0002] Hyaluronic acid is a linear polymeric polysaccharide formed by alternating bonds between β-DN-acetylglucosamine and β-D-glucuronic acid. Hyaluronic acid is relatively easy to obtain among mucopolysaccharides, and because it exhibits unique physicochemical and physiological properties, it is used in medicines and cosmetics, either by itself or in its various derivatives.

[0003] Hyaluronic acid has been synthesized into derivatives to improve its mechanical properties and to improve its properties to make it suitable for medical materials. One of these is sulfation. Hyaluronic acid is not sulfated in vivo, so sulfation has been studied to give it anticoagulant activity. It is generally known that the more hydroxyl groups in hyaluronic acid are sulfated, the higher the activity becomes. In addition, polysulfated hyaluronic acid is known to have inhibitory activity against the kallikrein-kinin system (Patent Document 1: JP-A-11-147901) and phospholipase A2 (Patent Document 2: JP-A-11-269077), and can be used as a therapeutic drug for allergic diseases (Patent Document 3: JP-A-11-335288), and to show a strong anti-inflammatory effect against selectin-mediated inflammation, which is one of the adhesion factors (Patent Document 4: JP-A-8-277224).

[0004] It has also been reported that low molecular weight polysulfated oligohyaluronic acid with a viscosity average molecular weight of 10,000 or less can be used as an active ingredient in cosmetics with excellent skin permeability (Patent Document 5: JP 10-195107 A), and that polysulfated hyaluronic acid oligosaccharides of 4 to 20 sugars have anticoagulant activity and antihyaluronidase activity and may be used as anticancer agents (Non-Patent Document 1: Glycobiology, vol. 11, No. 1, pp. 57-64, 2001).

[0005] Fibroblast growth factor (FGF) is one of the growth factors involved in angiogenesis, wound healing, and embryonic development. FGF is a heparin-binding protein that interacts with heparan sulfate, a type of proteoglycan on the cell surface, which results in the formation of FGF signal transduction. In addition, FGF plays an important role in the proliferation and differentiation process of a wide range of cells and tissues, including FGF-7, which promotes the proliferation and division of hair matrix cells. (Patent Document 6: JP2002-526026)

[0006] Since bFGF (FGF2) has a wide range of physiological activities, such as development, organogenesis, and homeostasis, treatment methods for diseases focusing on bFGF have been proposed in various fields. For example, in the field of skin, a treatment agent containing bFGF as an active ingredient for skin regeneration and skin ulcer treatment for wound healing has been released (Kaken Pharmaceutical Co., Ltd.: Fiblast Spray). It has also been proposed to use it as a treatment agent for bone diseases (Patent Document 7: JP Patent Publication No. 05-124975).

[0007] In the field of oral cavity, clinical trials are being conducted in which bFGF is administered directly to the gums of patients with periodontal disease during surgery, and its effectiveness as a drug for inducing periodontal tissue regeneration is becoming clear (Non-Patent Document 2: Murakami, Journal of the Japanese Society of Periodontology, 54, 38-45, 2012). (Non-Patent Document 3: M. Kitamura et al. PLoS One. 2008 Jul 2;3(7):e2611.)

[0008] It has been reported that certain types of sulfated polysaccharides have the effect of protecting FGF from decomposition, denaturation, inactivation, etc. For example, Patent Document 8 (WO92 / 13526) discloses that carrageenan stabilizes bFGF. Patent Document 9 (JP Patent Publication 02-138223) discloses a method for stabilizing FGF or its mutein, which is characterized by contacting FGF or its mutein with sulfated glucan in an aqueous medium. Non-Patent Document 4 (J. Cell. Physiol., 1986, 128, 475-484) describes that heparin or hexyuronyl hexosaminoglycan sulfate (HHS-4) protects bFGF from inactivation and enhances its physiological activity.

[0009] In addition, a medium and a culture method that can activate cells using sulfated hyaluronic acid and promote differentiation and maturation into nerve cells have been reported (Patent Document 10: JP 2009-278873 A). In this method, sulfated hyaluronic acid with a sulfation degree of 0.4 or 1.0 is used, and it is described that by adding a combination of sulfated hyaluronic acid and FGF-2, cells can be activated and differentiation and maturation into nerve cells can be promoted. [Prior art documents] [Patent documents]

[0010] [Patent Document 1] Patent Publication No. 11-147901 [Patent Document 2] Patent Publication No. 11-269077 [Patent Document 3] JP 11-335288 [Patent Document 4] Patent Publication No. 8-277224 [Patent Document 5] Patent Publication No. 10-195107 [Patent Document 6] Special table 2002-526026 [Patent Document 7] Japanese Patent Application Publication No. 05-124975 [Patent Document 8] WO92 / 13526 [Patent Document 9] Japanese Patent Publication No. 02-138223 [Patent Document 10] Patent Publication No. 2009-278873 [Non-patent literature]

[0011] [Non-Patent Document 1] Glycobiology, vol.11, No.1, pp.57-64, 2001 [Non-Patent Document 2] Murakami, Journal of the Japanese Society of Dentistry, 54, 38-45, 2012 [Non-Patent Document 3] M. Kitamura et al. PLoS One. 2008 Jul 2;3(7):e2611. [Non-Patent Document 4] J. Cell. Physiol., 1986, 128, 475-484 Summary of the Invention [Problem to be solved by the invention]

[0012] An object of the present invention is to provide a new use of sulfated hyaluronic acid. [Means for solving the problem]

[0013] As a result of intensive research, the inventors discovered that sulfated hyaluronic acid has the same protective effect on nutritional factors and growth factors, such as fibroblast growth factor (FGF), as heparin, but has an extremely weaker anticoagulant effect than heparin, and as a result, sulfated hyaluronic acid is useful even in applications where heparin cannot be used, thus completing the present invention. More specifically, the present invention includes the following: [1] A composition for reducing and / or preventing one or more side effects associated with exposure to radiation in a subject exposed to or at risk of radiation exposure, the composition comprising sulfated hyaluronic acid. [2] The composition according to [1] above, wherein the sulfated hyaluronic acid has a number of sulfate groups per hyaluronic acid disaccharide unit (hereinafter, the term "sulfation degree (number of sulfate groups / disaccharide unit)") of about 1.3 or more. [3] The composition according to [1] above, wherein the sulfated hyaluronic acid has a degree of sulfation (number of sulfate groups / disaccharide unit) of about 3.0 or more. [4] The composition according to any one of [1] to [3] above, wherein the molecular weight of the sulfated hyaluronic acid is about 30,000 or more. [5] The composition according to any one of [1] to [4] above, further comprising a growth factor. [6] The composition described in [5] above, wherein the growth factor is at least one growth factor selected from the group consisting of fibroblast growth factor (FGF), hepatic growth factor (HGF), heparin-binding epidermal growth factor (HBEGF), vascular endothelial growth factor (VEGF), midkine, cytokines, and bone morphogenetic proteins (BMPs). [7] The composition described in [5] above, wherein the growth factor is at least one growth factor selected from the group consisting of fibroblast growth factors FGF1, FGF2, FGF3, FGF4, FGF5, FGF6, FGF7, FGF8, and FGF9. [8] The composition according to any one of the above [1] to [7], wherein the composition is for use in any one of the administration forms selected from the group consisting of intravascular administration, intraperitoneal administration, and oral administration. [9] The composition according to any one of the above [1] to [8], wherein the radiation is at least one radiation selected from the group consisting of gamma radiation, X-ray radiation, solar radiation, cosmic radiation, electromagnetic radiation, bremsstrahlung radiation, ultraviolet radiation, and particulate radiation.

[10] The composition according to any one of the above [1] to [9], wherein the side effect associated with exposure to radiation is at least one side effect selected from the group consisting of acute nausea, vomiting, abdominal pain, diarrhea, dizziness, headache, fever, cutaneous radiation syndrome, low blood count, infection due to leukopenia, bleeding due to thrombocytopenia, anemia due to erythrombocytopenia, and death.

[11] Use of the composition according to any one of [1] to

[10] above for protecting internal organs of a subject from radiation.

[12] The use according to

[11] above, wherein the organ to be protected is at least one selected from the group consisting of bone marrow, liver, spleen, kidney, lung and gastrointestinal tract.

[13] The composition according to any one of [1] to

[10] above, which is used in combination with one or more antibiotics.

[14] A composition for use in an affected area where maintenance of blood coagulation function is required, the composition containing a growth factor as an active ingredient and further containing sulfated hyaluronic acid to protect the growth factor.

[15] The composition described in

[14] above, wherein the affected area in which maintenance of blood coagulation function is required is a wound.

[16] The composition according to

[14] above, wherein the affected area in which maintenance of blood coagulation function is required is an affected area of ​​periodontal disease.

[17] The composition described in

[14] above, wherein the affected area in which maintenance of blood coagulation function is required is an affected area for Platelet Rich Plasma (PRP) hair growth therapy.

[18] The composition according to any one of the above

[14] to

[17] , wherein the sulfated hyaluronic acid has a degree of sulfation (number of sulfate groups / disaccharide unit) of about 1.3 or more.

[19] The composition according to any one of the above

[14] to

[18] , wherein the molecular weight of the sulfated hyaluronic acid is about 30,000 or more.

[20] The composition according to any one of the above

[14] to

[19] , wherein the growth factor is at least one growth factor selected from the group consisting of fibroblast growth factor (FGF), hepatic growth factor (HGF), heparin-binding epidermal growth factor (HBEGF), vascular endothelial growth factor (VEGF), midkine, cytokines, and bone morphogenetic proteins (BMPs).

[21] The composition according to any one of the above

[14] to

[19] , wherein the growth factor is at least one growth factor selected from the group consisting of fibroblast growth factors FGF1, FGF2, FGF3, FGF4, FGF5, FGF6, FGF7, FGF8, and FGF9.

[22] Use of sulfated hyaluronic acid having a degree of sulfation (number of sulfate groups / disaccharide unit) of about 1.3 or more for stabilizing growth factors.

[23] The use described in

[22] above, wherein the growth factor is at least one growth factor selected from the group consisting of fibroblast growth factor (FGF), hepatic growth factor (HGF), heparin-binding epidermal growth factor (HBEGF), vascular endothelial growth factor (VEGF), midkine, cytokines, and bone morphogenetic proteins (BMPs).

[24] The use according to

[22] above, wherein the growth factor is at least one growth factor selected from the group consisting of fibroblast growth factors FGF1, FGF2, FGF3, FGF4, FGF5, FGF6, FGF7, FGF8, and FGF9.

[25] The use according to any one of the above

[22] to

[24] , wherein the molecular weight of the sulfated hyaluronic acid is 30,000 or more. Effect of the Invention

[0014] The present invention provides a new use of sulfated hyaluronic acid. [Brief description of the drawings]

[0015] [Figure 1] FIG. 1 shows the structure of the sulfated hyaluronic acid used in the present invention. [Diagram 2] FIG. 2 shows the results of confirming the bioavailability of fibroblast growth factor by sulfated hyaluronic acid using C2C12 cells (fibroblasts derived from mouse striated muscle cells). [Diagram 3] FIG. 3 shows the results of confirming the protective effect of sulfated hyaluronic acid on fibroblast growth factor 2 (FGF2). [Figure 4] FIG. 4 shows the results of confirming the protective effect of sulfated hyaluronic acid on fibroblast growth factor 1 (FGF1). [Diagram 5] FIG. 5 shows the results of confirming the protective effect of sulfated hyaluronic acid on fibroblast growth factor 7 (FGF7). [Figure 6] FIG. 6 shows the results of confirming the skin irritation caused by sulfated hyaluronic acid. [Figure 7] Figure 7 shows the results of measuring blood clotting time after adding sugar chains at various concentrations. HA is hyaluronic acid, HA-LS is low-sulfated hyaluronic acid, and HA-HS is high-sulfated hyaluronic acid. The figure on the right shows the results of blood clotting time when 10 μg / mL of each sugar chain was added. Each value is the mean ± standard deviation (n=3-6). ns not significant; *P<0.05; **P<0.01; ***P<0.001 [Figure 8] Figure 8 shows the results of confirming the antithrombin activity of sulfated hyaluronic acid. HA is hyaluronic acid, HA-LS is low-sulfated hyaluronic acid, and HA-HS is high-sulfated hyaluronic acid. Each value is the mean ± standard deviation (n=3). ns: not significant; *P < 0.05; **P < 0.01; ***P < 0.001 [Figure 9] Figure 9 shows the results of cryptoassay in mice administered only highly sulfated hyaluronic acid (HA-HS) 24 hours before whole-body irradiation with gamma rays. Each value is the mean ± standard deviation (n=3). *P < 0.05 [Figure 10] Figure 10 shows the results of cryptoassays in mice administered each glycan 24 hours before whole-body irradiation with gamma rays. HP is heparin, HA is hyaluronic acid, HA-LS is low-sulfated hyaluronic acid, and HA-HS is high-sulfated hyaluronic acid. Each value is the mean ± standard deviation (n = 3-9). ns not significant; *P < 0.05; **P < 0.01; ***P < 0.001. [Figure 11] Figure 11 shows the results of cryptoassays in mice administered with each glycan 24 hours after whole-body irradiation with gamma rays. Each value is the mean ± standard deviation (n = 6-12). ns non-significant; *P < 0.05; **P < 0.01; ***P < 0.001. [Figure 12]Figure 12 shows the results of administering each glycan 24 hours before whole-body γ-irradiation of mice, and measuring apoptosis by TUNEL staining 24 hours after γ-irradiation. Each value is the mean ± standard deviation (n = 3-4). ns non-significant; *P < 0.05; **P < 0.01; ***P < 0.001. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0016] The present invention will be described below with exemplary embodiments as examples, together with preferred methods and materials that can be used in the practice of the present invention. Unless otherwise specified in the text, all technical and scientific terms used in the present specification have the same meaning as commonly understood by those skilled in the art to which the present invention belongs. In addition, any materials and methods equivalent or similar to those described in the present specification can be used in the practice of the present invention. In addition, all publications and patents cited in the present specification in relation to the invention described in the present specification are incorporated as part of the present specification, for example, as a description of methods, materials, etc. that can be used in the present invention.

[0017] In the present specification, the expression "X to Y" means that X is the lower limit and Y is the upper limit. In the present specification, the term "about" means that ±10% is allowed. In the present specification, the terms "growth factor" and "growth factor" are interchangeable terms having the same meaning.

[0018] In the present invention, "sulfated hyaluronic acid" refers to a substance in which some or all of the hydroxyl groups of hyaluronic acid, a linear polymeric polysaccharide formed by alternating bonds of β-DN-acetylglucosamine and β-D-glucuronic acid, are replaced with sulfate groups. The structure of sulfated hyaluronic acid is shown in Figure 1. In sulfated hyaluronic acid, some or all of the ORs are replaced with sulfate groups. The degree of sulfation is expressed as the "sulfation degree". In this specification, the sulfation degree is expressed as the sulfate content (S content) (%) in a sulfated hyaluronic acid molecule, or the number of sulfate groups introduced (0 to 4) in a disaccharide unit, which is a constituent unit.

[0019] The sulfate content (turbidimetric method) can be measured using the Dogson-Price turbidimetric method (Dogson, KS and Price, RG, A note on the determination of the ester sulphate content of sulphated polysaccharides, Biochem J. vol 84(1),106-110, 1962) with potassium sulfate as the standard. The sulfate content (gravimetric method) can be measured using a method conforming to JIS JB. 2.17.3 Sulfate-gravimetric method. The sulfation degree can also be expressed as the average number of sulfate groups introduced. 0 to 4 sulfate groups can be introduced per disaccharide unit. When expressed as the average number, the average number of sulfate groups in a disaccharide unit can be calculated from the S content based on the molecular weight of sulfated hyaluronic acid. In this specification, when it is described that the "sulfation degree (number of sulfate groups / disaccharide unit) is 1.3", it means that an average of 1.3 sulfate groups are introduced per disaccharide unit.

[0020] The sulfated hyaluronic acid used in the present invention (hereinafter, sometimes simply referred to as "sulfated hyaluronic acid") is hyaluronic acid with a relatively high degree of sulfation. As a specific compound of sulfated hyaluronic acid, hyaluronic acid with a degree of sulfation of 1.0 or less as the number of sulfate groups / disaccharide unit has been reported (for example, see JP 2009-278873 A). The S content of the sulfated hyaluronic acid used in the present invention is about 7% or more by turbidimetric method, and in one aspect, it is preferably about 8% or more, more preferably about 9% or more, and even more preferably about 10% or more. In addition, the sulfated hyaluronic acid used in the present invention has a number of sulfate groups / disaccharide unit of about 1.3 or more, and in one aspect, it is preferably about 1.5 or more, more preferably about 1.6 or more, even more preferably about 1.8 or more, and even more preferably about 1.9 or more.

[0021] The sulfated hyaluronic acid used in the present invention is available from Tokyo Chemical Industry Co., Ltd., and sulfated hyaluronic acid with different degrees of sulfation is available. In addition, sulfated hyaluronic acid can be obtained by dissolving hyaluronic acid and sulfating agent in a suitable solvent and reacting them under heating using a known sulfation reaction.In addition, the sulfation degree can be adjusted by referring to a known method and appropriately changing the amount of sulfating agent used.The measurement of sulfation degree can be performed, for example, by the above-mentioned method, but can also be performed by quantitatively determining the sulfate ion produced by acid hydrolysis using ion chromatography.

[0022] The solvent used in the sulfation reaction may be any solvent that is generally usable in the art, without any restrictions. Examples of the solvent include N,N-dimethylformamide, dimethylsulfoxide, N-methylpyrrolidone, N,N-dimethylacetamide, pyridine, N,N-dimethylacrylamide, or a mixture thereof.

[0023] As the sulfating agent, any sulfating agent that can be generally used in the technical field can be used without limitation, and examples thereof include complexes of sulfuric anhydride with pyridine, picoline, 2,6-lutidine, trimethylamine, triethylamine, N,N-dimethylformamide, dioxane, etc., or sulfuric acid-dicyclohexylcarbodiimide, chlorosulfone, etc. In addition, in the sulfation reaction, an acid catalyst such as trifluoroacetic acid or trifluoromethanesulfonic acid may be added to the reaction system.

[0024] The reaction temperature and reaction time for the sulfation are not particularly limited, but examples include a temperature of 0 to 100° C. and a time of 5 minutes to 5 days.

[0025] The sulfated hyaluronic acid used in the present invention can be used without any particular limitation as long as it is of high molecular weight, but specifically, the molecular weight of the sulfated hyaluronic acid used in the present invention is, for example, about 20,000 or more, preferably about 30,000 or more, more preferably about 50,000 or more, and even more preferably about 100,000 or more.Preferably, as shown in the following examples, for example, sulfated hyaluronic acid with average molecular weight of about 30,000-70,000 is exemplified.When referring to the molecular weight of sulfated hyaluronic acid in the present invention, it means the molecular weight shown by any of the display methods generally used in the technical field, and is not limited thereto, but for example, the molecular weight calculated from the dynamic viscosity of raw material hyaluronic acid can be used.

[0026] The growth factors that can be used in the present invention are preferably cell growth factors, such as fibroblast growth factor (FGF), hepatic growth factor (HGF), heparin-binding epidermal growth factor (HBEGF), vascular endothelial growth factor (VEGF), midkine, cytokines, and bone morphogenetic proteins (BMPs). In the present specification, these are collectively referred to simply as growth factors, unless it is clear from the context that a specific growth factor is meant.

[0027] The fibroblast growth factor that can be used in the present invention is not particularly limited, and any fibroblast growth factor can be used, for example, but not limited to, FGF1, FGF2, FGF3, FGF4, FGF5, FGF6, FGF7, FGF8, and FGF9, preferably FGF1, FGF2, and FGF7, and more preferably FGF1 and FGF2. FGF can be prepared as appropriate by referring to known literature. Either natural FGF or recombinant FGF can be used as FGF, but recombinant FGF is preferred and is commercially available.

[0028] Other growth factors that can be used in the present invention can also be used without particular limitation. Cytokines include, for example, chemokines, interferons, interleukins, lymphokines, and tumor necrosis factors, preferably IL-6. Examples of bone morphogenetic factors include BMP-1, BMP-2, BMP-3, BMP-4, BMP-5, BMP-6, BMP-7, BMP-8a, BMP-8b, BMP-10, BMP-11, BMP-15, preferably BMP-2 or BMP-7. Other growth factors such as HGF, HBEGF, VEGF, NFG, cytokines, and BMP can be similarly prepared as appropriate by referring to known literature. Either natural or recombinant forms can be used, although recombinant forms are preferred and are commercially available.

[0029] One aspect of the present invention is the use of sulfated hyaluronic acid to reduce one or more side effects associated with radiation exposure in a subject who has been exposed to radiation or is at risk of radiation exposure, and the composition is administered to the subject for radiation protection, comprising sulfated hyaluronic acid in an amount effective to reduce radiation-related side effects.Sulfated hyaluronic acid does not show blood anticoagulant effect like heparin, so it can also be used for the damage of internal organs, such as gastrointestinal tract, caused by radiation damage.

[0030] Another aspect of the present invention is the use of sulfated hyaluronic acid and growth factor to reduce one or more side effects associated with radiation exposure in a subject exposed to radiation or at risk of radiation exposure, and the composition administered to the subject for radiation protection comprises sulfated hyaluronic acid and growth factor in an amount effective to reduce radiation-related side effects.Since growth factor such as fibroblast growth factor and sulfated hyaluronic acid each have radiation protection effect, their combined use can provide excellent radiation protection effect.In addition, sulfated hyaluronic acid has a protective effect, for example a stabilizing effect, of growth factor such as fibroblast growth factor, and can enhance the radiation protection effect of growth factor such as fibroblast growth factor.

[0031] Examples of the growth factors that can be used in the above aspects of the present invention include fibroblast growth factor (FGF), hepatic growth factor (HGF), heparin-binding epidermal growth factor (HBEGF), vascular endothelial growth factor (VEGF), midkine, a cytokine (preferably IL-6), and bone morphogenetic protein (BMP), and preferably FGF, HGF, HBEGF, or VEGF, and more preferably FGF.

[0032] The composition of the present invention can be used to increase the survival rate of a subject exposed to a radiation dose that is likely to kill the subject, and the composition contains sulfated hyaluronic acid or sulfated hyaluronic acid and a growth factor (preferably FGF, HGF, HBEGF, or VEGF) in an amount effective to increase the survival rate of a subject exposed to a radiation dose that is likely to kill the subject. The composition of the present invention is preferably administered to each subject intravascularly, intraperitoneally, or orally. The amount of radiation that is likely to kill a subject is known from the literature, and for example, the LD50 / 60d (i.e., the dose that causes 50% mortality in 60 days of exposure) in humans due to acute whole-body radiation exposure is greater than 250 rads (2.5 Gy), usually about 400 rads to 500 rads (4 Gy to 5 Gy).

[0033] The composition of the present invention can be used to reduce damage to a subject exposed to an amount of radiation likely to cause severe or mild damage to the subject, and contains sulfated hyaluronic acid or sulfated hyaluronic acid and a growth factor in an amount effective for reducing damage to the subject. The growth factor is preferably FGF, HGF, HBEGF, VEGF, midkine, cytokine, or BMP, more preferably FGF, HGF, HBEGF, or VEGF, and even more preferably FGF.

[0034] In such an embodiment, the sulfated hyaluronic acid of the present invention can be prepared from the above-mentioned sources or methods, and the degree of sulfation of the sulfated hyaluronic acid (number of sulfate groups / disaccharide unit) is about 1.3 or more, preferably about 1.5 or more, more preferably about 1.8 or more, and even more preferably about 3.0 or more.

[0035] The dosage of sulfated hyaluronic acid to a subject is 0.01mg-10mg, preferably 0.1mg-1mg per kg of body weight of the subject. In addition, when sulfated hyaluronic acid and fibroblast growth factor are administered to a subject in combination, the dosage of sulfated hyaluronic acid is 0.01mg-10mg, preferably 0.1mg-1mg, and the dosage of fibroblast growth factor is 0.01mg-100mg, preferably 0.1mg-10mg per kg of body weight of the subject.

[0036] The composition of the present invention may be administered to a subject before, during, or after the subject is exposed to radiation, but is preferably administered to the subject before the subject is exposed to radiation. The composition may be administered to a subject before the subject is exposed to radiation, but is preferably administered several hours to about 3 days before, more preferably half a day to 2 days before, and even more preferably half a day to 1 day before. It is thus preferable to administer the composition of the present invention in advance to a subject at risk of radiation exposure.

[0037] The form of the composition of the present invention when administered to a subject is not particularly limited, and may be any form such as tablet, powder, granule, liquid, injection, suspension, or other forms, or may be prepared at the time of use.When preparing at the time of use, the lyophilized or powdered composition of the present invention can be dissolved or suspended in water or a liquid for preparation at the time of use, and administered to a subject.When the composition of the present invention contains sulfated hyaluronic acid and fibroblast growth factor and is a composition for preparation at the time of use, it may be a powder (for example, a lyophilized agent) in which the two are mixed, or each may be a separate powder (for example, a lyophilized agent).

[0038] When administering the composition of the present invention to a subject, any administration route can be used, for example, intravascular administration, e.g., intravenous administration, intraperitoneal administration, oral administration, transdermal administration, etc., and intravascular administration is preferred.

[0039] The composition of the present invention preferably protects one or more internal organs of a subject from radiation. The protected organs are preferably one or more organs selected from the group consisting of bone marrow, liver, spleen, kidneys, lungs and gastrointestinal tract. The gastrointestinal tract is particularly preferably the small intestine.

[0040] Radiation-related side effects may be one or more of nausea, vomiting, abdominal pain, diarrhea, dizziness, headache, fever, skin radiation syndrome, low blood count, infection due to leukopenia, bleeding due to thrombocytopenia, anemia due to erythrombocytopenia, or death. Administering the composition of the present invention to a subject reduces one or more of these side effects. Also, in one embodiment, administering the composition of the present invention to a subject preferably increases the subject's chance of survival after exposure to radiation. In yet another embodiment, the radiation dose received by a subject may be lethal to the subject in the absence of radiation protection, and administering the composition of the present invention to a subject can protect the subject against higher radiation doses.

[0041] Administration of the compositions of the invention can also result in a reduced likelihood that an exposed subject will develop cancer as a result of long-term chronic radiation exposure.

[0042] The subject can be any animal. Preferably, the subject is a mammal, more preferably a human.

[0043] The radiation may include ionizing radiation. Ionizing radiation is sufficiently energetic to ionize atoms. The radiation may be, for example, one or more of gamma radiation, x-ray radiation, bremsstrahlung radiation, ultraviolet radiation, and particulate radiation (e.g., alpha radiation and beta radiation). The source of radiation may be a medical isotope. Preferably, the ionizing radiation is gamma radiation, alpha radiation, or beta radiation. Preferably, the radiation is from an artificial source. For example, the source of radiation may be radiation therapy used to treat disease (such as radiotherapy), radiation from medical imaging devices (such as CT scanners), radiosurgery (e.g., stereotactic radiosurgery), radiation used in nuclear weapons or nuclear reactors, such as reactors in power plants or submarines, or high-altitude radiation received, for example, during civilian or military flights or space flights. In one embodiment, the high-altitude radiation is natural ionizing radiation received at altitudes above 20,000 feet. The source of radiation may also result from a terrorist attack. Thus, artificial sources of radiation can include those derived from naturally occurring radioisotopes but applied to artificial therapies, sources or devices.

[0044] The composition of the present invention can be used to reduce and / or prevent one or more side effects associated with exposure of a subject to radiation, and the composition includes sulfated hyaluronic acid or sulfated hyaluronic acid and fibroblast growth factor in an amount effective to reduce and / or prevent one or more side effects associated with exposure of a subject to radiation.In one embodiment, the subject has received, is receiving, or will receive a single radiation exposure of 10mGy or more, 20mGy or more, 50mGy or more, 100mGy or more, 500mGy or more, 1Gy or more, 1.5Gy or more, 2Gy or more, 5Gy or more, 7.5Gy or more, 10Gy or more, or more than 10Gy.In humans, whole body exposure to high energy radiation of 5Gy or more in one dose usually causes death within 14 days.

[0045] In one embodiment, the composition of the present invention is administered to a subject together with one or more antibiotics. The antibiotics administered to a subject together with the composition of the present invention are not particularly limited, and any currently used antibiotics can be appropriately selected and used. The type of antibiotic is appropriately selected by a doctor, taking into consideration the patient's condition and other factors.

[0046] Another aspect of the present invention is a composition containing a growth factor as an active ingredient for use in affected areas where blood coagulation ability needs to be maintained (e.g., hemostasis), and further containing sulfated hyaluronic acid to protect the growth factor. Sulfated hyaluronic acid has a protective effect on growth factors and has a very low blood anticoagulant effect compared to heparin, so it is suitable for application to affected areas where blood coagulation ability needs to be maintained (e.g., hemostasis). Growth factors have effects such as angiogenesis and wound healing, and are used or developed as therapeutic agents for wound treatment and periodontal disease treatment. Therefore, the preferred use of the composition of the present invention containing a growth factor and sulfated hyaluronic acid for use in affected areas where blood coagulation ability needs to be maintained (e.g., hemostasis) includes treatment subjects where effects accompanied by cell proliferation are expected and where it is desired that the composition does not show an inhibitory effect on blood coagulation, such as wound treatment, periodontal disease treatment, and PRP hair growth therapy, are expected to have an effect accompanied by cell proliferation and do not show an inhibitory effect on blood coagulation.

[0047] Examples of the growth factors that can be used in the above aspects of the present invention include fibroblast growth factor (FGF), hepatic growth factor (HGF), heparin-binding epidermal growth factor (HBEGF), vascular endothelial growth factor (VEGF), midkine, a cytokine (preferably IL-6), and bone morphogenetic protein (BMP), and preferably FGF, HGF, HBEGF, or VEGF, and more preferably FGF.

[0048] In such an embodiment, the sulfated hyaluronic acid of the present invention can be prepared from the above-mentioned sources or methods, and the degree of sulfation of the sulfated hyaluronic acid (number of sulfate groups / disaccharide unit) is about 1.3 or more, preferably about 1.5 or more, more preferably about 1.8 or more, and even more preferably about 3.0 or more.

[0049] The composition of the present invention containing sulfated hyaluronic acid or sulfated hyaluronic acid and growth factors such as fibroblast growth factor can be used in medicines, quasi-drugs, and cosmetics.The formulation form can be selected from various forms, such as lotion, cream, milky lotion, foundation, pack, dusting powder, ointment, cataplasm, paste, plaster, essence, powder, pill, tablet, injection, suppository, emulsion, capsule, granule, liquid (including tincture, liquid extract, spirit, suspension, lemonade, etc.).In addition, for oral use, toothpaste, mouthwash, troche, etc. can be used.

[0050] The composition of the present invention can also contain other components used in medicines, quasi-drugs, and cosmetics, within the range that does not impair the effects of growth factors such as sulfated hyaluronic acid or fibroblast growth factor.The other components may be solid, liquid, or semi-solid, and can contain, for example, abrasives, foaming agents, humectants, moisturizing agents, binders, other medicinal ingredients, etc.In addition to vitamins, minerals, amino acids, etc., it can also contain excipients such as lactose, starch, cellulose, maltitol, dextrin, etc., surfactants such as glycerin fatty acid esters and sucrose fatty acid esters, coating agents such as gelatin, pullulan, shellac, and zein, fats and oils such as wheat germ oil, rice germ oil, and safflower oil, waxes such as beeswax, rice bran wax, and carnauba wax, sweeteners such as sucrose, glucose, fructose, stevia, saccharin, and sucralose, and acidulants such as citric acid, malic acid, and gluconic acid, etc. EXAMPLES

[0051] The present invention will be specifically described below with reference to examples, but the present invention is not limited to the following examples.

[0052] Example 1: Preparation of sulfated hyaluronic acid Sulfated hyaluronic acid was prepared by sulfating hyaluronic acid. Hyaluronic acid was sulfated using a known sulfation reaction, and various sulfation degrees were produced by appropriately adjusting the conditions. The hyaluronic acid used for sulfation had a molecular weight of 100,000. The sulfated hyaluronic acid was prepared first at a small scale (amount of hyaluronic acid used: 1 g) and then at a scaled-up scale (amount of hyaluronic acid used: 20 g). The sulfation degree of the prepared sulfated hyaluronic acid was confirmed. The results are shown in the table below. "S content (turbidimetric method)" is the result of measurement using the Dogson-Price turbidimetric method, and "S content (gravimetric method)" is the result of measurement using a method based on JIS JB. 2.17.3 sulfate-gravimetric method. "Sulfation degree" is the number of sulfate groups introduced, expressed as the average number per disaccharide unit.

[0053] [Table 1] Hereinafter, low sulfated hyaluronic acid will be abbreviated as HA-LS and high sulfated hyaluronic acid will be abbreviated as HA-HS.

[0054] The sulfated hyaluronic acid used in the present invention has a relatively high degree of sulfation, and has a high degree of sulfation that has not been reported before.In this specification, the one that has a relatively high degree of sulfation, but has a low degree of sulfation, is called low-sulfated hyaluronic acid, specifically, the one that has a degree of sulfation (number of sulfate groups / 2 monosaccharides) of about 1.9 is called low-sulfated hyaluronic acid, and the one that has a high degree of sulfation is called high-sulfated hyaluronic acid, specifically, the one that has a degree of sulfation (number of sulfate groups / 2 monosaccharides) of 3.0 or more is called high-sulfated hyaluronic acid.The highly sulfated hyaluronic acid shown in the above table indicates that most of the hydroxyl groups that can be replaced with sulfate groups are sulfated.

[0055] Example 2: Bioavailability of Fibroblast Growth Factors Mixed with Sulfated Hyaluronic Acid The bioavailability of sulfated hyaluronic acid mixed with fibroblast growth factors was confirmed as follows. C2C12 cells (fibroblasts derived from mouse striated muscle) were cultured in DMEM medium containing 10% bovine serum, and then the medium was changed to completely serum-free medium (1:1 mixture of DMEM / Ham's F10 medium, EGF: 50 ng / mL, bFGF: 5 ng / mL, 1μM dexamethasone, 0.12IU / mL insulin, 0.5% BSA) and the cells were seeded at 7,000 cells / mL on fibronectin-coated dishes, and heparin or sulfated hyaluronic acid was added and cultured for 3 days. Cell proliferation was then measured using the crystal violet method. Sulfated hyaluronic acid was added to completely serum-free medium at 2μg / mL, and medium without any addition and medium with heparin (2μg / mL) were used as controls. Since the addition of bFGF and heparin is effective for serum-free culture of C2C12 cells, if good cell proliferation is observed, it can be determined that the biological activity of FGF is being utilized. The results are shown in Figure 2. As shown in the figure, the addition of HA-LS was able to confirm significant cell proliferation similar to that of heparin.

[0056] Example 3: Protection of fibroblast growth factors by sulfated hyaluronic acid-1 The protective effect of sulfated hyaluronic acid on fibroblast growth factor was confirmed as follows: fibroblast growth factor and sulfated hyaluronic acid were mixed in PBS, pH 7.4, and the mixture was left to stand in an incubator at 37°C for 1 or 3 days, and then electrophoresis was performed to confirm the presence of undecomposed fibroblast growth factor. Fibroblast growth factors were used, FGF2 (bFGF), FGF1 (aFGF), and FGF7 (KGF), each of which was added to the medium at a final concentration of 0.1 mg / mL. Sulfated hyaluronic acid was used, and low sulfated hyaluronic acid (HA-LS) and high sulfated hyaluronic acid (HA-HS) prepared in Example 1 were mixed to a final concentration of 0.1 mg / mL. As controls, a medium containing nothing was added, and non-sulfated hyaluronic acid (HA) was added to a final concentration of 0.1 mg / mL. The results are shown in Figures 3 to 5. Figure 3 shows the results for FGF2 (bFGF), Figure 4 shows the results for FGF1 (aFGF), and Figure 5 shows the results for FGF7 (KGF), showing the band intensity of FGF after electrophoresis. In addition, FGF1 and FGF7 were prepared immediately after addition to the medium (only FGF added, 0 hours), and electrophoresis was performed together. As a result, it was found that the degradation of FGF2, FGF1, and FGF7 was significantly suppressed by sulfated hyaluronic acid, confirming the protective effect of sulfated hyaluronic acid on fibroblast growth factors.

[0057] Example 4: Effects of sulfated hyaluronic acid on the skin The safety of sulfated hyaluronic acid on skin cells was confirmed as follows, following the method described in OECD Test Guideline 439, a guideline for the safety of chemical substances. Using a human 3D cultured epidermis LabCyte EPI-MODEL (Japan Tissue Engineering Co., Ltd.), the effect of sulfated hyaluronic acid on epidermal cells was confirmed according to the manufacturer's instructions. Water (negative control) and SDS (positive control) were used as controls. The results are shown in Figure 6. The cell viability (%) was calculated using the following formula: Cell viability (%) = average measured value of test substance / average measured value of negative control x 100 The standard for evaluating safety is that a viable cell rate of 50% or more is confirmed to be safe. The test was conducted twice, and the results showed that the viability rate was close to 100% regardless of which sulfated hyaluronic acid was added, confirming the product's safety (non-irritation).

[0058] Example 5: Effect of sulfated hyaluronic acid on blood clotting The anticoagulant activity of sulfated hyaluronic acid was confirmed as follows. (1) Blood coagulation time measurement Blood clotting time was measured by a modified Lee-White test (see Sutton GC, Circulation, 2, 271-277, 1950). Specifically, 100 μL of fresh whole blood from BALB / c or C57BL / 6 mice was dispensed into 2 mL polypropylene tubes containing heparin, hyaluronic acid, low sulfated hyaluronic acid (HA-LS), or high sulfated hyaluronic acid (HA-HS), and heated to 37°C in a heat block. The final concentration of each sugar chain was 1, 10, 100, or 1000 μg / mL. Two negative control tubes were prepared, and one of them was gently tilted every 30 seconds until complete clotting. After confirming complete clotting, all tubes were tilted in the same way, and the time until clotting was determined. Measurements were continued for up to 3 hours. The results are shown in Figure 7. The left figure shows the blood clotting time at each concentration, and the right figure shows the blood clotting time when added at 10 μg / mL. Heparin extended the blood clotting time at 1 μg / mL, and blood clotting was not observed for more than 3 hours at 10 μg / mL. On the other hand, blood clotting was not observed for more than 3 hours at 100 μg / mL of highly sulfated hyaluronic acid (HA-HS) and 1000 μg / mL of low sulfated hyaluronic acid (HA-LS). Even at 1000 μg / mL of hyaluronic acid (HA), blood coagulated quickly. When each sugar chain was added at 10 μg / mL, blood clotting was not observed with heparin, indicating anticoagulant ability, but blood coagulated with highly sulfated hyaluronic acid (HA-HS), low sulfated hyaluronic acid (HA-LS), and hyaluronic acid (HA).

[0059] (2) Antithrombin activity measurement Antithrombin activity was measured using a Berrichrome Antithrombin III Auto B (Sysmex, Kobe, Japan). That is, human plasma was added as an antithrombin source to an excess amount of thrombin, and the amount of remaining thrombin after the reaction was measured using a chromogenic synthetic substrate to estimate the antithrombin activity. The thrombin provided was dissolved in a buffer solution, and the substrate was dissolved in distilled water to prepare a thrombin solution and a substrate solution, respectively. 1.4 μL of standard human plasma for blood coagulation tests was added to 10 μL of sugar chain diluted with distilled water. 84.6 μL of 37°C thrombin solution was added thereto, mixed well, and reacted at 37°C for 3 minutes. 14 μL of 37°C substrate was added thereto and stirred quickly, and the initial absorbance at 405 nm was measured within 30 seconds, and the absorbance was measured 120 seconds later. Meanwhile, the thrombin solution was diluted and measured in the same manner to prepare a calibration curve, and the amount of remaining thrombin was calculated from the absorbance. The results are shown in Figure 8. Antithrombin activity in human plasma in the presence of various concentrations of glycans was shown by the amount of residual thrombin. As the concentration of heparin increased, the amount of residual thrombin decreased and antithrombin activity in human plasma increased. On the other hand, low sulfated hyaluronic acid (HA-LS) and hyaluronic acid (HA) had significantly higher residual thrombin than heparin at concentrations of 10 μg / mL or more, and antithrombin activity in human plasma was low. High sulfated hyaluronic acid (HA-HS) had a lower amount of residual thrombin at low concentrations and showed no significant difference from heparin, but there was no decrease in the amount of residual thrombin with increasing concentration. Rather, at 1000 μg / mL, the amount of residual thrombin increased significantly and antithrombin activity in human plasma decreased. Neither sulfated hyaluronic acid nor hyaluronic acid showed any effect of enhancing antithrombin activity.

[0060] Example 6: Effect of sulfated hyaluronic acid on small intestinal crypt regeneration (1) Cryptoassay Intestinal regeneration was evaluated by measuring the number of crypts in the jejunum of 8-week-old BALB / c mice weighing 24 to 28 g (Winthers HR et al., Int J Radiat Biol Relat Stud Phys Chem Med, 17, 261-267, 1970). 10 μg of FGF1 was added to 0.5 mL of saline containing 5 μg / mL or 50 μg / mL of heparin, hyaluronic acid (HA), low-sulfated hyaluronic acid (HA-LS), and highly sulfated hyaluronic acid (HA-HS). The mice were intraperitoneally injected with 0.5 mL of saline. The mice were administered 24 hours before or after 10 Gy total body irradiation, and the mice were dissected 3.5 days after irradiation. The jejunal tissue was fixed in 10% neutral buffered formalin, embedded in paraffin, and then cut into cross sections for HE staining. Crypts consisting of 10 or more cells were counted under a microscope as viable crypts, and the total number of crypts was determined in 10 locations and averaged. The average number of crypts in each irradiated group was standardized by dividing it by the average number of crypts in the non-irradiated saline-treated control group.

[0061] Crypt assay was performed by administering only highly sulfated hyaluronic acid (HA-HS) 24 hours before whole-body irradiation of mice with 10 Gy of gamma rays. Highly sulfated hyaluronic acid (HA-HS) was diluted in 0.5 mL of saline to concentrations of 0 μg / mL, 5 μg / mL, and 50 μg / mL and injected intraperitoneally into mice (0 μg / mouse, 2.5 μg / mouse, 25 μg / mouse). The results are shown in Figure 9. In non-irradiated mice, no change was observed in the number of jejunal crypts even after administration of highly sulfated hyaluronic acid (HA-HS). A significant decrease in the number of crypts was observed after irradiation, but a significant increase in the number of crypts was observed in the 2.5 μg administration group, and the crypt regeneration effect was higher than that of the 25 μg administration group. This result confirmed that highly sulfated hyaluronic acid alone has a crypt regeneration effect. From now on, the crypt assay was performed with glycans at 5 μg / mL (2.5 μg / mouse).

[0062] (2) Crypt assay using pre-irradiation dosing 0.5 mL of physiological saline containing 10 μg FGF1 and 5 μg / mL glycan was prepared and injected intraperitoneally into mice 24 hours before 10 Gy total body irradiation with gamma rays, and a crypt assay was performed. The results are shown in Figure 10. Administration of FGF1 increased the number of crypts and promoted regeneration from radiation damage in the small intestine. On the other hand, heparin also increased the number of crypts, and the combination of FGF1 and heparin increased the number of crypts the most. On the other hand, highly sulfated hyaluronic acid (HA-HS) showed the same effect as heparin in increasing the number of crypts, and the combined effect with FGF1 was also significantly shown. Hyaluronic acid (HA) also increased the number of crypts, but no combined effect with FGF1 was observed. Low-sulfated hyaluronic acid (HA-LS) showed the same tendency as highly sulfated hyaluronic acid (HA-HS).

[0063] (3) Crypt assay using post-irradiation administration 0.5mL of saline containing 10μg FGF1 and 5μg / mL glycan was prepared and injected intraperitoneally into mice 24 hours after 10Gy total body irradiation with gamma rays, and a crypt assay was performed. The results are shown in Figure 11. FGF1 administration increased the number of crypts and promoted regeneration of the small intestine from radiation damage. On the other hand, heparin alone did not increase the number of crypts, and the combination of FGF1 and heparin significantly increased the number of crypts compared to the saline group, but no significant difference was observed when compared to the FGF1 alone group. On the other hand, low sulfated hyaluronic acid (HA-LS) and high sulfated hyaluronic acid (HA-HS) alone significantly increased the number of crypts. However, no effect was observed in combination with FGF1, and no significant difference in the number of crypts was observed between the FGF1 alone group and the glycan and FGF1 combined group.

[0064] Example 7: Effect of sulfated hyaluronic acid on apoptosis Radiation-induced apoptosis was evaluated in 8-week-old BALB / c mice weighing 24 to 28 g using the terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick end labeling (TUNEL) assay (see Gavrieli Y et al., Cell Biol., 119, 493-501, 1992). 10 μg of FGF1 and 5 μg / mL each of heparin, hyaluronic acid (HA), low-sulfated hyaluronic acid (HA-LS), and highly sulfated hyaluronic acid (HA-HS) were added to 0.5 mL of physiological saline and injected intraperitoneally into the mice. The mice were administered 24 hours before and dissected 24 hours after whole-body irradiation with 12 Gy gamma rays. Jejunal tissue was fixed in 10% neutral buffered formalin, embedded in paraffin, and then cut into thin cross sections and stained using the ApopTag Plus Peroxidase In Situ Apoptosis Detection Kit (Chemicon, Temecula, CA, USA). After deparaffinization, the tissue sections were treated with 20 μg / mL proteinase K and 3% hydrogen peroxide in PBS, equilibrated with the attached buffer, and reacted with terminal deoxynucleotidyl transferase (TdT) enzyme to label the 3'-OH termini of DNA strand breaks with digoxigenin. The tissue sections were reacted with peroxidase-labeled anti-digoxigenin antibody at room temperature for 30 minutes, and stained with DAB. The number of TUNEL-positive cells was determined for each crypt, and the average was calculated from measurements in 10 fields of more than 10 crypts.

[0065] Radiation-induced apoptosis in the jejunum was evaluated by measuring the number of TUNEL-positive cells in the jejunal crypts. FGF1 and glycans were administered 24 hours before irradiation and evaluated 24 hours after irradiation. The results are shown in Figure 12. FGF1 and heparin each alone reduced the number of TUNEL-positive cells, demonstrating an anti-apoptotic effect. Although the combined effect was also observed, there was no significant difference from the FGF1 alone group. On the other hand, highly sulfated hyaluronic acid also showed an anti-apoptotic effect similar to that of heparin, and the combined effect was significantly different from the FGF1 alone group and was more effective than heparin. Low-sulfated hyaluronic acid showed an anti-apoptotic effect almost the same as heparin, but hyaluronic acid showed no anti-apoptotic effect at all.

[0066] The above detailed description is merely illustrative of the objects and subject matter of the present invention and is not intended to limit the scope of the appended claims. Various modifications and substitutions to the described embodiments will be apparent to those skilled in the art from the teachings set forth herein without departing from the scope of the appended claims. [Industrial Applicability]

[0067] The composition of the present invention provides new uses for sulfated hyaluronic acid, such as a radiation protection agent, a wound healing agent, a hair growth promoter, or a periodontal disease treatment agent.

Claims

1. A composition for reducing and / or preventing one or more side effects associated with exposure to radiation, including at least a bleeding side effect, in a subject exposed to radiation or at risk of radiation exposure, the composition comprising sulfated hyaluronic acid (unless the composition comprises an in situ gelling polymer).

2. A composition for reducing and / or preventing one or more side effects associated with exposure to radiation in a subject who has been systemically exposed to radiation or who is at risk of systemic exposure to radiation, the composition comprising sulfated hyaluronic acid.

3. The composition according to claim 1 or 2, wherein the sulfated hyaluronic acid has a number of sulfate groups per hyaluronic acid disaccharide unit ("degree of sulfation (number of sulfate groups / disaccharide unit)") of 1.3 or more.

4. The composition according to claim 1 or 2, wherein the sulfated hyaluronic acid has a degree of sulfation (number of sulfate groups / disaccharide unit) of 3.0 or more.

5. The composition according to any one of claims 1 to 4, wherein the molecular weight of the sulfated hyaluronic acid is 30,000 or more.

6. The composition according to any one of claims 1 to 5, further comprising a growth factor.

7. The composition according to claim 6, wherein the growth factor is at least one growth factor selected from the group consisting of fibroblast growth factor (FGF), hepatic growth factor (HGF), heparin-binding epidermal growth factor (HBEGF), vascular endothelial growth factor (VEGF), nerve growth factor (e.g., NFG-2), cytokines, and bone morphogenetic proteins (BMPs).

8. The composition according to claim 6, wherein the growth factor is at least one growth factor selected from the group consisting of fibroblast growth factors FGF1, FGF2, FGF3, FGF4, FGF5, FGF6, FGF7, FGF8, and FGF9.

9. The composition according to any one of claims 1 to 8, wherein the composition is for administration in any one of the following dosage forms: intravascular administration, intraperitoneal administration, and oral administration.

10. The composition according to any one of claims 1 to 9, wherein the radiation is at least one radiation selected from the group consisting of gamma radiation, X-ray radiation, solar radiation, cosmic radiation, electromagnetic radiation, bremsstrahlung radiation, ultraviolet radiation, and particulate radiation.

11. 11. The composition of any one of claims 1 to 10, wherein the side effects associated with exposure to radiation include at least one side effect selected from the group consisting of acute nausea, vomiting, abdominal pain, diarrhea, dizziness, headache, fever, cutaneous radiation syndrome, low blood counts, infections due to leukopenia, bleeding due to thrombocytopenia, anemia due to erythrombocytopenia, and death.

12. Use of a composition according to any one of claims 1 to 11 for protecting the internal organs of a subject from radiation.

13. 13. The use according to claim 12, wherein the organ to be protected is at least one selected from the group consisting of bone marrow, liver, spleen, kidney, lung and gastrointestinal tract.

14. A composition according to any one of claims 1 to 11, used in combination with one or more antibiotics.

15. A composition for use in an affected area where maintenance of blood coagulation function is required, the composition containing a growth factor as an active ingredient and further containing sulfated hyaluronic acid for protecting the growth factor; Here, the affected area in which maintenance of blood coagulation ability is required is an affected area selected from the group consisting of an affected area of ​​a wound, an affected area of ​​periodontal disease, and an affected area of ​​Platelet Rich Plasma (PRP) hair growth therapy.

16. The composition according to claim 15, wherein the affected area in which maintenance of blood coagulation function is required is a wound.

17. The composition according to claim 15, wherein the affected area in which maintenance of blood coagulation function is required is an affected area of ​​periodontal disease.

18. The composition according to claim 15, wherein the affected area in which maintenance of blood coagulation function is required is an affected area for Platelet Rich Plasma (PRP) hair growth therapy.

19. The composition according to any one of claims 15 to 18, wherein the sulfated hyaluronic acid has a degree of sulfation (number of sulfate groups / disaccharide unit) of 1.3 or more.

20. The composition according to any one of claims 15 to 19, wherein the molecular weight of the sulfated hyaluronic acid is 30,000 or more.

21. The composition according to any one of claims 15 to 20, wherein the growth factor is at least one growth factor selected from the group consisting of fibroblast growth factor (FGF), hepatic growth factor (HGF), heparin-binding epidermal growth factor (HBEGF), vascular endothelial growth factor (VEGF), nerve growth factor (NFG), cytokines, and bone morphogenetic proteins (BMPs).

22. The composition according to any one of claims 15 to 20, wherein the growth factor is at least one growth factor selected from the group consisting of fibroblast growth factors FGF1, FGF2, FGF3, FGF4, FGF5, FGF6, FGF7, FGF8, and FGF9.

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