WO2015156426A1

WO2015156426A1 - Polymeric hydrogel labeled with radionuclide and loading angiogenesis-promoting protein or peptide, method for preparing same, and pharmaceutical composition for preventing or treating ischemic disease containing same as active ingredient

Info

Publication number: WO2015156426A1
Application number: PCT/KR2014/003004
Authority: WO
Inventors: 정환정; 이창문
Original assignee: 전북대학교 산학협력단
Priority date: 2014-04-07
Filing date: 2014-04-07
Publication date: 2015-10-15

Abstract

The present invention relates to a polymeric hydrogel labeled with radionuclide and loading an angiogenesis-promoting protein or peptide, a method for preparing the same, and a pharmaceutical composition for preventing or treating ischemic disease containing the same as an active ingredient. In the polymeric hydrogel according to the present invention, a biodegradable polymer is directly labeled with radionuclide and an angiogenesis-promoting protein or peptide is loaded thereon at the time of forming the hydrogel, so that the polymeric hydrogel can slowly release the angiogenesis-promoting protein or peptide while staying at the polymeric hydrogel-integrated lesion site, thereby preventing the waste of a large quantity of angiogenesis-promoting protein, and can confirm the in vivo distribution using a gamma camera, thereby quantifying the amount of particles staying at the site of ischemic disease and allowing the prediction of the treatment effect. In addition, the polymeric hydrogel according to the present invention can be favorably used in the treatment of disease, such as ischemic disease, by emitting radiation to the lesion site. Therefore, the polymeric hydrogel according to the present invention enables the expectation of treatment effects by both the radionuclide and the angiogenesis-promoting protein or peptide and can quantitatively estimate the accumulation of therapeutic agents, thereby increasing the treatment effect on the lesion.

Description

【Specification】

[Name of invention]

Polymeric hydrogel labeled with a radionuclide and supported by angiogenesis-promoting protein or peptide, a method for preparing the same, and a pharmaceutical composition for preventing or treating ischemic disease comprising the same as an active ingredient

Technical Field

The present invention relates to a polymer hydrogel labeled with a radionuclide and containing an angiogenic-promoting protein or peptide, a method for preparing the same, and a pharmaceutical composition for preventing or treating ischemic disease comprising the same as an active ingredient.

Background Art

<2> Ischemia refers to a condition where localized necrosis of tissue occurs due to cell damage caused by bleeding, embolism, and infarction to the tissues. Representative diseases of ischemia include ischemic stroke and ischemic heart disease, which can cause serious after-effects or death. In particular, ischemic cardiovascular diseases, such as acute myocardial infarction or severe angina pectoris, are increasing rapidly and deaths from these diseases are high worldwide. Treatments for such ischemic diseases include angiogenesis-promoting protein therapy and coronary angioplasty, which uses a stent to expand narrow blood vessels. In the case of the heart, arterial bypass surgery may be performed. However, if they fail to respond or fail, such as angiogenesis-promoting protein treatment or surgical treatment, _{there is} no further treatment and the patient is required to cut the diseased area or die, requiring a new treatment strategy. Recently, as the mechanism of angiogenesis has been revealed, attempts have been made to treat ischemic diseases by increasing side blood flow by inducing angiogenesis by administering genes or proteins of factors related to angiogenesis on the ischemic area. have. VEGF Vascular endothelial growth factor is a protein that specifically differentiates vascular endothelial cells and is known to be involved in general angiogenesis. o> The method of administering an angiogenic recombinant protein (pept ide) such as VEGF has the following problems. 1) Because the injected angiogenic proteins move to other tissues or lose their activity, it requires a large amount of high-purity protein to see the effects of angiogenesis. 2) It is preferable to continuously administer a small amount of protein for formation of side vessels, but there is a problem of frequent injection. The above problem can be solved by a delivery system that can slowly release angiogenic proteins at the injection site. And in vivo distribution of such an injection delivery system. There is a need for a follow-up strategy to accurately monitor the fabric and to predict the effectiveness of treatment.

Therefore, there is an urgent need for the development of a therapeutic agent using a radionuclide capable of releasing angiogenic proteins while remaining in an ischemic disease site after administration in the body.

[Detailed Description of the Invention]

[Technical problem]

<5> While the present inventors are studying a therapeutic agent using a radionuclide capable of releasing angiogenic proteins while staying in an ischemic disease site after administration to the body, the present inventors directly label radionuclides with biodegradable polymers. Promoting proteins or peptides were prepared with a supported polymer hydrogel, and the prepared polymer hydrogel was excellent in labeling efficiency and labeling stability of radionuclides so that the polymer hydrogel was directly injected into the ischemic disease site. Hydrogel stays in the ischemic diseased tissue area as it is, almost no leakage to the outside, it can be seen that the duration of the injected hydrogel stays in the disease site can predict the therapeutic effect of the therapeutic agent, and completed the present invention .

Accordingly, the present invention is to provide a polymer hydrogel labeled with a radionuclide and supported by angiogenesis-promoting protein or peptide, and a method for preparing the same.

In addition, the present invention is to provide a pharmaceutical composition for the prevention or treatment of ischemic disease, wherein the radionuclide is labeled, and angiogenesis-promoting protein or peptide is contained, and the polymer hydrogel is supported as an active ingredient.

In addition, the present invention is to provide a method for preparing a polymer-chelator hydrogel carrying a contrast dye and angiogenesis-promoting protein or peptide.

Technical Solution

The present invention provides a polymer hydrogel labeled with a radionuclide and containing an angiogenic-promoting protein or peptide, and a method for preparing the same.

In addition, the present invention provides a pharmaceutical composition for the prevention or treatment of ischemic diseases, wherein the radionuclide is labeled with an angiogenesis-promoting protein or peptide and the polymer hydrogel is supported as an active ingredient.

<1 1> The present invention also provides contrast dyes and angiogenesis-promoting proteins or peptides.

Provided is a method for preparing a polymer-chelator hydrogel.

Advantageous Effects <12> The polymer hydrogel according to the present invention is characterized by direct labeling of radionuclides on biodegradable polymers and angiogenesis-promoting proteins or peptides upon formation of hydrogels. -Promote proteins or peptides can be released to prevent waste of large amounts of angiogenesis-proteins or peptides, and in vivo distribution can be checked using gamma cameras to determine the amount of particles staying in the ischemic disease site. It can be quantified and the therapeutic effect can be predicted. Therefore, the polymer hydrogel according to the present invention can be expected at the same time the therapeutic effect by the radionuclide and angiogenesis-promoting protein or peptide, can increase the therapeutic effect of the lesion can be quantitatively assess the accumulation of the therapeutic agent.

[Brief Description of Drawings]

99 m ―

1 is a diagram showing the radiolabeling efficiency and stability of a polymeric hydrogel labeled with technicium (Tc) and supported with angiogenesis-promoting proteins or peptides.

99m

Fig. 2 shows the injection of a polymeric hadrogel labeled with technicium (Tc) and angiogenesis-promoting proteins or peptides into arteries of lower limb ischemic rats (Rat), and then accumulating in lesions by gamma imaging. Shows a gamma image of a rat.

99αι

Figure 3 is a diagram showing the therapeutic effect of the ischemic animal model after injection of chitosan-DTPA hydrogel labeled with Technicum (Tc) and loaded with angiogenesis-promoting protein VEGF.

Figure 4 is a graph evaluating the therapeutic effect of the ischemic animal model of the lower limb ischemia after injection of angiogenesis-promoting protein VEGF-loaded chitosan DTPA hydrogel through per fus i on gamma imaging.

<1 7>

[Form for implementation of invention]

The present invention provides a polymer hydrogel labeled with a radionuclide and supported by angiogenesis-promoting proteins or peptides.

<19> In addition, the present invention

1) preparing a polymer-chelator by reacting a biodegradable polymer with a chelator having a functional group capable of labeling radionuclides;

2) reacting the polymer-chelator prepared in step 1) with an activator for labeling the radionuclide and the radionuclide to produce a polymer-chelator labeled with the radionuclide; And

3) Angiogenesis to the radionuclide-labeled polymer-chelator prepared in step 2) Adding and stirring a bio-promoting protein or peptide and an anionic crosslinking material to prepare a polymer-chelator hydrogel labeled with a radionuclide and supported by angiogenesis-promoting protein or peptide;

A method for producing a polymer hydrogel containing a radionuclide labeled with an angiogenesis-promoting protein or peptide, wherein the radionuclide is a radionuclide capable of gamma imaging and PET imaging.

111 18 64 68 131 125 124 186 188 90, 166 „,. _ _^ ^

Radionuclides are labeled and characterized in that the radionuclide is selected from the group consisting of In, F, Cu, Ga, I, I, I, Re, Re, Y and Ho, and angiogenesis-promoting proteins or peptides, It provides a method for preparing a supported polymer hydrogel. Hereinafter, the present invention will be described in detail.

The polymer hydrogel according to the present invention is characterized in that the biodegradable polymer is directly labeled with radionuclides and supported on angiogenesis-promoting proteins or peptides during hydrogel formation.

Referring to the step-by-step detailed description of the method for producing a polymer hydrogel according to the present invention.

Step 1) is a step of preparing a polymer-chelator, the biodegradable polymer is dissolved in an aqueous solution of acetic acid, an organic solvent is added, the chelator is added and stirred to obtain a polymer-chelator. The obtained polymer-chelator is purified by dialysis with distilled water and freeze-dried.

The biodegradable polymer is preferably chitosan and its derivatives, polyglutamic acid, heparin, hyaluronic acid, alginic acid, pectin, carboxymethyl salose, protein, and the like, but is not limited thereto.

The chelator is a compound having a functional group capable of labeling a radionuclide, and SHPP (yV-succ inimi dy 1-3- [4-hydroxyoxyy 1] r op i onat e), DTPA (di ethyl enetri amine pentaacetic) acid), DOTA (1, 4, 7, 10-t et r aazacyc 1 ododecane-1, 4,7, 10-t et r aacet ic acid), NOTA (l, 4,7-triazacyclononane-l, 4, 7-triacet ic acid), TETA (1,4,8, 11-tetraazacyc lotetradecane-1, 4, 8, 11-tetraacet ic acid), histidine, tyrosine, tyrosine, including but not limited to . The chelator may vary depending on the radionuclide.

The organic solvent for dissolving the chelator is dimethylformamide (DMF), dimethyl sulfoxide Seeds (DMSO), 1,4-dioxane, tetrahydrofuran (THF), acetone, acetonitrile, methanol and the like.

The biodegradable polymer and the chelator have a weight ratio of 1: 0.01 to 1, preferably 1:

0. It is good to mix with a weight ratio of 0.5. If the chelator is excessively bound to the biodegradable polymer, it is difficult to form the mixture in an aqueous solution, which causes a problem in that it is difficult to form a hydrogel.

Step 2) is a step of preparing a polymer-chelator labeled with a radionuclide. The polymer-chelator is dissolved in an acetic acid solution, and then reacted with an activator for labeling the radionuclide and the radionuclide. A polymer-chelator labeled with a nuclide is obtained. The labeling efficiency of the radionuclide in the polymer-chelator labeled with the radionuclide is 99%.

_{_{Ί, ■, · ,, ,, Λ}} 99m 111 18 64 68 131 125 124 186 188

The radionuclide is Tc, In, F, Cu, Ga, I, I, I, Re, Re,

⁹ ° Y or ¹⁶⁶ Ho, and the like.

In step 3), the radionuclide is labeled with an angiogenesis-promoting protein or peptide, and a supported polymer-chelator hydrogel is prepared. A polymer hydrogel is prepared by reacting with a promoter protein or peptide and an anionic crosslinking material.

The angiogenesis-promoting protein or peptide may include VEGF (Vascular endothelial growth factor), IGF (Insul in— ike growth factor), Fibroblast growth factor (FGF), P1 ate let-derived growth factor (PDGF), and It may be one or more selected from the group consisting of PGF (placental growth factor), but is not limited thereto.

The anionic crosslinking material is used to form a hydrogel, which increases the labeling stability of the radionuclide when preparing the polymer hydrogel, thereby preventing the release of the radionuclide to the normal tissue outside the lesion site. have. In addition, the size of the polymer hydrogel particles can be adjusted according to the concentration of the anion of the crosslinking material, thereby widening the application range of the lesion. Anionic crosslinking materials include, but are not limited to, TPP (tripolyphosphate), alginic acid, pectin, carboxymethyl cellulose, polyglutamic acid, protein, DNA, RNA, and the like.

The polymer hydrogel may be prepared by adding angiogenic-promoting protein or peptide to a radionuclide-labeled polymer solution, followed by stirring by adding a negative crosslinking material, followed by electrospinning and electrospraying. It can be prepared using a spinning method or an emulsion dog method. In the electrospray method, angiogenic-promoting proteins or peptides are added to a radionuclide-labeled polymer-chelator, and the mixed solution is injected into the anionic crosslinker solution under high voltage through a pump. By spraying by an electrospray method can be prepared a polymer hydrogel.

When using an electrospinning method, angiogenesis-promoting protein or peptide is added to a polymer-chelator labeled with a radionuclide, and a negative pump crosslinking material is 1-20 kV through a syringe pump. The polymer can be prepared by electrospinning under the voltage of.

The radionuclide prepared by the above method is labeled with an angiogenesis-promoting protein or peptide, and the polymer-chelator hydrogel carrying the radionuclide has a labeling efficiency of at least 99¾ and a label stability of at least 40 hours. Do. In addition, the content and supporting efficiency of the angiogenesis-promoting protein or peptide in the polymer hydrogel can be controlled according to the amount of the polymer-chelator and the amount of the angiogenesis-promoting protein or peptide.

<44>

In addition, the present invention provides a pharmaceutical composition for the prevention or treatment of ischemic diseases in which the radionuclide is labeled and angiogenesis-promoting protein or peptide is contained as an active ingredient.

When the radionuclide is labeled and the angiogenesis-promoting protein or peptide is supported, the polymer-chelator hydrogel is directly injected into the ischemic lesion site of the ischemic animal model. As it remains in the tissue area, it accumulates with little leakage to the outside, and the improvement of the ischemic blood flow is observed to restore over 80% of the normal lower limb blood flow. In addition, it is possible to confirm the distribution in vivo using a gamma camera to quantify the amount of particles staying in the ischemic disease site, to predict the therapeutic effect, and to emit radiation at the site of the disease to treat diseases such as ischemic disease. It can be usefully used. Therefore, the polymer hydrogel labeled with the radionuclide according to the present invention and supported by angiogenesis-promoting protein or peptide can be usefully used for the prevention or treatment of ischemic diseases.

The ischemic diseases include but are not limited to lower limb ischemia, myocardial ischemia and cerebral infarction.

The composition of the present invention may contain one or more known active ingredients having a therapeutic effect on ischemic disease, together with a polymer hydrogel labeled with a radionuclide and supported by an angiogenesis-promoting protein or peptide. The polymer hydrogel according to the present invention may be administered in the form of an injection for the treatment of ischemic disease. Specifically, the polymer hydrogel of the present invention may be directly injected into a lesion, or injected into an artery or vein in close proximity to the lesion and delivered to the lesion. Compositions of the invention for parenteral administration include sterile aqueous or non-aqueous solutions, dispersants, suspensions, or emulsions, as well as sterile powders which are reconstituted just prior to use as sterile solutions or suspensions. Examples of suitable sterile aqueous and non-aqueous carriers, diluents, solvents or vehicles include water, physiological saline, ethane, polyols (e.g. glycerol, propylene glycol, polyethylene glycol, etc.), and mixtures thereof, vegetable oils (E.g., rib oil), injectable organic esters (e.g. ethyl oleate). For example, in the case of dispersing agents and suspending agents, use a coating material such as lecithin and maintain proper specific size, using a surface active agent can maintain proper fluidity ol _^.

The composition of the present invention may be used alone or in combination with methods using surgery, hormonal therapy, drug treatment and biological response modifiers for the treatment of ischemic diseases.

<51>

In addition, the present invention

1) preparing a polymer-chelator by reacting the biodegradable polymer and the chelator;

2) preparing a polymer-chelator carrying the contrast dye by reacting the polymer-chelator prepared in step 1) with a contrast dye (dye); And

3) Angiogenesis-promoting proteins or peptides and anionic crosslinkers are added and stirred to the polymer-chelator to which the contrast dye prepared in step 2) is carried or chemically bound, and the contrast dye and blood vessel Preparing a polymer-chelator hydrogel carrying angiogenesis-promoting protein or peptide;

A method for preparing a high molecular-chelator hydrogel carrying a contrast dye and an angiogenesis-promoting protein or peptide, comprising:

The angiogenesis-promoting protein or peptide may include a vesicular endothelial growth factor (VEGF), an insulfur growth factor (IGF), a fibroblast growth factor (FGF), a platelet derived growth factor (PDGF), and a PGF (PGF). A contrast dye and angiogenesis-promoting protein or peptide, characterized in that at least one selected from the group consisting of placental growth factor) provides a method for producing a polymer-chelator hydrogel supported. The polymer hydrogel prepared by the method for producing a polymer-chelator hydrogel, in which the contrast dye and angiogenesis-promoting protein or peptide are supported, is characterized in that the radionuclide is labeled and the angiogenesis-promoting protein or peptide is It can be used for the prevention and treatment of ischemic diseases in the same manner as the supported polymer hydrogel.

<59>

As described above, the polymer hydrogel according to the present invention directly label the radionuclide on the biodegradable polymer and carry the angiogenesis-promoting protein or peptide upon formation of the hydrogel, whereby the site of the polymer hydrogel is integrated. It is possible to release angiogenesis-promoting proteins or peptides slowly while staying in the body to prevent waste of large amounts of angiogenesis-proteins, and to check the distribution in vivo by using gamma cameras to stay in ischemic diseases. The amount of can be quantified and the therapeutic effect can be predicted. In addition, by radiating radiation at the lesion site it can be usefully used for the treatment of diseases such as ischemic disease. Therefore, the polymer hydrogel according to the present invention can be expected at the same time the therapeutic effect by the radionuclide and angiogenesis-promoting protein or peptide, and can increase the therapeutic effect of the lesion can be quantitatively evaluated the accumulation of the therapeutic agent.

<61>

Hereinafter, preferred embodiments will be presented to aid in understanding the present invention. However, the following examples are merely provided to more easily understand the present invention, and the contents of the present invention are not limited by the examples.

<63>

99m

Example 1: Technisium (ᅳ Tc) is Labeled and Supports Angiogenesis-Promoting Protein VEGF

Of prepared chitosan-DTPA hydrogel

<65> 1-1. Preparation of Chitosan-DTPA

<66>

In the reaction scheme, n is an integer of 100 ~ 800.

100 mg of chitosan was dissolved in 90 ml of 10% (v / v) acetic acid aqueous solution, 10 ml of methanol was added, and 10 mg of anhydrous diethylenetriamine pentaacetic acid (DTPA) was added thereto. Thereafter, the mixture was stirred at room temperature for 24 hours, dialyzed and purified using distilled water, and lyophilized to obtain chitosan-DTPA.

2. Labeling of Technium ("Tc") in Chitosan-DTPA

2.5 mg of chitosan-DTPA prepared in Example 1-1 was dissolved in 5 ml of 0 · 5% (ν / ν) acetic acid aqueous solution, and then 0.1 ml of SnCl ₂ solution (2.5 mg / ml 0.04 N HC1). And technium

99m

(Tc) ol was added and stirred for 10 minutes. After the reaction was completed, the chitosan-DTPA was labeled with Tec. <7 i> 1-3. Chitosan Labeled with Technicium (" ^m Tc) and Supported Angiogenesis-Promoting Protein-

Preparation of DTPA Hydrogels

<72>

<73> Chitosan Labeled with 2.5 mg / ml of Technicium (Tc) Obtained in Example 1-2

To 1 ml of DTPA, add 0.01 mg of angiogenesis-promoting protein VEGF (Vascular endothelial growth factor), add 1 ml of 2 mg / ml TPP tr ipolyphosphate (Ag) with stirring In contrast, a hydrogel was prepared. At this time, the ratio of angiogenesis-promoting protein to chitosan-DTPA is a weight ratio of 1: 0.001 to 2, preferably a weight ratio of 1: 0.001 to 1. The size and strength of the hydrogel may vary depending on the concentration of TPP added.

<74>

Experimental Example 1 Determination of Labeling Efficiency of Technisium on Chitosan-DTPA Hydrogel Labeled with Technicium (^ Tc)

99m

<76> The chitosan-DTPA labeled with Technicum (Tc) prepared in Example 1-2 was tested.

99m

In order to confirm the labeling efficiency of Knee (Tc), the following experiment was performed. Specifically, using Gelman's ITLC-SG as the stationary phase,

^QQ m

Chromatography of the technicium (Tc) -labeled chitosan-DTPA hydrogel prepared in Example 1-2 using salt ine was carried out to confirm the labeling efficiency of technicium ( ⁹ Tc), Label stability was assessed for 40 hours. The results are shown in FIG.

99m

As shown in FIG. 1, the technique of chitosan-DTPA labeled with technicium (Tc)

99m

The labeling efficiency of Ni (Tc) was over 99% and stable for over 40 hours.

<79>

9 m

<80> Experimental Example 2 :. Chitosan-DTPA Hydroxide with TECHNIP (一 Tc) Labeled and VEGF

Evaluation of Treatment Effect of Lower Limb Ischemia in Animal Model

<8i> Acute lower limb ischemia animal model was made by blocking the lower limb artery of rat with silk, and after 24 hours, the ischemic disease was confirmed by perfusion gamma image. Chitosan-DTPA hydrogel labeled with Technicium (Tc) prepared in 1 and loaded with VEGF was injected. The gamma image was used to confirm the accumulation of hydrogel at the disease site, and the amount of VEGF loaded at the disease site was quantitatively evaluated using the region of interest analysis method. A week later, blood f low was confirmed by perfusion gamma imaging and compared with the opposite normal leg. Ischemic animal model was created in the same way and evaluated by dividing the group injected with chitosan hydrogel only and the group treated with nothing other than surgery.

99m

Specifically, chitosan-DTPA high labeled with technicium (Tc) and supported with VEGF. Drogel was injected into the lower extremity artery of the disease and the accumulation was confirmed by gamma imaging. The results are shown in FIGS. 2 and 3.

99m

As shown in FIG. 2, the chitosan-DTPA hydrogel labeled with technicium (Tc) and VEGF accumulated at the lower limb ischemia disease site, and the degree of leakage to the outside was hardly observed.

99m

As shown in FIG. 3, after injection of chitosan-DTPA hydrogel labeled with technicium (Tc) and VEGF, the blood flow of the lower limb ischemia was observed and normal. He recovered to more than 80% of the blood flow (f low) of the lower extremity.

Industrial Applicability

The present invention is applicable to the industry for the manufacture of a pharmaceutical composition for the prevention and treatment of ischemic diseases.

<86>

Claims

【Scope of Claim】

【Claim 1】

2) producing a radionuclide-labeled polymer-chelator by reacting the polymer-chelator prepared in step 1) with a radionuclide and an activator for labeling the radionuclide; and

3) An angiogenesis-promoting protein or peptide, and an anionic cross-linking material are added to the radionuclide-labeled polymer-chelator prepared in step 2) above and stirred to label the radionuclide and form an angiogenesis-promoting protein or peptide. A. Preparing a supported polymer-chelator hydrogel;

A method for producing a polymer hydrogel labeled with a radionuclide and carrying an angiogenesis-promoting protein or peptide, including, wherein the radionuclide is a radionuclide capable of gamma imaging and PET imaging, including Tc,

111 18„ 64^ 68„ 131 125 _τ 124 _τ 186 188^ 90, _r „. 166, , _ ,„ . . _ _Λ _ „

A radionuclide-labeled and angiogenesis-promoting protein or peptide, characterized in that it is one or more selected from the group consisting of In, F, Cu, Ga, I, I, I, Re, Re, Y and Ho, Method for producing supported polymer hydrogel.

【Claim 2】

The method of claim 1, wherein the angiogenesis-promoting protein or peptide is VEGF Vascular endothelial growth factor), IGF (Insul in 1 ike growth factor), FGF (Fibroblast growth factor), PDGF (Platelet-der ived growth factor) A method for producing a polymer hydrogel labeled with a radionuclide and carrying an angiogenesis-promoting protein or peptide, characterized in that it is one or more selected from the group consisting of PGF (lacental growth factor).

【Claim 3】

The method of claim 1, wherein the biodegradable polymer is one or more selected from the group consisting of chitosan and its derivatives, polyglutamic acid, heparin, hyaluronic acid, alginic acid, pectin, carboxymethyl salolose, and protein. A method of producing a polymer hydrogel labeled with a radionuclide and loaded with an angiogenesis-promoting protein or peptide.

[Claim 4]

The method of claim 1, wherein the chelator having a functional group capable of labeling the radionuclide is SHPP (V-succinimidyl-3-[4-hydr oxypheny 1] pr op i ona te), DTPA. (diethylenetriamine pentaacet ic acid), DOTA (1, 4, 7, 10-tetr aazacyc 1 ododecane- 1,4,7, 10-tetraacet ic acid), NOTA (1, 4, 7-tri azacyc 1 ononane-1, 4, 7-tri acet ic acid), TETA (1,4,8, 11-tetraazacyc lotetradecane-1,4,8, 11-tet raacet ic acid), histidine, tyrosine and a protein containing tyrosine A method for producing a polymer hydrogel labeled with a radionuclide and carrying an angiogenesis-promoting protein or peptide, characterized in that it is one or more selected from the group,

【Claim 5】

The polymer of claim 1, wherein in step 1), the biodegradable polymer and the chelator are mixed at a weight ratio of 1:0.01-1, and the radionuclide is labeled and the angiogenesis-promoting protein or peptide is supported. Method for producing hydrogel.

【Claim 6】

In claim 1, the anionic cross-linking material in step 3) is at least one selected from the group consisting of TPP (tripolyphosphate), alginic acid, pectin, carboxymethyl cellulose, polyglutamic acid, protein, DNA, and RNA. A method of producing a polymer hydrogel labeled with a radionuclide and loaded with an angiogenesis-promoting protein or peptide.

[Claim 7]

A polymer hydrogel prepared according to claim 1, labeled with a radionuclide and loaded with an angiogenesis-promoting protein or peptide.

【Claim 8】

The radionuclide of Paragraph 7 is labeled and the angiogenesis-promoting protein or peptide is. A pharmaceutical composition for the prevention or treatment of ischemic disease containing a supported polymer hydrogel as an active ingredient.

[Claim 9]

The pharmaceutical composition for the prevention or treatment of ischemic disease according to claim 8, wherein the ischemic disease is at least one selected from the group consisting of lower extremity ischemia, myocardial ischemia, and cerebral infarction.

[Claim 10]

The pharmaceutical composition for preventing or treating ischemic disease according to claim 9, wherein the composition is in an injectable formulation.

【Claim 111

1) A process for producing a polymer-chelator by reacting a biodegradable polymer with a chelator. total;

2) reacting the polymer-chelator prepared in step 1) with a contrast dye to produce a polymer-chelator carrying the contrast dye; and

3) Add angiogenesis-promoting protein or peptide and negative cross-linking material to the polymer-chelator prepared in step 2) or chemically bound to the contrast dye and stir to promote contrast dye and angiogenesis. Preparing a polymer-chelator hydrogel loaded with protein or ^J peptide;

A method for producing a polymer-chelator hydrogel loaded with contrast dye and angiogenesis-promoting protein or peptide, including:

The angiogenesis-promoting proteins or peptides include VEGF Vascular endothelial growth factor (IGF), Insul in-like growth factor (IGF), Fibroblast growth factor (FGF), Platelet-derived growth factor (PDGF), and placental growth factor (PGF). A method for producing a polymer-chelator hydrogel loaded with a contrast dye and an angiogenesis-promoting protein or peptide, characterized in that it is one or more selected from the group consisting of.