Sulfonated saccharide hydrogel for treating myocardial infarction and preparation method thereof

Abstract

The invention discloses an application of sulfonated saccharide hydrogel containing sulfonated saccharide and a hydrogel matrix material in prevention and / or treatment of myocardial infarction. Specifically, the invention provides application of sulfonated saccharide hydrogel containing sulfonated saccharide and a hydrogel matrix material, and the sulfonated saccharide hydrogel is used for preparing a pharmaceutical composition for preventing and / or treating myocardial infarction. Wherein the molecular weight of the sulfonated saccharide is 1000Da-300000Da, the sulfur content is 0.5-30%, and the hydrogel matrix material comprises an extracellular matrix material and a derivative thereof. The sulfonated saccharide hydrogel can effectively promote neovascularization and cardiac function recovery after myocardial infarction under the condition that exogenous cells or growth factors are not introduced, ventricular hypertrophy and fibrosis degree in the later stage of infarction are remarkably inhibited, and a new thought is provided for clinical myocardial infarction transformation treatment.

Description

Technical Field

[0001] The invention belongs to the field of biomedical materials, and in particular relates to a sulfonated sugar hydrogel for treating myocardial infarction and a preparation method thereof. Background Art

[0002] With the development of social economy and the changes in people's lifestyles, especially the aging of the population and the acceleration of urbanization, the unhealthy lifestyles of residents have become increasingly prominent, and the impact of cardiovascular risk factors on residents' health has become more significant, and the incidence of cardiovascular disease continues to increase. According to the results of the "China Cardiovascular Health and Disease Report" released in 2020, there are currently 330 million people with cardiovascular disease in my country, including 11.39 million patients with coronary heart disease, 5 million patients with cor pulmonale, 8.9 million patients with heart failure, 4.87 million patients with atrial fibrillation, 2.5 million patients with rheumatic heart disease, and 2 million patients with congenital heart disease. Although the application of traditional drug therapy, stent intervention and bypass surgery has greatly improved the survival rate of patients with myocardial infarction, it is impossible to save dead myocardial cells, reverse ventricular remodeling and subsequent heart failure. Exogenous growth factors have greatly limited their application due to their short half-life, high cost and induction of abnormal blood vessel growth. Stem cell therapy has not been widely used in clinical practice due to limited sources, donor safety and potential fibrosis.

[0003] After myocardial infarction, autologous cardiomyocytes and circulating macrophages are stimulated by the surrounding injury environment to secrete a small amount of endogenous growth factors to participate in angiogenesis and myocardial repair. However, the secretion of endogenous growth factors is low and the repair driving force is insufficient, making it impossible to achieve complete repair of in vivo tissues. Therefore, how to regulate the myocardial injury microenvironment and increase the secretion of endogenous repair factors to mediate myocardial function recovery is a major challenge in the treatment of myocardial infarction.

[0004] Hydrogel materials have the advantages of good biocompatibility and cell adhesion, but they are often used only as carriers for cell proliferation, migration and growth, and cannot improve the ischemic microenvironment to increase the secretion of endogenous growth factors and induce angiogenesis and myocardial function repair. Therefore, it is an urgent problem to study hydrogel materials that can effectively activate the myocardial injury microenvironment, induce arterial angiogenesis and restore ventricular remodeling. Summary of the invention

[0005] The purpose of the present invention is to provide a sulfonated carbohydrate hydrogel for use in treating myocardial infarction.

[0006] In a first aspect, the present invention provides a use of a sulfonated carbohydrate hydrogel containing a sulfonated carbohydrate and a hydrogel matrix material for preparing a pharmaceutical composition for preventing and / or treating myocardial infarction.

[0007] In another preferred embodiment, the myocardial infarction is type 1 myocardial infarction, type 2 myocardial infarction, type 3 myocardial infarction, type 4 myocardial infarction or type 5 myocardial infarction.

[0008] In another preferred embodiment, the myocardial infarction includes myocardial infarction caused by coronary artery intervention, coronary artery surgery, coronary artery bypass grafting, or coronary artery plaque secondary to thrombosis.

[0009] In another preferred embodiment, the prevention and / or treatment of myocardial infarction has one or more characteristics selected from the following group:

[0010] (1) Increased wall thickness in the infarcted area;

[0011] (2) reduce myocardial infarction area;

[0012] (3) Reduce myocardial scar hyperplasia;

[0013] (4) Increase the amount of cardiac muscle;

[0014] (5) restore ventricular wall motion at the site of myocardial infarction;

[0015] (6) Reduce the degree of fibrosis at the site of myocardial infarction;

[0016] (7) Promote myocardial arterial angiogenesis; and

[0017] (8) Improve heart contractile function.

[0018] In another preferred embodiment, the sulfonated sugar is selected from the group consisting of sulfonated chitosan, sulfonated chito-oligosaccharide, sulfonated dextran, chondroitin sulfate, heparin, heparan sulfate, or a combination thereof.

[0019] In another preferred embodiment, the sulfonated sugar is sulfonated chitosan oligosaccharide.

[0020] In another preferred embodiment, the molecular weight of the sulfonated sugar is 1000-300000 Da.

[0021] In another preferred embodiment, the molecular weight of the sulfonated sugar is 1000-20000 Da.

[0022] In another preferred embodiment, the molecular weight of the sulfonated sugar is 1000-15000 Da.

[0023] In another preferred embodiment, the molecular weight of the sulfonated sugar is 1500 to 100000 Da.

[0024] In another preferred embodiment, the molecular weight of the sulfonated sugar is 1000Da, 2000Da, 3000Da, 4000Da, 5000Da, 6000Da, 8000Da, 8000Da, 9000Da, 10000Da, 20000Da, 30000Da, 40000Da, 50000Da, 60000Da, 70000Da, 80000Da, 90000Da or 100000Da.

[0025] In another preferred embodiment, the molecular weight of the sulfonated chitosan oligosaccharide is 1000-10000 Da, preferably 1000-5000 Da.

[0026] In another preferred embodiment, the molecular weight of the sulfonated chitosan is 5000-50000 Da, preferably 5000-30000 Da, and more preferably 5000-20000 Da.

[0027] In another preferred embodiment, the molecular weight of the chondroitin sulfate is 10,000-100,000 Da, for example, 70,000 Da.

[0028] In another preferred embodiment, the molecular weight of the heparin is 8000-200000 Da, preferably 10000-150000 Da.

[0029] In another preferred embodiment, the molecular weight of the sulfonated dextran is 20,000 to 200,000 Da, preferably 50,000 to 150,000 Da.

[0030] In another preferred embodiment, the sulfur content of the sulfonated sugar is 0.5% to 30%.

[0031] In another preferred embodiment, the sulfur content of the sulfonated sugar is 1% to 25%.

[0032] In another preferred embodiment, the sulfur content of the sulfonated sugar is 5% to 20%.

[0033] In another preferred embodiment, the sulfur content of the sulfonated sugar is 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19% or 20%.

[0034] In another preferred embodiment, the sulfonated sugar hydrogel further contains water.

[0035] In another preferred embodiment, the hydrogel matrix material is selected from extracellular matrix molecular materials or their derivatives.

[0036] In another preferred embodiment, the hydrogel matrix material is selected from the following group: collagen, fibrinogen, Matrigel matrix glue, methacryloyl hyaluronic acid (HAMA), methacryloyl collagen (GelMA), or a combination thereof.

[0037] In another preferred embodiment, the sulfonated sugar hydrogel is prepared by cross-linking and curing a sulfonated sugar solution with a solution of a hydrogel matrix material.

[0038] In another preferred embodiment, the sulfonated carbohydrate hydrogel is a sulfonated chitosan oligosaccharide hyaluronic acid hydrogel.

[0039] In another preferred embodiment, the sulfonated chitosan oligosaccharide hyaluronic acid hydrogel is prepared by curing a sulfonated chitosan solution with a methacrylylated hyaluronic acid (HAMA) solution in the presence of a photoinitiator through ultraviolet light.

[0040] In another preferred embodiment, the sulfonated carbohydrate hydrogel is prepared by the following method, which comprises the following steps:

[0041] (a) mixing a sulfonated sugar with a first solvent to obtain a sulfonated sugar solution;

[0042] (b) mixing the hydrogel matrix material with a second solvent to obtain a hydrogel matrix precursor solution;

[0043] (c) mixing the sulfonated sugar solution and the hydrogel matrix precursor solution and optionally cross-linking and curing them to obtain the sulfonated sugar hydrogel.

[0044] In another preferred embodiment, in step (a), the first solvent is water, preferably deionized water or physiological saline.

[0045] In another preferred embodiment, in step (a), the concentration of the sulfonated sugar solution is 0.01-500 μg / mL.

[0046] In another preferred embodiment, in step (a), the concentration of the sulfonated sugar solution is 0.1-400 μg / mL.

[0047] In another preferred embodiment, in step (a), the concentration of the sulfonated sugar solution is 0.5-300 μg / mL.

[0048] In another preferred embodiment, in step (a), the concentration of the sulfonated sugar solution is 1-200 μg / mL, preferably 50-200 μg / mL, such as 50 μg / mL, 100 μg / mL, 150 μg / mL, 200 μg / mL.

[0049] In another preferred embodiment, in step (b), the second solvent is water, preferably deionized water or physiological saline.

[0050] In another preferred embodiment, in step (b), the mass volume fraction of the hydrogel matrix material in the hydrogel matrix precursor solution is 0.1% to 30%.

[0051] In another preferred embodiment, in step (b), the mass volume fraction of the hydrogel matrix material in the hydrogel matrix precursor solution is 0.5% to 5%.

[0052] In another preferred embodiment, in step (b), the mass volume fraction of the hydrogel matrix material in the hydrogel matrix precursor solution is 1% to 10%.

[0053] In another preferred embodiment, in step (b), when the hydrogel matrix material is selected from methacryloyl hyaluronic acid (HAMA), methacryloyl collagen (GelMA), or a combination thereof, the mass volume fraction of the hydrogel matrix material in the hydrogel matrix precursor solution is 1% to 20%, preferably 5% to 20%.

[0054] In another preferred embodiment, in step (b), when the hydrogel matrix material is selected from collagen, fibrinogen, or a combination thereof, the mass volume fraction of the hydrogel matrix material in the hydrogel matrix precursor solution is 0.1% to 2%, for example 0.2%, 0.5%, or 1%.

[0055] In another preferred embodiment, in step (c), the volume ratio of the sulfonated sugar solution to the hydrogel matrix precursor solution is 1:0.1 to 1:10.

[0056] In another preferred embodiment, in step (c), the volume ratio of the sulfonated sugar solution to the hydrogel matrix precursor solution is 1:0.5 to 1:8.

[0057] In another preferred embodiment, in step (c), the volume ratio of the sulfonated sugar solution to the hydrogel matrix precursor solution is 1:0.5 to 1:5.

[0058] In another preferred embodiment, in step (c), the volume ratio of the sulfonated sugar solution to the hydrogel matrix precursor solution is 1:0.5 to 1:2.

[0059] In another preferred embodiment, in step (c), the volume ratio of the sulfonated sugar solution to the hydrogel matrix precursor solution is 1:1.

[0060] In another preferred embodiment, when the hydrogel matrix material is selected from methacryloyl hyaluronic acid (HAMA), methacryloyl collagen (GelMA), or a combination thereof, the step (c) comprises: mixing the sulfonated sugar solution and the hydrogel matrix precursor solution in the presence of a photoinitiator, and curing them by irradiation with ultraviolet light to obtain a cross-linked sulfonated sugar hydrogel.

[0061] In another preferred embodiment, the photoinitiator is IC2959.

[0062] In another preferred embodiment, the photoinitiator is IC2959 solution, and its concentration is 0.1 mg / mL to 5 mg / mL.

[0063] In another preferred embodiment, the wavelength of the ultraviolet lamp is 405 nm.

[0064] In another preferred embodiment, the UV light irradiation time is 10-30 seconds, for example, 20 seconds.

[0065] In another preferred embodiment, when the hydrogel matrix material is fibrinogen, the step (c) comprises: mixing the sulfonated sugar solution and the hydrogel matrix precursor solution, and then adding thrombin for solidification to obtain a cross-linked sulfonated sugar hydrogel.

[0066] In another preferred embodiment, the final concentration of thrombin is 0.5-5 U / mL, such as 1 U / mL.

[0067] In another preferred embodiment, after adding thrombin, the mixture is mixed and allowed to stand to solidify.

[0068] In another preferred embodiment, the time for adding thrombin and standing to solidify is 2-15 minutes, such as 10 minutes.

[0069] In another preferred embodiment, when the hydrogel matrix material is collagen, the step (c) comprises: mixing the sulfonated sugar solution and the hydrogel matrix precursor solution, then adjusting the pH to above 7, and standing to solidify to obtain a cross-linked sulfonated sugar hydrogel.

[0070] In another preferred embodiment, the pH is adjusted to 7-7.5, such as 7.2.

[0071] In another preferred embodiment, the curing time after pH adjustment is 10-45 min, such as 30 min.

[0072] In another preferred embodiment, when the hydrogel matrix material is Matrigel, curing is not required in step (c).

[0073] In another preferred embodiment, when the hydrogel matrix material is Matrigel, the step (c) comprises: mixing the sulfonated sugar solution and the hydrogel matrix precursor solution, and letting the mixture stand to obtain a cross-linked sulfonated sugar hydrogel.

[0074] In another preferred embodiment, the sulfonated carbohydrate hydrogel is prepared by the following method, which comprises the following steps:

[0075] (s1) providing a hydrogel prepolymerization solution, wherein the hydrogel prepolymerization solution comprises a sulfonated sugar solution, a hydrogel matrix precursor solution, and an optional auxiliary agent; wherein the auxiliary agent is selected from a photoinitiator, thrombin, or a pH regulator; and

[0076] Optionally (s2) solidifying the hydrogel prepolymer solution to obtain a cross-linked sulfonated carbohydrate hydrogel.

[0077] In another preferred embodiment, the sulfonated sugar solution and the hydrogel matrix precursor solution are as described above.

[0078] In another preferred embodiment, the auxiliary agent is selected from a photoinitiator, thrombin or a pH regulator.

[0079] In another preferred embodiment, when the hydrogel matrix material in the hydrogel prepolymerization solution is selected from methacryloyl hyaluronic acid (HAMA), methacryloyl collagen (GelMA), or a combination thereof, the auxiliary agent is a photoinitiator.

[0080] In another preferred embodiment, when the hydrogel matrix material in the hydrogel prepolymerization solution is fibrinogen, the auxiliary agent is thrombin.

[0081] In another preferred embodiment, when the hydrogel matrix material in the hydrogel prepolymerization solution is collagen, the auxiliary agent is a pH regulator.

[0082] In another preferred embodiment, when the hydrogel matrix material in the hydrogel prepolymerization solution is Matrigel, the cross-linked sulfonated carbohydrate hydrogel can be prepared without step (s2).

[0083] In another preferred embodiment, when the auxiliary agent is a photoinitiator, the curing is photocuring, preferably UV light curing, more preferably 405 nm UV light curing.

[0084] In another preferred embodiment, when the auxiliary agent is thrombin or a pH regulator, the solidification is static solidification.

[0085] In another preferred embodiment, the sulfonated carbohydrate hydrogel is prepared by the following method, which comprises the following steps:

[0086] (i) mixing the sulfonated sugar with a first solvent to obtain a sulfonated sugar solution;

[0087] (ii) mixing a hydrogel matrix material, a photoinitiator and a second solvent to obtain a hydrogel solution;

[0088] (iii) mixing the sulfonated sugar solution and the hydrogel solution to obtain a hydrogel prepolymer solution;

[0089] (iv) irradiating the hydrogel prepolymerization solution with a 405 nm ultraviolet curing lamp for 10-60 seconds to obtain the sulfonated sugar hydrogel.

[0090] In another preferred embodiment, the HAMA gel matrix material is prepared using the following method, comprising the steps of:

[0091] (a) dissolving hyaluronic acid in ultrapure water to obtain a hyaluronic acid solution;

[0092] (b) adding methacrylic anhydride dropwise to the hyaluronic acid solution under ice-water bath conditions, and adjusting the pH of the solution system with sodium hydroxide to maintain the pH at 8.5, and continuing stirring under this condition for 4 hours; stirring the hyaluronic acid solution system at room temperature overnight; dialyzing the hyaluronic acid solution in ultrapure water at room temperature for 7 days using a 14000Da dialysis bag to remove incompletely reacted methacrylic anhydride;

[0093] (c) adjusting the pH to 7.2 and freeze-drying to obtain methacrylated hyaluronic acid (HAMA) powder.

[0094] In another preferred embodiment, the pharmaceutical composition is in the form of a hydrogel.

[0095] In another preferred embodiment, the pharmaceutical composition is an implant.

[0096] In another preferred embodiment, the pharmaceutical composition is administered by dripping into the surface of the heart.

[0097] In another preferred embodiment, the pharmaceutical composition is administered by applying on the surface of the heart.

[0098] In another preferred embodiment, in the pharmaceutical composition, the content of the sulfonated carbohydrate hydrogel is 50wt% to 99wt%.

[0099] In another preferred embodiment, the pharmaceutical composition further comprises a pharmaceutically acceptable excipient, and the excipient is selected from: one or more of a solvent, a diluent, a disintegrant, a precipitation inhibitor, a surfactant, a glidant, a binder, a lubricant, a dispersant, a suspending agent, an isotonic agent, a thickener, an emulsifier, a preservative, a stabilizer, a hydrating agent, an emulsification accelerator, a buffer, an absorbent, a flavoring agent, a sweetener, an ion exchanger, a release agent, a coating agent, a flavoring agent, an antioxidant, a carbohydrate, a fat, a vitamin, an amino acid, a trace element or a protein.

[0100] In the second aspect of the present invention, a use of a hydrogel prepolymerization solution is provided for preparing a pharmaceutical composition for preventing and / or treating myocardial infarction; wherein the hydrogel prepolymerization solution comprises: a sulfonated sugar solution, a hydrogel matrix precursor solution and optionally an auxiliary agent.

[0101] In another preferred embodiment, the sulfonated sugar solution, the hydrogel matrix precursor solution and the auxiliary agent are independently as described in the first aspect of the present invention.

[0102] In another preferred embodiment, the pharmaceutical composition is administered by dripping into the surface of the heart.

[0103] In a third aspect of the present invention, there is provided a method for preventing and / or treating myocardial infarction, the method comprising administering a sulfonated saccharide hydrogel containing a sulfonated saccharide and a hydrogel matrix material, or a hydrogel prepolymerization solution to a subject in need thereof;

[0104] The hydrogel prepolymerization solution comprises: a sulfonated sugar solution, a hydrogel matrix precursor solution and optionally an auxiliary agent.

[0105] In another preferred embodiment, the sulfonated sugar solution, the hydrogel matrix precursor solution and the auxiliary agent are independently as described in the first aspect of the present invention.

[0106] In another preferred embodiment, the subject is a human or non-human mammal, such as rats, mice, dogs, rabbits, pigs, sheep, and cows.

[0107] It should be understood that within the scope of the present invention, the above-mentioned technical features of the present invention and the technical features specifically described below (such as embodiments) can be combined with each other to form a new or preferred technical solution. Due to space limitations, they will not be described one by one here. BRIEF DESCRIPTION OF THE DRAWINGS

[0108] Figure 1 The results show that sulfonated chitosan-based Matrigel induces vascularized sprouts.

[0109] Figure 2 The sprouting-inducing behavior of heparin (100,000 Da)-based collagen hydrogel on arterial circle cells is shown.

[0110] Figure 3 The figures show the cardiac ultrasound results of mice in the sham operation group (Sham group), hyaluronic acid hydrogel group (HA group), and sulfonated chitosan oligosaccharide / hyaluronic acid hydrogel (SCOS HA group) 28 days after myocardial infarction.

[0111] Figure 4 The graphs show the results of Masson's trichrome staining of the mouse hearts in the sham operation group (Sham group), hyaluronic acid hydrogel group (HA group), and sulfonated chitosan oligosaccharide / hyaluronic acid hydrogel (SCOS HA group) 28 days after myocardial infarction.

[0112] Figure 5The figures show the results of immunofluorescence staining of α-SMA and CD31 in the hearts of mice in the sham operation group (Sham group), hyaluronic acid hydrogel group (HA group), and sulfonated chitosan oligosaccharide / hyaluronic acid hydrogel (SCOS HA group) 28 days after myocardial infarction. DETAILED DESCRIPTION

[0113] After extensive and in-depth research, the inventors of the present application unexpectedly discovered that sulfonated sugars have the effect of promoting arterial angiogenesis, and the hydrogels loaded by them can significantly promote functional reconstruction after myocardial infarction. On this basis, the present invention was completed.

[0114] the term

[0115] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.

[0116] As used herein, the terms "comprise", "include", and "contain" are used interchangeably and include not only closed definitions, but also semi-closed and open definitions. In other words, the terms include "consisting of", "consisting essentially of".

[0117] Sulfonated sugars

[0118] The sulfonated sugars involved in the present invention include sulfonated polysaccharides and sulfonated oligosaccharides.

[0119] Polysaccharide is a polymeric high molecular carbohydrate composed of sugar chains linked by glycosidic bonds, at least 10 monosaccharides. Polysaccharides composed of the same monosaccharide structure are called homopolysaccharides, and polysaccharides composed of different monosaccharide structures are called heteropolysaccharides. Polysaccharides can be further hydrolyzed to produce a series of intermediate products, and the final complete hydrolysis product is a monosaccharide.

[0120] Oligosaccharide, also known as oligosaccharide, refers to a compound composed of 2-10 glycosidic bonds. The glycosidic bond is formed by the shrinkage of the glycoside hydroxyl group of a monosaccharide and a hydroxyl group of another monosaccharide.

[0121] Sulfonated sugars refer to products obtained by sulfonation of polysaccharides or oligosaccharides.

[0122] The sulfonated sugar in the present invention is mainly prepared by using chitosan, chitosan oligosaccharide and dextran as raw materials, selecting chlorosulfonic acid, concentrated sulfuric acid and sulfur trioxide pyridine complex as sulfonation reagents in an anhydrous and oxygen-free reaction system. The molecular weight of the sulfonated sugar is determined by aqueous gel permeation chromatography (Viscotek 270max, Malvern Company, UK).

[0123] The sulfur content of the sulfonated sugars was determined by an elemental analyzer (Elementar, Vario Macro, Germany).

[0124] The sulfonated sugars of the present invention include sulfonated chitosan, sulfonated chitosan oligosaccharide, sulfonated dextran, chondroitin sulfate, heparin and heparan sulfate, with a molecular weight of 1000-300000 Da and a sulfur content of 0.5-30%.

[0125] In another preferred embodiment, the molecular weight of the sulfonated sugar is 1500-10000 Da, and the sulfur content is 0.5%-20%.

[0126] In another preferred embodiment, the molecular weight of the sulfonated polysaccharide is 7000-100000 Da, and the sulfur content is 0.5-20%.

[0127] In another preferred embodiment, the molecular weight of the sulfonated oligosaccharide is 1500-3000 Da, and the sulfur content is 1%-20%.

[0128] Sulfonated carbohydrate hydrogel and its preparation

[0129] The present invention provides a sulfonated carbohydrate hydrogel, comprising a hydrogel matrix material and a sulfonated carbohydrate. The hydrogel raw materials include collagen, fibrinogen, Matrigel matrix glue, methacryloyl hyaluronic acid (HAMA), and methacryloyl collagen (GelMA). The sulfonated carbohydrates include sulfonated chitosan, sulfonated chito-oligosaccharide, sulfonated dextran, chondroitin sulfate, heparin, and heparan sulfate. The two are mixed to obtain an aqueous solution of a certain proportion, and cross-linked and cured by ultraviolet light initiation. The hydrogel preparation process is as described in the preparation of GelMA hydrogel (Nature Protocols, 11: 727-746).

[0130] The mass ratio of the sulfonated sugar to the hydrogel matrix material in the sulfonated sugar hydrogel of the present invention is 1:2 to 1:4000.

[0131] In another preferred embodiment, the mass ratio of the sulfonated sugar to the hydrogel matrix material in the sulfonated sugar hydrogel is 1:100 to 1:1000.

[0132] In another preferred embodiment, the mass ratio of the sulfonated sugar to the hydrogel matrix material is 1:2 to 1:500.

[0133] In another preferred embodiment, the mass ratio of the sulfonated sugar to the hydrogel matrix material is 1:500 to 1:2.

[0134] In the present invention, the "sulfonated sugar solution" refers to a solution obtained by mixing the sulfonated sugar and the first solvent.

[0135] In the present invention, the "hydrogel matrix precursor solution" refers to a solution obtained by mixing the hydrogel matrix material and the second solvent.

[0136] In the present invention, "hydrogel solution" refers to a solution obtained by mixing a hydrogel matrix material, a second solvent and an optional auxiliary agent.

[0137] In the present invention, the "hydrogel prepolymerization solution" refers to a solution obtained by mixing a sulfonated sugar solution and a hydrogel solution, which contains sulfonated sugar, a hydrogel matrix material, an optional auxiliary agent, and a solvent.

[0138] The present invention also provides a method for preparing a sulfonated carbohydrate hydrogel, comprising the following steps:

[0139] (a) mixing a sulfonated sugar with a first solvent to obtain a sulfonated sugar solution;

[0140] (b) mixing the hydrogel matrix material with a second solvent to obtain a hydrogel matrix precursor solution;

[0141] (c) mixing the sulfonated sugar solution and the hydrogel matrix precursor solution and optionally cross-linking and curing them to obtain the sulfonated sugar hydrogel.

[0142] In another preferred embodiment, the steps (a), (b) and (c) are independently as described above.

[0143] More specifically, the method comprises the following steps:

[0144] i) providing IC2959 photoinitiator;

[0145] ii) dissolving the hydrogel matrix material in the IC2959 solution to obtain a hydrogel solution;

[0146] iii) dissolving the sulfonated sugar in the hydrogel solution to obtain a hydrogel prepolymer solution;

[0147] iv) irradiating the prepolymerized solution with a 405 nm ultraviolet curing lamp for 10-60 seconds to obtain the sulfonated sugar hydrogel.

[0148] In another preferred embodiment, the concentration of IC2959 solution in step i) is 0.1 mg / mL to 5 mg / mL.

[0149] In another preferred embodiment, the mass ratio of the sulfonated sugar to the hydrogel matrix material in step iii) is 1:1000 to 1:100.

[0150] In another preferred embodiment, the mass ratio of the sulfonated sugar to the hydrogel matrix material in step iii) is 1:500 to 1:2.

[0151] Hydrogel prepolymerization solution, kit

[0152] The present invention also provides a medicine box, comprising:

[0153] (a) a first kit comprising a sulfonated sugar solution;

[0154] (b) A second kit comprising a hydrogel matrix precursor solution and optional auxiliary agents.

[0155] The present invention also provides a hydrogel prepolymerization solution, comprising a sulfonated sugar solution, a hydrogel matrix precursor solution and optionally an auxiliary agent.

[0156] In another preferred embodiment, during the transportation process, the sulfonated sugars are not prevented from directly cross-linking with the hydrogel matrix material under the action of the auxiliary agent, and they are transported in different medicine boxes.

[0157] When in use, the liquids in the first medicine box and the second medicine box are directly mixed to obtain a hydrogel prepolymer solution, which is then quickly dripped onto the surface of the heart to obtain the sulfonated sugar hydrogel of the present invention under ultraviolet light irradiation or self-crosslinking.

[0158] use

[0159] The invention provides a sulfonated sugar hydrogel containing sulfonated sugar and a hydrogel matrix material, a hydrogel prepolymerization solution and a medicine kit for preparing a pharmaceutical composition for preventing and / or treating myocardial infarction.

[0160] In another preferred embodiment, the pharmaceutical composition is also used to increase the thickness of the infarcted ventricular wall, reduce the area of ​​myocardial infarction, alleviate myocardial tissue scar hyperplasia, prevent myocardial infarction, increase the number of myocardium and improve cardiac contractile function.

[0161] In another preferred embodiment, the use is to prepare an implant for treating myocardial infarction.

[0162] In another preferred embodiment, the use is to prepare an implant for inducing new blood vessel growth in myocardial infarction and repairing myocardial function.

[0163] The research results of the present invention show that the sulfonated sugar hydrogel has a good therapeutic effect on arterial angiogenesis and myocardial function recovery during myocardial infarction.

[0164] The main advantages of the present invention are:

[0165] (1) The sulfonated carbohydrate hydrogel for treating myocardial infarction of the present invention does not contain exogenous cells or growth factors, has a long half-life and is highly safe.

[0166] (2) The sulfonated carbohydrate hydrogel for treating myocardial infarction of the present invention can effectively promote the growth of new blood vessels and recovery of cardiac function after myocardial infarction without introducing exogenous cells or growth factors, and significantly inhibit ventricular hypertrophy and fibrosis in the late stage of infarction.

[0167] The present invention will be further described below in conjunction with specific examples. It should be understood that these examples are only used to illustrate the present invention and are not intended to limit the scope of the present invention. The experimental methods in the following examples without specifying specific conditions are usually based on conventional conditions or the conditions recommended by the manufacturer. Unless otherwise stated, percentages are weight percentages.

[0168] Material

[0169] Photoinitiator IC2959, chemical name 2-hydroxy-2-methyl-1-[4-(2-hydroxyethoxy)phenyl]-1-propanone

[0170] HAMA, chemical name methacrylated hyaluronic acid hydrogel

[0171] GelMA, chemical name is methacrylated gelatin hydrogel

[0172] Matrigel matrix gel is a commercial gel precursor solution. It is a basement membrane matrix extracted from EHS mouse tumors rich in extracellular matrix proteins. Its main components include laminin, type IV collagen, heparin sulfate glycoprotein, nestin, etc.

[0173] Comparative Example 1

[0174] Preparation of cross-linked hydrogels

[0175] Deionized water and HAMA with a molecular weight of 60,000 Da were used to prepare a hydrogel matrix precursor solution with a mass volume fraction of 5%, wherein the content of photoinitiator IC2959 was 0.5%. The prepolymerization solution was irradiated with a 405 nm wavelength ultraviolet curing lamp for 20 seconds to obtain a cross-linked hydrogel.

[0176] Comparative Example 2

[0177] Preparation of cross-linked hydrogels

[0178] Deionized water and GelMA with a molecular weight of 100,000 Da were used to prepare a hydrogel matrix precursor solution with a mass volume fraction of 10%, wherein the content of photoinitiator IC2959 was 0.1%, and a 405 nm wavelength ultraviolet curing lamp was used to irradiate the prepolymer solution for 10 seconds to obtain a cross-linked hydrogel.

[0179] Comparative Example 3

[0180] Preparation of cross-linked hydrogels

[0181] The Matrigel gel precursor solution was taken out from the -20°C refrigerator and placed in a 4°C environment in advance to be fully melted. The melted Matrigel gel precursor solution was placed in a 37°C cell culture incubator for 30 minutes to obtain a cross-linked hydrogel.

[0182] Comparative Example 4

[0183] Preparation of cross-linked hydrogels

[0184] The pH of the type I collagen solution with a final concentration of 1 mg / mL was adjusted to a neutral pH of 7.2 and transferred to a 37° C. cell culture incubator for 30 minutes to obtain a cross-linked hydrogel.

[0185] Example 1

[0186] Preparation of cross-linked sulfonated chitosan oligosaccharide hydrogel

[0187] Sulfonated chitosan oligosaccharide with a molecular weight of 1900Da and a sulfur content of 12% was dissolved in deionized water to prepare a sulfonated chitosan oligosaccharide solution with a concentration of 100μg / mL. Deionized water and HAMA with a molecular weight of 60000Da were used to prepare a hydrogel matrix precursor solution with a mass volume fraction of 10%, wherein the content of photoinitiator IC2959 was 0.5%. The sulfonated chitosan oligosaccharide solution and the hydrogel matrix precursor solution were uniformly mixed in a volume ratio of 1:1, and the prepolymer solution was irradiated with a wavelength of 405nm ultraviolet curing lamp for 20 seconds to obtain a cross-linked sulfonated chitosan oligosaccharide hydrogel.

[0188] Example 2

[0189] Preparation of cross-linked sulfonated chitosan oligosaccharide hydrogel

[0190] Sulfonated chitosan oligosaccharide with a molecular weight of 2500Da and a sulfur content of 15% was dissolved in deionized water to prepare a sulfonated chitosan oligosaccharide solution with a concentration of 50μg / mL. Deionized water and GelMA with a molecular weight of 100000Da were used to prepare a hydrogel matrix precursor solution with a mass volume fraction of 10%, wherein the content of photoinitiator IC2959 was 0.1%. The sulfonated chitosan oligosaccharide solution and the hydrogel matrix precursor solution were uniformly mixed in a volume ratio of 1:1, and the prepolymerized solution was irradiated with a wavelength of 405nm ultraviolet curing lamp for 20 seconds to obtain a cross-linked sulfonated chitosan oligosaccharide hydrogel.

[0191] Example 3

[0192] Preparation of cross-linked sulfonated chitosan hydrogel

[0193] Sulfonated chitosan with a molecular weight of 10000Da and a sulfur content of 20% was dissolved in deionized water to prepare a sulfonated chitosan solution with a concentration of 200μg / mL. A hydrogel matrix precursor solution with a mass volume fraction of 0.5% was prepared using physiological saline and fibrinogen. The sulfonated chitosan solution and the hydrogel matrix precursor solution were uniformly mixed in a volume ratio of 1:1, thrombin was added to the mixture to make the final concentration of 1U / mL, and the mixture was mixed and left to stand for 10 minutes to obtain a cross-linked sulfonated chitosan hydrogel.

[0194] Example 4

[0195] Preparation of cross-linked sulfonated chitosan hydrogel

[0196] Sulfonated chitosan with a molecular weight of 8000Da and a sulfur content of 8% was dissolved in deionized water to prepare a sulfonated chitosan solution with a concentration of 100μg / mL. Deionized water and HAMA with a molecular weight of 60000Da were used to prepare a hydrogel matrix precursor solution with a mass volume fraction of 5%, wherein the content of photoinitiator IC2959 was 0.2%. The sulfonated chitosan solution and the hydrogel matrix precursor solution were uniformly mixed in a volume ratio of 1:1, and the prepolymer solution was irradiated with a wavelength of 405nm ultraviolet curing lamp for 20 seconds to obtain a cross-linked sulfonated chitosan hydrogel.

[0197] Example 5

[0198] Preparation of cross-linked sulfonated chitosan hydrogel

[0199] Sulfonated chitosan with a molecular weight of 18000Da and a sulfur content of 13% was dissolved in deionized water, and the sulfonated chitosan concentrate (80 μg / mL) was mixed into the Matrigel precursor solution at a volume ratio of 1:100, and placed in a 37°C cell culture incubator for 30 minutes to obtain a cross-linked sulfonated chitosan hydrogel.

[0200] Example 6

[0201] Preparation of cross-linked chondroitin sulfate hydrogel

[0202] Chondroitin sulfate with a molecular weight of 70000Da and a sulfur content of 8% was dissolved in deionized water to prepare a chondroitin sulfate solution with a concentration of 100μg / mL. A hydrogel matrix precursor solution with a mass volume fraction of 0.5% was prepared using physiological saline and fibrinogen. The chondroitin sulfate solution and the hydrogel matrix precursor solution were uniformly mixed in a volume ratio of 1:1, thrombin was added to the mixture to make the final concentration of 1U / mL, and the mixture was mixed and left to stand for 10 minutes to obtain a cross-linked chondroitin sulfate hydrogel.

[0203] Example 7

[0204] Preparation of cross-linked heparin hydrogel

[0205] Heparin with a molecular weight of 100,000 Da and a sulfur content of 10% was dissolved in deionized water to prepare a heparin solution with a concentration of 50 μg / mL. A hydrogel matrix precursor solution with a mass volume fraction of 0.2% was prepared using physiological saline and collagen. The heparin solution and the hydrogel matrix precursor solution were evenly mixed in a volume ratio of 1:1, and the system pH was calibrated to 7.2. After fully mixing, the cross-linked heparin hydrogel was obtained by standing at 37°C for 30 minutes.

[0206] Example 8

[0207] Preparation of cross-linked heparin hydrogel

[0208] Heparin with a molecular weight of 10000Da and a sulfur content of 15% was dissolved in deionized water to prepare a heparin solution with a concentration of 10μg / mL. Deionized water and GelMA with a molecular weight of 100000Da were used to prepare a hydrogel matrix precursor solution with a mass volume fraction of 5%, in which the content of photoinitiator IC2959 was 0.5%. The heparin solution and the hydrogel matrix precursor solution were uniformly mixed in a volume ratio of 1:1, and the prepolymer solution was irradiated with a wavelength of 405nm ultraviolet curing lamp for 20 seconds to obtain a cross-linked heparin hydrogel.

[0209] Example 9

[0210] Preparation of cross-linked heparin hydrogel

[0211] Heparin with a molecular weight of 100,000 Da and a sulfur content of 10% was dissolved in deionized water to prepare a heparin solution with a concentration of 20 μg / mL. An equal volume of Matrigel was added, and after being fully mixed, the solution was allowed to stand at 37° C. for 30 minutes to obtain a cross-linked heparin hydrogel.

[0212] Example 10

[0213] Preparation of cross-linked sulfonated dextran hydrogel

[0214] Sulfonated dextran with a molecular weight of 150,000 Da and a sulfur content of 20% was dissolved in deionized water to prepare a sulfonated dextran solution with a concentration of 1 μg / mL. Deionized water and GelMA with a molecular weight of 100,000 Da were used to prepare a hydrogel matrix precursor solution with a mass volume fraction of 5%, in which the content of photoinitiator IC2959 was 0.5%. The sulfonated dextran solution and the hydrogel matrix precursor solution were uniformly mixed in a volume ratio of 1:1, and the prepolymer solution was irradiated with a wavelength of 405 nm ultraviolet curing lamp for 20 seconds to obtain a cross-linked sulfonated dextran hydrogel.

[0215] Embodiment 11

[0216] Preparation of cross-linked sulfonated dextran hydrogel

[0217] Sulfonated dextran with a molecular weight of 50,000 Da and a sulfur content of 15% was dissolved in deionized water to prepare a sulfonated dextran solution with a concentration of 100 μg / mL. A hydrogel matrix precursor solution with a mass volume fraction of 0.2% was prepared using physiological saline and collagen. The sulfonated dextran solution and the hydrogel matrix precursor solution were evenly mixed in a volume ratio of 1:1, and the system pH was calibrated to 7.2. After fully mixing, the cross-linked sulfonated dextran hydrogel was obtained by standing at 37°C for 30 minutes.

[0218] Example 12

[0219] The hydrogel described in Comparative Example 3 and the sulfonated chitosan Matrigel hydrogel described in Example 5 were subjected to relevant experiments to evaluate the effect of the sulfonated carbohydrate hydrogel on the annulation ability of endothelial cells, as follows:

[0220] The sulfonated chitosan concentrate (80 μg / mL) with a molecular weight of 18000Da and a sulfur content of 13% was mixed into the Matrigel precursor solution at a volume ratio of 1:100. After thorough mixing, 50 μL of the Matrigel precursor solution containing sulfonated chitosan was added to a 96-well plate. The control group was a simple Matrigel precursor solution of the same volume. The plate was placed in a 37°C cell culture incubator for 30 minutes to allow complete gelation. The pre-cultured human umbilical vein endothelial cells (HUVECs) were digested and centrifuged, and 1×10 4 The cells / well were seeded at a density of 100 μg / well in a 96-well plate covered with gel. After the well plate was transferred to a cell culture incubator and cultured for 4 hours, it was fixed with 4% paraformaldehyde, and the cytoskeleton and cell nucleus were stained with FITC-labeled phalloidin stain and DAPI stain, respectively, and recorded with a laser confocal fiber microscope.

[0221] The results showed that HUVECs were more likely to form ring structures in Matrigel hydrogels containing sulfonated chitosan ( Figure 1 ). Endothelial cell ring formation is considered to be a necessary process in angiogenesis. This result reflects that Matrigel hydrogel containing sulfonated chitosan is beneficial to promote the vascularization behavior of endothelial cells.

[0222] Embodiment 13

[0223] The hydrogel described in Comparative Example 4 and the heparin hydrogel described in Example 7 were subjected to relevant experiments to evaluate the effect of the sulfonated carbohydrate hydrogel on vascular sprouting, as follows:

[0224] Four-week-old male SD rats were killed by cervical dislocation and the epidermis was fully sterilized by soaking in 75% ethanol for 15 minutes. The thoracic cavity of the SD rat was opened with dissecting scissors, the thoracic aorta was carefully separated, and it was rinsed several times with normal saline. The adipose tissue on the surface of the arterial tissue was peeled off, and the artery was cut into 1-1.5 mm arterial rings under a microscope. It was placed in a 24-well plate and 500 μL of heparin hydrogel was added. 1 mL of DMEM medium containing 10% fetal bovine serum was added and cultured for 7 days, and fresh medium was replaced every 2 days. After obvious vascular sprouting behavior was observed under an optical microscope, it was fixed with 4% paraformaldehyde, and the endothelial cells were immunofluorescently stained with CD31 antibody, and observed and recorded under a laser confocal microscope.

[0225] The results showed that in collagen hydrogels containing heparin, arterial rings could stimulate sprouting to form more CD31-positive microvascular structures ( Figure 2 ). Vascular sprouting is considered to be an essential process in angiogenesis. This result reflects that collagen hydrogel containing heparin is conducive to promoting the sprouting behavior of angiogenesis.

[0226] Embodiment 14

[0227] The hydrogel described in Comparative Example 1 and the sulfonated chitosan oligosaccharide hyaluronic acid hydrogel described in Example 1 were subjected to relevant experiments to evaluate the therapeutic effect of the sulfonated carbohydrate hydrogel on myocardial infarction, as follows:

[0228] 1. Experimental Grouping and Drug Administration

[0229] Twenty-four 10-week-old male SPF C57BL / 6 mice were housed in a barrier environment animal laboratory at a temperature of 24±2°C, a relative humidity of 40-70%, and a 12-hour light-dark cycle. After 2 days of adaptive feeding, the mice were randomly divided into 3 groups, 8 mice in each group, namely:

[0230] (1) Sham group: negative control group, except that coronary artery ligation was not performed, other operations were consistent with the construction method of the myocardial infarction animal model, and no hydrogel was applied.

[0231] (2) Hyaluronic acid hydrogel group (HA group): After successful modeling, a hydrogel without sulfonated sugar was administered.

[0232] (3) Sulfonated chitosan oligosaccharide / hyaluronic acid hydrogel group (SCOS HA group): After successful modeling, hydrogel containing sulfonated chitosan was applied.

[0233] 2. Establishment of the mouse myocardial infarction model

[0234] 10-week-old male C57BL / 6 mice were taken and anesthetized with 0.8% sodium pentobarbital at an injection dose of 80g / kg. The mice were fixed on a mouse fixator in a supine position, intubated and connected to a small animal ventilator to maintain breathing. After shaving and disinfecting the left chest, the chest cavity was opened between the 4th and 5th ribs to expose the complete heart of the mouse. The left anterior descending branch was ligated near the left coronary artery using a 7-0 non-absorbable nylon suture with a needle. After coronary artery ligation, the myocardium below the ligature line was pale and the contraction movement of the anterior wall was weakened, indicating that the model was successful.

[0235] After successful modeling, 20 μL of hydrogel containing or not containing sulfonated sugars was dropped on the surface of the infarcted heart and completely gelled by ultraviolet cross-linking or self-cross-linking.

[0236] The opening was sutured again, and the mice were kept in an SPF environment for further observation after they woke up.

[0237] 28 days after myocardial infarction, mice were anesthetized with isoflurane inhalation and the precordial skin was prepared. Cardiac ultrasound was performed using a small animal ultrasound system, and the probe was adjusted to obtain clear left ventricular long-axis two-dimensional images for arterial monitoring of cardiac function changes in mice.

[0238] Cardiac ultrasound results showed that the sulfonated chitosan oligosaccharide hydrogel treatment could effectively restore the wall motion and myocardial contraction of the left ventricular infarction site ( Figure 3 ).

[0239] 4. Histological Evaluation

[0240] 28 days after myocardial infarction, mice were killed by cervical dislocation and the intact heart was carefully removed. They were fixed with 4% paraformaldehyde for 72 hours, dehydrated step by step, embedded in paraffin, sectioned and stained with Masson's trichrome. After staining, they were photographed and recorded using an optical microscope.

[0241] The results of Masson's trichrome staining of cardiac tissue showed that red represents normal myocardial tissue. Myocardial tissue necrosis will cause scar invasion. Scars with high collagen expression are easily stained blue. The arrows represent blood vessels. Compared with the simple hydrogel HA group, the sulfonated chitosan oligosaccharide hydrogel group (SCOS HA) had a lower degree of fibrosis and fewer necrotic areas (see Figure 4 ).

[0242] 5. Immunofluorescence Staining

[0243] 28 days after myocardial infarction, mice were killed by cervical dislocation and the intact heart was carefully removed. They were fixed with 4% paraformaldehyde for 4 hours, embedded in freezing embedding solution and sliced, and the peripheral smooth muscle cells and endothelial cells were stained and marked with antibodies α-SMA and CD31, respectively. After staining, they were photographed and recorded using a laser confocal microscope.

[0244] Immunofluorescence staining results showed that sulfonated chitosan oligosaccharide hydrogel can effectively promote arterial angiogenesis in the infarcted area (see Figure 5 ).

[0245] The above results indicate that sulfonated carbohydrate hydrogel has a good therapeutic effect on arterial angiogenesis and myocardial function recovery during myocardial infarction.

[0246] In summary, the preparation method of the sulfonated carbohydrate hydrogel provided in this application is simple to operate and suitable for industrial production. The prepared sulfonated carbohydrate hydrogel has good biocompatibility and can effectively promote the growth of new blood vessels and recovery of cardiac function after myocardial infarction without the introduction of exogenous cells or growth factors, and significantly inhibit the degree of ventricular hypertrophy and fibrosis in the late stage of infarction, providing a new idea for the transformative treatment of clinical myocardial infarction.

[0247] All documents mentioned in the present invention are cited as references in this application, just as each document is cited as reference individually. In addition, it should be understood that after reading the above teachings of the present invention, those skilled in the art can make various changes or modifications to the present invention, and these equivalent forms also fall within the scope defined by the claims attached to this application.

Claims

1. A use of a sulfonated sugar hydrogel containing a sulfonated sugar and a hydrogel matrix material, characterized in that: Used for preparing a pharmaceutical composition for preventing and / or treating myocardial infarction.

2. The use according to claim 1, characterized in that The prevention and / or treatment of myocardial infarction has one or more characteristics selected from the following group: (1) Increased wall thickness in the infarcted area; (2) reduce myocardial infarction area; (3) Reduce myocardial scar hyperplasia; (4) Increase the amount of cardiac muscle; (5) restore ventricular wall motion at the site of myocardial infarction; (6) Reduce the degree of fibrosis at the site of myocardial infarction; (7) promoting myocardial arterial angiogenesis; and (8) Improve heart contractile function.

3. The use according to claim 1, characterized in that The sulfonated sugar is selected from the group consisting of sulfonated chitosan, sulfonated chito-oligosaccharide, sulfonated dextran, chondroitin sulfate, heparin, heparan sulfate, or a combination thereof; The hydrogel matrix material is selected from the group consisting of collagen, fibrinogen, Matrigel, methacryloyl hyaluronic acid, methacryloyl collagen, or a combination thereof.

4. The use according to claim 1, characterized in that The molecular weight of the sulfonated sugar is 1000-300000Da, and the sulfur content is 5-30%.

5. The use according to claim 1, characterized in that: The sulfonated carbohydrate hydrogel is prepared by the following method, which comprises the following steps: (a) mixing a sulfonated sugar with a first solvent to obtain a sulfonated sugar solution; (b) mixing the hydrogel matrix material with a second solvent to obtain a hydrogel matrix precursor solution; (c) mixing the sulfonated sugar solution and the hydrogel matrix precursor solution and optionally cross-linking and curing them to obtain the sulfonated sugar hydrogel.

6. The use according to claim 5, characterized in that In step (a), the concentration of the sulfonated sugar solution is 0.01 to 500 μg / mL; In step (b), the mass volume fraction of the hydrogel matrix material in the hydrogel matrix precursor solution is 0.1 to 30%.

7. The use according to claim 5, characterized in that When the hydrogel matrix material is selected from methacryloyl hyaluronic acid (HAMA), methacryloyl collagen (GelMA), or a combination thereof, the step (c) comprises: mixing the sulfonated sugar solution and the hydrogel matrix precursor solution in the presence of a photoinitiator, and curing by irradiating with ultraviolet light to obtain a cross-linked sulfonated sugar hydrogel; When the hydrogel matrix material is fibrinogen, the step (c) comprises: mixing the sulfonated sugar solution and the hydrogel matrix precursor solution, and then adding thrombin for solidification to obtain a cross-linked sulfonated sugar hydrogel; When the hydrogel matrix material is collagen, the step (c) comprises: mixing the sulfonated sugar solution and the hydrogel matrix precursor solution, then adjusting the pH to above 7, and standing to solidify to obtain a cross-linked sulfonated sugar hydrogel; When the hydrogel matrix material is Matrigel, the step (c) comprises: mixing the sulfonated sugar solution and the hydrogel matrix precursor solution, and letting the mixture stand to obtain a cross-linked sulfonated sugar hydrogel.

8. The use according to claim 5, characterized in that In step (c), the volume ratio of the sulfonated sugar solution to the hydrogel matrix precursor solution is 1:0.01 to 1:10, preferably 1:0.5 to 1:5, and more preferably 1:0.5 to 1:

2.

9. The use according to claim 1, characterized in that The pharmaceutical composition is administered by applying to the surface of the heart.

10. A use of a hydrogel prepolymer solution for preparing a pharmaceutical composition for preventing and / or treating myocardial infarction; wherein: The hydrogel prepolymerization solution comprises: a sulfonated sugar solution, a hydrogel matrix precursor solution and optionally an auxiliary agent; The auxiliary agent is selected from a photoinitiator, thrombin or a pH regulator.

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