Functional hydrogel and its preparation and application in the preparation of drugs for treating glaucoma

Abstract

The present invention belongs to the field of ophthalmic drugs, and specifically discloses a functional hydrogel, which comprises a hydrogel matrix and pharmaceutically effective amounts of component A and component B dispersed therein; the component A is a compound of formula 1 and its pharmaceutically acceptable salts; the component B is exosomes derived from bone marrow mesenchymal stem cells. The present invention also includes the preparation of the functional hydrogel and its application in the preparation of glaucoma. The research of the present invention shows that by using the functional hydrogel, synergy can be achieved, and then based on the idea of protecting the optic nerve and visual function, the therapeutic effect of glaucoma can be improved and the treatment time effect can be improved.

Description

Technical Field

[0001] The present invention belongs to the field of biomedical materials, and particularly relates to the field of treating glaucoma optic nerve and visual function protection. Background Art

[0002] Glaucoma is a chronic degenerative eye disease characterized by progressive damage to the optic nerve, which can lead to irreversible vision loss. Its characteristic pathological change is the selective damage of retinal ganglion cells (RGCs) and their axons. Glaucoma affects tens of millions of people worldwide, making it a major public health problem. Currently, the treatment options for glaucoma mainly focus on reducing intraocular pressure to slow down the progression of the disease. However, the efficacy of intraocular pressure-lowering interventions is often limited and cannot prevent further damage to the optic nerve and visual function. Therefore, simply reducing intraocular pressure is not sufficient to ensure a good prognosis for glaucoma, and the development of neuroprotective drugs is a key breakthrough in the drug treatment of glaucoma. However, there is still a lack of effective drugs for protecting the optic nerve and visual function in clinical practice.

[0003] The currently clinically used glaucoma treatment drugs have the following problems. Firstly, the effectiveness of the drugs is difficult to guarantee. The drugs cannot directly reach the retina, and after passing through multiple intraocular barriers, the effectiveness of the drugs is greatly reduced and they cannot play a good role. Secondly, some eye drops need to be administered frequently, which will bring some problems to the ocular surface. There are also some drugs that can achieve good effects, but have great toxic and side effects on other organs of the body and cannot be well applied. The stem cell treatment for glaucoma is still in the animal experiment stage. Although many breakthroughs and phased results have been achieved, there are still many problems, and there is still a long way to go from experiment to application. In addition, most patients lack a full understanding of the progression of glaucoma, so low compliance is a common problem in treatment. Applying new drug delivery methods that replace eye drops, such as ocular surface implants and intraocular drug delivery, may improve patient compliance. Therefore, developing a new drug delivery system is an urgent need for the current clinical treatment of glaucoma and can open up a new way for the drug treatment of glaucoma.

[0004] Some prior art has reported some drugs for treating glaucoma. For example, the Chinese patent document with the publication number CN111166870A discloses the application of exenatide in the preparation of drugs for treating ocular ischemic diseases by eye drops and improving ocular blood circulation, indicating that exenatide can be used to treat ocular ischemic diseases such as acute glaucoma attacks and improve ocular blood circulation. Another example is the Chinese patent document with the publication number CN111632025A, which discloses a bimatoprost ophthalmic thermosensitive in-situ gel and its preparation method and application, indicating that bimatoprost is suitable for ophthalmic preparations for primary open-angle glaucoma and ocular hypertension with a relatively high incidence.

[0005] Although the treatment effect of glaucoma continues to improve with the continuous improvement of glaucoma treatment methods, the existing treatment ideas mainly focus on the regulation of intraocular pressure, and there are few reports on effective technical treatments for glaucoma based on neuroprotection. Summary of the invention

[0006] In view of the shortcomings of existing glaucoma treatment methods, the first purpose of the present invention is to provide a functional hydrogel, which aims to improve the treatment effect of glaucoma based on the protection mechanism of optic nerve and visual function, and improve the duration of drug action.

[0007] The second purpose of the present invention is to provide the preparation of the functional hydrogel and its application in the preparation of drugs for treating glaucoma.

[0008] The third object of the present invention is to provide a drug for treating glaucoma comprising the functional hydrogel.

[0009] A functional hydrogel comprising a hydrogel matrix and pharmaceutically effective amounts of component A and component B dispersed therein;

[0010] The component A comprises a compound of formula 1 and a pharmaceutically acceptable salt thereof;

[0011]

[0012] The component B is exosomes derived from bone marrow mesenchymal stem cells.

[0013] Previous studies have shown that the use of components A and B alone has a certain alleviating effect on glaucoma, but still has drawbacks such as rapid metabolism and poor efficacy. To address this deficiency, the present invention innovatively combines components A and B and further disperses and embeds them in a hydrogel matrix. This achieves a synergistic effect and allows for slow release, achieving long-term protective effects. This can reduce the frequency of dosing, mitigate the toxic side effects and poor patient compliance associated with repeated drug use, and significantly improve the treatment of glaucoma by protecting the optic nerve and visual function.

[0014] In the present invention, the exosomes derived from bone marrow mesenchymal stem cells are one of the keys to the synergistic effect with component A.

[0015] In the present invention, the diameter of the exosomes is ≤150 nm.

[0016] In the present invention, as a typical embodiment, the hydrogel matrix is a chitosan-based hydrogel.

[0017] In the present invention, in the functional hydrogel, the concentration of component A is above 15uM, preferably 15 to 50uM, and more preferably 20 to 30uM;

[0018] The concentration of Component B is above 100 μg / mL, preferably 150 - 300 μg / mL, and more preferably 190 - 210 μg / mL.

[0019] The present invention also provides a method for preparing the functional hydrogel, which is obtained by mixing and co-incubating the Component A, Component B and the hydrogel matrix.

[0020] In the present invention, the step for obtaining exosomes is: culturing bone marrow mesenchymal stem cells and harvesting the cell supernatant; filtering, separating and resuspending the precipitate of the cell supernatant to obtain exosomes.

[0021] The present invention also provides an application of the functional hydrogel in the preparation of a medicament for treating glaucoma.

[0022] The research of the present invention shows that based on the combination of the Component A and Component B, further combined with the combined control of the dispersion matrix of the hydrogel matrix, synergism can be unexpectedly achieved, which can improve the therapeutic effect of glaucoma, improve the protection effect of glaucoma optic nerve and visual function. In addition, it can also achieve sustained release synergistically and improve the action time effect.

[0023] Preferably, the functional hydrogel is used in the preparation of a glaucoma medicament for improving the protection of optic nerve and visual function.

[0024] The present invention also provides a medicament for treating glaucoma, which contains the functional hydrogel.

[0025] The medicament for treating glaucoma according to the present invention may further optionally contain pharmaceutically acceptable excipients as needed.

[0026] The medicament for treating glaucoma according to the present invention can be an external preparation or an injection preparation for local administration to the eye; further, it is a vitreous cavity injection preparation.

[0027] The functional hydrogel of the present invention has an excellent ROS response effect, which can solve the problems of retinal ganglion cell loss and visual function damage in glaucoma.

[0028] Beneficial effects

[0029] The functional hydrogel provided by the present invention, based on the combination of its components, can achieve synergism, and can significantly improve the therapeutic effect of glaucoma based on a new idea of protecting optic nerve and visual function. In addition, due to the synergistic combination of the components of the functional hydrogel, sustained release of the components can also be achieved, which can greatly reduce the administration frequency of current glaucoma drug treatments, relieve the toxic and side effects caused by multiple drug administrations, and solve the problem of poor patient compliance. Brief description of the drawings

[0030] Figure 1 Injectability, ROS responsiveness, and structural identification of the hydrogel composite (EL-Gel) prepared in Example 1;

[0031] Figure 2 Extraction and identification of exosomes derived from bone marrow mesenchymal stem cells in Example 1;

[0032] Figure 3 Mass spectrum and quantification chart of Release 1 of EL-Gel prepared in Example 1;

[0033] Figure 4 Structural observation of the retina after 3 days and 28 days of intravitreal injection of EL-Gel prepared in Example 1 for evaluating the safety of EL-Gel;

[0034] Figure 5 Retinal ganglion cell protective effect of EL-Gel prepared in Example 1 on glaucoma-damaged mice;

[0035] Figure 6 Visual function protective effect of EL-Gel prepared in Example 1 on glaucoma-damaged mice;

[0036] Figure 7 Protective effect of EL-Gel prepared in Example 1 on R28 cell injury induced by OGD / R model. Detailed implementation manners

[0037] To make the objectives, technical solutions, and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings in the embodiments of the present invention. Obviously, the described embodiments are some, but not all, of the embodiments of the present invention. All other embodiments obtained by those of ordinary skill in the art based on the embodiments of the present invention without creative efforts shall fall within the protection scope of the present invention.

[0038] Preparation Example 1 - Functional hydrogel:

[0039] Step 1. Collect bone marrow samples and culture bone marrow mesenchymal stem cells, specifically including: centrifuging blood samples at 1000 rcf for 5 minutes and discarding the supernatant. Treat the precipitated cells with red blood cell lysis buffer. Resuspend the lysed cells and seed them in a culture dish. After 24 hours, wash away the floating cells and change the culture medium after 48 hours. The cell culture medium consists of 85% F12 medium, 15% fetal bovine serum, and 1% penicillin-streptomycin solution.

[0040] Step 2. Continue to culture the bone marrow mesenchymal stem cells, and then collect the cell supernatant, specifically including: culturing the cells in a medium containing normal serum until the cells reach 80% confluence; removing the original medium, and then continuing to culture the cells in an exosome-free medium for 48 hours, and collecting the cell supernatant.

[0041] Step 3. Filter, separate and resuspend the precipitate of the cell supernatant to obtain exosomes; the final concentration is 1.6×10 11 particles / mL.

[0042] Step 4. Perform particle size analysis and TEM observation on the extracted exosomes, and characterize the marker expression of the exosomes by Western blotting technology.

[0043] Step 5. Mix the formula 1, exosomes and chitosan hydrogel, and perform co-incubation to obtain a formula 1, exosome hydrogel mixture system (labeled as EL-Gel). The concentrations of formula 1 and exosomes can be adjusted as needed. In the following cases, if not otherwise specified, the EL-Gel mentioned refers to the functional hydrogel with the concentration of formula 1 being 25 uM and the concentration of exosomes being 200 ug / mL.

[0044] Example 1

[0045] Injectability, ROS responsiveness and structural characteristics of the hydrogel (functional hydrogel of Preparation Example 1)

[0046] As Figure 1 shown, A. The composite hydrogel provided by the present invention has injectability; B. The composite hydrogel provided by the present invention has ROS responsiveness, and after treatment with H2O2, it changes from a gel state to a water state; C. The hydrogel mixture system provided by the present invention has the electron microscopy characteristics of the hydrogel; D. The composite hydrogel provided by the present invention is in a gel state.

[0047] Example 2

[0048] Mass spectrometry and quantification diagram of the release of formula 1 by EL-Gel in the retina

[0049] Animal experiments: Wild-type C57BL / 6J mice (8 weeks old, 18 - 20 g) were used to establish an acute ocular hypertension (AOH) mouse model by increasing intraocular pressure (120 mmHg for 60 minutes). Briefly, the mydriatic and corneal anesthetized mice were placed on a heated working table. A 30-gauge needle connected to a normal saline infusion device was carefully inserted into the anterior chamber of the mouse, and the anterior chamber pressure was gradually increased to 120 mmHg; the pressure was maintained for 60 minutes. The control group underwent sham surgery without increasing the pressure in the left eye. For intravitreal administration, intravitreal injection was performed before the start of the AOH model using a Hamilton syringe equipped with a 30-gauge glass micro-needle under a dissecting microscope. 48 hours before the AOH model was established, the drug (0.5 μL of EL-Gel in Example 1) was slowly injected into the vitreous cavity of the eyeball. All animal experiments were reviewed and approved by the Animal Care and Use Committee of the Experimental Animal Research Center of Xiangya School of Medicine, Central South University.

[0050] A solution of Formula 1 at 2.00 mg / mL was prepared with methanol and then successively diluted to standard curve working solutions of 5000, 2000, 500, 200, 50, 20, 5, and 2 ng / mL, and detected using a chromatograph and a mass spectrometer. With the peak area as the ordinate and the concentration of Formula 1 as the abscissa, the standard curve was calculated. The sample data was calculated according to the standard curve to obtain the final content of Formula 1 in the retina.

[0051] As Figure 2 shown: A. Formula 1 could not be detected in the retina of normal mice; B. After the retina was treated with EL-Gel for 7 days, the content of Formula 1 in the retina was detected by a chromatograph and a mass spectrometer, and the peak emergence time was around 1.7 min; C. After the retina was treated with EL-Gel for 28 days, the content of Formula 1 in the retina was detected by a chromatograph and a mass spectrometer, and the peak emergence time was around 1.7 min; D. The standard curve for the quantification of Formula 1; E. Quantification of the content of Formula 1 in the retina at 7 days and 28 days through the standard curve. Regarding the pharmacokinetics of Formula 1, it has been demonstrated that when Formula 1 was simply injected, the half-life was only 4.6 h, and multiple administrations were required to achieve a better effect. Our results show that a single treatment with EL-Gel can allow Formula 1 to remain in the retina for 28 days, greatly reducing the dosing frequency and alleviating the side effects caused by multiple administrations.

[0052] Example 3

[0053] Observation of the retinal structure 3 days and 28 days after intravitreal injection of EL-Gel (0.5 μL) was used to evaluate the safety of EL-Gel;

[0054] After anesthesia, mice were administered intravitreal medication. Topical anesthesia was then administered with oxybuprocaine hydrochloride eye drops (Santen Pharmaceutical, Japan). Mydriasis was dilated with compound tropicamide eye drops (Santen Pharmaceutical, Japan), and the ocular surface was disinfected with levofloxacin eye drops (Bausch and Lomb, Shandong, China) and povidone-iodine (Yintao Pharmaceutical, Jiangxi, China). After adequate pupil dilation, the mice were immobilized under a microscope, and 0.5 μl of the solution was injected into the vitreous cavity of both eyes using a 10 μL microsyringe with a 33-gauge needle (Hamilton, Switzerland). A control group of mice received the same volume of solvent. After injection, the needle was slowly withdrawn, and the ocular surface of the mice was covered with tobramycin-dexamethasone ophthalmic ointment (ALCON, Spain). The mice were then placed in cages to recover from anesthesia. Retinal morphology was observed by HE analysis on days 3 and 28 after modeling.

[0055] HE staining: The eyeball tissue was fixed and embedded in paraffin, and cut into 10-μm thick sections. The sections were stained with hematoxylin-eosin (HE). Each sample was photographed under a microscope. The retinal staining results, including retinal thickness and the number of ganglion cell layer cells, were analyzed blindly. Figure 3 As shown, there was no difference in retinal structure between the mice and normal mice 3d and 28d after intraocular injection of EL-Gel.

[0056] Example 4

[0057] EL-Gel promotes the protection of retinal ganglion cells in mice with glaucomatous damage;

[0058] Intravitreal administration: Mice were weighed and anesthetized with an intraperitoneal injection of 1% sodium pentobarbital (prepared in saline) at a dose of 10 ml / kg. Topical anesthesia was then administered with oxybuprocaine hydrochloride eye drops (Santen Pharmaceutical, Japan). Mydriasis was dilated with compound tropicamide eye drops (Santen Pharmaceutical, Japan), and the ocular surface was disinfected with levofloxacin eye drops (Bausch and Lomb, Shandong, China) and povidone-iodine (Yintao Pharmaceutical, Jiangxi, China). After adequate pupil dilation, the mice were immobilized under a microscope, and 0.5 μl of each solution was injected into the vitreous cavity of both eyes using a 10 μL microsyringe with a 33-gauge needle (Hamilton, Switzerland). A control group of mice received the same volume of solvent. After the injection, the needle was slowly withdrawn, and the mice were treated with tobramycin-dexamethasone ophthalmic ointment (ALCON, Spain) and placed in their cages to recover from anesthesia. Relevant phenotypes were detected on the 1st and 7th days after modeling.

[0059] Specific groups: Ctr: Normal control group; AOH-1d: Acute ocular hypertension 1-day model group; AOH-1d+EL-Gel: Acute ocular hypertension 1-day + EL-Gel treatment group; AOH-7d: Acute ocular hypertension 7-day model group; AOH-7d+EL: Acute ocular hypertension 7-day + Component A + Component B treatment group; AOH-7d+EL-Gel: Acute ocular hypertension 7-day + EL-Gel treatment group.

[0060] Retinal flat mount: Enucleate the mouse eyeballs, fix them in 4% paraformaldehyde (m / v) at room temperature for 2 hours, then remove the cornea and lens. After refixing in 4% PFA (m / v) at room temperature for 1 hour, isolate the retina, transfer it to a glass slide, and cut it into four equal quadrants. The retina is permeabilized in PBS containing 0.3% Triton X-100 (v / v) for 10 minutes, then blocked in blocking solution (5% BSA) (m / v) for 1 hour, and then stained with anti-Brn3a (1:500) overnight at 4°C. After rinsing 3 times with PBS, incubate the retina with Alexa Fluor 488-conjugated goat anti-rabbit secondary antibody at room temperature for 2 hours, and then mount the slides. Images are acquired using a fluorescence microscope (Leica, Wetzlar, Germany). Each retina is divided into 4 quadrants (sharing a central quadrant), and Brn3a-positive cells are counted using Image J to determine the RGCs density (RGCs per square millimeter) of that retina. Calculate the average of the mean densities of all retinas in each group. Using the control sample as a reference, calculate the percentage of the relative RGCs density in each group compared to the average value. Statistical analysis of the relative Brn3a+ cells in each group is performed using GraphPad Prism software.

[0061] As Figure 4 shown, after intravitreal injection of El-Gel for 1 day and 7 days, the results of retinal flat mount showed that compared with the simple AOH group, the number of Brn3a+ cells was significantly restored. And the long-acting protective effect of the sustained-release system constructed by EL-Gel is better than that of the simple drug combination (EL group).

[0062] Example 5

[0063] Protective effect of EL-Gel on visual function in glaucoma-damaged mice

[0064] Flash VEP (FVEP) detection

[0065] After the intervention was completed, the mice were anesthetized with 1% sodium pentobarbital, the pupils were fully dilated, the mice were fixed prone on the operating table, and the heating pad was maintained at 37°C. Carboxymethylcellulose eye drops were instilled into both eyes to keep the corneas moist. Three needle electrodes were inserted into the subcutaneous tissue of the mice as follows: the recording electrode was inserted at the midpoint between the two ear roots, contacting the surface of the occipital bone; the reference electrode was inserted subcutaneously into the nose; the ground electrode was inserted subcutaneously into the tail. One eye was completely covered, and the detection was performed according to the international standard procedure. After the detection was completed, the above steps were repeated for the other eye. The N1-P1 amplitude values of each group were analyzed.

[0066] Flash electroretinogram (FERG) detection

[0067] After the intervention was completed and before the test, the mice were allowed to adapt to the dark environment for more than 12 hours. Other preparation steps were the same as those for VEP, keeping the test environment dark and using red light illumination. Two corneal loop electrodes and three needle electrodes were prepared. The specific electrode placement methods were as follows: the bilateral recording electrodes contacted the corneas, and hydroxyethyl cellulose eye drops were instilled into both eyes to increase current conduction and keep the corneas moist. The reference electrode was inserted subcutaneously on both sides of the nose wings, and the ground electrode was inserted subcutaneously into the tail. After the detection was completed, the amplitudes of the a-wave and b-wave of each group were analyzed.

[0068] As Figure 5 shown, A: FERG detection showed that the amplitudes of both the a-wave and b-wave in AOH model mice were significantly decreased, and after treatment with EL-Gel, the amplitude values of the a-wave and b-wave were significantly restored compared with those in simple AOH mice; B. FVEP detection showed that the N1-P1 amplitude value in AOH model mice was significantly decreased compared with the normal group, and after treatment with EL-Gel, the N1-P1 amplitude value was significantly restored compared with that in simple AOH mice.

[0069] Example 6

[0070] Protective effect of EL-Gel on OGD / R model-induced R28 cell damage

[0071] Cell culture:

[0072] R28 cells are a retinal progenitor cell line with differentiation potential and are commonly used for in vitro studies of the neuroprotection and pathological mechanisms of RGCs. R28 cells were cultured in DMEM low-glucose medium supplemented with 10% fetal bovine serum and 1% penicillin-streptomycin solution. The oxygen-glucose deprivation (OGD / R) model was established as follows: first, the cell culture medium was replaced with serum-free and sugar-free medium and cultured in an anaerobic incubator for 4 hours, then replaced with normal medium (serum-free) and cultured under normal conditions until the experiment was completed. EL-Gel was added before the two medium replacements, and the final concentration in the medium was diluted 1000 times.

[0073] Cell DNA damage detection: Prepare coverslips in advance in a culture dish. After treating OGD / R-damaged cells with EL-Gel, place the cell coverslips on glass slides, fix them with 4% paraformaldehyde for 20 minutes, and then incubate them with 0.3% Triton solution for 5 minutes. Then rinse them 5 times with PBS, block them with 5% FBS for 30 minutes, incubate them overnight at 4°C with γ-H2AX primary antibody (a classic protein marker for DNA damage). The next day, incubate the cells with fluorescent secondary antibody at room temperature for 1 hour and counterstain with DAPI. Collect images using a microscope. Quantify the staining intensity using Image J.

[0074] Cell viability and cytotoxicity staining: Seed R28 cells into 24-well plates. Remove the culture medium of each group, rinse once with PBS, prepare a cell viability and cytotoxicity staining reagent calcein AM / PI (indicating cell death) and low-glucose basal medium at a ratio of 1:2000, incubate at 37°C in the dark for 30 minutes, then take out the staining solution and place it in PBS, and observe and photograph the staining conditions of each group under a fluorescence microscope.

[0075] As Figure 6 shown, (A) both the fluorescence intensity and quantity of γ-H2AX staining in the OGD / R group cells increased significantly. After treating the cells with EL-Gel, the fluorescence intensity and quantity of γ-H2AX staining decreased significantly. This indicates that EL-Gel can improve OGD / R-induced DNA damage in R28 cells; (B) the number of PI-positive cells in the OGD / R group increased significantly. After EL-Gel treatment, the number of PI-positive cells decreased significantly, indicating that EL-Gel reduces OGD / R-induced R28 cell death.

[0076] Example 7

[0077] EL-Gel has a better protective effect on R28 cells when used in combination than when used alone with either of its components.

[0078] Construct an OGD / R model as in the cell culture of Example 6, and add drugs to each group before constructing the model. Ctr group: normal control group; OGD / R group: oxygen-glucose deprivation model group; OGD / R + EL-Gel group: group treated with EL-Gel while constructing the OGD / R model; OGD / R + Component B group: group treated with the same dose of Component B alone while constructing the OGD / R model; OGD / R + Component A group: group treated with the same dose of Component A alone while constructing the OGD / R model; OGD / R + EL group: group treated with the same dose of Component A and Component B in combination while constructing the OGD / R model; OGD / R + Gel group: group treated with the same dose of chitosan hydrogel while constructing the OGD / R model.

[0079] As Figure 7(A) The cell morphology of the OGD / R group cells changed significantly under white light. After treatment with EL-Gel, most cells returned to normal morphology. (B) Intervention with component A alone, component B alone, the combination of component A and B, and chitosan hydrogel alone in R28 cells did not achieve better protective effects compared with the OGD / R group.

Claims

1. A functional hydrogel, characterized in that, It comprises a hydrogel matrix and a pharmaceutically effective amount of Component A and Component B dispersed therein; The said Component A is a compound of Formula 1 and its pharmaceutically acceptable salts; Formula 1 The said Component B is exosomes derived from bone marrow mesenchymal stem cells; In the said functional hydrogel, the concentration of the said Component A is 15 - 50 uM; The concentration of Component B is 150 - 300 μg / mL.

2. The functional hydrogel according to claim 1, wherein The diameter of the exosomes ≤ 150 nm.

3. The functional hydrogel according to claim 1, wherein The said hydrogel matrix is a chitosan-based hydrogel.

4. A method for preparing the functional hydrogel according to any one of claims 1 to 3, characterized in that, Mix and co-incubate the said Component A, Component B and the hydrogel matrix to obtain it.

5. The preparation method of the functional hydrogel according to claim 4, characterized in that, The steps for obtaining exosomes are: culturing bone marrow mesenchymal stem cells and harvesting the cell supernatant; filtering, separating and resuspending the precipitate of the said cell supernatant to obtain exosomes.

6. Use of the functional hydrogel according to any one of claims 1 - 3 or the functional hydrogel prepared by the preparation method according to any one of claims 4 - 5 in the preparation of a medicament for treating glaucoma.

7. A drug for treating glaucoma, characterized in that, It contains the functional hydrogel according to any one of claims 1 - 3 or the functional hydrogel prepared by the preparation method according to any one of claims 4 - 5.

8. The medicament for treating glaucoma according to claim 7, characterized in that, It further contains pharmaceutically acceptable excipients.

9. The medicament for treating glaucoma according to claim 7 or 8, characterized in that, It is an external preparation or an injection preparation for local ocular administration.

10. The drug for treating glaucoma according to claim 9, wherein, It is an intravitreal injection preparation.

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