A targeted hydrogel with chemotherapeutic and anti-tumor immune effects and preparation method and use thereof

Abstract

The present application provides a kind of targeted hydrogel with chemotherapy and anti-tumor immune effect, it contains the following weight ratio of raw materials: pluronics 50-76 parts, hyaluronic acid 151-226 parts, Cu (NO3) 2320-480 parts, doxorubicin hydrochloride 1.6-2.4 parts.The present application also provides the preparation method and use of targeted hydrogel.The present application is embedded in the reticular framework structure formed by pluronics and hyaluronic acid and doxorubicin hydrochloride and copper sulfate, self-assembly forms hydrogel.In the hydrogel system, the reticular framework structure composed of pluronics and hyaluronic acid can resist strong acid environment and alkaline environment, and slowly target release drug under the weak acid microenvironment of tumor degradation;After drug release, copper ion can cause tumor cell copper death procedure, cause cell apoptosis, and synergistic doxorubicin hydrochloride activates innate immunity in vivo, realize longer anti-tumor response, enhance the therapeutic effect of CRC mouse model, improve the bioavailability of DOX, and play the role of chemotherapy and anti-tumor immunity.

Description

Technical Field

[0001] This invention relates to a targeted hydrogel with chemotherapeutic and antitumor immunomodulatory effects, its preparation method, and its uses. Background Technology

[0002] Colorectal cancer (CRC) is one of the leading causes of cancer-related death worldwide, and traditional treatments—including surgery, chemotherapy, and immunotherapy—are often limited by systemic toxicity, drug resistance, and the immunosuppressive tumor microenvironment (TME). Oral drug delivery systems, designed to maximize local drug concentrations while minimizing off-target effects, have emerged as promising alternatives for CRC treatment. However, their clinical translation faces obstacles such as poor bioavailability, insufficient tumor specificity, and the inability to dynamically modulate the TME. Recent advances in cancer biology have revealed a copper-dependent cell death mechanism—cuproptosis—a cell death mechanism driven by mitochondrial protein toxicity, which may represent a potential therapeutic pathway. Unlike ferroptosis, cuproptosis utilizes copper ions to disrupt cellular homeostasis through the accumulation of lipid-acylated mitochondrial enzymes and subsequent proteotoxic stress. However, utilizing this pathway to treat CRC requires precise spatiotemporal copper ion delivery and synergistic integration with existing therapies to overcome tumor heterogeneity and immunosuppression.

[0003] The physiological complexities of the gastrointestinal tract—including enzymatic degradation, pH variations, and mucosal barriers—pose unique challenges to oral nanomedicine. While intravenously administered nanocarriers have dominated oncology research, reliance on enhanced permeability and retention (EPR) effects is particularly unreliable in colorectal cancer due to the dense matrix and heterogeneous vascularization of the tumor. Systemic administration of copper ions, while inducing copper death, typically fails to achieve therapeutic intratumoral concentrations without inducing off-target toxicity. Recent efforts in chemokinetic therapy (CDT) using endogenous metal ions such as iron or copper have shown promise, but these approaches primarily rely on intratumoral injection or off-target systemic delivery, limiting their clinical feasibility in colorectal cancer. In contrast, oral nanoplatforms that overcome the gastrointestinal challenges and utilize colorectal cancer-specific biomarkers such as CD44 may revolutionize local therapy by combining tumor-selective accumulation with minimized systemic exposure. This strategy aligns with the growing emphasis on immunogenic cell death (ICD) in colorectal cancer, where dying tumor cells stimulate dendritic cell maturation and cytotoxic T cell infiltration, thereby counteracting immunosuppressive signals from the tumor mechanotherapy (TME). However, a few systems are able to integrate dual pro-death and immunostimulatory functions in a single oral delivery vehicle, highlighting the urgent need for multifunctional designs that address the multifaceted pathophysiology of colorectal cancer.

[0004] Application No. 202210330522.2, Invention Title: Application of Copper Ion-Tanonic Acid Co-assembled Antibacterial Nanosheets as Antitumor Drug Carriers, discloses the application of copper ion-tannic acid co-assembled antibacterial nanosheets as carriers for antitumor drugs. The method involves ultrasonically dispersing antibacterial nanosheets in deionized water, adding antitumor drugs, and thoroughly stirring under dark conditions to load the antitumor drugs onto the antibacterial nanosheets. After centrifugation and drying, antibacterial and antitumor nanomedicines are obtained. This application mainly relies on intratumoral injection or non-targeted systemic delivery, with the main mechanism focusing on dual cell death promotion, namely copper death and chemotherapy. Ma Jia et al., Preparation of Doxorubicin Liposomes by Copper Acetate Gradient Method, Chinese Journal of Pharmaceutics (Online Edition), November 2022, Vol. 20, No. 6, discloses the preparation of doxorubicin liposomes using a copper acetate gradient and explores its drug loading mechanism. Currently reported literature has not used this method for the treatment of colorectal cancer. Summary of the Invention

[0005] This invention provides a targeted hydrogel with chemotherapy and anti-tumor immunomodulatory effects, and also provides a method for preparing the targeted hydrogel and its uses.

[0006] This invention provides a targeted hydrogel with chemotherapeutic and antitumor immunomodulatory effects, comprising the following raw materials in the indicated weight ratios:

[0007] 50-76 parts of Prunic, 151-226 parts of hyaluronic acid, 320-480 parts of Cu(NO3)2, and 1.6-2.4 parts of doxorubicin hydrochloride.

[0008] Preferably, it contains the following raw materials in the indicated weight ratios:

[0009] 63 parts of Pronicotinic Acid, 189 parts of Hyaluronic Acid, 2400 parts of Cu(NO3), and 2 parts of Doxorubicin Hydrochloride.

[0010] The targeted hydrogel of this invention is formed by uniformly embedding Cu(NO3)2 and doxorubicin hydrochloride in a network framework structure assembled from Plunkic acid and hyaluronic acid, and then self-assembling to form a hydrogel.

[0011] This invention also provides a method for preparing the targeted hydrogel with chemotherapy and anti-tumor immunomodulatory effects, which includes the following steps:

[0012] a. Weigh the raw materials according to their respective weight ratios;

[0013] b. Dissolve Prönkel in deionized water to form Prönkel solution, and then dissolve hyaluronic acid in Prönkel solution to make solution A;

[0014] c. Prepare a Cu(NO3)2 solution by dissolving Cu(NO3)2 in deionized water;

[0015] d. Add Cu(NO3)2 solution to solution A to obtain gel carrier solution; sonicate the gel carrier solution and mix evenly; add doxorubicin hydrochloride (DOX) powder to the gel carrier solution and mix thoroughly to form solution B;

[0016] e. Place solution B in a metal bath at 40°C for 3 hours, and then solidify it overnight in a refrigerator at 4°C. The solidified product is the hydrogel GEL@DOX.

[0017] The present invention also provides the use of the targeted hydrogel in the preparation of drugs with chemotherapeutic and antitumor immunomodulatory effects.

[0018] The drug mentioned is a drug for treating colorectal cancer.

[0019] The present invention provides a pharmaceutical composition for treating colon cancer, comprising the aforementioned targeted hydrogel.

[0020] The pharmaceutical composition of the present invention is prepared into a commonly used oral formulation by adding pharmaceutically acceptable excipients or auxiliary ingredients, with the targeted hydrogel as the main component.

[0021] This invention relates to a targeted hydrogel, GEL@DOX, composed of Pluronic acid, hyaluronic acid, copper sulfate, and doxorubicin hydrochloride (DOX). Copper sulfate and doxorubicin hydrochloride are uniformly embedded within a network framework structure assembled from Pluronic acid and hyaluronic acid, self-assembling to form a hydrogel. In this hydrogel system, the network framework structure composed of Pluronic acid and hyaluronic acid resists both strongly acidic and alkaline environments, and degrades in the weakly acidic tumor microenvironment, resulting in slow, targeted drug release. After drug release, copper ions induce copper death in tumor cells, leading to apoptosis, and synergistically activate innate immunity with doxorubicin hydrochloride, achieving a longer-lasting anti-tumor response and enhancing the therapeutic effect in a CRC mouse model. This novel hydrogel system improves the bioavailability of DOX and exerts chemotherapeutic and anti-tumor immunomodulatory effects. Attached Figure Description

[0022] Figure 1 Schematic diagram of the structure of GEL@DOX oral nanogel;

[0023] Figure 2 Schematic diagram of GEL@DOX oral nanogel preparation;

[0024] Figure 3 Schematic diagram of the gel texture formed by different ratios of GEL@DOX oral nanogel raw materials;

[0025] Figure 4 GEL@DOX oral nanogel UV absorption spectrum and zata potential diagram;

[0026] Figure 5GEL@DOX oral nanogel scanning electron microscopy, elemental analysis and stress-strain mapping;

[0027] Figure 6 GEL@DOX oral nanogel release diagrams in vitro under different buffer media;

[0028] Figure 7 Schematic diagram of GEL@DOX oral nanogel in vitro simulated sustained-release drug delivery;

[0029] Figure 8 IC50 assay and cytotoxicity results of different cell lines using GEL@DOX oral nanogels;

[0030] Figure 9 GEL@DOX oral nanogel subcutaneous residence and organ metabolism in nude mice;

[0031] Figure 10 Results of tumor suppression in a subcutaneous colorectal cancer model using GEL@DOX oral nanogels;

[0032] Figure 11 Safety evaluation of GEL@DOX oral nanogel subcutaneous colorectal cancer model;

[0033] Figure 12 Results of tumor suppression in a GEL@DOX oral nanocollagen model of colorectal cancer. Detailed Implementation

[0034] Example 1: Preparation method of the targeted hydrogel system (GEL@DOX) of the present invention

[0035] 1. Synthesis of GEL@DOX:

[0036] Step 1: First, dissolve 140 mg of Prönache in 10 mL of deionized water to form a 140 mg / mL Prönache solution. Then, dissolve 420 mg of hyaluronic acid in the Prönache solution to prepare solution A. Dissolve 800 mg of Cu(NO3)2 in 1 mL of deionized water to prepare an 800 mg / mL Cu(NO3)2 solution. Finally, add 500 μL of Cu(NO3)2 solution to 4.5 mL of solution A to obtain the gel carrier solution. Sonicate the gel carrier solution for 30 minutes to ensure uniform mixing. Then, add 2 mg of DOX powder to the gel carrier solution and mix thoroughly to form solution B. Place solution B in a 40°C metal bath for 3 hours, then solidify overnight in a 4°C refrigerator. The solidified product is labeled GEL@DOX. See Figure 1-2 .

[0037] Solution A was prepared with a near-distribution ratio of 1:1, 1:2, 1:3, and 1:4 for Pranic acid and hyaluronic acid. It was found that at a 1:1 ratio, no gel formed; at a 1:2 ratio, gelation was incomplete; and at a 1:4 ratio, the gel solidified severely, becoming excessively hard and unable to pass through a syringe, significantly impacting subsequent experiments. Therefore, a 1:3 ratio (140 mg Pranic acid and 420 mg hyaluronic acid per 10 mL of deionized water) was adopted as the optimal ratio for further research. Figure 3 .

[0038] 2. Characterization of GEL@DOX:

[0039] The zeta potential, morphology, and elemental composition of the GEL@DOX hydrogel system were analyzed using a nanoparticle tracker (NTA, PTM ZetaView) and a scanning electron microscope (Hitachi HT7800, SEM, Sichuan University). The UV-Vis absorption spectra of both GEL@DOX and DOX were measured using a UV-Vis spectrophotometer (UV-3600, Shimadzu, Japan).

[0040] like Figure 4 As stated in -A, compared with DOX (480nm), the UV-Vis absorption spectrum of GEL@DOX shifts to 520nm, further indicating the successful synthesis of GEL@DOX.

[0041] In addition, the zeta potentials of GEL and GEL@DOX were evaluated. Figure 4 -B), and according to scanning electron microscopy, the system exhibits a stable and uniform mesh structure. Figure 5 -A / B) indicates that the hydrogel system has successfully self-assembled, and the elements within the system are analyzed and located, such as Figure 5 As shown in Figures -C to 5-E, the system exhibits a uniform distribution, indicating good uniformity in its slow-release process. Further tests were conducted on the system's physical properties, including its pressure-strain curves, as shown below. Figure 5 -F.

[0042] The following efficacy tests demonstrate the beneficial effects of this invention.

[0043] Experimental Example 1: In vitro release of DOX from GEL@DOX of the present invention:

[0044] 1 ml of GEL@DOX (2 mg / ml based on DOX addition) was added to a pre-expanded dialysis bag with a molecular weight cutoff (MWCO) of 3000 Da. The dialysis bag was then immersed in four different solutions (9 ml): GSH-free phosphate-buffered saline (PBS) (pH 5.0 and 7.4) and GSH-containing phosphate-buffered saline (PBS) (pH 5.0 and 7.4). The apparatus was placed in a culture medium at 37°C and shaken at 200 rpm. 1 ml of the test solution was extracted daily from the extradialysis bag solution for 6 days. The sample contained DOX diffused from the dialysis bag. Immediately afterward, an equal volume (1 ml) of the corresponding fresh buffer solution was added to the dialysis buffer to maintain the system. The cumulative release percentage (Er) was calculated as follows. This method is from the National Center for Nanoscience and Technology.

[0045] E

[0046] E r Cumulative release percentage V e : Buffer replacement volume

[0047] V0: Total buffer volume (m) drug Total mass of gel

[0048] Ci:T Concentration of the released liquid during the i-th displacement sampling

[0049] n: Number of times solution is taken

[0050] like Figure 6 As shown, after incubation at 37°C for 144 hours, the release behavior of DOX in the GEL@DOX hydrogel system was measured. Better release results were observed at pH 5.0, indicating that the GEL@DOX hydrogel system has the ability to deliver pH-responsive drugs. Furthermore, calculations show that GSH-containing phosphate-buffered saline (PBS) at pH 5.0 can theoretically simulate the tumor microenvironment. Tumor cells, due to their active metabolism, often lead to a decrease in local pH (acidic environment), and the tumor microenvironment typically has a high GSH level to protect tumor cells from oxidative stress. Under this environment, the final release rate achievable by the GEL@DOX hydrogel system is approximately 75%. Therefore, 75% was used as a coefficient, multiplied by the amount of DOX added to the gel system, to obtain the actual drug loading for subsequent drug dosing in experiments. That is, when the DOX added to the GEL@DOX hydrogel system is 2 mg / mL, the concentration of GEL@DOX is recorded as 1.5 mg / mL.

[0051] Experimental Example 2: In vitro cytotoxicity test

[0052] The sustained-release toxicity of the GEL@DOX hydrogel system was determined using the MTT assay. Simply put, 5000 cells were added to each well of a 96-well plate and cultured overnight. Cells were treated with different concentrations of DOX for 24 hours. Then, 10 μL (5 mg / mL) of MTT solution was added and incubated for 4 hours. After removing the culture medium, 150 μL of dimethyl sulfoxide (DMSO) was added, and the absorbance at 570 nm was measured to calculate the IC50 for each cell line. 50， Subsequently according to 1 / 3 IC 50 Concentration administration, IC 50 The results are as follows Figure 8 As shown in -A.

[0053] Add 2000 cells to each well of a 96-well plate and incubate overnight. Treat the cells with DOX and GEL@DOX as follows. Figure 7 As shown. Subsequent procedures are the same as above. Cell viability was measured on day 5, and the viability results for different cell types are shown below. Figure 8 -B to 8-D.

[0054] Experimental Example 3: In vivo / ex vivo drug retention fluorescence imaging analysis

[0055] BALB / c nude mice (≈25g) were purchased from Chengdu Yaokang Biotechnology Co., Ltd. All in vivo studies were conducted in accordance with the "Guidelines for Animal Experimentation" and were approved by the Ethics Committee of Sichuan University School of Medicine. A subcutaneous colorectal cancer model was established to study the in vivo retention capacity of the gel. This study used RKO cells to establish a subcutaneous colorectal cancer model in BALB / c nude mice and used bioluminescence imaging technology to compare the drug's targeting and retention time. When the subcutaneous tumor reached 100mm... 3 Tumor-bearing mice were subcutaneously injected with DOX and the GEL@DOX hydrogel system (n=3 per group; DOX dose: 1 mg / kg body weight; GEL@DOX dose: 0.75 mg / kg body weight). The mice were then monitored and photographed for 5 days using a small animal in vivo imaging system to obtain the drug distribution in the body. Organs were removed 5 days after injection for stereofluorescence imaging. Figure 9 As shown in Figure -A, fluorescence imaging revealed that GEL@DOX had a long residence time in the tumor region, exhibiting better tumor accumulation capacity and a good sustained-release effect. Five days after injection, organs and tumor tissues were obtained for off-topic imaging, such as... Figure 9 As shown in -C, the prolonged accumulation time of GEL@DOX within tumor tissue further indicates enhanced drug aggregation, suggesting that the drug's anti-tumor effect is sustained.

[0056] Figure 9 -A Real-time in vivo imaging of subcutaneous injection of DOX and GEL@DOX in an RKO tumor-bearing BALB / c nude mouse model. Figure 9 B and Figure 9Analysis of the radiation intensity of major organs and tumors 5 days after administration of C.

[0057] Experimental Example 4: Antitumor Analysis of Subcutaneous Tumor Model

[0058] A subcutaneous colorectal tumor model was established in Balb / c mice by subcutaneous injection of CT26 cells using the above method. When the tumor volume reached 80 mm... 3 Tumor-bearing mice were randomly divided into four groups. On day one, each group received a subcutaneous injection of 50 μL of saline, DOX, GEL (containing hyaluronic acid, pranoxetine, and copper nitrate), or GEL@DOX. The DOX group received 1 mg / kg, and the GEL@DOX group received 0.75 mg / kg. The GEL group received the same dose as the GEL@DOX group (n=5). The DOX group received a repeat dose on day 7 (DOX group received two doses, GEL@DOX group received one dose). Tumor volume and body weight were recorded daily. Mice were euthanized on day 14, and the results were analyzed.

[0059] Figure 10 The results showed that the treatment effect of the GEL@DOX group was significantly better than that of other treatment groups. Figure 10 -A showed that after treatment, the tumor volume in the GEL@DOX group was significantly smaller than that in the other groups. Calculations of tumor weight and inhibition rate revealed (see...) Figure 10 -B), the GEL@DOX group showed reduced tumor weight and a tumor inhibition rate of 81.35±2.29%. Notably, the GEL@DOX group demonstrated superior treatment efficacy with lower actual DOX dosage and fewer treatment sessions. Furthermore, no significant changes in body weight were observed between the different treatment groups. Figure 10 -C), and after collecting blood supernatant for biochemical analysis, there were no significant differences in ALT, AST, UR, and CR among the different treatment groups in mice. Figure 11 The system is said to have good biocompatibility. Figure 11 ).

[0060] Experimental Example 5: Antitumor Analysis of the Orthotopic Tumor Model

[0061] An orthotopic colorectal tumor model was established by injecting CT26 cells into specific locations in the intestinal wall (e.g., colon) of Balb / c mice. On day 7 post-surgery, tumors were fluorescently located, and mice with similar tumor fluorescence intensity and area were selected for subsequent experiments. The tumor-bearing mice were divided into four groups, each receiving 50 μL of saline, DOX, GEL (containing hyaluronic acid, phenylethylamine, and copper nitrate), or GEL@DOX via gavage on day 1. The DOX group received 1 mg / kg, and the GEL@DOX group received 0.75 mg / kg. The GEL group received the same dosage as the GEL@DOX group (n=5) to simulate targeted therapy for colorectal cancer using oral medication. The DOX group received a repeat dose on day 7, meaning the DOX group received two doses, and the GEL@DOX group received one dose. Mice were euthanized on day 10 after administration, and the results were analyzed.

[0062] Figure 12 The results showed that the treatment effect of GEL@DOX in the orthotopic colorectal cancer group was significantly better than that in other treatment groups. Figure 12 -A used bioluminescence to detect tumor size. The fluorescence area and intensity of the GEL@DOX group were significantly smaller than those of the DOX group. After euthanasia of mice, autopsy revealed tumors in the colorectal region. The results were consistent with the bioluminescence detection results. Figure 12 -B). The experimental results also suggest that the treatment effect of the GEL@DOX group may be best around day 7, which corresponds to the result in Experiment 1 where the release of GEL@DOX reached its peak after day 5 after simulating the tumor microenvironment, further verifying the sustained-release efficacy of the hydrogel system.

[0063] The above experiments demonstrate that the oral nanogel platform of this invention, targeting CD44, synergistically exerts effects of copper apoptosis, chemotherapy, and innate immune activation. The nanogel possesses tunable physicochemical properties and stimulus-responsive drug release, making it an ideal matrix for combination therapy. Hyaluronic acid (HA), as a natural ligand for the overexpressed CD44 receptor in colorectal cancer, plays a dual role: it both promotes tumor targeting and binds to copper ions (Cu). 2+ The copolymers of Cu and Pluronic form a structurally stable nanocolloid. Pluronic further enhances the colloidal stability and mucosal permeability, while Cu... 2+ Not only as a therapeutic agent, but also as a dynamic cross-linking agent, it ensures pH-responsive drug release in an acidic TME. A multifunctional oral nanogel (HA / Pluronic / Cu) is proposed. 2+ HA-mediated CD44 targeting ensures selective accumulation in colorectal cancer cells, bypassing the gastrointestinal and stromal barriers. In the acidic TME, Cu... 2+Released to trigger a Fenton-like reaction, generating hydroxyl radicals (·OH), which amplify copper death through mitochondrial protein toxicity. This copper-driven mitochondrial disruption not only directly kills tumor cells but also releases mitochondrial DNA (mtDNA) into the cytoplasm, activating the cGAS-STING pathway to recruit anti-tumor immune cells. Simultaneously, DOX enhances cytotoxicity and immunogenic cell death (ICD), promoting a pro-inflammatory TME characterized by enhanced dendritic cell activation and T cell infiltration. By integrating chemotherapy, copper death, and STING-dependent immune guidance into an oral delivery platform, HPC nanogels bypass multidrug resistance and reprogram the immunosuppressive microenvironment.

[0064] This invention pioneers an orally deliverable nanoplatform that combines metal ion-mediated cell death pathways with immune modulation, bringing a paradigm shift to the treatment of colorectal cancer. Rigorous in vitro and in vivo validation demonstrated that HPC nanogels overcome gastrointestinal degradation, enhance tumor-specific biodistribution, and stimulate potent anti-tumor immunity. By addressing the physiological, chemical, and immunological barriers of colorectal cancer in a single system, this design establishes a transformative strategy for precision oncology, bridging the critical gap between local oral therapy and systemic anti-tumor immunity.

[0065] This invention provides an oral nanogel platform targeting CD44. The oral drug delivery system aims to maximize local drug concentration while minimizing off-target effects. This design addresses issues such as systemic toxicity, drug resistance, and the immunosuppressive tumor microenvironment (TME) in the clinical treatment of colorectal cancer. This application aims to design a mechanism that combines dual cell death promotion and immune stimulation to address the multifaceted pathophysiology of colorectal cancer. This strategy aligns with the increasing emphasis on immunogenic cell death (ICD) in colorectal cancer.

[0066] In summary, this invention successfully designed a pH-sensitive hydrogel system with sustained-release properties, enhancing the bioavailability of DOX, triggering ICD, and activating a systemic immune response. In vivo experimental results show that GEL@DOX treatment effectively inhibits the growth of colorectal cancer cells and exhibits excellent sustained-release activity. Further research indicates that GEL@DOX effectively activates the immune system in vivo, reshaping the immunosuppressive microenvironment within tumors. The current design has three advantages. GEL@DOX solves the problems of poor DOX bioavailability and the need for long-term use. Due to the excellent sustained-release effect of the hydrogel system, the dosage of DOX required for the same inhibitory effect on tumors is much higher than that of GEL@DOX. Furthermore, GEL@DOX can lyse in an acidic tumor environment, releasing lethal ROS, effectively leading to mitochondrial dysfunction and intrinsic apoptosis in tumor cells.

Claims

1. A targeted hydrogel with chemotherapeutic and antitumor immunomodulatory effects, characterized in that: It contains the following ingredients in the following weight ratio: 50-76 parts of Prunic, 151-226 parts of hyaluronic acid, 320-480 parts of Cu(NO3)2, and 1.6-2.4 parts of doxorubicin hydrochloride.

2. The targeted hydrogel with chemotherapy and anti-tumor immunomodulatory effects according to claim 1, characterized in that: It contains the following ingredients in the following weight ratio: 63 parts of Pronicotinic Acid, 189 parts of Hyaluronic Acid, 2400 parts of Cu(NO3), and 2 parts of Doxorubicin Hydrochloride.

3. The targeted hydrogel with chemotherapy and anti-tumor immunomodulatory effects according to claim 1 or 2, characterized in that: It is formed by uniformly embedding Cu(NO3)2 and doxorubicin hydrochloride in a network framework structure assembled from Plunkic acid and hyaluronic acid, and then self-assembling to form a hydrogel.

4. The method for preparing the targeted hydrogel with chemotherapy and anti-tumor immunomodulatory effects according to any one of claims 1-3, characterized in that: It includes the following steps: a. Weigh the raw materials according to their respective weight ratios; b. Dissolve Prönkel in deionized water to form Prönkel solution, and then dissolve hyaluronic acid in Prönkel solution to make solution A; c. Prepare a Cu(NO3)2 solution by dissolving Cu(NO3)2 in deionized water; d. Add Cu(NO3)2 solution to solution A to obtain gel carrier solution; sonicate the gel carrier solution and mix evenly; add doxorubicin hydrochloride (DOX) powder to the gel carrier solution and mix thoroughly to form solution B; e. Place solution B in a metal bath at 40°C for 3 hours, and then solidify it overnight in a refrigerator at 4°C. The solidified product is the hydrogel GEL@DOX.

5. Use of the targeted hydrogel according to any one of claims 1-3 in the preparation of a medicament with chemotherapeutic and antitumor immunomodulatory effects.

6. The use according to claim 5, characterized in that: The drug mentioned is a medication for treating colorectal cancer.

7. A pharmaceutical composition for treating colon cancer, characterized in that: It contains the targeted hydrogel as described in any one of claims 1-3.

8. The pharmaceutical composition for treating colon cancer according to claim 7, characterized in that: It is a commonly used oral pharmaceutical preparation made by adding pharmaceutically acceptable excipients or auxiliary ingredients to the main component of targeted hydrogel.

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