Drug-loaded nanocomposite hydrogel and preparation method and application thereof

Abstract

The present application relates to a kind of drug-loaded nanocomposite hydrogel and its preparation method and application.A kind of drug-loaded nanocomposite hydrogel, the drug-loaded nanocomposite hydrogel is the drug-loaded hollow manganese dioxide nanoparticle modified by inulin hydrogel on surface;The drug is selected from: the drug with intestinal tract as target point;The hollow manganese dioxide nanoparticle is loaded at least one described drug.The drug-loaded nanocomposite hydrogel of the present application can realize colon site targeted drug delivery, increase the retention time and accumulation amount of drug in colon, also can regulate intestinal flora, stimulate the immune function of organism, and then improve the prevention and treatment effect of drug.Moreover, since the hollow manganese dioxide nanoparticle in the present application has super large hollow volume, can load double drug or multiple drugs, reaches the effect of comprehensive treatment of intestinal disease.

Description

Technical Field

[0001] This invention relates to the pharmaceutical field, and in particular to a drug-loaded nanocomposite hydrogel, its preparation method, and its application. Background Technology

[0002] In recent years, a growing body of research has demonstrated a causal relationship between gut microbiota dysbiosis and the pathogenesis of colorectal cancer. Several bacteria that accumulate in the gut and induce tumor proliferation, activate IL-6 / STAT3 signaling, promote inflammation, cause DNA damage, and protect tumors from immune attack have been identified as contributing factors to colorectal cancer. These bacteria include *Fusobacterium nucleatum*, *Alistipes*, *Peptostreptococcus anaerobius*, *Helicobacter pylori*, and the enterotoxin-producing *Bacteroides fragilis*. On the other hand, some probiotics, including *Lachnospiraceae*, *Bifidobacterium*, *Lactobacillus*, *Roseburia*, and *Streptococcus thermophilus*, have been found to be significantly reduced in colorectal cancer patients. These probiotics are believed to have a protective effect on colorectal cancer patients because their metabolites, short-chain fatty acids (acetic acid, propionic acid, butyric acid, etc.), play an important role in reducing intestinal inflammation, protecting the integrity of the intestinal epithelial barrier, and stimulating the body's immune function.

[0003] Current research on nanomedicines for colorectal cancer treatment primarily focuses on how to precisely deliver drugs to the intestines to exert their effects and how to maximize the inhibition of colorectal cancer tumor cell proliferation. However, for solid tumors like colorectal cancer that are easily influenced by gut microbiota, effective inhibition is difficult to achieve. Therefore, there is an urgent need to develop a drug that can prevent and treat colorectal cancer by regulating gut microbiota. Summary of the Invention

[0004] Therefore, it is necessary to provide a drug-loaded nanocomposite hydrogel to address the problem that current nanomedicines are unable to effectively inhibit solid tumors such as colorectal cancer that are susceptible to the influence of gut microbiota.

[0005] A drug-loaded nanocomposite hydrogel, wherein the drug-loaded nanocomposite hydrogel is a drug-loaded hollow manganese dioxide nanoparticle with a surface modified by inulin hydrogel; the drug is selected from drugs whose target is the intestine; and the hollow manganese dioxide nanoparticles are loaded with at least one of the drugs.

[0006] The aforementioned drug-loaded nanocomposite hydrogel consists of internal drug-loaded hollow manganese dioxide nanoparticles and an externally modified inulin hydrogel "shell." Through the synergistic effect of the inulin hydrogel, hollow manganese dioxide nanoparticles (H-MnO2), and drugs, it can achieve targeted drug delivery to the colon, increase drug retention time and accumulation in the colon, regulate intestinal flora, and stimulate the body's immune function, thereby enhancing the preventive and therapeutic effects of the drugs. Moreover, because the hollow manganese dioxide nanoparticles in this invention have an ultra-large hollow volume, they can carry two or more drugs to achieve a comprehensive therapeutic effect on intestinal diseases.

[0007] In one embodiment, the drug is selected from: anti-intestinal tumor drugs and / or anti-enteritis drugs.

[0008] In one embodiment, the inulin concentration in the inulin hydrogel is 40-45%.

[0009] In one embodiment, the drug-loaded nanocomposite hydrogel is oxaliplatin-loaded hollow manganese dioxide nanoparticles with a surface modified by inulin hydrogel.

[0010] The aforementioned drug-loaded nanocomposite hydrogel consists of an inner oxaliplatin-loaded hollow manganese dioxide nanoparticle (Oxa / H-MnO2) and an outer modified inulin hydrogel "shell". Through the synergistic effect of inulin hydrogel, hollow manganese dioxide nanoparticle (H-MnO2) and oxaliplatin, it can achieve targeted drug delivery to the colon, increase the drug concentration and retention time of oxaliplatin in the colon, regulate the intestinal flora and the flora in intestinal tumors, stimulate the body's immune function, and thus improve the therapeutic effect of oxaliplatin.

[0011] The inulin hydrogel "shell" protects the internal nanomedicine—oxaliplatin-loaded hollow manganese dioxide nanoparticles—from the acidic environment of the stomach and upper digestive tract during its journey to the colon, ensuring the drug concentration of the oxaliplatin-loaded hollow manganese dioxide nanoparticles in the colon. After the antitumor nanocomposite hydrogel (Oxa / H-MnO2-IN) reaches the colon, the inulin hydrogel "shell" is fermented and degraded by probiotics in the colon, exposing the internal oxaliplatin-loaded hollow manganese dioxide nanoparticles. After being absorbed by the colon, the oxaliplatin-loaded hollow manganese dioxide nanoparticles aggregate in the colorectal cancer tumor microenvironment through the EPR (high permeability and retention effect of solid tumors) effect, exerting their antitumor efficacy.

[0012] Inulin hydrogel and oxaliplatin have a synergistic effect on immunomodulation. The high-concentration inulin hydrogel "shell" in this invention essentially meets the daily dietary fiber standard (inulin is a type of dietary fiber), reaching a dosage capable of regulating probiotics and harmful bacteria. Therefore, after being degraded by probiotics in the colon, it can upregulate the number of probiotics in the intestine and intestinal tumors that can metabolize and produce short-chain fatty acids, and downregulate the number of harmful bacteria in the intestine and intestinal tumors that induce colorectal cancer. By upregulating the number of probiotics, it promotes an increase in the content of butyrate (a short-chain fatty acid), a metabolite of probiotics in the intestine. Butyrate can promote CD8+ metabolism through the HDAC / ID2 / IL-12 receptor pathway. + T cell infiltration into tumor tissue and promotion of CD8 + T cells secrete IFN-γ in tumor tissue, which greatly enhances the body's anti-tumor CD8+. + T-cell immune response. Oxaliplatin has high immunogenicity, and inulin hydrogel synergizes with oxaliplatin in terms of immune function, thus improving the therapeutic effect of oxaliplatin in colorectal cancer.

[0013] Meanwhile, the inulin hydrogel "shell" can prolong gastric emptying time through its own thickening effect. It can also adsorb the anti-tumor nanocomposite hydrogel onto the intestinal mucosa layer through its own thickening effect, thereby increasing the retention time of oxaliplatin-loaded hollow manganese dioxide nanoparticles in the colon. This, in turn, increases the accumulation of oxaliplatin-loaded hollow manganese dioxide nanoparticles in the colon, enabling sustained release and action against colorectal cancer tumors. This significantly improves the drug efficiency and efficacy of oxaliplatin-loaded hollow manganese dioxide nanoparticles in the colon. Synergistic antitumor effect of hollow manganese dioxide nanoparticles and oxaliplatin. Hollow manganese dioxide nanoparticles can catalyze the decomposition of excess H2O2 in the tumor microenvironment through the Fenton reaction to generate active O2, alleviating tumor hypoxia and achieving a certain antitumor effect. This, in turn, synergizes with the chemotherapy drug oxaliplatin, improving the therapeutic effect of oxaliplatin in the treatment of colorectal cancer.

[0014] In one embodiment, the drug-loaded nanocomposite hydrogel has a particle size of 210-260 nm, the oxaliplatin-loaded hollow manganese dioxide nanoparticles have a particle size of 170-210 nm, and the hollow manganese dioxide nanoparticles have a particle size of 150-190 nm.

[0015] This invention also provides a method for preparing the drug-loaded nanocomposite hydrogel, comprising the following steps: (1) Preparation of silica nanospheres: Silica nanospheres were prepared by microemulsion method; (2) Preparation of hollow manganese dioxide nanoparticles: After reducing the silica nanospheres with potassium permanganate, chemical etching is performed to obtain the hollow manganese dioxide nanoparticles. (3) Preparation of drug-loaded nanocomposite hydrogel: The drug is loaded onto the surface of the hollow manganese dioxide nanoparticles and then modified with inulin to obtain the drug.

[0016] In one embodiment, in step (3), the drug-loaded nanocomposite hydrogel is prepared by the following method: the drug is loaded onto the surface of the hollow manganese dioxide nanoparticles, and then inulin is added to the drug-loaded hollow manganese dioxide nanoparticle solution to make the concentration of inulin reach 35~45%; after stirring at 60~70°C, it is allowed to stand at 3~5°C to obtain the product.

[0017] In one embodiment, the method for preparing the drug-loaded nanocomposite hydrogel includes the following steps: (1) Preparation of silica nanospheres: ① Mix Triton X-100, cyclohexane, n-hexanol, ammonia, and water to obtain solution a; ② Add tetraethyl orthosilicate and 3-aminopropyltriethoxysilane to solution a obtained in step ① to obtain the silica nanospheres; (2) Preparation of hollow manganese dioxide nanoparticles: ① The silica nanospheres obtained in step (1) are mixed with potassium permanganate solution to obtain H-MnO2@sSiO2; ② The H-MnO2@sSiO2 obtained in step ① is mixed with sodium carbonate solution to obtain the hollow manganese dioxide nanoparticles; (3) Preparation of drug-loaded nanocomposite hydrogels: ① The hollow manganese dioxide nanoparticles obtained in step (2) are mixed with the drug solution to obtain the drug-loaded hollow manganese dioxide nanoparticles; ② Dissolve the drug-loaded hollow manganese dioxide nanoparticles obtained in step ① and add inulin to make the inulin concentration reach 35~45%. Stir at 60~70℃ and let stand at 3~5℃ to obtain the final product.

[0018] In one embodiment, the method for preparing the drug-loaded nanocomposite hydrogel includes the following steps: (1) Preparation of silica nanospheres: ① Mix 10-11 mL of Triton X-100 (99-100% concentration), 40-50 mL of cyclohexane (99-100% concentration), and 10-11 mL of n-hexanol (98-100% concentration) and stir for 4-6 min; then add 1.3-1.7 mL of ammonia (25-28% concentration) and 1.8-2.2 mL of water, and stir for 25-35 min to obtain solution a; ② Add 0.8~1.2 mL of tetraethyl orthosilicate with a concentration of 99.99~100% and 0.1~0.3 mL of 3-aminopropyltriethoxysilane with a concentration of 98~100% to solution a obtained in step ①, stir for 22~26 h, centrifuge for 15~25 min, wash, and dry to obtain the silica nanospheres; (2) Preparation of hollow manganese dioxide nanoparticles: ① Dissolve 90-110 mg of the silica nanospheres obtained in step (1) in 90-110 mL of water; dissolve 675-825 mg of the potassium permanganate in 67.5-82.5 mL of water; add the potassium permanganate aqueous solution to the silica nanosphere aqueous solution to obtain a mixture; sonicate the mixture at 15-25℃ for 0.8-1.2 h; stir for 15-17 h, centrifuge for 10-20 min, wash, and obtain a solution of H-MnO2@sSiO2; ② Dissolve 19-23 g of sodium carbonate in 70-90 mL of water and let it cool to room temperature; combine two portions of the H-MnO2@sSiO2 solution obtained in step ①, resuspend them in 18-22 mL, and mix them with the sodium carbonate solution; heat at 55-65℃ for 10-14 h, centrifuge for 10-20 min, wash, and obtain a solution of hollow manganese dioxide particles; (3) Preparation of drug-loaded nanocomposite hydrogels: ① Mix the hollow manganese dioxide nanoparticle solution obtained in step (2) with the drug solution, stir at room temperature for 22-26 h, centrifuge, wash, and obtain a drug-loaded hollow manganese dioxide nanoparticle solution. ② Add inulin to the solution of drug-loaded hollow manganese dioxide nanoparticles obtained in step ①, so that the inulin concentration reaches 35~45%; then stir at 60~70℃ for 5~7 h, and let stand at 3~5℃ for 15~17 h to obtain the final product.

[0019] The present invention also provides the application of the drug-loaded nanocomposite hydrogel described above in the preparation of drugs for treating solid tumors, enteritis, or for the comprehensive treatment of intestinal diseases.

[0020] In one embodiment, the solid tumor is colorectal cancer.

[0021] Compared with the prior art, the present invention has the following beneficial effects: The drug-loaded nanocomposite hydrogel of this invention consists of internal drug-loaded hollow manganese dioxide nanoparticles and an externally modified inulin hydrogel "shell." Through the synergistic effect of the inulin hydrogel, hollow manganese dioxide nanoparticles (H-MnO2), and drugs, it can achieve targeted drug delivery to the colon, increase the retention time and accumulation of drugs in the colon, regulate intestinal flora, and stimulate the body's immune function, thereby enhancing the preventive and therapeutic effects of drugs. Moreover, because the hollow manganese dioxide nanoparticles in this invention have an ultra-large hollow volume, they can carry two or more drugs to achieve a comprehensive therapeutic effect on intestinal diseases.

[0022] When the drug-loaded nanocomposite hydrogel of the present invention carries oxaliplatin, the drug-loaded nanocomposite hydrogel is composed of internal oxaliplatin-loaded hollow manganese dioxide nanoparticles (Oxa / H-MnO2) and an externally modified inulin hydrogel "shell". Through the synergistic effect of inulin hydrogel, hollow manganese dioxide nanoparticles (H-MnO2) and oxaliplatin, it can achieve targeted drug delivery to the colon, increase the drug concentration and retention time of oxaliplatin in the colon, regulate the intestinal flora and the flora in intestinal tumors, stimulate the body's immune function, and thus improve the therapeutic effect of oxaliplatin, especially the effect of preventing and treating solid tumors such as colorectal cancer that are susceptible to the influence of flora.

[0023] The preparation method of the drug-loaded nanocomposite hydrogel of the present invention is simple and easy to operate, and is suitable for commercial promotion. Attached Figure Description

[0024] Figure 1 Images of the blank inulin hydrogel and the Oxa / H-MnO2-IN prepared in Example 1; Figure 2 The average particle size of H-MnO2, Oxa / H-MnO2, and Oxa / H-MnO2-IN solutions was determined using a Malvern laser particle size analyzer. Figure 3 This is a scanning electron microscope image of Oxa / H-MnO2-IN prepared in Example 1; Figure 4 The elemental analysis diagram of the corresponding region of Oxa / H-MnO2-IN obtained in Example 1 is shown below. Figure 5 The changes in particle size, PDI, and potential of Oxa / H-MnO2-IN prepared in Example 1 when incubated in simulated gastric juice (SGF), small intestinal juice (SIF), and colonic juice (SCF) are shown. Figure 6Fluorescent images of the colon of mice treated with H-MnO2, prepared for drug administration Example 1; Figure 7 A comparative graph showing the regulatory effect (relative abundance) of Oxa / H-MnO2-IN prepared in Example 1 on the colonic microbiota; Figure 8 The therapeutic effect of Oxa / H-MnO2-IN prepared in Example 1 on colorectal cancer tumors is shown in Figure A, which is a physical image of colon tumors in mice 14 days after administration. Figures B and C are CD8+ tumors 14 days after administration. + The proportion of T cells infiltrating tumor tissue and the tumor infiltrating CD8 + IFN-γ secretion by T cells. Detailed Implementation

[0025] To facilitate understanding of the present invention, a more complete description will be given below with reference to the accompanying drawings. Preferred embodiments of the invention are shown in the drawings. However, the invention can be implemented in many different forms and is not limited to the embodiments described herein. Rather, these embodiments are provided to provide a thorough and complete understanding of the disclosure of the invention.

[0026] Unless otherwise defined, 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 pertains. The terminology used herein in the description of the invention is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. The term "and / or" as used herein includes any and all combinations of one or more of the associated listed items.

[0027] Unless otherwise specified, all reagents used in the following examples are commercially available; and all methods used in the following examples are conventional methods.

[0028] Example 1 Preparation of drug-loaded nanocomposite hydrogels.

[0029] (1) Preparation of the silica nanospheres: ① Mix 10.6 mL of 100% Triton X-100, 45 mL of 99.5% cyclohexane, and 10.8 mL of 98% n-hexanol and stir for 5 min; then add 1.5 mL of 25-28% ammonia and 2 mL of deionized water, and stir for 30 min to obtain solution a; ② Add 1 mL of 99.99% tetraethyl orthosilicate (TEOS) and 0.2 mL of 98% 3-aminopropyltriethoxysilane (APTES) to solution a obtained in step ①. After stirring for 24 h, centrifuge at 12500 rpm for 20 min and remove the supernatant. Wash with ethanol 3 times and deionized water 3 times in sequence. After centrifugation to remove water, vacuum dry for one day. After drying, 300~400 mg of silica nanospheres are obtained.

[0030] (2) Preparation of the hollow manganese dioxide nanoparticles: ① Dissolve 100 mg of silica nanospheres obtained in step (1) in 100 mL of deionized water and sonicate to dissolve; dissolve 750 mg of potassium permanganate in 75 mL of deionized water and sonicate to dissolve. Potassium permanganate solution was added dropwise to an aqueous solution of silica nanospheres under ultrasonic conditions to obtain a mixture; the mixture was ultrasonicated for 1 h at 20 °C; after stirring overnight (16 h), it was centrifuged at 12500 rpm for 15 min and the supernatant was removed; after washing three times with deionized water, an aqueous solution of H-MnO2@sSiO2 was obtained. ② Dissolve 21.198 g of anhydrous sodium carbonate in 80 mL of deionized water, sonicate to dissolve, and then let it cool to room temperature; combine the two portions of H-MnO2@sSiO2 aqueous solution obtained in step ① and resuspend them in 20 mL, mix with sodium carbonate solution, sonicate slightly to homogenize, heat in an oil bath at 60℃ for 12 h, centrifuge for 15 min, remove the supernatant, wash with deionized water 3 times, and obtain an aqueous solution of hollow manganese dioxide particles (H-MnO2); (3) Preparation of the drug-loaded nanocomposite hydrogel: ① Mix the aqueous solution of hollow manganese dioxide nanoparticles obtained in step (2) with an appropriate amount of oxaliplatin aqueous solution, stir at room temperature for 24 h, remove the supernatant after centrifugation, and wash once with deion to remove the residual free oxaliplatin, to obtain an aqueous solution of oxaliplatin-loaded hollow manganese dioxide nanoparticles (Oxa / H-MnO2). ② Add an appropriate amount of inulin to the aqueous solution of oxaliplatin-loaded hollow manganese dioxide nanoparticles obtained in step ① to make the inulin concentration reach 40%; then stir at 70℃ for 6 h, and place the mixed solution in a refrigerator at 4℃ and let it stand overnight (16 h) to obtain the drug-loaded nanocomposite hydrogel (Oxa / H-MnO2-IN).

[0031] Experimental Example 1 The Oxa / H-MnO2-IN prepared in Example 1 was characterized.

[0032] (1) Following conventional methods, the average particle sizes of H-MnO2, Oxa / H-MnO2, and Oxa / H-MnO2-IN solutions were measured using a Malvern laser particle size analyzer (model: Malvern Nano ZS), and were found to be 171 nm, 189 nm, and 237 nm, respectively. (See...) Figure 2 ) The average particle size of H-MnO2, Oxa / H-MnO2, and Oxa / H-MnO2-IN solutions gradually increases, indicating that oxaliplatin (Oxa) was successfully loaded onto H-MnO2 nanoparticles and inulin was successfully modified onto the Oxa / H-MnO2 surface.

[0033] (2) Scanning electron microscopy observation of antitumor nanocomposite hydrogels, such as Figure 3 As shown, the antitumor nanocomposite hydrogel prepared in Example 1 has a loose and porous sheet-like sponge structure.

[0034] (3) Elemental analysis of the antitumor nanocomposite hydrogel prepared in Example 1 was performed using an energy dispersive spectrometer (model: Oxford Xplore 30), such as... Figure 4 As shown, C, O, Mn, and Pt elements were detected in the corresponding regions, indirectly proving the successful synthesis of the nanocomposite hydrogel.

[0035] Experimental Example 2 Observe the changes in particle size, PDI, and potential of Oxa / H-MnO2-IN incubated in simulated gastric juice (SGF), small intestinal juice (SIF), and colonic juice (SCF).

[0036] Oxa / H-MnO2-IN nanohydrogel is an inulin-modified nanohydrogel loaded with oxaliplatin. Since particle size, PDI, and potential cannot be detected in the gel state, a low concentration of inulin was used to synthesize Oxa / H-MnO2-IN nanosolution to detect particle size, PDI, and potential in simulated gastrointestinal fluid.

[0037] Experimental Methods: The effect of gastrointestinal fluids (SGF, pH 1.2), simulated intestinal fluid (SIF, pH 7.4), and simulated colonic fluid (SCF, pH 6.8) on the physical stability of Oxa / H-MnO2-IN nanosolution was evaluated. All three simulated fluids were commercially available. Following standard procedures, 1 mL of Oxa / H-MnO2-IN nanosolution was added to 9 mL of simulated gastrointestinal fluid and incubated for a specified time (2 hours for SGF, 4 hours for SIF, and 8 hours for SCF). This is because oral formulations take approximately 4-6 hours to reach the colon, with the first 1-2 hours in the stomach and 2-4 hours in the small intestine. At the last time point of each simulated culture medium, the average particle size (nm), PDI, and Zata potential (mV) of the nanoparticles were measured and evaluated against the initial measurements before incubation.

[0038] Test results: such as Figure 5 As shown in Figure A, the particle size of the Oxa / H-MnO2-IN solution remained essentially unchanged after incubation in simulated gastric, small intestinal, and colonic fluids. This indicates that the Oxa / H-MnO2-IN nanosolution is unaffected by the acidic environment of the stomach and upper digestive tract and maintains good stability in the gastrointestinal tract. Similarly, this also demonstrates that the Oxa / H-MnO2-IN nanogel is unaffected by the acidic environment of the stomach and upper digestive tract and maintains good stability in the gastrointestinal tract. Figure 5 As shown in Figure B, the particle size of the Oxa / H-MnO2-IN solution decreased after incubation with different concentrations of inulinase. When the inulinase concentration was 20 U / mL, the particle size was basically the same as that of the Oxa / H-MnO2 nanoparticles. This proves that the outer inulin shell of the Oxa / H-MnO2-IN nanosolution was degraded by inulinase secreted by specific bacteria in the colon, allowing the internal nanomedicine Oxa / H-MnO2 to be released and exert its therapeutic effect.

[0039] The particle size, PDI (polymer dispersibility index), and potential of the antitumor nanocomposite hydrogel showed no significant changes, indicating that it can be protected from the acidic environment of the stomach and upper digestive tract and maintain good stability in the gastrointestinal tract.

[0040] Experimental Example 3 Observe the accumulation and retention time of Oxa / H-MnO2-IN in the colon of balb / c mice.

[0041] Experimental methods: Six commercially available Balb / c mice were divided into two groups of three mice each. The mice in each group were administered Cy5.5 / H-MnO2 nanosolution and Cy5.5 / H-MnO2-IN nanohydrogel by gavage (Cy5.5 was administered at a dose of 20 mg / kg of mouse body weight, and the gavage volumes of Cy5.5 / H-MnO2 and Cy5.5 / H-MnO2-IN were 0.2 ml and 0.2 ml, respectively). After 24 hours of gavage, the colons of the mice were dissected, and the colons of the two groups of mice were photographed using a small animal in vivo imaging instrument. The fluorescence intensity in the colons of the two groups of mice was observed and compared.

[0042] Test results: such as Figure 6 As shown, compared with the cy5.5 / H-MnO2 nanosolution administered by gavage, the fluorescence intensity of the cy5.5 / H-MnO2-IN nanohydrogel was stronger after 24 h of gavage, indicating that the cy5.5 / H-MnO2-IN nanohydrogel accumulated more in the colon and had a longer retention time due to its thickening effect and gel strength-dependent mechanism.

[0043] Experiment Example 4 An evaluation experiment on the regulatory effect of Oxa / H-MnO2-IN on colonic flora.

[0044] Experimental Methods: Thirty-six commercially available Balb / c mice with orthotopic colon cancer were randomly divided into six groups of six mice each. The treatment regimens for the six groups were as follows: Control group (administered PBS by gavage), H-MnO2 group (administered H-MnO2 by gavage), Inulin group (administered Inulin by gavage), Oxa group (administered Oxa by gavage), Oxa / H-MnO2 group (administered Oxa / H-MnO2 by gavage), and Oxa / H-MnO2-IN group (administered Oxa / H-MnO2-IN hydrogel by gavage). The medications were administered by gavage once daily for 14 consecutive days (Oxa dosage was based on 20 mg / mL). The mice were administered the drug at a ratio of g / kg body weight. The gavage volumes of PBS, H-MnO2, Inulin, Oxa, Oxa / H-MnO2, and Oxa / H-MnO2-IN were 0.2 ml, 0.1 ml, 0.2 ml, 0.2 ml, 0.2 ml, and 0.2 ml, respectively. After the last administration, feces from each group of mice were collected. The abundance of each bacterial community in the feces of each group of mice was detected by 16S RNA sequencing (primer sequence: 468 bp, primer information: 338F806R) according to standard methods, and the differences of each bacterial community in the feces of each group of mice were compared.

[0045] Test results: such as Figure 7The results showed that, compared with the control group, the Oxa / H-MnO2-IN nanohydrogel group increased the abundance of beneficial bacteria Lactobacillus and Bifidobacterium, and decreased the abundance of harmful bacteria Alistipes and Helicobacter pylori.

[0046] Experimental Example 5 Evaluation trial of the therapeutic effect of Oxa / H-MnO2-IN on colorectal cancer tumors.

[0047] Experimental Methods: Thirty-six commercially available Balb / c mice with orthotopic colon cancer were randomly divided into 6 groups, with 6 mice in each group. The treatment regimens for the 6 groups were as follows: Control group (administered PBS by gavage), H-MnO2 group (administered H-MnO2 by gavage), Inulin group (administered Inulin by gavage), Oxa group (administered Oxa by gavage), Oxa / H-MnO2 group (administered Oxa / H-MnO2 by gavage), and Oxa / H-MnO2-IN group (administered Oxa / H-MnO2-IN hydrogel by gavage). The medications were administered by gavage once daily for 14 consecutive days. The dosage of xa was 20 mg / kg of mouse body weight. The gavage volumes for PBS, H-MnO2, Inulin, Oxa, Oxa / H-MnO2, and Oxa / H-MnO2-IN were 0.2 ml, 0.1 ml, 0.2 ml, 0.2 ml, 0.2 ml, and 0.2 ml, respectively. After the last administration, all mice were sacrificed, and their colons were removed. The size and weight of tumors in the colon were observed and compared using standard methods. Flow cytometry was used to measure CD8+ levels in the orthotopic colon tumors of the mice after administration. + The content of T cells was determined by IFN-γ in the serum of mice after administration via ELISA.

[0048] Test results: such as Figure 8 As shown in Figure A, the in situ tumor size of the Oxa / H-MnO2-IN nanohydrogel group was significantly smaller than that of the other material groups, indicating that the Oxa / H-MnO2-IN nanohydrogel has an extremely strong anti-tumor effect. Figure 8 As shown in B and 8C, CD8 of the Oxa / H-MnO2-IN nanohydrogel group + The proportion of T cell infiltration into tumors and the amount of IFN-γ secretion were significantly higher than those of other material groups, indicating that Oxa / H-MnO2-IN nanohydrogel can promote CD8+ secretion. + T cell infiltration into tumor tissue and promotion of CD8 + T cells secrete IFN-γ in tumor tissue, which greatly enhances the body's anti-tumor CD8+. + T-cell immune response.

[0049] The technical features of the above embodiments can be combined in any way. For the sake of brevity, not all possible combinations of the technical features in the above embodiments are described. However, as long as there is no contradiction in the combination of these technical features, they should be considered to be within the scope of this specification.

[0050] The embodiments described above are merely illustrative of several implementations of the present invention, and while the descriptions are relatively specific and detailed, they should not be construed as limiting the scope of the invention patent. It should be noted that those skilled in the art can make various modifications and improvements without departing from the concept of the present invention, and these all fall within the protection scope of the present invention. Therefore, the protection scope of this invention patent should be determined by the appended claims.

Claims

1. A drug-loaded nanocomposite hydrogel, characterized in that, The drug-loaded nanocomposite hydrogel is a drug-loaded hollow manganese dioxide nanoparticle with a surface modified by inulin hydrogel. The drug in question is oxaliplatin; The hollow manganese dioxide nanoparticles are loaded with at least one of the drugs; The preparation method of the drug-loaded nanocomposite hydrogel includes the following steps: (1) Preparation of silica nanospheres: Silica nanospheres were prepared by microemulsion method; (2) Preparation of hollow manganese dioxide nanoparticles: After reducing the silica nanospheres with potassium permanganate, chemical etching is performed to obtain the hollow manganese dioxide nanoparticles. (3) Preparation of drug-loaded nanocomposite hydrogel: The drug is loaded onto the surface of the hollow manganese dioxide nanoparticles, and then inulin is added to the drug-loaded hollow manganese dioxide nanoparticle solution to make the concentration of inulin reach 35-45%; after stirring at 60-70°C, it is allowed to stand at 3-5°C to obtain the product.

2. The drug-loaded nanocomposite hydrogel according to claim 1, characterized in that, The inulin hydrogel has an inulin concentration of 40-45%.

3. The drug-loaded nanocomposite hydrogel according to claim 1, characterized in that, The drug-loaded nanocomposite hydrogel has a particle size of 210-260 nm, the oxaliplatin-loaded hollow manganese dioxide nanoparticles have a particle size of 170-210 nm, and the hollow manganese dioxide nanoparticles have a particle size of 150-190 nm.

4. The method for preparing the drug-loaded nanocomposite hydrogel according to any one of claims 1 to 3, characterized in that, Includes the following steps: (1) Preparation of silica nanospheres: Silica nanospheres were prepared by microemulsion method; (2) Preparation of hollow manganese dioxide nanoparticles: After reducing the silica nanospheres with potassium permanganate, chemical etching is performed to obtain the hollow manganese dioxide nanoparticles. (3) Preparation of drug-loaded nanocomposite hydrogel: The drug is loaded onto the surface of the hollow manganese dioxide nanoparticles, and then inulin is added to the drug-loaded hollow manganese dioxide nanoparticle solution to make the concentration of inulin reach 35-45%; after stirring at 60-70°C, it is allowed to stand at 3-5°C to obtain the product.

5. The preparation method according to claim 4, characterized in that, Includes the following steps: (1) Preparation of silica nanospheres: ① Mix Triton X-100, cyclohexane, n-hexanol, ammonia, and water to obtain solution a; ② Add tetraethyl orthosilicate and 3-aminopropyltriethoxysilane to solution a obtained in step ① to obtain the silica nanospheres; (2) Preparation of hollow manganese dioxide nanoparticles: ① The silica nanospheres obtained in step (1) are mixed with potassium permanganate solution to obtain H-MnO2@sSiO2; ② The H-MnO2@sSiO2 obtained in step ① is mixed with sodium carbonate solution to obtain the hollow manganese dioxide nanoparticles; (3) Preparation of drug-loaded nanocomposite hydrogels: ① The hollow manganese dioxide nanoparticles obtained in step (2) are mixed with the drug solution to obtain the drug-loaded hollow manganese dioxide nanoparticles; ② Dissolve the drug-loaded hollow manganese dioxide nanoparticles obtained in step ① and add inulin to make the inulin concentration reach 35~45%. Stir at 60~70℃ and let stand at 3~5℃ to obtain the final product.

6. The preparation method according to claim 5, characterized in that, Includes the following steps: (1) Preparation of silica nanospheres: ① Mix 10-11 mL of Triton X-100 (99-100% concentration), 40-50 mL of cyclohexane (99-100% concentration), and 10-11 mL of n-hexanol (98-100% concentration) and stir for 4-6 min; then add 1.3-1.7 mL of ammonia (25-28% concentration) and 1.8-2.2 mL of water, and stir for 25-35 min to obtain solution a; ② Add 0.8~1.2 mL of tetraethyl orthosilicate with a concentration of 99.99~100% and 0.1~0.3 mL of 3-aminopropyltriethoxysilane with a concentration of 98~100% to the solution a obtained in step ①. After stirring for 22~26 h, centrifuge for 15~25 min, wash, and dry to obtain the silica nanospheres. (2) Preparation of hollow manganese dioxide nanoparticles: ① Dissolve 90-110 mg of the silica nanospheres obtained in step (1) in 90-110 mL of water; dissolve 675-825 mg of the potassium permanganate in 67.5-82.5 mL of water; add the potassium permanganate aqueous solution to the silica nanosphere aqueous solution to obtain a mixture; sonicate the mixture at 15-25℃ for 0.8-1.2 h; stir for 15-17 h, centrifuge for 10-20 min, wash, and obtain a solution of H-MnO2@sSiO2; ② Dissolve 19-23 g of sodium carbonate in 70-90 mL of water and let it cool to room temperature; combine two portions of the H-MnO2@sSiO2 solution obtained in step ①, resuspend them in 18-22 mL, and mix them with the sodium carbonate solution; heat at 55-65℃ for 10-14 h, centrifuge for 10-20 min, wash, and obtain a solution of hollow manganese dioxide particles; (3) Preparation of drug-loaded nanocomposite hydrogels: ① Mix the hollow manganese dioxide nanoparticle solution obtained in step (2) with the drug solution, stir at room temperature for 22-26 h, centrifuge, wash, and obtain a drug-loaded hollow manganese dioxide nanoparticle solution. ② Add inulin to the solution of drug-loaded hollow manganese dioxide nanoparticles obtained in step ①, so that the inulin concentration reaches 35~45%; then stir at 60~70℃ for 5~7 h, and let stand at 3~5℃ for 15~17 h to obtain the final product.

7. The use of the drug-loaded nanocomposite hydrogel according to any one of claims 1 to 3 in the preparation of drugs for treating intestinal tumors.

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