Ophiopogonin D and oxaliplatin co-assembled nano-drug delivery system as well as preparation method and application thereof

Abstract

The invention provides application of combined use of ophiopogonin D and oxaliplatin in preparation of a medicine for treating colorectal cancer. The invention also provides a nano-drug delivery system containing co-assembly of ophiopogonin D and oxaliplatin, and the nano-drug delivery system is prepared by the following steps: by taking ZIF-8, ophiopogonin D and oxaliplatin as raw materials, synthesizing ZIF-8 (at) OPHamp, taking the ZIF-8 (at) OPHamp as a carrier, and taking the ZIF-8 (at) OPHamp as a carrier to prepare the nano-drug delivery system containing co-assembly of ophiopogonin D and oxaliplatin. The invention relates to an OXA nanoparticle (ZOX NPs). The invention also provides a co-assembled nano-drug delivery system containing the hyaluronic acid. According to the invention, ophiopogonin D and oxaliplatin are co-assembled by using a nano traditional Chinese medicine delivery system, and the nano preparation HZOX NPs is successfully synthesized, so that the drug can be more enriched at a tumor site, and the problems of poor stability, poor water solubility, low bioavailability and the like of ophiopogonin D are solved. The nanoparticles are uniform in particle size, high in stability and good in biocompatibility, the nano traditional Chinese medicine delivery system can resist colorectal cancer chemical resistance, and a promising strategy is provided for colorectal cancer treatment.

Description

Technical Field

[0001] This invention relates to a nanomedicine delivery system containing ophiopogonin D and oxaliplatin co-assembled. Background Technology

[0002] Colorectal cancer, a malignant tumor that seriously threatens human health, has attracted widespread attention in the medical community due to its epidemiological characteristics. As a serious malignant disease, it is recognized as one of the most prevalent cancers in clinical practice, posing a significant challenge to public health security. While existing clinical interventions for colorectal cancer can partially control disease progression, they still face many limitations. Currently, the main treatment involves a combination of surgical resection, radiotherapy, and chemotherapy, but postoperative recurrence and metastasis remain common. Given this situation, there is an urgent need for in-depth research and development of innovative therapeutic drugs and comprehensive treatment plans to optimize clinical treatment pathways and develop new drugs with greater safety and efficacy advantages, thereby effectively inhibiting tumor progression and reducing the risk of recurrence. In recent years, breakthroughs have been made in the field of nanotechnology, greatly promoting the development and application of novel nanoparticles. Studies have shown that nanoparticles exhibit unique advantages in cancer prevention and treatment, not only intervening in the occurrence and development of cancer through targeted action but also significantly reducing multidrug resistance in cancer cells, thereby improving treatment outcomes.

[0003] Metal-organic frameworks (MOFs), a novel class of porous materials, are characterized by the self-assembly of metal ions / clusters and organic ligands through coordination bonds. MOFs, with their precisely tunable morphology and size, excellent drug loading capacity, and superior biocompatibility, have become ideal carrier platforms for developing novel drug delivery systems and cancer treatment strategies. Zeoliticimidazolate framework-8 (ZIF-8), one of the most commonly used MOFs, is particularly advantageous as a drug carrier for tumor therapy due to its high porosity, tunable pore size, ease of preparation, efficient encapsulation / loading of goods from small molecules to biomacromolecules, and pH-responsive biodegradability. Furthermore, the introduction of nanocarriers not only helps prolong drug delivery performance but also improves the bioavailability of natural extracts in vivo. While ZIF-8 offers unique advantages as a drug carrier (such as high specific surface area, pH-responsive solubility, and ease of functionalization), it still faces formulation challenges. For example, ZIF-8 has low affinity for certain drugs, leading to reduced drug loading and potential premature drug release. High concentrations (>30 μg / mL) of ZIF-8 nanoparticles may cause toxicity, necessitating dosage limitation. ZIF-8 may also experience structural instability or degradation under acidic environments or enzymatic action, resulting in unpredictable drug release kinetics and difficulty in maintaining effective therapeutic concentrations. To improve biocompatibility and targeting, functional modifications to the ZIF-8 surface (such as coating with polydopamine or polyethylene glycol) are often required. However, this may block pores, reduce drug loading capacity, and the modification process is complex, costly, and difficult to scale up.

[0004] Oxaliplatin, a commonly used chemotherapy drug, can cause excessive oxidative stress in tumor cells, leading to a large accumulation of reactive oxygen species (ROS) and ultimately tumor cell death. Oxaliplatin resistance is a key bottleneck in the treatment of colorectal cancer, severely restricting the improvement of clinical efficacy. Nanoparticle formulations can reduce the dosage of oxaliplatin to below the conventional dose level while maintaining equivalent antitumor activity. Nanoparticle co-delivery systems, with their precise control over drug distribution and multi-target synergistic intervention capabilities, can simultaneously achieve the triple core goals of dose optimization, chemotherapy sensitization, and resistance reversal. Based on this, nanotechnology achieves a synergistic breakthrough in the bottleneck of colorectal cancer treatment through multiple mechanisms, including precise delivery to reduce dosage requirements, targeted inhibition to overcome key resistance pathways, and combination with sensitizing drugs to enhance synergistic efficacy.

[0005] Traditional Chinese medicine Ophiopogon japonicus and its phytochemicals can be used alone or in combination with chemotherapy drugs, making them safe and reliable anticancer drug candidates. Ophiopogonin D, the main active ingredient extracted from the tuberous root of Ophiopogon japonicus, belongs to the steroidal saponin class of compounds. In recent years, its antitumor activity has received widespread attention, becoming one of the hot topics in natural product antitumor research. Studies have shown that Ophiopogonin D exerts its antitumor effect through multi-pathway cross-regulation, possessing the dual advantages of directly killing tumor cells and regulating the immune microenvironment, making it one of the representative molecules in the modernization of traditional Chinese medicine research. It has been proven that Ophiopogonin D can exert its anticancer effect by inhibiting proliferation and inducing cell cycle arrest, cell death, and autophagy, as well as inhibiting metastasis and angiogenesis in vitro and in vivo. Ophiopogonin D can activate pro-apoptotic proteins (such as Bax and Caspase-3) while inhibiting anti-apoptotic proteins (such as Bcl-2 and Survivin), overcoming chemotherapy resistance in colorectal cancer. Ophiopogonin D can block key autophagy pathways (such as PI3K / AKT / mTOR) and weaken the drug resistance of cancer cells. Ophiopogonin D is used in combination with the chemotherapy drug oxaliplatin to improve efficacy, reduce toxicity, and regulate multidrug resistance. However, Ophiopogonin D as an anti-tumor drug still has the following limitations:

[0006] 1. Poor water solubility

[0007] Ophiopogonin D has low water solubility, which may affect its distribution and absorption in vivo. Low water solubility may lead to insufficient accumulation of the drug in tumor tissue, thereby reducing therapeutic efficacy.

[0008] 2. Low bioavailability

[0009] Ophiopogonin D has low bioavailability. This means that in practical applications, higher doses are required to achieve the desired therapeutic effect, which may increase the risk of side effects.

[0010] While current literature has addressed the solubility issue of Ophiopogonin D, its efficacy in treating colorectal cancer remains limited. Summary of the Invention

[0011] This invention provides a nanomedicine delivery system containing ophiopogonin D and oxaliplatin co-assembled, its preparation method, and its uses.

[0012] The use of Ophiopogonin D in combination with oxaliplatin in the preparation of a drug for treating colorectal cancer. Specifically, the drug is a drug that reverses chemotherapy resistance in colorectal cancer.

[0013] The mass ratio of Ophiopogon japonicus saponin D to oxaliplatin is 1:1.

[0014] This invention also provides a nanomedicine delivery system containing ophiopogonin D and oxaliplatin co-assembled, which is synthesized from ZIF-8@OPH&OXA nanoparticles (ZOX NPs) using ZIF-8, ophiopogonin D and oxaliplatin as raw materials.

[0015] The mass ratio of ZIF-8, Ophiopogon japonicus saponin D, and oxaliplatin is 2:1:1.

[0016] ZIF-8 is prepared from 2-methylimidazole and zinc nitrate in a mass ratio of 10:1; the preparation method is as follows:

[0017] a. Weigh out the 2-methylimidazole and zinc nitrate in the specified mass ratios;

[0018] b. First, 2-methylimidazole was dissolved in deionized water and magnetically stirred at 300 rpm / min for 5 min at room temperature. Then, zinc nitrate hexahydrate dissolved in deionized water was added dropwise. The solution was stirred at 1200 rpm / min under magnetic stirring for 30 min. The precipitate was obtained by centrifugation (12000 rpm / min, 20 min) and then washed three times with deionized water.

[0019] This invention also provides a method for preparing the aforementioned Ophiopogon japonicus saponin D and oxaliplatin co-assembled nanomedicine delivery system, which includes the following steps:

[0020] a. Weigh the raw materials according to the specified mass ratio;

[0021] b. Dissolve ZIF-8 in deionized water and sonicate the mixture to prepare solution A; then dissolve Ophiopogonin D in methanol to form solution B and sonicate it; dissolve oxaliplatin in deionized water and sonicate it to form solution C.

[0022] c. Dissolve solution A by stirring at 1000 rpm for 5 min; during this period, add solution B dropwise while stirring magnetically; then stir the mixture at 1000 rpm for 12 h at room temperature and cover with tin foil; methanol is then removed by rotary evaporation; then add solution C dropwise while stirring magnetically, and stir the mixture at 1000 rpm for 12 h at room temperature to obtain ZIF-8@OPH&OXA nanoparticles (ZOX NPs).

[0023] This invention provides a co-assembled nanomedicine delivery system containing hyaluronic acid. It involves further coating the ZOX NPs co-assembled nanomedicine delivery system containing ophiopogonin D and oxaliplatin with hyaluronic acid (HA) to prepare HA / ZIF-8@OPF&OXA nanoparticles (HZOX NPs), wherein the mass ratio of ZIF-8, ophiopogonin D, oxaliplatin, and HA is 2:1:1:1. This invention also provides a method for preparing the aforementioned co-assembled nanomedicine delivery system containing hyaluronic acid. HA is dissolved in deionized water, and the prepared HA / ZIF-8@OPF&OXA nanoparticles are slowly added dropwise to the HA-containing deionized water solution. The mixture is stirred at 1000 rpm for 30 min in a 25 ml round-bottom flask to obtain the HZOX NP nanoparticles. After modification with hyaluronic acid, the nanoparticles exhibit enhanced tumor targeting and stronger tumor growth inhibition.

[0024] The present invention also provides the use of the aforementioned co-assembled nanomedicine delivery system in the preparation of drugs for reversing chemotherapy resistance in colorectal cancer and for treating colorectal cancer.

[0025] The drug formulation is an oral or injectable formulation.

[0026] The present invention utilizes Ophiopogonin D. Ophiopogonin D is a steroidal saponin compound isolated from the traditional Chinese medicine Ophiopogon japonicus. It possesses rich biological activity, particularly in antitumor, anti-inflammatory, and neuroprotective applications, and has been extensively studied. Current in vitro cell experiments and animal model studies show that Ophiopogonin D exhibits significant inhibitory effects on various solid tumors and hematological malignancies, with its antitumor activity primarily targeting colorectal cancer, lung cancer, liver cancer, and breast cancer. The structure of Ophiopogonin D is shown in the diagram; it is a white solid with the molecular formula C0. 44 H 70 O 16 Its relative molecular weight is 855.02.

[0027]

[0028] This invention relates to a nanomedicine delivery system, HA / ZIF-8@OPH&OXA (HZOX NPs), composed of ZIF-8, OPH, OXA, and HA. OPH and OXA are encapsulated within ZIF-8, and HA is modified onto the surface of the ZOX NPs, thereby synthesizing HA / ZIF-8@OPH&OXA nanoparticles (HZOX NPs). HZOX NPs can specifically accumulate in tumor tissues through enhanced permeability and retention rate (EPR) and HA-mediated targeting mechanisms. When the prepared nanoplatform reaches the tumor site, the weakly acidic intracellular microenvironment promotes the rapid degradation of HZOX NPs. This degradation ultimately leads to the complete structural collapse of the nanoparticles, releasing the OPH and OXA loaded within the ZIF-8 into the tumor site.

[0029] This invention utilizes a nano-medicine delivery system to co-assemble Ophiopogon japonicus saponin D with oxaliplatin, successfully synthesizing the nano-formulation HZOX NPs. This not only allows for greater drug accumulation at tumor sites but also overcomes the problems of Ophiopogon japonicus saponin D's instability, poor water solubility, and low bioavailability. The nanoparticles exhibit uniform particle size, high stability, and good biocompatibility. Preliminary experiments show that HZOX NPs possess good antitumor activity and resistance to chemoresistant colorectal cancer, indicating their potential clinical application value in cancer treatment. The nano-medicine delivery system demonstrates synergistic effects and the ability to significantly reverse multidrug resistance, suggesting its significant potential for clinical translation and application in the treatment of colorectal cancer. Attached Figure Description

[0030] Figure 1 Preparation of HZOX NPs;

[0031] Figure 2 Particle size distribution of ZIF-8;

[0032] Figure 3 Electron micrograph of ZIF-8;

[0033] Figure 4 Particle size distribution of ZOX NPs;

[0034] Figure 5 Electron micrographs of ZOX NPs;

[0035] Figure 6 Particle size distribution of HZOX NPs;

[0036] Figure 7 Electron micrographs of HZOX NPs;

[0037] Figure 8 Zeta potential of ZIF-8, ZOX NPs and HZOX NPs;

[0038] Figure 9 Size distribution of HZOX NPs in H2O, PBS and DMEM solutions;

[0039] Figure 10 Screening and comparison test of Ophiopogon japonicus saponins D+ and oxaliplatin;

[0040] Figure 11 Cytotoxicity of HZOX NPs;

[0041] Figure 12 HZOX NPs inhibit the proliferation of colorectal cancer cells;

[0042] Figure 13 Toxicity analysis of HZOX NPs on normal cells. Detailed Implementation

[0043] This invention provides a nanomedicine delivery system of HA / ZIF-8@OPF&OXA nanoparticles (HZOX NPs), composed of ZIF-8, Ophiopogonin D (OPH), oxaliplatin (OXA), and hyaluronic acid (HA). ZIF-8@OPH&OXA nanoparticles (ZOX NPs) were synthesized using a thin-film dispersion method. Subsequently, the surface of ZOX NPs was modified with HA to enhance their tumor-targeting ability, thus synthesizing HA / ZIF-8@OPF&OXA nanoparticles (HZOX NPs). HZOX NPs can specifically accumulate in tumor tissues through enhanced permeability and retention rate (EPR) and HA-mediated targeting mechanisms. When the prepared nanoplatform reaches the tumor site, the weakly acidic intracellular microenvironment promotes the rapid degradation of HZOX NPs. This degradation ultimately leads to the complete structural collapse of the nanoparticles, releasing the OPH and OXA loaded within the nanoparticles to the tumor site.

[0044] Example 1: Preparation of the nano-medicine delivery system (HZOX NPs) of the present invention

[0045] Synthesis flowchart as follows Figure 1 As shown:

[0046] 1. ZIF-8 nanoparticles were synthesized according to the reported method.

[0047] 2-Methylimidazole was dissolved in deionized water and magnetically stirred (300 rpm / min) for 5 minutes at room temperature. Zinc nitrate hexahydrate (dissolved in deionized water) was then added dropwise. The solution was stirred magnetically (1200 rpm / min) for 30 minutes. The precipitate was obtained by centrifugation (12000 rpm / min, 20 minutes) and then washed three times with deionized water. Analysis was performed using dynamic light scattering and transmission electron microscopy. Figure 2 , Figure 3 The successful preparation of ZIF-8 nanoparticles has been confirmed.

[0048] 2. ZOX NPs were prepared by thin-film dispersion method.

[0049] ZIF-8 was dissolved in deionized water, and the mixture was sonicated for 2 min to prepare solution A. OPH was then dissolved in methanol to form solution B, which was also sonicated for 2 min. Similarly, oxaliplatin was dissolved in deionized water and sonicated for 2 min to form solution C. Next, solution A was stirred at 1000 rpm for 5 min, during which solution B was added dropwise with magnetic stirring. The mixture was then stirred at 1000 rpm for 12 h at room temperature and covered with aluminum foil. Methanol was subsequently removed by rotary evaporation. Solution C was then added dropwise with magnetic stirring, and the mixture was stirred at 1000 rpm for 12 h at room temperature to obtain ZOX NPs. Dynamic light scattering (DLS) analysis showed that the particle size of the ZOX NPs was 142.7 nm. Figure 4 Furthermore, transmission electron microscopy images showed morphological changes in ZOX NPs, clearly indicating that OPH and OXA were successfully encapsulated. Figure 5 This size range is advantageous for drug delivery systems to utilize the EPR effect for tumor targeting.

[0050] 3. Hyaluronic acid modification

[0051] HA was dissolved in deionized water and sonicated for 2 min. Afterwards, the resulting ZOX NPs were dispersed in the HA aqueous solution and stirred at 1000 rpm / min for 30 min in a round-bottom flask to obtain HZOX NP nanoparticles. DLS analysis and transmission electron microscopy confirmed the successful synthesis of uniform nanoparticles with an average particle size of 153.8 nm. Furthermore, the zeta potential changes of ZIF-8, ZOX NPs, and HZOX NPs confirmed the successful encapsulation of OPH and OXA by ZIF-8 and the successful modification of HA. Figure 6-8 ).

[0052] 4. Stability assessment of HZOX NPs

[0053] The stability of nanoformulations is a key factor in their therapeutic efficacy, and stability can be assessed by monitoring changes in the morphology and particle size distribution. DLS analysis was used to evaluate the stability of HZOX NPs. The size distribution of HZOX NPs in MilliQ water was measured for 7 consecutive days, and the fluctuations were negligible. Furthermore, when dispersed in NaCl or DMEM with 10% FBS added, no significant change in particle size was detected by DLS after 7 days at 4°C. This indicates that HZOX NPs exhibit good long-term stability in different media. Figure 9 ).

[0054] Example 2: Screening and preparation of the mass ratio of OPH and OXA in the nano-medicine delivery system (HZOX NPs) of the present invention.

[0055] To improve the efficiency of the nanoplatform, a nanoparticle size analyzer was used to evaluate the dispersion ability of OPH and OXA at ​​different mass ratios. For example... Figure 10 As shown, a better particle size was observed at a mass ratio of 1:1, consistent with the parameter characterization of the EPR effect of nanoparticles. Differences in solubility were observed at different ratios, with significant precipitation observed in nanoparticles at ratios of 1:2 and 2:1.

[0056] The following pharmacodynamic tests demonstrate the beneficial effects of this invention.

[0057] Experimental Example 1: In vitro cytotoxicity experiment

[0058] Experimental objective: To verify whether HZOX NPs have tumor-killing effects using the CCK8 cytotoxicity assay.

[0059] Experimental procedure: After digesting and counting LoVo, LoVo / OXA, and 116 cells in the logarithmic growth phase, 1 × 10⁻⁶ cells were distributed per well. 4 Cells were seeded at a density of 100 μL in 96-well plates, with sterile saline added to the peripheral wells to maintain humidity. The plates were incubated at 37°C and 5% CO2 for 24 h. The next day, the old medium was replaced with medium containing different concentrations of the drug. The experimental groups were designed as follows: ZIF-8, OPH, OXA, ZOX NPs, and HZOX NPs in drug-containing medium, and incubated for 4 h. After incubation, the supernatant was discarded, and 100 μL of fresh medium was added. Incubation continued for 24 h. Then, 10 μL of CCK8 solution was added, and the plates were incubated at 37°C for 2 h. After incubation was terminated, the absorbance of each well was measured at 450 nm using a microplate reader.

[0060] The experimental results are shown in Figure 11 .

[0061] Summary of experimental results: Figure 11It can be seen that HZOX NPs have a significant killing effect on colorectal cancer cells, and at the same dose, HZOX NPs induce tumor cell death significantly more strongly than the OPH, OXA, and ZOX NPs groups. Furthermore, CCK8 assay results showed that the combination of Ophiopogon japonicus saponins and oxaliplatin had a stronger inhibitory effect on drug-resistant colorectal cancer cells LoVo / OXA. This indicates that HZOX NPs can be effectively internalized by colorectal cancer cells, thereby reversing drug resistance, and this effect has been well validated in drug-resistant cell lines.

[0062] Experimental Example 2: This nanomaterial inhibits the proliferation of colorectal cancer cells.

[0063] Experimental objective: To verify whether HZOX NPs have tumor-killing effects by performing an EdU cell proliferation assay.

[0064] Experimental Procedure: LoVo, LoVo / OXA, and 116 cells in logarithmic growth phase were digested and seeded at an appropriate density in 96-well plates. Cells were cultured overnight at 37°C to allow adherence. The following day, drug treatment was performed. After treatment, the old culture medium was discarded, and the cells were exposed to diluted EdU solution and incubated at 37°C for 4 hours. After incubation, the cells were washed three times with PBS buffer, and fixed with 4% paraformaldehyde fixative at room temperature for 30 minutes. The paraformaldehyde solution in the wells was washed away, and glycine solution was added to each well for 5 minutes to neutralize the fixative effect. The cells were then rinsed 1-2 times with PBS solution, each time for 5 minutes on a shaker. Apollo staining was performed under dark conditions for 30 minutes. Staining was terminated by rinsing 1-2 times with 0.5% Triton X-100 PBS solution, each time for 10 minutes. The cells were then treated with 100 μL / well Hoechst... Stain with 33342 for 30 min; after staining, wash three times with PBS. Immediately observe and acquire images under a fluorescence microscope.

[0065] The experimental results are shown in Figure 12 .

[0066] Summary of experimental results: HZOX NPs can effectively inhibit the proliferation of colorectal cancer cells. For example... Figure 12 As shown, treatment with HZOX NPs significantly reduced the positive rate of EdU markers. The results of the EdU assay were consistent with those of the MTT assay. In conclusion, this study demonstrates that HZOX NPs have a good anti-tumor effect on colorectal cancer cells and can effectively reverse drug resistance.

[0067] Experimental Example 3: Toxicity Analysis of the Nanomaterial to Normal Cells

[0068] The cytotoxicity of HZOX NPs in normal cells was assessed using the CCK8 assay. Human kidney epithelial cells (293T cells) and immortalized human hepatocytes (LO2 cells) were selected as normal cell models to evaluate the cytotoxicity of HZOX NPs. Specific experimental procedures are detailed in Experiment Example 1.

[0069] The experimental results are shown in Figure 13 :

[0070] Experimental results summary: HZOX NPs can selectively destroy tumor cells, and their cytotoxicity to normal cells is negligible, indicating that they have good in vitro biocompatibility.

[0071] In summary, this invention successfully designed a pH-sensitive nanoplatform to deliver OPH and OXA to colorectal cancer cells. HZOX NPs can specifically accumulate in tumor tissues through enhanced permeability and retention rate (EPR) and HA-mediated targeting mechanisms. The nanoplatform enhances the bioavailability of OPH and OXA. In vitro experimental results show that HZOX NP treatment effectively inhibits the growth of colorectal cancer cells. HZOX NPs solve the problems of poor solubility and bioavailability of OPH. Significant antitumor effects were observed in both chemotherapy-resistant and non-drug-resistant colorectal cancer cells, and HZOX NPs exhibited good in vitro biocompatibility. Therefore, this study experimentally confirms that HZOX NPs are a highly promising antitumor drug candidate. Based on its excellent performance, we innovatively utilized nanoparticle assembly technology to construct a co-delivery therapeutic system targeting drug-resistant colorectal cancer. The synergistic effect exhibited by this system and its significant effect in overcoming drug resistance provide a novel, more effective, and safer clinical strategy for colorectal cancer treatment, highlighting its important value in the field of cancer therapy.

Claims

1. The use of Ophiopogonin D in combination with oxaliplatin in the preparation of drugs for the treatment of colorectal cancer.

2. The use according to claim 1, characterized in that: The drug described is a drug that reverses chemotherapy resistance in colorectal cancer.

3. The use according to claim 1 or 2, characterized in that: The mass ratio of Ophiopogon japonicus saponin D to oxaliplatin is 1:

1.

4. A nanomedicine delivery system containing ophiopogonin D and oxaliplatin co-assembled, characterized in that: It is synthesized from ZIF-8@OPH&OXA nanoparticles (ZOX NPs) containing zeolite imidazole ester skeleton 8 (ZIF-8), ophiopogonin D and oxaliplatin.

5. The nanomedicine delivery system containing ophiopogonin D and oxaliplatin co-assembled according to claim 4, characterized in that: The mass ratio of ZIF-8, Ophiopogon japonicus saponin D, and oxaliplatin is 2:1:

1.

6. The nanomedicine delivery system containing ophiopogonin D and oxaliplatin co-assembled according to claim 4 or 5, characterized in that: The zeolite imidazole ester skeleton 8 (ZIF-8) is prepared from 2-methylimidazolium and zinc nitrate in a mass ratio of 10:1; the preparation method is as follows: a. Weigh out the 2-methylimidazole and zinc nitrate in the specified mass ratios; b. First, 2-methylimidazole was dissolved in deionized water and magnetically stirred at 300 rpm / min for 5 min at room temperature. Then, zinc nitrate hexahydrate dissolved in deionized water was added dropwise. The solution was stirred magnetically at 1200 rpm / min for 30 min. The precipitate was obtained by centrifugation (12000 rpm / min, 20 min) and then washed three times with deionized water.

7. The preparation method of the nanomedicine delivery system co-assembled with Ophiopogon japonicus saponin D and oxaliplatin according to any one of claims 4-6, characterized in that: It includes the following steps: a. Weigh the raw materials according to the specified mass ratio; b. Dissolve ZIF-8 in deionized water and sonicate the mixture to prepare solution A; then dissolve Ophiopogonin D in methanol to form solution B and sonicate it; dissolve oxaliplatin in deionized water and sonicate it to form solution C. c. Stir solution A at 1000 rpm for 5 min; during this period, add solution B dropwise with magnetic stirring; then stir the mixture at 1000 rpm for 12 h at room temperature and cover with tin foil; methanol is then removed by rotary evaporation; then add solution C dropwise with magnetic stirring, and stir the mixture at 1000 rpm for 12 h at room temperature to obtain ZIF-8@OPH&OXA nanoparticles (ZOX NPs).

8. A co-assembled nanomedicine delivery system containing hyaluronic acid, characterized in that: It is prepared by further modifying the ZOX NPs nanoparticles containing ophiopogonin D and oxaliplatin co-assembled nanodrug delivery system as described in any one of claims 4-6 with hyaluronic acid (HA) to form HA / ZIF-8@OPF&OXA nanoparticles (HZOX NPs), wherein the mass ratio of ZIF-8, ophiopogonin D, oxaliplatin and HA is 2:1:1:

1.

9. A method for preparing the hyaluronic acid-containing co-assembled nanomedicine delivery system according to claim 8, characterized in that: The process involves dissolving HA in deionized water, slowly adding the prepared HA / ZIF-8@OPF&OXA nanoparticles dropwise into the HA-containing deionized water, and stirring at 1000 rpm / min for 30 min in a 25 ml round-bottom flask to obtain HZOX nanoparticles.

10. Use of the co-assembled nanomedicine delivery system according to any one of claims 4-6 or claim 8 in the preparation of a medicament for treating colorectal cancer; preferably, the medicament is a medicament for reversing chemotherapy resistance in colorectal cancer; the formulation of the medicament is an oral formulation or an injectable formulation.

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