Metal organic coordination network material as well as preparation method and application thereof

Abstract

A preparation method of the metal organic coordination network comprises the following steps: respectively dissolving copper acetate and polyphenol in deionized water or dimethyl sulfoxide to form a copper acetate solution and a polyphenol solution, then dropwise adding the copper acetate solution into the polyphenol solution, introducing nitrogen, and carrying out a heating reaction to obtain a reaction mixture; centrifuging the reaction mixture to obtain a precipitation product metal organic coordination network; the metal organic coordination network is applied to preparation of a material for targeted killing of hypoxic tumor cells. The metal organic coordination network disclosed by the invention is good in dispersity, high in biocompatibility and good in hypoxic tumor cell selective toxicity. After being degraded by glutathione, the glutathione derivative has good peroxidase activity, can catalyze hydrogen peroxide to generate hydroxyl free radicals with stronger cytotoxicity, and can be used for converting tumors into oxidative phosphorylation metabolic pathways by inhibiting hypoxic tumor glycolysis and remodeling hypoxic tumor metabolism, promoting tumor cells to generate ROS (reactive oxygen species), and further improving the activity of glutathione. And the sensitivity of hypoxic tumors to copper death is enhanced.

Description

Technical Field

[0001] The invention relates to the technical field of metal nanomaterials, and in particular to an anti-tumor metal-polyphenol glycolysis inhibitor nanomedicine and a preparation method and application thereof. Background Art

[0002] Hypoxia is one of the characteristics of solid tumors, and the oxygen partial pressure in solid tumors is usually less than 10 mmHg. The rapid proliferation of tumor cells, high-level metabolism and abnormal structure of tumor blood vessels are the main causes of tumor hypoxia. Tumor cells in hypoxic areas further mediate tolerance to radiotherapy and chemotherapy by upregulating hypoxia-inducible factors (HIF) and downstream gene transcriptional activation, thereby leading to the failure of current mainstream clinical treatment methods and poor prognosis of radiotherapy and chemotherapy for tumor patients.

[0003] At present, the treatment strategies for hypoxic tumors mainly include using hypoxia to kill tumors and alleviating tumor hypoxia. Using hypoxia to kill tumors mainly includes hypoxia-activated small molecule prodrugs and hypoxia-responsive nano-delivery systems; alleviating tumor hypoxia mainly includes inducing tumor vascular normalization, exogenous oxygen delivery, and tumor in situ reaction to generate oxygen. However, due to factors such as low delivery efficiency, insufficient oxygen production efficiency, narrow treatment time window, and off-target toxic side effects, the effectiveness of the above treatment strategies is still greatly limited. Therefore, there is an urgent need for a safe, effective, and precise treatment system to kill hypoxic tumors.

[0004] Metabolic shift is a prominent feature of hypoxic tumors. In order to meet the nutritional needs of rapid tumor cell proliferation, the energy metabolism of hypoxic tumors shifts from aerobic respiration to glycolysis. In addition, hypoxia reduces the sensitivity of tumor cells to copper death. Copper death is a copper-dependent programmed cell death. Copper ions oligomerize the tricarboxylic acid cycle (TCA)-related protein DLAT and induce cell death. Inhibiting the glycolysis pathway of hypoxic tumors, forcing the metabolic pathway to shift toward the tricarboxylic acid cycle, enhancing mitochondrial oxidative phosphorylation, increasing reactive oxygen species, and enhancing the sensitivity of hypoxic tumor cells to copper death may effectively kill hypoxic tumors.

[0005] Metal-organic network (MON) is a porous material formed by metal ions and organic ligands connected by coordination bonds. As a new type of material, MON has a wide range of application potential in the field of medical biomaterials due to its highly adjustable composition, porosity, large internal surface area, and sustainable development. A variety of natural polyphenols have the ability to regulate cell metabolism and energy supply, and can chelate with metal ions to prepare MON or coordination polymers.

[0006] Based on the above background, the inventors studied the use of CuMON, a metal organic coordination network nanomedicine assembled by coordination of polyphenols and copper ions with glycolysis inhibition function, for the treatment of hypoxic tumors. Currently, there is no report on the application of CuMON in the treatment of hypoxic tumors. Summary of the invention

[0007] The present invention provides a metal organic coordination network CuMON for anti-tumor treatment and a preparation method and application thereof, aiming to solve the problems existing in the above-mentioned background technology.

[0008] In order to achieve the above technical objectives, the present invention mainly adopts the following technical solutions: One of the purposes of the present invention is to provide a method for preparing a metal organic coordination network material CuMON, comprising the following steps: S1 reaction: copper acetate [Cu(OAc) 2 ·H 2 O] and polyphenols with glycolysis inhibitory function such as epigallocatechin gallate (EGCG), quercetin (QUE) or epigallocatechin (EGC) were dissolved in deionized water or dimethyl sulfoxide (DMSO) to form a copper acetate solution and a polyphenol solution. Then, the copper acetate solution was added dropwise to the polyphenol solution under continuous stirring and nitrogen (N) was passed through the solution. 2 ) and simultaneously heating the reaction until the reaction is completely completed to obtain a reaction mixture; S2 Purification: The reaction mixture prepared in step S1 is transferred into a centrifuge tube, centrifuged, washed (washed twice with deionized water or washed twice with methanol and then washed twice with deionized water for purification), and finally centrifuged to obtain the precipitated product metal organic coordination network CuMON.

[0009] In step S1, the mass ratio of the copper acetate hydrate to the polyphenol is 30-80:50-250, the concentration of the copper acetate solution is 3 mg / ml-8 mg / ml, and the concentration of the polyphenol solution is 3.125 mg / ml-15.625 mg / ml; the heating reaction temperature is 85°C, and the reaction time is 12-24 h.

[0010] In step S1, the mass of the copper acetate hydrate [Cu(OAc)2·H2O] is 30-80 mg; the mass of the epigallocatechin gallate (EGCG) is 50-250 mg, the mass of quercetin (QUE) is 50-250 mg, and the mass of epigallocatechin (EGC) is 50-250 mg.

[0011] In step S1, the heating reaction temperature is 85° C. and the reaction time is 12 to 24 h.

[0012] In the present invention, preferably, in step S2, the centrifugal speed is 12000 g to 20000 g, and the centrifugal time is 5 to 10 minutes.

[0013] In step S3, the tumor cell membrane is a CT26 cell membrane.

[0014] The second objective of the present invention is to provide a metal organic coordination network material CuMON that selectively kills hypoxic tumor cells. The metal organic coordination network CuMON is mainly prepared by the preparation method of the metal organic coordination network CuMON that selectively kills hypoxic tumors as described above.

[0015] Preferably, the particle size of the metal organic coordination network CuMON is 80-100 nm.

[0016] At the same time, the present invention also provides a metal organic coordination network coating, which is obtained by coating the metal organic coordination network with the tumor cell membrane.

[0017] Preparation method of metal organic coordination network coating: extract tumor cell membrane (use cell membrane protein and cytoplasm protein extraction kit manufactured by Shanghai Biotime Biotechnology Co., Ltd., product number: P0033, extract according to the product manual), then mix CuMON with the cell membrane, ultrasonicate and stir in an ice bath, co-extrude using a membrane extruder, and then centrifuge to obtain the coating product CuMON@CCM.

[0018] The third object of the present invention is to provide a metal-organic coordination network CuMON that selectively kills hypoxic tumor cells prepared by the preparation method of the metal-organic coordination network CuMON that selectively kills hypoxic tumor cells as described above, or the use of the metal-organic coordination network CuMON that selectively kills hypoxic tumor cells as described above in the preparation of materials for targeted killing of hypoxic tumor cells.

[0019] Compared with the prior art, the present invention mainly has the following beneficial effects: (1) The present invention adopts a hydrothermal method to synthesize a metal organic coordination network CuMON that selectively kills hypoxic tumors. The particle size is 80-100 nm, the dispersibility is good, the biocompatibility is high, and the selective toxicity of hypoxic tumor cells is good. After being degraded by glutathione (GSH), the nanoparticles have good peroxidase (POD) activity and can catalyze hydrogen peroxide (H 2 O 2 ) can produce more cytotoxic hydroxyl free radicals. In addition, it can inhibit hypoxic tumor glycolysis, reshape hypoxic tumor metabolism, transform the tumor to the oxidative phosphorylation metabolic pathway, promote tumor cells to produce ROS, and enhance the sensitivity of hypoxic tumors to copper death.

[0020] (2) The preparation process disclosed in the present invention is simple, the reaction conditions are mild, and the synthesis is easy, thus being suitable for popularization and use. BRIEF DESCRIPTION OF THE DRAWINGS

[0021] Figure 1 This is the result of the CuEGCG MON metal organic coordination network cytotoxicity experiment; Figure 2 This is the result of the CuQUE MON metal organic coordination network cytotoxicity experiment; Figure 3 This is the result of the CuEGC MON metal organic coordination network cytotoxicity experiment; Figure 4 This is the transmission electron microscopy image of the CuEGCG MON metal organic coordination network; Figure 5 This is the hydrated particle size distribution diagram of CuEGCG MON metal organic coordination network; Figure 6 This is the experimental result of the degradation of CuEGCG MON metal organic coordination network by GSH; Figure 7 This is the result of the POD activity experiment after the CuEGCG MON metal organic coordination network was degraded by GSH; Figure 8 This is the flow cytometry result of CuEGCG MON metal organic coordination network promoting the increase of ROS in hypoxic tumor cells; Fig. 9 The statistical diagram of CuEGCG MON metal organic coordination network inhibiting glucose uptake and lactate efflux in tumor cells; Fig.10 This is a statistical diagram of the degradation of CuEGCG MON metal organic coordination network by intracellular GSH; Fig.11 The results of a protein immunoblotting experiment showing that CuEGCG MON metal organic coordination network leads to copper death of tumor cells; Fig.12 This is a physical picture of the anti-tumor efficacy of CuEGCG MON metal organic coordination network based on subcutaneous transplanted tumors in mice; Fig.13 This is a statistical graph of tumor weight after CuEGCG MON metal organic coordination network treated subcutaneous tumors in mice; Fig.14 These are H&E and Ki67 staining images of the two groups of mice after tumor treatment. DETAILED DESCRIPTION

[0022] The present invention is described in detail below with reference to the accompanying drawings and specific embodiments, but the present invention is not limited to these embodiments. Embodiment 1:

[0023] Synthesis of various metal-organic coordination networks 50 mg of copper acetate [Cu(OAc) 2 ·H 2 O] and 120 mg epigallocatechin gallate (EGCG) were dissolved in 10 ml and 16 ml deionized water, respectively, to form a solution. Then, the copper acetate solution was added dropwise to the EGCG solution under continuous stirring. The mixture was purged with nitrogen (N 2 ) and simultaneously heated the reaction until the reaction was completely completed (reaction temperature was 85°C, reaction time was 12 h), and a reaction mixture was obtained; the reaction mixture was transferred into a centrifuge tube, centrifuged (centrifugal speed was 20000 g, centrifugal time was 8 minutes) and washed twice with deionized water for purification, and finally centrifuged to obtain the precipitated product CuEGCG MON metal organic coordination network.

[0024] 50 mg of copper acetate [Cu(OAc) 2 ·H 2 O] and 90 mg of quercetin (QUE) were dissolved in 10 ml of deionized water and 16 ml of dimethyl sulfoxide (DMSO) respectively to form a solution, and then the copper acetate solution was added dropwise to the QUE solution under continuous stirring, and nitrogen (N2) was passed and heated to react until the reaction was completely completed (reaction temperature was 85°C, reaction time was 22 h), to obtain a reaction mixture; the obtained reaction mixture was transferred into a centrifuge tube, centrifuged (centrifugal speed was 12000 g, centrifugal time was 10 minutes) and washed twice with methanol, washed twice with deionized water for purification, and finally centrifuged to obtain the precipitated product CuQUE MON metal organic coordination network.

[0025] 50 mg of copper acetate [Cu(OAc)2·H2O] and 110 mg of epigallocatechin (EGC) were dissolved in 10 ml of deionized water and 16 ml of dimethyl sulfoxide (DMSO) to form a solution. Then, the copper acetate solution was added dropwise to the EGC solution under continuous stirring. The mixture was purged with nitrogen (N 2 ) and simultaneously heated the reaction until the reaction was completely completed (reaction temperature was 85°C, reaction time was 18 h), to obtain a reaction mixture; the reaction mixture was transferred into a centrifuge tube, centrifuged (centrifugal speed was 15000 g, centrifugal time was 5 minutes) and washed twice with methanol, washed twice with deionized water for purification, and finally centrifuged to obtain a precipitated product CuEGC MON metal organic coordination network.

[0026] Preparation of metal organic coordination network coatings: extract tumor cell membranes (using the cell membrane protein and cytoplasm protein extraction kit manufactured by Shanghai Biotime Biotechnology Co., Ltd., catalog number: P0033, extracted according to the product instructions), and then mix the above-mentioned CuEGCG MON metal organic coordination network, CuQUE MON metal organic coordination network or CuEGC MON metal organic coordination network with the cell membrane at a mass ratio of 1:1, with a concentration of 0.1 mg / ml, ultrasonically stirred in an ice bath, co-extruded using a film extruder, and then centrifuged to obtain the coating products CuEGCG@CCM, CuQUE@CCM or CuEGC@CCM. Embodiment 2:

[0027] CuEGCG MON metal organic coordination network cytotoxicity assay First, mouse CT26 colon cancer cells were inoculated in a 96-well plate, with the number of cells inoculated being 5×10 per well. 3 24 hours after cell inoculation, the cells were treated with 0, 6.25, 12.5, 25, 50, and 100 μg / ml CuEGCG MON in normoxic and hypoxic environments, and CCK8 was used to detect cell viability 48 hours later. The results are shown in the attached figure. Figure 1 As shown, the CuEGCG MON metal-organic coordination network has significant selective toxicity against hypoxic tumor cells.

[0028] CuQUE MON Metal-Organic Coordination Network Cytotoxicity Assay First, mouse CT26 colon cancer cells were inoculated in a 96-well plate, with the number of cells inoculated being 5×10 per well. 324 hours after cell inoculation, cells were treated with 0, 6.25, 12.5, 25, 50, and 100 μg / ml CuQUE MON in normoxic and hypoxic environments, and CCK8 was used to detect cell viability 48 hours later. The results are shown in the attached figure. Figure 2 As shown, the CuQUE MON metal-organic coordination network has significant selective toxicity against hypoxic tumor cells.

[0029] CuEGC-MON metal-organic coordination network cytotoxicity assay First, mouse CT26 colon cancer cells were inoculated in a 96-well plate, with the number of cells inoculated being 5×10 per well. 3 24 hours after cell inoculation, the cells were treated with 0, 6.25, 12.5, 25, 50, and 100 μg / ml CuEGC MON in normoxic and hypoxic environments, and CCK8 was used to detect cell viability 48 hours later. The results are shown in the attached figure. Figure 3 As shown, the CuEGC MON metal-organic coordination network has significant selective toxicity against hypoxic tumor cells. Embodiment 3:

[0030] Characterization of the CuEGCG MON metal organic coordination network prepared by the present invention 1) Transmission electron microscopy experiment of CuEGCG MON metal organic coordination network Take 20 μL of CuEGCG MON metal organic coordination network liquid in a 1.5 mL centrifuge tube, add 980 μL of deionized water to dilute, and then take 10 μL of the diluted sample and drop it on the carbon support film copper grid. The copper grid is placed in an electronic drying oven to dry overnight to prepare the transmission electron microscope (TEM) sample.

[0031] The results observed by TEM are shown in the attached Figure 4 As shown, the CuEGCG MON metal organic coordination network is a uniform spherical nanoparticle with a real particle size of about 80 to 100 nm.

[0032] 2) Determination of hydrated particle size of CuEGCG MON metal-organic coordination network Take the synthesized CuEGCG MON metal organic coordination network and dilute it to 2mL with deionized water. Take 1mL of it to measure the hydrated particle size. Water bath ultrasonic for 15min, each group of samples is measured three times in parallel, and the changes in the whole process are monitored. The hydrated particle size distribution diagram of the CuEGCG MON metal organic coordination network is shown in the attached figure. Figure 5 As shown, the prepared CuEGCG MON metal organic coordination network is well dispersed and the average hydrated particle size is 270 nm. Embodiment 4:

[0033] Degradation of GSH by the CuEGCG MON metal organic coordination network prepared by the present invention The synthesized CuEGCG MON metal organic coordination network was mixed with GSH to make the final concentration of CuEGCG MON metal organic coordination network 0, 25, 50, 100, 200 μg / ml, and the final concentration of GSH 10 mM. After reacting for 30 minutes, the remaining GSH concentration was detected using a glutathione detection kit. The results are shown in the attached figure. Figure 6 As shown, the prepared CuEGCG MON metal-organic coordination network has the ability to rapidly degrade GSH. Embodiment 5:

[0034] Determination of POD enzyme activity after the CuEGCG MON metal organic coordination network prepared by the present invention is degraded by GSH Take an appropriate amount of CuEGCG MON metal organic coordination network with a concentration of 400μg / ml, add GSH to make the final concentration of GSH 20mM, react for 15min, centrifuge at 12000rpm for 10min and take the supernatant. Add 2.5mM 3,3',5,5'-tetramethylbenzidine (TMB) and 80mM H 2 O 2 PBS with a pH of 7.4 was used as the reaction buffer. The results are shown in the attached Figure 7 As shown, the CuEGCG MON metal-organic coordination network had good POD enzyme activity after being degraded by GSH. Embodiment 6:

[0035] The CuEGCG MON metal organic coordination network prepared by the present invention promotes the determination of ROS levels in hypoxic tumor cells The DCF method is used to measure intracellular ROS. DCFH-DA itself has no fluorescence and can freely pass through the cell membrane. After entering the cell, it can be hydrolyzed by the esterase in the cell to produce DCFH. However, DCFH cannot penetrate the cell membrane, so the probe can be easily loaded into the cell. The reactive oxygen ROS in the cell can oxidize the non-fluorescent DCFH to produce fluorescent DCF. Therefore, the level of reactive oxygen in the cell can be known by detecting the fluorescence of DCF.

[0036] First, mouse CT26 colon cancer cells were inoculated in a 6-well plate, with the number of cells inoculated being 1×10 per well. 5The cells were divided into normoxic control group, normoxic CuEGCG MON 50μg / ml group, hypoxic control group and hypoxic CuEGCG MON 50μg / ml group. After 24 hours of cell inoculation, they were treated according to the above groupings. After 24 hours, they were washed twice with sterile PBS solution, and DCFH-DA (Sigma) with a final concentration of 10μM was added and incubated in an incubator for 30 min. After staining, the intracellular fluorescence level was detected by the FITC channel of the flow cytometer. The results are shown in the attached figure. Figure 8 As shown, the CuEGCG MON metal-organic coordination network can significantly increase the ROS level in hypoxic tumor cells. Embodiment 7:

[0037] Test on the ability of the CuEGCG MON metal organic coordination network prepared by the present invention to inhibit tumor cell glucose uptake and lactate secretion First, mouse CT26 colon cancer cells were inoculated in a 6-well plate, with the number of cells inoculated being 1×10 per well. 5 The cells were divided into normoxic control group, normoxic CuEGCG MON 12.5μg / ml group, hypoxic control group and hypoxic CuEGCG MON 12.5μg / ml group. After 24 hours of cell inoculation, they were treated according to the above grouping. After 24 hours, the cell culture medium and cells of each group were collected respectively. The BCA reagent was used to detect the cell protein concentration in each group. The glucose detection kit and lactate detection kit were used to detect the content of glucose and lactate in the supernatant of each group. The difference method was used to calculate the glucose uptake of cells in each group. The results are shown in the attached figure. Fig. 9 As shown, the CuEGCG MON metal-organic coordination network can significantly inhibit the glucose uptake and lactate secretion of hypoxic tumor cells.

[0038] Embodiment 8:

[0039] CuEGCG MON metal-organic coordination network degrades intracellular GSH First, mouse CT26 colon cancer cells were inoculated in a 6-well plate, with the number of cells inoculated being 1×10 per well. 5 The cells were divided into normoxic control group, normoxic CuEGCG MON 25μg / ml group, hypoxic control group and hypoxic CuEGCG MON 25μg / ml group. After 24 hours of cell inoculation, the cells were treated according to the above grouping. After 24 hours of treatment, the cells in each group were collected and lysed with RIPA lysis buffer. The cell protein concentration in each group was detected using BCA reagent, and the GSH level in the cell lysate of each group was detected using glutathione detection kit. The results are shown in the attached figure. Fig.10 As shown, the CuEGCG MON metal-organic coordination network effectively degrades intracellular GSH. Embodiment 9:

[0040] Western blotting experiment of CuEGCG-MON metal organic coordination network promoting copper death of tumor cells First, mouse CT26 colon cancer cells or mouse 4T1 breast cancer cells were inoculated in 6-well plates, with the number of cells inoculated being 1×10 per well. 5 There were three groups of cells: hypoxic control group and hypoxic CuEGCG MON 50μg / ml group. Cells were treated according to the above grouping 24 hours after inoculation. Cells in each group were collected 24 hours later, and the cells were lysed with RIPA lysis buffer. The cell protein concentration in each group was detected using BCA reagent. A 7.5% gel was prepared using the Yase Quick Gel Kit. After electrophoresis at 140mV for 40 minutes, the membrane was transferred at 400mA for 40 minutes. The obtained PVF membrane was blocked with 5% milk powder and incubated with DLAT (ABclonal) primary antibody at 4°C overnight. Then, it was exposed to light after incubation with secondary antibody for 1h. The results are shown in the attached figure. Fig.11 As shown, the CuEGCG MON metal-organic coordination network significantly promoted the DLAT aggregation in hypoxic tumors. Embodiment 10:

[0041] Experimental study on the antitumor activity of the CuEGCG@CCM metal organic coordination network based on mouse subcutaneous transplanted tumors prepared by the present invention 1) Establishment of mouse subcutaneous tumor model In order to study the therapeutic effect of CuEGCG@CCM on tumors in mice, a subcutaneous transplanted tumor model was first established in Balb / c mice. The skin of the mouse armpit was prepared and hair removed for 3×3 cm. 2×10 6 CT26 tumor cells were injected into the subcutaneous tissue of mice. After 7 days, the tumor of the mice grew to about 100 mm. 3 , the model was built successfully.

[0042] 2) CuEGCG MON and CuEGCG@CCM metal organic coordination network for the treatment of subcutaneous transplanted tumors in mice During the modeling process, mice were randomly divided into 4 groups (Control group, commercial hypoxia-activated drug tiazolamine group, CuEGCG MON group, CuEGCG@CCM group), with 6 mice in each group. The control group was injected with 0.2 ml PBS through the tail vein, and the commercial hypoxia-activated drug tiazolamine group was injected with 0.2 mL of commercial tiazolamine (1 mg / ml, CHIOGELTM, medical device registration number: 20143642114, Shanghai Qisheng Biotechnology Co., Ltd.) through the tail vein. Each mouse in the CuEGCG MON group was injected with 0.2 mL of 1 mg / mL CuEGCG MON metal organic coordination network solution. Each mouse in the CuEGCG@CCM group was injected with 0.2 mL of CuEGCG@CCM metal organic coordination network solution containing 1 mg / mL CuEGCG MON. Injections were given every other day until 4 treatments were completed. During this period, the long and short diameters of the tumors were measured every day. After 10 days, the mouse tumors were dissected and tumor photos were taken, as shown in the attached figure. Fig.12 The tumor mass of each mouse was measured using a scale. Fig.13 The statistical results showed that the tumor mass of mice in the CuEGCG@CCM group was significantly different from that in the Control group, the commercial hypoxia-activated drug Tizalamine group, and the CuEGCG MON group. Fig.14 The H&E and Ki67 staining images of the tissues are shown. H&E is mainly used to observe the pathological morphology of tissues, and Ki67 staining is used to observe the proliferation of tumors. The above results reflect that CuEGCG@CCM can significantly inhibit tumor size and is superior to the commercial hypoxia-activated drug tiazolamine group.

[0043] It should be noted that, in this article, relational terms such as first and second, etc. are only used to distinguish one entity or operation from another entity or operation, and do not necessarily require or imply any such actual relationship or order between these entities or operations. Moreover, the terms "include", "comprise" or any other variants thereof are intended to cover non-exclusive inclusion, so that a process, method, article or device including a series of elements includes not only those elements, but also other elements not explicitly listed, or also includes elements inherent to such process, method, article or device. In the absence of further restrictions, the elements defined by the sentence "comprise a ..." do not exclude the existence of other identical elements in the process, method, article or device including the elements.

[0044] Although embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that various changes, modifications, substitutions and variations may be made to the embodiments without departing from the principles and spirit of the present invention, and that the scope of the present invention is defined by the appended claims and their equivalents.

Claims

1. A method for preparing a metal organic coordination network, comprising the following steps: S1, dissolving copper acetate hydrate and polyphenols having glycolysis inhibition function in deionized water or dimethyl sulfoxide to form a copper acetate solution and a polyphenol solution, respectively, then adding the copper acetate solution dropwise to the polyphenol solution under continuous stirring, passing nitrogen gas and heating the reaction at the same time until the reaction is completely completed, to obtain a reaction mixture; S2. The reaction mixture prepared in step S1 is transferred into a centrifuge tube, centrifuged, washed, and finally centrifuged to obtain a precipitated product, a metal organic coordination network CuMON.

2. The preparation method according to claim 1, characterized in that: The polyphenol having the function of inhibiting glycolysis is epigallocatechin gallate, quercetin or epigallocatechin.

3. The preparation method according to claim 1 or 2, characterized in that: In step S1, the mass ratio of the copper acetate hydrate to the polyphenol is 30-80:50-250, the concentration of the copper acetate solution is 3 mg / ml-8 mg / ml, and the concentration of the polyphenol solution is 3.125 mg / ml-15.625 mg / ml; the heating reaction temperature is 85°C, and the reaction time is 12-24 h.

4. The preparation method according to claim 1 or 2, characterized in that: In step S2, the centrifugation and washing refers to first centrifuging the reaction mixture in a centrifuge tube, and then washing the product after centrifugation twice with deionized water or first washing it twice with methanol and then washing it twice with deionized water for purification; the centrifugal speed is 12000 g to 20000 g, and the centrifugation time is 5 to 10 minutes.

5. The metal organic coordination network material prepared by the preparation method according to any one of claims 1 to 4.

6. The metal organic coordination network material according to claim 5, characterized in that: The diameter of the metal organic coordination network is 80-100 nm.

7. A metal organic coordination network coating, obtained by coating a tumor cell membrane with the metal organic coordination network according to claim 5 or 6.

8. The metal-organic coordination network coating according to claim 7, characterized in that: The tumor cell membrane is a CT26 cell membrane.

9. Use of the metal organic coordination network material according to claim 5 or 6 in the preparation of a drug for targeted killing of hypoxic tumor cells.

10. Use of the metal organic coordination network coating according to claim 7 or 8 in the preparation of a drug for targeted killing of hypoxic tumor cells.

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