Copper coordination polymer as well as preparation method and application thereof

Abstract

The invention discloses a copper coordination polymer as well as a preparation method and application thereof, and belongs to the technical field of antitumor drugs. The structure of the copper coordination polymer is shown as a formula I in the specification. The copper coordination polymer is used as a carrier to carry the responsive nanoparticles formed by hexaacetonitrile and ferrous tetrafluoroborate, so that the self-tracing of copper death in tumor cells can be realized.

Description

Technical Field

[0001] The present invention relates to the technical field of anti-tumor drugs, and in particular to a copper coordination polymer and a preparation method and application thereof. Background Art

[0002] Currently, cancer treatments mainly include surgery, radiotherapy, and chemotherapy. Among them, chemotherapy is a very important means of cancer treatment. Studies have found that the use of a single chemotherapy drug can easily induce the development of drug resistance in tumor cells, and has disadvantages such as large toxic side effects and limited treatment options, which greatly reduces the therapeutic effect. Combination therapy, that is, the simultaneous use of two or more drugs with different anti-cancer mechanisms, can effectively overcome the shortcomings of single chemotherapy drug treatment, reduce the development of cancer cell resistance, reduce toxic side effects, and enhance the therapeutic effect through the synergistic effect of different drugs. Therefore, the development of new chemotherapy drug combination therapies has important research and clinical practical significance.

[0003] Copper is a trace element in the human body and is closely associated with various signaling pathways and even tumor-related biological behaviors. Excess copper can cause cell death, and the mechanism and specific form of copper-induced cell death have long been unclear. A study in early 2022 demonstrated that copper ionization is an independent form of cell death. Copper ionophores, lipid-soluble molecules that reversibly bind copper ions, have played a crucial role in the discovery and research of copper ionization. Furthermore, as anti-tumor drugs, copper ionophores can modulate intracellular copper ion concentrations to induce copper ionization. The anti-tumor applications of various copper ionophores have been explored, but their current animal application and clinical development are limited by two major issues. First, the precise mechanism of copper ionization remains undefined, and the drug action process is not visualizable, making it difficult to effectively track the distribution and release of copper ionization drugs. Second, the stability of existing copper ionophores is problematic. Traditionally used copper complexes often exhibit extreme stability. Some complexes are prone to dissociation, potentially causing systemic toxicity after administration, while others are overly stable, resulting in slow release and poor therapeutic efficacy.

[0004] Therefore, it is urgent to develop a copper-loaded drug that has both long-term stability, sensitive release ability, and imaging tracing ability. Summary of the Invention

[0005] In view of this, the technical problem to be solved by the present invention is to provide a copper coordination polymer and its preparation method and application. The copper coordination polymer is used as a carrier to support responsive nanoparticles formed by hexaacetonitrile and ferrous tetrafluoroborate, which can achieve self-tracing of copper death in tumor cells.

[0006] In order to achieve the above purpose, the technical solution adopted by the present invention is as follows:

[0007] The present invention provides a copper coordination polymer, the structure of which is shown in Formula I:

[0008]

[0009] The present invention also provides a method for preparing the above-mentioned copper coordination polymer, comprising the following steps:

[0010] The complex represented by formula III and the monopolyethylene glycol monomethyl ether phosphate represented by formula IV are ultrasonically reacted to prepare the copper coordination polymer represented by formula I;

[0011]

[0012] The copper coordination polymer represented by the above formula I is prepared by surface-modifying copper 5,5',5"-(benzene-1,3,5-triyltris(acetylene-2,1-diyl))triisophthalate with monopolyethylene glycol monomethyl ether phosphate.

[0013] Preferably, the mass ratio of the complex represented by Formula III to the monopolyethylene glycol monomethyl ether phosphate represented by Formula IV is 1:(0.5-20); more preferably 1:(1-10). In some specific embodiments of the present invention, the mass ratio is preferably 1:1, 1:3, 1:5, or 1:10.

[0014] Preferably, the ultrasonic power of the ultrasonic reaction is 5 to 50 W; more preferably 25 W.

[0015] Preferably, the temperature of the ultrasonic reaction is 0-20°C; more preferably 10-20°C.

[0016] Preferably, the ultrasonic reaction time is 5 to 96 hours, more preferably 6 hours.

[0017] Preferably, the method for preparing the complex represented by formula III of the present invention comprises the following steps:

[0018] Copper nitrate dihydrate, 5,5',5"-(benzene-1,3,5-triyltris(acetylene-2,1-diyl))triisophthalic acid and an acidic end-capping agent are mixed and reacted to prepare a complex represented by formula III.

[0019] Preferably, the molar ratio of the 5,5',5"-(benzene-1,3,5-triyltris(acetylene-2,1-diyl))triisophthalic acid to copper nitrate disesquihydrate is 1:(1-30); more preferably 1:(2-20). In some specific embodiments of the present invention, the molar ratio is preferably 1:8.29 or 1:4.15 or 1:2.08 or 1:16.58.

[0020] Preferably, the acidic end-capping agent is selected from one or more of tetrafluoroboric acid, phosphoric acid, trifluoroacetic acid, acetic acid, and benzoic acid; more preferably, it is one or more of tetrafluoroboric acid, phosphoric acid, acetic acid, and benzoic acid; further preferably, it is tetrafluoroboric acid and phosphoric acid, or phosphoric acid, or acetic acid, or tetrafluoroboric acid, or benzoic acid.

[0021] Preferably, the temperature of the mixing reaction is 80°C-120°C; more preferably 80°C or 120°C.

[0022] In the method for preparing the complex represented by the above formula III, the mixing reaction can be a direct reaction or a reaction under microwave power irradiation.

[0023] The power of the microwave irradiation is preferably 30-50W.

[0024] The preparation method of the monopolyethylene glycol monomethyl ether phosphate represented by the above formula IV comprises the following steps:

[0025] Phosphorus oxychloride and polyethylene glycol monomethyl ether shown in formula VI are mixed and reacted under Lewis acid catalysis to obtain phosphoric acid monopolyethylene glycol monomethyl ether ester shown in formula IV.

[0026]

[0027] The solvent for the reaction is selected from low-temperature solvents, including but not limited to dichloromethane, chloroform, acetonitrile, ethyl acetate, and the like.

[0028] The Lewis acids include, but are not limited to, aluminum trichloride, gallium trichloride, indium trichloride, titanium tetrachloride, and the like. The copper coordination polymer of the present invention, as a carrier, has the advantages of high porosity and high specific surface area. Its internal pore diameter is 2 to 3 nm, enabling it to effectively load small molecule catalysts while isolating interference from macromolecules such as polysaccharides and proteins.

[0029] The present invention also provides a responsive nanoparticle, which is composed of a carrier and hexaacetonitrile ferrous tetrafluoroborate supported thereon;

[0030] The carrier is the copper coordination polymer mentioned above or the copper coordination polymer prepared by the above preparation method.

[0031] Preferably, the hexaacetonitrile ferrous tetrafluoroborate is prepared by reacting reduced iron powder and nitrosyl tetrafluoroborate under an inert atmosphere.

[0032] The present invention also provides an anti-tumor drug comprising the above-mentioned responsive nanoparticles.

[0033] Since the responsive nanoparticles have a very strong intrinsic Raman signal, and the signal is in the range of 1800-2500 cm -1The bioorthogonal region of the nanoparticles allows the nanoparticles to be used as Raman tracers. Furthermore, the Raman signal of the nanoparticles changes with the release of copper ions, enabling self-tracing of drug release via Raman imaging.

[0034] The responsive nanoparticles can introduce copper ions into tumor cells and, under the highly oxidative environment within the tumor cells, oxidatively break the acetylenic groups in the 5,5',5"-(benzene-1,3,5-triyltris(acetylene-2,1-diyl))triisophthalic acid in the inner layer of the responsive nanoparticles, thereby degrading the inner layer complex, releasing copper ions, mediating the copper death of the tumor cells, and achieving self-tracing of copper death.

[0035] Compared with the prior art, the copper coordination polymer provided by the present invention has a structure as shown in Formula I. The copper coordination polymer is used as a carrier to support responsive nanoparticles formed by hexaacetonitrile and ferrous tetrafluoroborate, which can achieve self-tracing of copper death in tumor cells. BRIEF DESCRIPTION OF THE DRAWINGS

[0036] Figure 1 This is a schematic diagram of the structure of the self-tracing copper death nanoparticles of the present invention;

[0037] Figure 2 The X-ray diffraction pattern (measured) and computer-simulated diffraction pattern (simulated) of 5,5',5"-(benzene-1,3,5-triyltris(acetylene-2,1-diyl))triisophthalate copper prepared in Example 1 are shown.

[0038] Figure 3 The X-ray diffraction pattern (measured) and computer-simulated diffraction pattern (simulated) of 5,5',5"-(benzene-1,3,5-triyltris(acetylene-2,1-diyl))triisophthalate copper prepared in Example 2;

[0039] Figure 4 The X-ray diffraction pattern (measured) and computer-simulated diffraction pattern (simulated) of 5,5',5"-(benzene-1,3,5-triyltris(acetylene-2,1-diyl))triisophthalate copper prepared in Example 3 of the present invention are shown;

[0040] Figure 5 The X-ray diffraction pattern (measured) and computer-simulated diffraction pattern (simulated) of 5,5',5"-(benzene-1,3,5-triyltris(acetylene-2,1-diyl))triisophthalate copper prepared in Example 4;

[0041] Figure 6The X-ray diffraction pattern (measured) and computer-simulated diffraction pattern (simulated) of 5,5',5"-(benzene-1,3,5-triyltris(acetylene-2,1-diyl))triisophthalate copper prepared in Example 5;

[0042] Figure 7 The X-ray diffraction pattern (measured) and computer-simulated diffraction pattern (simulated) of 5,5',5"-(benzene-1,3,5-triyltris(acetylene-2,1-diyl))triisophthalate copper prepared in Example 6;

[0043] Figure 8 This is the hydrogen nuclear magnetic resonance spectrum of the monopolyethylene glycol monomethyl ether phosphate prepared in Example 11;

[0044] Figure 9 This is the nuclear magnetic resonance phosphorus spectrum of monopolyethylene glycol monomethyl ether phosphate prepared in Example 11;

[0045] Figure 10 This is the H NMR spectrum of hexaacetonitrile ferrous tetrafluoroborate prepared in Example 14;

[0046] Figure 11 This is the thermogravimetric loss curve of hexaacetonitrile ferrous tetrafluoroborate prepared in Example 14;

[0047] Figure 12 This is the X-ray photoelectron spectrum of hexaacetonitrile ferrous tetrafluoroborate prepared in Example 16;

[0048] Figure 13 This is a transmission electron micrograph of the self-tracing copper death nanomedicine prepared in Example 3;

[0049] Figure 14 Dynamic light scattering data for some of the products in Examples 17-66, specifically Examples 17, 19, 21, 30, 31, 39, 40, 43, 48, 49, 52, 57, 58, 61, and 66;

[0050] Figure 15 Cytotoxicity test data of the self-tracing copper-death nanomedicine prepared in Example 17 (left) and Example 18 (right) after incubation with 4T1 cells for 24 hours;

[0051] Figure 16 This is a Raman spectrum of copper 5,5',5"-(benzene-1,3,5-triyltris(acetylene-2,1-diyl))triisophthalate prepared in Example 1, Example 19, Example 22, and Example 30;

[0052] Figure 17Western Blot images of Example 17 (No. 1), Example 1 (No. 2), Example 14 (No. 3), and PBS control (No. 4);

[0053] Figure 18 Microscopic images of the self-tracing copper nanoparticle drug prepared in Example 17, where the left image is an optical microscope image and the right image is a Raman image of the same position;

[0054] Figure 19 This is a graph showing the percentage of copper ion release of the self-tracing copper death nanodrug prepared in Example 17 in different simulated microenvironments and time points. DETAILED DESCRIPTION

[0055] To further illustrate the present invention, the copper coordination polymer provided by the present invention, its preparation method and application are described in detail below with reference to the examples.

[0056] The structure of the following 5,5',5"-(benzene-1,3,5-triyltris(acetylene-2,1-diyl))triisophthalate copper is shown in Formula III:

[0057]

[0058] The structure of the following hexaacetonitrile ferrous tetrafluoroborate is shown in Formula II:

[0059]

[0060] Example 1

[0061] Weigh 1 g of 5,5',5"-(benzene-1,3,5-triyltris(acetylene-2,1-diyl))triisophthalic acid and 3 g of copper nitrate dihydrate into a reaction flask. Add 300 mL of N,N-dimethylformamide (DMF) and stir vigorously until completely dissolved. Raise the temperature to 80°C, then add 0.8 g of phosphoric acid and 0.8 g of tetrafluoroboric acid. Continue stirring for 72 hours before cooling to room temperature. Centrifuge the resulting mixture at high speed, discard the supernatant, and re-disperse the solid with methanol. Stir vigorously for 1 hour, centrifuge again, discard the supernatant, and re-disperse the precipitate with methanol. Repeat this process five or more times until no DMF is detected in the supernatant. Dry the precipitate in a vacuum at 80°C for 3 hours. This yields a blue powdery copper complex with the structure of Formula III.

[0062] The copper 5,5',5"-(benzene-1,3,5-triyltris(acetylene-2,1-diyl))triisophthalate obtained above was tested. Figure 2This is the X-ray diffraction pattern of 5,5',5"-(benzene-1,3,5-triyltris(acetylene-2,1-diyl))trisisophthalate copper prepared in Example 1. The results show that 5,5',5"-(benzene-1,3,5-triyltris(acetylene-2,1-diyl))trisisophthalate copper was successfully synthesized and has the structure of Formula III.

[0063] Example 2

[0064] Weigh 1 g of 5,5',5"-(benzene-1,3,5-triyltris(acetylene-2,1-diyl))triisophthalic acid and 3 g of copper nitrate dihydrate into a reaction flask. Add 300 mL of N,N-dimethylformamide (DMF) to the mixture. Stir vigorously until completely dissolved. Raise the temperature to 80°C, then add 1.6 g of phosphoric acid. Continue stirring for 72 hours before cooling to room temperature. Centrifuge the resulting mixture at high speed, discard the supernatant, and re-disperse the solid with methanol. Stir vigorously for 1 hour, centrifuge again, discard the supernatant, and re-disperse the precipitate with methanol. Repeat this process five or more times until no DMF is detected in the supernatant. Dry the precipitate under vacuum at 80°C for 3 hours. This yields a blue powdery copper complex with the structure of Formula III.

[0065] The copper 5,5',5"-(benzene-1,3,5-triyltris(acetylene-2,1-diyl))triisophthalate obtained above was tested. Figure 3 This is the X-ray diffraction pattern of 5,5',5"-(benzene-1,3,5-triyltris(acetylene-2,1-diyl))trisisophthalate copper prepared in Example 2. The results show that the 5,5',5"-(benzene-1,3,5-triyltris(acetylene-2,1-diyl))trisisophthalate copper has the structure of Formula III.

[0066] Example 3

[0067] Weigh 1 g of 5,5',5"-(benzene-1,3,5-triyltris(acetylene-2,1-diyl))triisophthalic acid and 3 g of copper nitrate dihydrate into a reaction flask. Add 300 mL of N,N-dimethylformamide (DMF) to the mixture. Stir vigorously until completely dissolved. Raise the temperature to 80°C, then add 1.6 g of acetic acid. Continue stirring for 72 hours before cooling to room temperature. Centrifuge the resulting mixture at high speed, discard the supernatant, and re-disperse the solid with methanol. Stir vigorously for 1 hour, centrifuge again, discard the supernatant, and re-disperse the precipitate with methanol. Repeat this process five or more times until no DMF is detected in the supernatant. Dry the precipitate under vacuum at 80°C for 3 hours. This yields a blue powdery copper complex with the structure of Formula III.

[0068] The copper 5,5',5"-(benzene-1,3,5-triyltris(acetylene-2,1-diyl))triisophthalate obtained above was tested. Figure 4 This is the X-ray diffraction pattern of 5,5',5"-(benzene-1,3,5-triyltris(acetylene-2,1-diyl))trisisophthalate copper prepared in Example 3. The results show that the 5,5',5"-(benzene-1,3,5-triyltris(acetylene-2,1-diyl))trisisophthalate copper has the structure of Formula III.

[0069] Example 4

[0070] Weigh 1 g of 5,5',5"-(benzene-1,3,5-triyltris(acetylene-2,1-diyl))triisophthalic acid and 3 g of copper nitrate dihydrate into a reaction flask. Add 300 mL of N,N-dimethylformamide (DMF) to the mixture. Stir vigorously until completely dissolved. Raise the temperature to 80°C, then add 1.6 g of tetrafluoroboric acid. Continue stirring for 72 hours before cooling to room temperature. Centrifuge the resulting mixture at high speed, discard the supernatant, and re-disperse the solid with methanol. Stir vigorously for 1 hour, centrifuge again, discard the supernatant, and re-disperse the precipitate with methanol. Repeat this process five or more times until no DMF is detected in the supernatant. Dry the precipitate under vacuum at 80°C for 3 hours. This yields a blue powdery copper complex with the structure of Formula III.

[0071] The copper 5,5',5"-(benzene-1,3,5-triyltris(acetylene-2,1-diyl))triisophthalate obtained above was tested. Figure 5 This is the X-ray diffraction pattern of 5,5',5"-(benzene-1,3,5-triyltris(acetylene-2,1-diyl))trisisophthalate copper prepared in Example 4. The results show that the 5,5',5"-(benzene-1,3,5-triyltris(acetylene-2,1-diyl))trisisophthalate copper has the structure of Formula III.

[0072] Example 5

[0073] Weigh 1 g of 5,5',5"-(benzene-1,3,5-triyltris(acetylene-2,1-diyl))triisophthalic acid and 3 g of copper nitrate dihydrate into a reaction flask. Add 300 mL of N,N-dimethylformamide (DMF) to the mixture. Stir vigorously until completely dissolved. Raise the temperature to 80°C, then add 1.6 g of benzoic acid. Continue stirring for 72 hours before cooling to room temperature. Centrifuge the resulting mixture at high speed, discard the supernatant, and re-disperse the solid with methanol. Stir vigorously for 1 hour, centrifuge again, discard the supernatant, and re-disperse the precipitate with methanol. Repeat this process five or more times until no DMF is detected in the supernatant. Dry the precipitate under vacuum at 80°C for 3 hours. This yields a blue powdery copper complex with the structure of Formula III.

[0074] The copper 5,5',5"-(benzene-1,3,5-triyltris(acetylene-2,1-diyl))triisophthalate obtained above was tested. Figure 6 This is the X-ray diffraction pattern of 5,5',5"-(benzene-1,3,5-triyltris(acetylene-2,1-diyl))trisisophthalate copper prepared in Example 5. The results show that the 5,5',5"-(benzene-1,3,5-triyltris(acetylene-2,1-diyl))trisisophthalate copper has the structure of Formula III.

[0075] Example 6

[0076] Weigh 0.1 g of 5,5',5"-(benzene-1,3,5-triyltris(acetylene-2,1-diyl))triisophthalic acid and 0.3 g of copper nitrate dihydrate into a reaction flask. Add 30 mL of N,N-dimethylformamide (DMF) and stir vigorously until completely dissolved. Raise the temperature to 80°C, then add 0.08 g of phosphoric acid and 0.08 g of tetrafluoroboric acid. Irradiate the reaction tube with a microwave power of 40 W and heat to 120°C for 2 h. Cool to room temperature. Centrifuge the resulting mixture, discard the supernatant, and re-disperse the solid with methanol. Stir vigorously for 1 h, centrifuge again, discard the supernatant, and re-disperse the precipitate with methanol. Repeat this process five or more times until no DMF is detected in the supernatant. Dry the precipitate in a vacuum at 80°C for 3 h. A blue powder of the copper complex with the structure of Formula III is obtained.

[0077] The copper 5,5',5"-(benzene-1,3,5-triyltris(acetylene-2,1-diyl))triisophthalate obtained above was tested. Figure 7 This is the X-ray diffraction pattern of 5,5',5"-(benzene-1,3,5-triyltris(acetylene-2,1-diyl))trisisophthalate copper prepared in Example 6. The results show that the 5,5',5"-(benzene-1,3,5-triyltris(acetylene-2,1-diyl))trisisophthalate copper has the structure of Formula III.

[0078] Example 7

[0079] Weigh 0.1 g of 5,5',5"-(benzene-1,3,5-triyltris(acetylene-2,1-diyl))triisophthalic acid and 0.3 g of copper nitrate dihydrate into a reaction flask. Add 30 mL of N,N-dimethylformamide (DMF) and stir vigorously until completely dissolved. Raise the temperature to 80°C, then add 0.16 g of phosphoric acid. Irradiate the reaction tube with a 40 W microwave power and raise the temperature to 120°C for 2 hours. Cool to room temperature. Centrifuge the resulting mixture, discard the supernatant, and re-disperse the solid with methanol. Stir vigorously for 1 hour, centrifuge again, discard the supernatant, and re-disperse the precipitate with methanol. Repeat this process five or more times until no DMF is detected in the supernatant. Dry the precipitate in a vacuum at 80°C for 3 hours to obtain a blue powdery copper complex with the structure of Formula III.

[0080] Example 8

[0081] Weigh 0.1 g of 5,5',5"-(benzene-1,3,5-triyltris(acetylene-2,1-diyl))triisophthalic acid and 0.3 g of copper nitrate dihydrate into a reaction flask. Add 30 mL of N,N-dimethylformamide (DMF) and stir vigorously until completely dissolved. Raise the temperature to 80°C, then add 0.16 g of tetrafluoroboric acid. Irradiate the reaction tube with a microwave power of 40 W and heat to 120°C for 2 h. Cool to room temperature. Centrifuge the resulting mixture, discard the supernatant, and re-disperse the solid with methanol. Stir vigorously for 1 h, centrifuge again, discard the supernatant, and re-disperse the precipitate with methanol. Repeat this process five or more times until no DMF is detected in the supernatant. Dry the precipitate in a vacuum at 80°C for 3 h. This yields a blue powdery copper complex with the structure of Formula III.

[0082] Example 9

[0083] Weigh 0.1 g of 5,5',5"-(benzene-1,3,5-triyltris(acetylene-2,1-diyl))triisophthalic acid and 0.3 g of copper nitrate dihydrate into a reaction flask. Add 30 mL of N,N-dimethylformamide (DMF) and stir vigorously until completely dissolved. Raise the temperature to 80°C, then add 0.16 g of acetic acid. Irradiate the reaction tube with a microwave power of 40 W and heat to 120°C for 2 h. Cool to room temperature. Centrifuge the resulting mixture, discard the supernatant, and re-disperse the solid with methanol. Stir vigorously for 1 h, centrifuge again, discard the supernatant, and re-disperse the precipitate with methanol. Repeat this process five or more times until no DMF is detected in the supernatant. Dry the precipitate in a vacuum at 80°C for 3 h. This yields a blue powdery copper complex with the structure of Formula III.

[0084] Example 10

[0085] Weigh 0.1 g of 5,5',5"-(benzene-1,3,5-triyltris(acetylene-2,1-diyl))triisophthalic acid and 0.3 g of copper nitrate dihydrate into a reaction flask. Add 30 mL of N,N-dimethylformamide (DMF) and stir vigorously until completely dissolved. Raise the temperature to 80°C, then add 0.16 g of benzoic acid. Irradiate the reaction tube with a microwave power of 40 W and heat to 120°C for 2 h. Cool to room temperature. Centrifuge the resulting mixture, discard the supernatant, and re-disperse the solid with methanol. Stir vigorously for 1 h, centrifuge again, discard the supernatant, and re-disperse the precipitate with methanol. Repeat this process five or more times until no DMF is detected in the supernatant. Dry the precipitate in a vacuum at 80°C for 3 h. This yields a blue powdery copper complex with the structure of Formula III.

[0086] In the above Examples 1-10, 5,5',5"-(benzene-1,3,5-triyltris(acetylene-2,1-diyl))triisophthalate copper of Formula III was synthesized using different end-capping agents in different proportions and different synthesis methods.

[0087] The above-mentioned preparation of 5,5',5"-(benzene-1,3,5-triyltris(acetylene-2,1-diyl))trisisophthalate copper was performed using two different methods: a solvothermal method and a microwave synthesis method. The difference between the two methods is that the solvothermal method is more time-consuming, but the temperature is lower, and a larger amount of product, namely 5,5',5"-(benzene-1,3,5-triyltris(acetylene-2,1-diyl))trisisophthalate copper, can be synthesized in a single batch. The microwave synthesis method, on the other hand, has a faster synthesis rate and shorter preparation time, but is limited by the uniformity of microwave irradiation, and can only synthesize grams of product in a single batch. The amount of product synthesized corresponds to the amount of raw materials used, that is, when the amount of raw materials used is low, the amount of product synthesized is also low.

[0088] Example 11

[0089] Weigh 10 g of polyethylene glycol monomethyl ether (molecular weight 5000), use toluene-azeotropic dehydration treatment, add 200 mL of anhydrous dichloromethane to dissolve. Weigh 1.59 g of anhydrous indium trichloride, stir and disperse to obtain a mixture, and cool the entire reaction flask with an ice bath. Take another 0.92 g of phosphorus oxychloride, dissolve it in 50 mL of anhydrous dichloromethane, and add it dropwise to the dichloromethane solution of polyethylene glycol monomethyl ether in an ice bath. After stirring and reacting for 4 hours, the reaction flask is warmed to room temperature and the reaction is continued for 12 hours.

[0090] After the reaction is completed, the mixture is precipitated with ether and filtered to obtain a flocculent precipitate. The precipitate is redissolved with water, and the pH of the aqueous solution is adjusted to 7 with sodium bicarbonate. Indium hydroxide precipitate appears in the solution, and the precipitate is discarded by centrifugation. The supernatant is dialyzed with ultrapure water and freeze-dried to obtain a white powder product having a structure of formula IV, monopolyethylene glycol monomethyl ether phosphate.

[0091] The monopolyethylene glycol monomethyl ether phosphate obtained above was tested. Figure 8 This is the hydrogen nuclear magnetic resonance spectrum of the monopolyethylene glycol monomethyl ether phosphate prepared in Example 11. Figure 9 This is the nuclear magnetic resonance phosphorus spectrum of the monopolyethylene glycol monomethyl ether phosphate prepared in Example 11. The results show that the monopolyethylene glycol monomethyl ether phosphate was successfully synthesized and has the structure of Formula IV.

[0092] Example 12

[0093] Weigh 10 g of polyethylene glycol monomethyl ether (molecular weight 5000), use toluene-azeotropic dehydration treatment, add 200 mL of anhydrous dichloromethane to dissolve. Weigh 0.96 g of anhydrous aluminum chloride, stir and disperse to obtain a mixture, and cool the entire reaction flask with an ice bath. Take another 0.92 g of phosphorus oxychloride, dissolve it in 50 mL of anhydrous dichloromethane, and add it dropwise to the dichloromethane solution of polyethylene glycol monomethyl ether in an ice bath. Keep stirring and react for 4 hours, then raise the reaction flask to room temperature and continue the reaction for 12 hours.

[0094] After the reaction is completed, the mixture is precipitated with ether and filtered to obtain a flocculent precipitate. The precipitate is redissolved with water, and the pH of the aqueous solution is adjusted to 7 with sodium bicarbonate. Aluminum hydroxide precipitate appears in the solution, and the precipitate is discarded by centrifugation. The supernatant is dialyzed with ultrapure water and freeze-dried to obtain a white powder product having the structure of Formula IV, monopolyethylene glycol monomethyl ether phosphate.

[0095] Example 13

[0096] Weigh 10g of polyethylene glycol monomethyl ether (molecular weight 5000), treat with toluene azeotropic dehydration, and dissolve in 200ml of anhydrous dichloromethane. Weigh 1.27g of anhydrous gallium trichloride, stir and disperse to obtain a mixture, and cool the entire reaction flask in an ice bath. Separately, dissolve 0.92g of phosphorus oxychloride in 50ml of anhydrous dichloromethane, then add dropwise to the polyethylene glycol monomethyl ether dichloromethane solution in an ice bath. Stir and react for 4 hours, then warm the reaction flask to room temperature and continue the reaction for 12 hours.

[0097] After the reaction is completed, the mixture is precipitated with ether and filtered to obtain a flocculent precipitate. The precipitate is redissolved with water, and the pH of the aqueous solution is adjusted to 7 with sodium bicarbonate. Aluminum hydroxide precipitate appears in the solution, and the precipitate is discarded by centrifugation. The supernatant is dialyzed with ultrapure water, and the solution is dialyzed with ultrapure water and freeze-dried to obtain a white powder product having a structure of formula IV, monopolyethylene glycol monomethyl ether phosphate.

[0098] The above Examples 11-13 synthesized monopolyethylene glycol monomethyl ether phosphate of formula IV.

[0099] Example 14

[0100] Weigh 1.12g of reduced iron powder, add 100mL of acetonitrile to a container, disperse with mechanical stirring, cool the entire reaction flask with an ice bath, introduce nitrogen bubbling, slowly add 1.17g of nitrosyl tetrafluoroborate, maintain stirring and nitrogen bubbling for 12h, terminate the reaction, increase the nitrogen flow to blow away the exhaust gas. Use a magnet and centrifuge to remove all unreacted solids, vacuum dry the resulting liquid, and purify the resulting solid by recrystallization with ultrapure water to obtain a white powdery product having the structure of formula II, hexaacetonitrile combined with ferrous tetrafluoroborate.

[0101] The hexaacetonitrile ferrous tetrafluoroborate obtained above was tested. Figure 10 This is the H NMR spectrum of the hexaacetonitrile ferrous tetrafluoroborate prepared in Example 14. The results show that the hexaacetonitrile ferrous tetrafluoroborate has the structure of Formula II. Figure 11 This is the thermogravimetric curve of hexaacetonitrile ferrous tetrafluoroborate prepared in Example 14. The results show that the mass fractions of fluorine and iron in the product are consistent with the expected structure.

[0102] Example 15

[0103] Weigh 1.12g of reduced iron powder, add 50mL of acetonitrile and 50mL of tert-butyl alcohol to a container, disperse with mechanical stirring, cool the entire reaction flask with an ice bath, introduce nitrogen bubbling, slowly add 1.17g of nitrosyl tetrafluoroborate, maintain stirring and nitrogen bubbling for 12h, terminate the reaction, increase the nitrogen flow to blow away the exhaust gas. Use a magnet and centrifuge to remove all unreacted solids, vacuum dry the resulting liquid, and purify the resulting solid by recrystallization with ultrapure water to obtain a white powdery product having the structure of formula II: hexaacetonitrile combined with ferrous tetrafluoroborate.

[0104] Example 16

[0105] Weigh 1.12g of reduced iron powder, add 100mL of DMF and 10mL of acetonitrile to a container, disperse with mechanical stirring, cool the entire reaction flask with an ice bath, introduce nitrogen bubbling, slowly add 1.17g of nitrosyl tetrafluoroborate, maintain stirring and nitrogen bubbling for 12 hours, terminate the reaction, increase the nitrogen flow to blow away the exhaust gas. Use a magnet and centrifuge to remove all unreacted solids, precipitate the resulting liquid with methanol, and purify the resulting solid by recrystallization with ultrapure water to obtain a white powder product having the structure of formula II, hexaacetonitrile combined with ferrous tetrafluoroborate.

[0106] The hexaacetonitrile ferrous tetrafluoroborate obtained above was tested. Figure 12 This is the X-ray photoelectron spectrum of the hexaacetonitrile ferrous tetrafluoroborate prepared in Example 16. The results show that the hexaacetonitrile ferrous tetrafluoroborate has the structure of Formula II.

[0107] The above Examples 14-16 synthesized hexaacetonitrile ferrous tetrafluoroborate of formula II.

[0108] The following Examples 17-66 synthesized self-tracing copper death nanomedicines.

[0109] Example 17

[0110] Weigh 10 mg of 5,5',5"-(benzene-1,3,5-triyltris(acetylene-2,1-diyl))trisisophthalate copper salt synthesized in Example 1 and 30 mg of monopolyethylene glycol monomethyl ether phosphate synthesized in Example 11. Add 10 mL of ultrapure water, maintain ultrasonication at 20°C for 6 h at a power of 25 W, then add hexaacetonitrile ferrous tetrafluoroborate having the structure of Formula II synthesized in Example 14, and continue ultrasonication for 0.5 h. Dialyze and lyophilize to obtain a light blue powdery self-tracing copper apoptosis nanomedicine.

[0111] The self-tracing copper death nanomedicine obtained above was tested. Figure 13 This is a transmission electron microscope image of the self-tracing copper death nanodrug prepared in Example 17.

[0112] Example 18

[0113] Weigh 10 mg of 5,5',5"-(benzene-1,3,5-triyltris(acetylene-2,1-diyl))trisisophthalate copper salt synthesized in Example 1 and 10 mg of monopolyethylene glycol monomethyl ether phosphate synthesized in Example 11. Add 10 mL of ultrapure water, maintain ultrasonication at 20°C for 6 h, and add hexaacetonitrile ferrous tetrafluoroborate having the structure of Formula II synthesized in Example 14. Continue ultrasonication for 0.5 h. Dialyze and lyophilize to obtain a light blue powdery self-tracing copper apoptosis nanomedicine.

[0114] Example 19

[0115] Weigh 10 mg of 5,5',5"-(benzene-1,3,5-triyltris(acetylene-2,1-diyl))trisisophthalate copper salt synthesized in Example 1 and 50 mg of monopolyethylene glycol monomethyl ether phosphate synthesized in Example 11. Add 10 mL of ultrapure water, maintain ultrasonication at 20°C for 6 h, and add hexaacetonitrile ferrous tetrafluoroborate having the structure of Formula II synthesized in Example 14. Continue ultrasonication for 0.5 h. Dialyze and lyophilize to obtain a light blue powdery self-tracing copper apoptosis nanomedicine.

[0116] Example 20

[0117] Weigh 10 mg of 5,5',5"-(benzene-1,3,5-triyltris(acetylene-2,1-diyl))trisisophthalate copper salt synthesized in Example 1 and 100 mg of monopolyethylene glycol monomethyl ether phosphate synthesized in Example 11. Add 10 ml of ultrapure water, maintain ultrasonication at 20°C for 6 h at a power of 25 W. Then, add hexaacetonitrile ferrous tetrafluoroborate having the structure of Formula II synthesized in Example 14, and continue ultrasonication for 0.5 h. Dialyze and lyophilize to obtain a light blue powdery self-tracing copper apoptosis nanomedicine.

[0118] Example 21

[0119] Weigh 10 mg of 5,5',5"-(benzene-1,3,5-triyltris(acetylene-2,1-diyl))triisophthalate copper salt synthesized in Example 1, weigh 30 mg of monopolyethylene glycol monomethyl ether phosphate synthesized in Example 11, and weigh 1.9 mg of disodium hydrogen phosphate. Add 10 ml of ultrapure water, and sonicate for 6 hours at 25 W ultrasonic power and 20° C. Then, add hexaacetonitrile ferrous tetrafluoroborate having the structure of Formula II synthesized in Example 14, and sonicate for 0.5 hours. Dialyze and lyophilize to obtain a light blue powdery self-tracing copper apoptosis nanomedicine.

[0120] The above Examples 17-21 synthesized self-tracing copper death nanomedicines.

[0121] Examples 22 to 30

[0122] 10 mg of 5,5',5"-(benzene-1,3,5-triyltris(acetylene-2,1-diyl))triisophthalate copper synthesized in Examples 2, 3, 4, 5, 6, 7, 8, 9, and 10, respectively, and 30 mg of monopolyethylene glycol monomethyl ether phosphate synthesized in Example 12 were weighed, 10 ml of ultrapure water was added, and the ultrasonic reaction was continued for 6 hours at an ultrasonic power of 25 W and a temperature of 20 degrees Celsius. At this time, hexaacetonitrile tetrafluoroborate ferrous acid having the structure of Formula II synthesized in Example 14 was added, and ultrasonication was continued for 0.5 hours. Dialysis was performed and lyophilization was performed to obtain a light blue powdery product, a self-tracing copper death nanodrug.

[0123] Examples 31 to 39

[0124] Weigh 10 mg of 5,5',5"-(benzene-1,3,5-triyltris(acetylene-2,1-diyl))triisophthalate copper synthesized in Examples 2, 3, 4, 5, 6, 7, 8, 9, and 10, respectively, and weigh 10 mg of monopolyethylene glycol monomethyl ether phosphate synthesized in Example 12. Add 10 ml of ultrapure water, control the ultrasonic power at 25 W, and continue ultrasonic reaction for 6 hours under the condition of controlling the temperature at 20 degrees Celsius. At this time, add hexaacetonitrile tetrafluoroborate ferrous acid having the structure of Formula II synthesized in Example 14, and continue ultrasonication for 0.5 hours. Dialyze and freeze-dry to obtain a light blue powder product self-tracing copper death nanodrug.

[0125] Examples 40 to 48

[0126] Weigh 10 mg of 5,5',5"-(benzene-1,3,5-triyltris(acetylene-2,1-diyl))triisophthalate copper synthesized in Examples 2, 3, 4, 5, 6, 7, 8, 9, and 10, respectively, and weigh 50 mg of monopolyethylene glycol monomethyl ether phosphate synthesized in Example 12. Add 10 ml of ultrapure water, control the ultrasonic power at 25 W, and continue ultrasonic reaction for 6 hours under the condition of controlling the temperature at 20 degrees Celsius. At this time, add hexaacetonitrile tetrafluoroborate ferrous acid having the structure of Formula II synthesized in Example 14, and continue ultrasonication for 0.5 hours. Dialyze and freeze-dry to obtain a light blue powder product self-tracing copper death nanodrug.

[0127] Examples 49 to 57

[0128] Weigh 10 mg of 5,5',5"-(benzene-1,3,5-triyltris(acetylene-2,1-diyl))triisophthalate copper synthesized in Examples 2, 3, 4, 5, 6, 7, 8, 9, and 10, respectively, and weigh 100 mg of monopolyethylene glycol monomethyl ether phosphate synthesized in Example 12. Add 10 ml of ultrapure water, control the ultrasonic power at 25 W, and continue ultrasonic reaction for 6 hours under the condition of controlling the temperature at 20 degrees Celsius. At this time, add hexaacetonitrile tetrafluoroborate ferrous acid having the structure of Formula II synthesized in Example 14, and continue ultrasonication for 0.5 hours. Dialyze and freeze-dry to obtain a light blue powder product self-tracing copper death nanodrug.

[0129] Examples 58 to 66

[0130] Weigh 10 mg of 5,5',5"-(benzene-1,3,5-triyltris(acetylene-2,1-diyl))triisophthalate copper synthesized in Example 2, Example 3, Example 4, Example 5, Example 6, Example 7, Example 8, Example 9, and Example 10, weigh 30 mg of the monopolyethylene glycol monomethyl ether phosphate synthesized in Example 12, and weigh 1.9 mg of disodium hydrogen phosphate, add 10 ml of ultrapure water, control the ultrasonic power to 25 W, and maintain the temperature at 20 degrees Celsius. The reaction was ultrasonically reacted for 6 hours. Hexaacetonitrile-containing ferrous tetrafluoroborate (having the structure of Formula II) synthesized in Example 14 was then added, and ultrasonication was continued for 0.5 hours. Dialysis was performed and lyophilization was performed. A light blue powdery product, the self-tracing copper-death nanodrug, was obtained. The addition of disodium hydrogen phosphate in Examples 58-66 can regulate the modification level of monopolyethylene glycol monomethyl ether phosphate, thereby affecting the blood stability and drug release time of the self-tracing copper-death nanodrug. The self-tracing copper-death nanodrug prepared with the addition of disodium hydrogen phosphate resulted in faster release.

[0131] The synthesis of the self-tracing copper death nanoparticle drug of the present invention requires the addition of three types of components during the ultrasonic reaction, namely:

[0132] 1. 5,5',5"-(benzene-1,3,5-triyltris(acetylene-2,1-diyl))triisophthalate copper synthesized according to any one of Examples 1 to 10;

[0133] 2. Monopolyethylene glycol monomethyl ether phosphate synthesized in any one of Examples 11 to 13; 3. Hexaacetonitrile ferrous tetrafluoroborate having the structure of Formula II synthesized in any one of Examples 14 to 16;

[0134] Any combination of these three types of components belongs to the embodiments.

[0135] The mass ratio of the three components in any of the above combinations is preferably 5,5',5"-(benzene-1,3,5-triyltris(acetylene-2,1-diyl))triisophthalate copper:monopolyethylene glycol monomethyl ether phosphate:hexaacetonitrile ferrous tetrafluoroborate = 1:(1-10):(0.01-0.3).

[0136] The self-tracing copper death nanoparticles obtained above were tested. Figure 14 The dynamic light scattering data of some products in Examples 17 to 66 (specifically Examples 17, 19, 21, 30, 31, 39, 40, 43, 48, 49, 52, 57, 58, 61 and 66) prove that the particle size of the self-tracing copper death nanodrug can be effectively controlled by the above method. The particle size results of the examples not listed are similar to those of Example 17.

[0137] The antitumor activity of all the above self-tracing copper-death nanomedicines was tested by cytotoxicity experiments.

[0138] 4T1 cells were seeded in 96-well plates at a density of 5000 cells / well and cultured for 24 hours. 4T1 cells were incubated for 24 hours with CuCl2 and Examples 17-21 at concentrations ranging from 43.33, 20.83, 13.89, 10.42, 6.94, 5.21, 3.47, and 2.60 μmol / L (equivalent copper concentration). The drug-containing medium was discarded, and after washing with PBS, 10 μL of CCK-8 reagent and 90 μL of culture medium were added to each well. The cells were incubated in the dark at 37°C for 1 hour, and the absorbance at 450 nm was measured using a microplate reader.

[0139] Figure 15 Cytotoxicity experiments were conducted on the self-tracing copper-death nanoparticles prepared in Examples 17 and 18 after incubation with 4T1 cells for 24 hours. The results showed that the self-tracing copper-death nanoparticles prepared in Examples 17 and 18 were able to reduce cancer cell survival rates to 43±4% and 20±6%, respectively. The cytotoxicity data for the self-tracing copper-death nanoparticles prepared in other Examples 17 to 66 ranged from 43±4% to 20±6%, demonstrating similar anticancer effects.

[0140] Raman spectroscopy experiments were performed to test the Raman imaging performance of all the above self-tracing copper death nanomedicines. Figure 16 1 and 2 are Raman spectra of Example 1, Example 19, Example 22, and Example 30. The experimental results show that Example 1 has the strongest Raman intensity, while the Raman intensities of Examples 19, 22, and 30 are slightly weaker than that of Example 1.

[0141] Western blot experiments were performed to test the ability of all the self-tracing copper-death nanomedicines to induce copper-death in tumor cells. 4T1 cells were seeded in 6-well plates at a cell density of 10 6 4T1 cells were incubated with PBS, Example 1, Example 14, and Example 17 at 43.33 μmol / L for 12 hours, and the drug-containing supernatant was discarded. The cells were then treated with cell lysis buffer for 30 minutes. Cells were scraped off using a cell scraper and the supernatant was centrifuged. Protein concentration was determined using a BCA protein assay kit. Loading buffer was added and the total protein concentration was adjusted to the desired level. Each sample was then incubated at 100°C for 15 minutes.

[0142] Proteins were developed on a 10% Bis-Tris polyacrylamide gel, with 20 μg of protein sample loaded into each lane. Separation was performed by electrophoresis at 80 V for 30 min and 120 V for 2 h. Proteins were transferred to a PVDF membrane using a gel electrophoresis apparatus. Blocking was performed for 1 h with blocking buffer. The PVDF membrane was then incubated with primary antibodies against DLAT, LIAS, and FDX1 for 12 h, washed with TBST, and then incubated with β-tubulin for 12 h. Finally, ECL chemiluminescence reagent was added, and Western blot images were acquired using a gel imaging system.

[0143] Figure 17 These are Western Blot images of Example 17 (No. 1 in the figure), Example 1 (No. 2 in the figure), Example 14 (No. 3 in the figure), and PBS control (No. 4 in the figure). The experimental results show that Example 1, Example 14, and the blank control did not cause cell copper death, while Example 17 caused cell copper death.

[0144] The responsive release performance of all the self-tracing copper-death nanomedicines described above was tested by ICP-MS. PBS solutions containing hydrogen peroxide simulating different in vivo environments were prepared, with hydrogen peroxide concentrations of 10 mM, 2 mM, 1 mM, and 0 mM, respectively. Examples 17 to 22 were mixed with the various hydrogen peroxide-simulated environments and stored at 37°C. Samples were taken at different time points. The copper ion concentration of each sample at different time points was determined by ICP-MS.

[0145] Figure 19 The percentage of copper ion release in various simulated microenvironments and time points of the self-tracing copper-death nanodrug prepared in Example 17. The experimental results show that the self-tracing copper-death nanodrug can only be degraded and release copper ions under the catalysis of a certain concentration of hydrogen peroxide.

[0146] The above embodiments are only intended to help understand the method and core concept of the present invention. It should be noted that, without departing from the principles of the present invention, a number of improvements and modifications may be made to the present invention by those skilled in the art, and such improvements and modifications also fall within the scope of protection of the claims of the present invention.

Claims

1. A copper coordination polymer, characterized in that The structure is shown in Formula I:

2. The method for preparing the copper coordination polymer according to claim 1, wherein The following steps are involved: The complex represented by formula III and the monopolyethylene glycol monomethyl ether phosphate represented by formula IV are ultrasonically reacted to prepare the copper coordination polymer represented by formula I; 3. The preparation method according to claim 2, characterized in that The mass ratio of the complex represented by formula III to the monopolyethylene glycol monomethyl ether phosphate represented by formula IV is 1:(0.5-20).

4. The preparation method according to claim 2, characterized in that The ultrasonic power of the ultrasonic reaction is 5 to 50W; The temperature of the ultrasonic reaction is 0 to 20°C; The ultrasonic reaction time is 5 to 96 hours.

5. The preparation method according to claim 2, characterized in that The preparation method of the complex represented by formula III comprises the following steps: Copper nitrate dihydrate, 5,5',5"-(benzene-1,3,5-triyltris(acetylene-2,1-diyl))triisophthalic acid and an acidic end-capping agent are mixed and reacted to prepare a complex represented by formula III.

6. The preparation method according to claim 5, characterized in that The molar ratio of the 5,5',5"-(benzene-1,3,5-triyltris(acetylene-2,1-diyl))triisophthalic acid to copper nitrate disesquihydrate is 1:(1-30).

7. The preparation method according to claim 5, characterized in that The acidic end-capping agent is selected from one or more of tetrafluoroboric acid, phosphoric acid, trifluoroacetic acid, acetic acid, and benzoic acid; The temperature of the mixing reaction is 80°C-120°C.

8. A responsive nanoparticle, characterized in that: It consists of a carrier and hexaacetonitrile ferrous tetrafluoroborate supported thereon; The carrier is the copper coordination polymer according to claim 1 or the copper coordination polymer prepared by the preparation method according to any one of claims 2 to 7.

9. The responsive nanoparticle according to claim 8, characterized in that The hexaacetonitrile ferrous tetrafluoroborate is prepared by reacting reduced iron powder and nitrosyl tetrafluoroborate under an inert atmosphere.

10. An anti-tumor drug, characterized in that: Comprising the responsive nanoparticles according to claim 8 or 9.

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