Titanium-based nanocomposite for microwave thermal diagnosis and treatment integration and preparation method and application thereof

Abstract

The application provides a titanium-based nanocomposite for microwave thermal diagnosis and treatment integration, and a preparation method and application thereof.The titanium-based nanocomposite comprises a titanium-based metal organic framework material as a core, metal nanoparticles loaded on the surface and in the channel of the titanium-based metal organic framework material, and a covalent organic framework material as a shell.The titanium-based nanocomposite has good microwave heat conversion performance under microwave irradiation, can generate active oxygen under microwave irradiation, has good microwave dynamic performance, can be used for magnetic resonance imaging, can guide tumor treatment in real time, and can be effectively applied to the biomedical field.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of titanium-based nanocomposites, and particularly relates to a titanium-based nanocomposite with tumor microwave hyperthermia, microwave dynamic therapy and magnetic resonance imaging performance, and a preparation method and application thereof. BACKGROUND

[0002] In recent years, with the continuous deterioration of the living environment, the incidence and mortality of cancer are also increasing, which has become one of the major diseases endangering human health. How to effectively prevent and treat cancer is one of the important problems faced by scientific research and clinical work. In recent years, non-invasive treatment has become a research hotspot, such as phototherapy, radiotherapy, ultrasonic therapy, microwave hyperthermia and microwave therapy.

[0003] Compared with the non-invasive treatment methods such as phototherapy and ultrasonic therapy, microwave therapy has greater penetration depth and better biological safety, and has developed rapidly in clinical application in recent years. Microwave hyperthermia has the advantages of high heating efficiency, fast heating speed, large penetration depth and no interference from bones and gases, but when a large area of tumor is ablated, it will cause damage to the surrounding normal tissues, and the tumor ablation in the warm area is not complete, which will cause tumor residue and recurrence. Therefore, single microwave hyperthermia is not enough to effectively eliminate tumors, and needs to be combined with other treatment methods.

[0004] Microwave dynamics is a treatment method that uses microwaves as energy sources to produce reactive oxygen species (ROS) with cytotoxicity, and then kill tumor cells, destroy tumor blood vessels and stimulate immune responses. Microwave dynamic therapy has the advantages of high selectivity, short treatment time and less drug resistance, and its combination with microwave hyperthermia can effectively kill the tumor residue after microwave hyperthermia and further enhance the efficacy of microwave therapy.

[0005] Under the stimulation of microwaves, microwave sensitizers can absorb and convert the energy of microwaves to produce heat or reactive oxygen species in tumor areas, reduce the toxic and side effects on normal tissues, and make microwave therapy have better application prospects. At present, the microwave sensitizers used in microwave therapy include liquid metal, copper oxide nanoparticles, bismuth-manganese metal organic framework materials and manganese metal organic framework materials, but the safety of liquid metal, copper, bismuth or manganese is poor, which affects their further application in vivo.

[0006] In traditional treatment methods, diagnosis and treatment are two relatively independent processes. The delivery of contrast agents is a big problem, and it is difficult to accurately deliver them to the target area. Excessive contrast agents will cause damage to normal tissues to some extent. In addition, the time difference between diagnosis and treatment will delay the treatment opportunity to some extent, and cannot exert the best treatment efficacy, and the media used by the two may interact with each other. SUMMARY

[0007] In order to improve the deficiencies of the prior art, the purpose of the present application is to provide a titanium-based nanocomposite with tumor microwave hyperthermia, microwave dynamic therapy and magnetic resonance imaging performance, and a preparation method and application thereof. The titanium-based nanocomposite has the characteristics of high biological safety, microwave hyperthermia, microwave dynamic therapy and magnetic resonance imaging functions. As a microwave sensitizer, the titanium-based nanocomposite can be used for microwave heat-dynamic synergistic therapy of tumors while minimizing damage to the organism; in addition, the titanium-based nanocomposite can realize magnetic resonance imaging and other functions on the basis of microwave heat-dynamic synergistic therapy, and through the organic combination of multiple functional units, the synergistic therapy of tumors can be realized under the guidance of real-time imaging, which has important significance for the clinical treatment of tumors.

[0008] The purpose of the present application is achieved by the following technical solutions:

[0009] A titanium-based nanocomposite, comprising a titanium-based metal organic framework material, metal nanoparticles and a covalent organic framework material; the metal nanoparticles are distributed on the surface and / or in the channels of the titanium-based metal organic framework material, forming a titanium-based metal organic framework material loaded with metal nanoparticles; and the covalent organic framework material is coated on the surface of the titanium-based metal organic framework material loaded with metal nanoparticles.

[0010] According to an embodiment of the present application, the metal nanoparticles are selected from at least one of metal nickel nanoparticles, metal gadolinium nanoparticles, metal manganese nanoparticles and metal iron nanoparticles.

[0011] According to an embodiment of the present application, the titanium-based metal organic framework material is prepared by using a titanium source and an organic ligand in the presence of a regulator.

[0012] According to an embodiment of the present application, the titanium source for forming the titanium-based metal organic framework material is selected from at least one of titanium isopropoxide, titanium tetrachloride and titanium acid tetra-n-butyl ester.

[0013] According to an embodiment of the present application, the organic ligand for forming the titanium-based metal organic framework material is a carboxylic acid ligand, for example, selected from at least one of terephthalic acid, 2-amino terephthalic acid, 2,5-dihydroxy terephthalic acid, 4-pyridine carboxylic acid, fumaric acid and trimesic acid.

[0014] According to an embodiment of the present application, the regulator for forming the titanium-based metal organic framework material is selected from at least one of benzoic acid, acetic acid, mercaptoacetic acid and citric acid.

[0015] According to embodiments of the present application, the molar ratio of the titanium source forming the titanium-based metal organic framework material and the organic ligand forming the titanium-based metal organic framework material is 1:0.5-10, for example 1:0.5, 1:0.6, 1:0.7, 1:0.8, 1:0.9, 1:1, 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9 or 1:10.

[0016] According to embodiments of the present application, the molar ratio of the titanium source forming the titanium-based metal organic framework material and the modulator forming the titanium-based metal organic framework material is 1:1-50, for example 1:1, 1:2, 1:3, 1:5, 1:8, 1:10, 1:12, 1:15, 1:18, 1:20, 1:22, 1:25, 1:28, 1:30, 1:32, 1:35, 1:38, 1:40, 1:42, 1:45, 1:48 or 1:50.

[0017] According to embodiments of the present application, the covalent organic framework material is prepared from a covalent organic framework monomer containing an amino functional group and a covalent organic framework monomer containing an aldehyde functional group.

[0018] According to embodiments of the present application, the covalent organic framework monomer containing an amino functional group forming the covalent organic framework material is selected from at least one of p-phenylenediamine, 1,3,5-tris(4-aminophenyl)benzene and 2,5-diethoxyterephthalic dihydrazide.

[0019] According to embodiments of the present application, the covalent organic framework monomer containing an aldehyde functional group forming the covalent organic framework material is selected from at least one of benzaldehyde, trimesaldehyde and 2,5-dimethoxybenzene-1,4-dicarboxaldehyde.

[0020] According to embodiments of the present application, the mass ratio of the covalent organic framework monomer containing an amino functional group forming the covalent organic framework material and the covalent organic framework monomer containing an aldehyde functional group forming the covalent organic framework material is 1:0.5-2, for example 1:0.5, 1:0.6, 1:0.7, 1:0.8, 1:0.9, 1:1.0, 1:1.1, 1:1.2, 1:1.3, 1:1.4, 1:1.5, 1:1.6, 1:1.7, 1:1.8, 1:1.9 or 1:2.

[0021] According to an embodiment of the present application, the metal nanoparticles are distributed in a satellite-like manner on the surface and / or in the pores of the titanium-based metal organic framework material. The "satellite-like" refers to the growth of a plurality of smaller-sized metal nanoparticles on the surface and / or in the pores of the larger-sized titanium-based metal organic framework material, and the plurality of smaller-sized metal nanoparticles are referred to as satellites, which are distributed in a satellite-like manner on the surface and / or in the pores of the titanium-based metal organic framework material.

[0022] According to an embodiment of the present application, the covalent organic framework material is coated on the surface of the composite to form a shell layer with a thickness of 2-20 nm.

[0023] According to an embodiment of the present application, the titanium-based metal organic framework material has a particle size distribution of 50-400 nm, for example, 50 nm, 60 nm, 70 nm, 80 nm, 90 nm, 100 nm, 120 nm, 150 nm, 180 nm, 200 nm, 220 nm, 250 nm, 280 nm, 300 nm, 320 nm, 340 nm, 350 nm, 380 nm, or 400 nm.

[0024] According to an embodiment of the present application, the metal nanoparticles have a particle size distribution of 1-50 nm, for example, 1 nm, 2 nm, 3 nm, 4 nm, 5 nm, 6 nm, 7 nm, 8 nm, 10 nm, 12 nm, 15 nm, 18 nm, 20 nm, 22 nm, 25 nm, 28 nm, 30 nm, 32 nm, 35 nm, 38 nm, 40 nm, 45 nm, 48 nm, or 50 nm.

[0025] According to an embodiment of the present application, the titanium-based metal organic framework material has a disc, rounded cube, or regular octahedron morphology. The morphology of the titanium-based metal organic framework material can be adjusted by selecting different adjusting agents and organic ligands.

[0026] According to an embodiment of the present application, the titanium-based nanocomposite has a core-satellite-shell structure, wherein the core refers to the titanium-based metal organic framework material as the inner core; the satellite refers to the metal nanoparticles distributed in a satellite-like manner on the surface of the titanium-based metal organic framework material; and the shell refers to the covalent organic framework material as the outer shell.

[0027] According to an embodiment of the present application, the particle size distribution of the titanium-based nanocomposite is 50-500 nm, for example, 1 nm, 2 nm, 3 nm, 4 nm, 5 nm, 6 nm, 7 nm, 8 nm, 10 nm, 12 nm, 15 nm, 18 nm, 20 nm, 22 nm, 25 nm, 28 nm, 30 nm, 32 nm, 35 nm, 38 nm, 40 nm, 45 nm, 48 nm, 50 nm, 60 nm, 70 nm, 80 nm, 90 nm, 100 nm, 120 nm, 150 nm, 180 nm, 200 nm, 220 nm, 250 nm, 280 nm, 300 nm, 320 nm, 340 nm, 350 nm, 380 nm, 400 nm, 420 nm, 450 nm, 480 nm, or 500 nm.

[0028] According to an embodiment of the present application, the mass of the metal nanoparticles accounts for 0.5-30% of the total mass of the titanium-based nanocomposite, for example, 0.5%, 0.6%, 0.8%, 1%, 1.2%, 1.5%, 1.8%, 2%, 2.5%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 12%, 15%, 18%, 20%, 22%, 25%, 28%, or 30%.

[0029] According to an embodiment of the present application, the mass of the titanium-based metal organic framework material accounts for 60-85% of the total mass of the titanium-based nanocomposite, for example, 60%, 62%, 63%, 64%, 65%, 68%, 70%, 72%, 74%, 75%, 76%, 78%, 80%, 81%, 82%, 84%, or 85%.

[0030] According to an embodiment of the present application, the mass of the covalent organic framework material accounts for 10-16.5% of the total mass of the titanium-based nanocomposite, for example, 10%, 10.5%, 11%, 11.5%, 12%, 12.5%, 13%, 13.5%, 14%, 14.5%, 15%, 15.5%, 16%, or 16.5%.

[0031] According to an embodiment of the present application, the titanium-based nanocomposite has good microwave heat conversion performance under microwave irradiation; and can generate reactive oxygen under microwave irradiation, has good microwave dynamic performance; and can be used for magnetic resonance imaging, real-time guidance of tumor treatment, and can be effectively applied to the biomedical field.

[0032] The present application also provides a preparation method of the above titanium-based nanocomposite, which comprises the following steps:

[0033] 1) dispersing a titanium source, an organic ligand, and a regulator in a solvent to perform a solvothermal reaction, to obtain a titanium-based metal organic framework material;

[0034] 2) dispersing the titanium-based metal organic framework material and the metal salt obtained in step 1) in a solvent to prepare a metal ion-adsorbed titanium-based metal organic framework material;

[0035] 3) mixing and reacting the metal ion-adsorbed titanium-based metal organic framework material obtained in step 2) and a reducing agent to prepare a metal nanoparticle-loaded titanium-based metal organic framework material;

[0036] 4) mixing and reacting the metal nanoparticle-loaded titanium-based metal organic framework material obtained in step 3) and a covalent organic framework monomer containing an amino functional group;

[0037] 5) mixing and reacting the product obtained in step 4) and a covalent organic framework monomer containing an aldehyde functional group;

[0038] 6) mixing and reacting the product obtained in step 5), the covalent organic framework monomer containing an amino functional group in an amount equal to that in step 4), the covalent organic framework monomer containing an aldehyde functional group in an amount equal to that in step 5), and a catalyst to prepare the titanium-based nanocomposite material.

[0039] According to an embodiment of the present application, in step 1), the titanium source is at least one selected from the group consisting of titanium isopropoxide, titanium tetrachloride, and tetra-n-butyl titanate, and preferably is titanium isopropoxide.

[0040] According to an embodiment of the present application, in step 1), the organic ligand is at least one selected from the group consisting of carboxylic acid ligands, for example, at least one selected from the group consisting of terephthalic acid, 2-amino terephthalic acid, 2,5-dihydroxy terephthalic acid, 4-picolinic acid, fumaric acid, and trimesic acid, and preferably is 2-amino terephthalic acid.

[0041] According to an embodiment of the present application, in step 1), the adjusting agent is at least one selected from the group consisting of benzoic acid, acetic acid, mercaptoacetic acid, and citric acid.

[0042] According to an embodiment of the present application, in step 1), the solvent is at least one selected from the group consisting of N,N-dimethylformamide, methanol, and N,N-diethylformamide, and preferably is a mixture of N,N-dimethylformamide and methanol.

[0043] According to an embodiment of the present application, in step 1), the mass concentration of the titanium source is 1-30 mg / mL; the molar ratio of the titanium source to the organic ligand is 1:0.5-10; and the molar ratio of the titanium source to the adjusting agent is 1:1-50.

[0044] According to embodiments of the present application, in step 1), the temperature of the solvothermal reaction is 120-200 °C, for example 120 °C, 130 °C, 140 °C, 150 °C, 160 °C, 170 °C, 180 °C, 190 °C or 200 °C; the time of the solvothermal reaction is 2-48 h, for example 2 h, 4 h, 8 h, 10 h, 12 h, 16 h, 18 h, 24 h, 36 h or 48 h.

[0045] According to embodiments of the present application, in step 2), the metal is selected from at least one of nickel, gadolinium, manganese and iron.

[0046] According to embodiments of the present application, in step 2), the metal salt is selected from at least one of metal chloride, metal nitrate, metal sulfate, metal acetate and metal carbonate. Exemplarily, the metal salt is selected from at least one of nickel chloride, nickel nitrate, nickel acetate, manganese chloride, manganese nitrate, iron chloride, gadolinium nitrate.

[0047] According to embodiments of the present application, in step 2), the solvent is selected from at least one of N,N-dimethylformamide, methanol and N,N-diethylformamide, preferably a mixture of N,N-dimethylformamide and methanol.

[0048] According to embodiments of the present application, in step 2), the mass ratio of the titanium-based metal-organic framework to the metal salt is 1:0.1-10, for example 1:0.1, 1:0.2, 1:0.3, 1:0.4, 1:0.5, 1:0.6, 1:0.7, 1:0.8, 1:0.9, 1:1, 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9 or 1:10.

[0049] According to embodiments of the present application, in step 2), the temperature of the mixing is room temperature, and the time of the mixing is 1-6 h.

[0050] According to embodiments of the present application, in step 2), the mass concentration of the metal salt is 5-300 mg / mL; preferably 20-50 mg / mL.

[0051] According to the experimental scheme of the present application, in step 3), the mass ratio of the reducing agent to the titanium-based metal organic framework material adsorbing metal ions is 1:0.1-50, preferably 1:1-20, for example 1:0.1, 1:0.2, 1:0.3, 1:0.4, 1:0.5, 1:0.6, 1:0.7, 1:0.8, 1:0.9, 1:1, 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9, 1:10, 1:12, 1:15, 1:18, 1:20, 1:22, 1:25, 1:28, 1:30, 1:32, 1:35, 1:38, 1:40, 1:42, 1:45, 1:48 or 1:50.

[0052] According to the experimental scheme of the present application, in step 3), the reducing agent is selected from at least one of sodium borohydride, hydrazine hydrate and lithium aluminum hydride.

[0053] According to the experimental scheme of the present application, in step 3), the temperature of the reaction is room temperature, and the time of the reaction is 10-60 s.

[0054] According to the experimental scheme of the present application, in step 3), it specifically comprises the following steps:

[0055] The titanium-based metal organic framework material adsorbing metal ions obtained in step 2) is dispersed in a solvent to obtain a mixed system 1; the reducing agent is dispersed in the same solvent to obtain a mixed system 2; the mixed system 2 is added to the mixed system 1 at a speed of 3-5 drops / s, and stirred at room temperature for 10-60 s to prepare the titanium-based metal organic framework material loaded with metal nanoparticles.

[0056] According to the experimental scheme of the present application, the ratio of the volume of the solvent in the mixed system 1 to the volume of the solvent in the mixed system 2 is 1:0.1-1.

[0057] According to the embodiment of the present application, the solvent is selected from at least one of N,N-dimethylformamide, methanol and N,N-diethylformamide, preferably a mixture of N,N-dimethylformamide and methanol.

[0058] According to the experimental scheme of the present application, in step 4), the mass ratio of the titanium-based metal organic framework material loaded with metal nanoparticles to the covalent organic framework monomer containing an amino functional group is 5-30:1, for example 5:1, 6:1, 7:1, 8:1, 9:1, 10:1, 12:1, 14:1, 15:1, 16:1, 17:1, 18:1, 19:1, 20:1, 22:1, 23:1, 24:1, 25:1, 26:1, 28:1 or 30:1.

[0059] According to the experimental scheme of the present application, in step 4), the temperature of the reaction is room temperature, and the time of the reaction is 0.5-4h.

[0060] According to the experimental scheme of the present application, in step 4), the temperature of the reaction is room temperature, and the time of the reaction is 0.5-4h.

[0061] The metal nanoparticle-loaded titanium-based metal organic framework material obtained in step 3) is dispersed in a solvent to obtain a mixture 3; the covalent organic framework monomer containing an amino functional group is dispersed in the same solvent to obtain a mixture 4; the mixture 4 is added dropwise to the mixture 3 at a speed of 20-30 drops / min, and stirred at room temperature for 0.5-4h, and the product is collected and washed.

[0062] According to the experimental scheme of the present application, the volume ratio of the solvent in the mixture 3 to the volume of the solvent in the mixture 4 is 5-30:1, for example, 5:1, 6:1, 7:1, 8:1, 9:1, 10:1, 12:1, 14:1, 15:1, 16:1, 17:1, 18:1, 19:1, 20:1, 22:1, 23:1, 24:1, 25:1, 26:1, 28:1 or 30:1.

[0063] According to the experimental scheme of the present application, the solvent is selected from acetonitrile.

[0064] According to the experimental scheme of the present application, in step 5), the mass ratio of the covalent organic framework monomer containing an amino functional group to the covalent organic framework monomer containing an aldehyde functional group is 1:0.5-2, for example, 1:0.5, 1:0.6, 1:0.7, 1:0.8, 1:0.9, 1:1.0, 1:1.1, 1:1.2, 1:1.3, 1:1.4, 1:1.5, 1:1.6, 1:1.7, 1:1.8, 1:1.9 or 1:2.

[0065] According to the experimental scheme of the present application, in step 5), the temperature of the reaction is room temperature, and the time of the reaction is 0.5-4h.

[0066] According to the experimental scheme of the present application, in step 5), the temperature of the reaction is room temperature, and the time of the reaction is 0.5-4h.

[0067] The product obtained in step 4) is dispersed in a solvent to obtain a mixture 5; the covalent organic framework monomer containing an aldehyde functional group is dispersed in the same solvent to obtain a mixture 6; the mixture 6 is added dropwise to the mixture 5 at a speed of 20-30 drops / min, and stirred at room temperature for 0.5-4h, and the product is collected and washed.

[0068] According to the experimental scheme of the present application, the volume ratio of the solvent in the mixed system 5 to the volume of the solvent in the mixed system 6 is 5:1-30:1, for example, 5:1, 6:1, 7:1, 8:1, 9:1, 10:1, 12:1, 14:1, 15:1, 16:1, 17:1, 18:1, 19:1, 20:1, 22:1, 23:1, 24:1, 25:1, 26:1, 28:1 or 30:1.

[0069] According to the experimental scheme of the present application, the solvent is selected from acetonitrile.

[0070] According to the experimental scheme of the present application, in step 6), the temperature of the reaction is room temperature, and the time of the reaction is 2-12h.

[0071] According to the experimental scheme of the present application, in step 6), the catalyst is selected from acetic acid.

[0072] According to the experimental scheme of the present application, in step 6), the reaction is carried out in the presence of a solvent, and the solvent is selected from acetonitrile.

[0073] According to the experimental scheme of the present application, the mixing is carried out under the condition of magnetic stirring, and the stirring speed is 700-1200r / min.

[0074] The present application also provides the application of the above-mentioned titanium-based nanocomposite in microwave hyperthermia.

[0075] The present application also provides the application of the above-mentioned titanium-based nanocomposite in microwave hyperthermia and nuclear magnetic resonance imaging.

[0076] The present application also provides the application of the above-mentioned titanium-based metal organic framework material in microwave hyperthermia.

[0077] The present application also provides a diagnosis and treatment reagent, which comprises the above-mentioned titanium-based nanocomposite.

[0078] According to the embodiment of the present application, the diagnosis and treatment reagent is the above-mentioned titanium-based nanocomposite.

[0079] According to the embodiment of the present application, the diagnosis and treatment reagent is used for microwave hyperthermia and nuclear magnetic resonance imaging of tumors.

[0080] According to the embodiment of the present application, the diagnosis and treatment reagent is a nanoscale microwave-responsive tumor diagnosis and treatment reagent integrating microwave hyperthermia, microwave dynamic therapy and magnetic resonance imaging.

[0081] According to the experimental scheme of the present application, the titanium-based nanocomposite or titanium-based metal organic framework material is dispersed in physiological saline to prepare a dispersion liquid with a concentration of 0-10mg / mL, and the temperature change is recorded under microwave irradiation.

[0082] According to the experimental method of the present application, the titanium-based nanocomposite or titanium-based metal organic framework material is dispersed in a buffer solution to form a dispersion solution with a concentration of 0-5 mg / mL, a detection indicator is added with or without hydrogen peroxide, and after microwave irradiation, the solution is allowed to stand for 5-180 min, and the level of active oxygen generated is detected.

[0083] According to the experimental method of the present application, the titanium-based nanocomposite is dispersed in deionized water to form a dispersion solution with a concentration of 0-10 mg / mL, and magnetic resonance imaging is performed in vitro.

[0084] According to the embodiments of the present application, the titanium-based nanocomposite has good microwave heat conversion performance under microwave irradiation, is a good microwave heat sensitization agent, and can effectively kill tumor cells.

[0085] According to the embodiments of the present application, the titanium-based nanocomposite can generate active oxygen under microwave irradiation, induce tumor cell apoptosis, is a good microwave dynamic sensitization agent, and can synergistically kill tumor cells with microwave thermotherapy.

[0086] According to the embodiments of the present application, the titanium-based nanocomposite can be used for magnetic resonance imaging, is a good contrast agent, and can guide tumor treatment in real time.

[0087] The present application has the following advantages:

[0088] The present application provides a titanium-based nanocomposite for microwave heat diagnosis and treatment integration, a preparation method and application thereof, the titanium-based nanocomposite comprising a titanium-based metal organic framework material as a core, metal nanoparticles loaded on the surface and channels of the titanium-based metal organic framework material, and a covalent organic framework material as a shell. The titanium-based nanocomposite has the following advantages:

[0089] (1) The titanium-based nanocomposite of the present application takes a titanium-based metal organic framework material with microwave response performance as the core, loads metal nanoparticles with magnetic resonance imaging performance on the surface and / or channels of the titanium-based metal organic framework material, and coats the surface of the titanium-based metal organic framework material loaded with metal nanoparticles with a covalent organic framework material, further enhancing the microwave heat-dynamic performance of the titanium-based nanocomposite.

[0090] (2) The titanium-based nanocomposite is used in the field of microwave therapy for the first time. First, the titanium-based material has good biological safety and can be widely used in the body. Second, based on the microwave absorption performance of the titanium-based metal organic framework and its porous structure, it has high microwave heat conversion efficiency under microwave irradiation, and kills tumor cells by high temperature. At the same time, titanium as a variable valence metal can effectively produce reactive oxygen in the presence of hydrogen peroxide, and microwave stimulation can further enhance its reactive oxygen production performance, induce tumor cell apoptosis, and thus perform microwave thermotherapy, microwave dynamic therapy and synergistic therapy.

[0091] (3) The titanium-based nanocomposite has good biocompatibility and controllable size, can be easily targeted to the tumor area through the high permeability and retention effect of solid tumors, and selectively kills tumor cells through microwave heat-dynamic therapy under microwave stimulation. In addition, the magnetic resonance imaging performance can be used for real-time imaging to guide tumor treatment. Finally, a microwave-responsive tumor diagnosis and treatment reagent is obtained, and the elimination of tumors can also be realized under the guidance of real-time imaging, which has good clinical application prospect. BRIEF DESCRIPTION OF DRAWINGS

[0092] Figure 1 Transmission electron microscope characterization diagram of titanium-based metal organic framework material prepared for example 1.

[0093] Figure 2 Transmission electron microscope characterization diagram of titanium-based metal organic framework material loaded with nickel nanoparticles prepared for example 2.

[0094] Figure 3 Transmission electron microscope characterization diagram of titanium-based nanocomposite prepared for example 3.

[0095] Figure 4 Transmission electron microscope characterization diagram of titanium-based nanocomposite prepared for example 5.

[0096] Figure 5 Schematic diagram of microwave heating curve of titanium-based metal organic framework material prepared for example 1 in physiological saline.

[0097] Figure 6 Schematic diagram of microwave heating curve of titanium-based nanocomposite prepared for example 3 in physiological saline.

[0098] Figure 7 Comparison of heating effect of titanium-based metal organic framework material prepared for example 1 and titanium-based nanocomposite prepared for example 3.

[0099] Figure 8 Schematic diagram of characterization of microwave dynamic performance of titanium-based metal organic framework material prepared for example 1 and titanium-based nanocomposite prepared for example 3.

[0100] Figure 9 Magnetic resonance imaging characterization chart of the titanium-based nanocomposite prepared in Example 3 and standard curve.

[0101] Figure 10 In vitro tumor cell inhibition effect of the titanium-based metal organic framework material prepared in Example 1 and the titanium-based nanocomposite prepared in Example 3.

[0102] Figure 11 Intracellular reactive oxygen species generation performance of the titanium-based metal organic framework material prepared in Example 1 and the titanium-based nanocomposite prepared in Example 3.

[0103] Figure 12 In vivo tumor inhibition effect of the titanium-based metal organic framework material prepared in Example 1 and the titanium-based nanocomposite prepared in Example 3. DETAILED DESCRIPTION

[0104] The preparation method of the present application will be further described in detail below in combination with specific examples. It should be understood that the following examples are only illustratively and explain the present application, and should not be interpreted as limiting the scope of protection of the present application. Any technology realized based on the above description of the present application is covered within the scope intended to be protected by the present application.

[0105] The experimental methods used in the following examples are conventional methods unless otherwise specified; the reagents, materials, etc. used in the following examples can be obtained from commercial channels unless otherwise specified.

[0106] Example 1

[0107] 0.305 mL of titanium isopropoxide, 0.2744 g of 2-amino terephthalic acid, and 2.2 g of benzoic acid were added into 20 mL of N,N-dimethylformamide and 2.2 mL of methanol, and ultrasonically dispersed uniformly.

[0108] The mixed solution was placed in a reaction kettle and reacted in a 150°C oven for 12 h.

[0109] After cooling to room temperature, centrifugation was performed, and then washing with N,N-dimethylformamide and methanol was performed once, respectively, and finally the sample was dispersed in ethanol for storage, thereby preparing the titanium-based metal organic framework material.

[0110] The titanium-based metal organic framework material prepared in Example 1 was characterized by transmission electron microscopy, and the particle size observed by transmission electron microscopy was 80-90 nm (as shown in FIG. 1). Figure 1

[0111] ​The prepared titanium-based metal organic framework material has good microwave heat conversion performance, and the microwave sensitization performance thereof has time and concentration dependence. When the concentration is 2 mg / mL, the temperature can reach 50 DEG C, and the temperature is increased by 8.3 DEG C relative to the physiological saline control group, indicating that the titanium-based metal organic framework material can be used for tumor microwave heat therapy (as shown in Figure 5 ).

[0112] The prepared titanium-based metal organic framework material has good microwave dynamic performance. The active oxygen production is characterized by fluorescence intensity. The fluorescence intensity of the titanium-based metal organic framework material + microwave group is more than 10 times that of the control group and 3 times that of the simple microwave group, indicating that the titanium-based metal organic framework material can produce active oxygen under microwave stimulation and can be used for tumor microwave dynamic therapy (as shown in Figure 8 ).

[0113] Example 2

[0114] 1) Take 0.305 mL titanium isopropoxide, 0.2772 g 2-amino terephthalic acid, and 2.2 g benzoic acid into 20 mL N,N-dimethylformamide and 2.2 mL methanol, ultrasonically disperse uniformly, and then place in a reaction kettle, and react in a 150 DEG C oven for 12 h. After cooling to room temperature, centrifugal, and then wash once with N,N-dimethylformamide and methanol respectively.

[0115] 2) Take 20 mg of the product obtained in step 1) and 100 mg of NiCl2·6H2O into 3 mL of N,N-dimethylformamide, ultrasonically disperse uniformly, and then magnetically stir in a 85 DEG C water bath for 2 h to obtain a titanium-based metal organic framework material adsorbed with Ni 2+ ;

[0116] 3) Disperse the product in step 2) into 1 mL of methanol, and then stir vigorously at room temperature. Then take 2 mg of sodium borohydride into 1 mL of methanol, and then quickly add, so that the Ni 2+ is reduced to Ni element, thereby obtaining a titanium-based metal organic framework material loaded with Ni nanoparticles.

[0117] The titanium-based metal organic framework material loaded with metal nanoparticles obtained in the present embodiment is characterized by transmission electron microscopy. As can be seen from Figure 2 , the metal nickel nanoparticles are loaded on the surface of the titanium-based metal organic framework material, and the metal nickel nanoparticles present a satellite-like distribution feature.

[0118] The prepared titanium-based metal organic framework material loaded with metal nanoparticles can be used for magnetic resonance imaging due to the presence of nickel metal nanoparticles, and can be used for tumor diagnosis and treatment in combination with the microwave heat-dynamic performance of the titanium-based metal organic framework material.

[0119] Example 3

[0120] 1) Take 0.305 mL titanium isopropoxide, 0.2772 g 2-amino terephthalic acid, 2.2 g benzoic acid into 20 mL N,N-dimethylformamide and 2.2 mL methanol, ultrasonic dispersion uniformly, then placed in a reaction kettle, reacted in a 150°C oven for 12 h. After cooling to room temperature, centrifugation, then washed once with N,N-dimethylformamide and methanol respectively.

[0121] 2) Take 20 mg of the product obtained in step 1) and 100 mg of NiCl2·6H2O into N,N-dimethylformamide, ultrasonic dispersion uniformly, magnetic stirring in a 85°C water bath for 2 h, to obtain a titanium-based metal-organic framework material adsorbed with Ni 2+ ;

[0122] 3) Disperse the product in step 2) into 1 mL methanol, vigorously stirred at room temperature, then take 2 mg sodium borohydride into 1 mL methanol, quickly add (3-5 drops / s), reduce Ni 2+ to Ni element, thereby obtaining a titanium-based metal-organic framework material loaded with Ni nanoparticles;

[0123] 4) Take 60 mg of the product obtained in step 3) and 1.2 g of polyvinylpyrrolidone into 30 mL acetonitrile; then take 3 mg 1,3,5-tris(4-aminophenyl)benzene into 3 mL acetonitrile and 0.4 mL ethanol, drop it into the above solution at a very slow speed (20-30 drops / min), magnetic stirring at room temperature for 1 h, centrifugation and washing.

[0124] 5) Disperse the product obtained in step 4) into 30 mL acetonitrile and add 1.2 g of polyvinylpyrrolidone, then take 2.6 mg 2,5-dimethoxybenzene-1,4-dicarboxaldehyde into 3 mL acetonitrile, drop it into the above solution at a very slow speed (20-30 drops / min), magnetic stirring at room temperature for 1 h, centrifugation and washing.

[0125] 6) Disperse the product obtained in step 5) into 30 mL acetonitrile and add 1.2 g of polyvinylpyrrolidone, drop 3 mg 1,3,5-tris(4-aminophenyl)benzene dissolved in 3 mL acetonitrile into it at a very slow speed (20-30 drops / min), drop 2.6 mg 2,5-dimethoxybenzene-1,4-dicarboxaldehyde dissolved in 3 mL acetonitrile into it at a very slow speed (20-30 drops / min), at the same time add 200 μL acetic acid, magnetic stirring at room temperature for 12 h, centrifugation and washing, to obtain the final titanium-based nanocomposite material for microwave thermal diagnosis and treatment integration.

[0126] Transmission electron microscopy characterization proves that the titanium-based nanocomposite obtained in this embodiment for microwave thermal diagnosis and treatment integration has a core-satellite-shell structure, which can confirm the successful loading of nickel nanoparticles on the titanium-based metal organic framework material and the coating of covalent organic framework (such as Figure 3 ) on it.

[0127] The mass percentage of the titanium-based metal organic framework material in the titanium-based nanocomposite prepared is 82.5%, the mass percentage of the metal nanoparticles is 1%, and the mass percentage of the covalent organic framework material is 16.5%.

[0128] The titanium-based nanocomposite prepared has good microwave heat conversion performance, and its microwave sensitization performance has time and concentration dependence. When the concentration is 2 mg / mL, the temperature can reach 60℃, which is 14.3℃ higher than that of the physiological saline control group, and 6℃ higher than that of the titanium-based metal organic framework material under the same concentration, further improving the microwave heat conversion performance of the titanium-based nanocomposite, which can be more effectively used for tumor microwave thermal therapy (as shown in Figure 6 ).

[0129] The titanium-based nanocomposite prepared has good microwave dynamic performance. The amount of reactive oxygen produced is characterized by fluorescence intensity, and the fluorescence intensity of the titanium-based nanocomposite + microwave group is more than 20 times that of the control group and 5 times that of the simple microwave group. In addition, compared with the titanium-based metal organic framework material + microwave group, the fluorescence intensity is 1.6 times that of the titanium-based metal organic framework material + microwave group, further improving the microwave dynamic performance of the titanium-based nanocomposite, which can be more effectively used for tumor microwave dynamic therapy (as shown in Figure 8 ).

[0130] The titanium-based nanocomposite prepared can be used for magnetic resonance imaging due to the presence of nickel metal nanoparticles, and has greater potential for tumor diagnosis and treatment practical application due to the more excellent microwave heat-dynamic performance of the titanium-based nanocomposite.

[0131] Example 4

[0132] 1) Take 0.305 mL of titanium isopropoxide, 0.2772 g of 2-amino terephthalic acid, and 2.2 g of benzoic acid into 20 mL of N,N-dimethylformamide and 2.2 mL of methanol, ultrasonically disperse uniformly, and then place in a reaction kettle, react in a 150℃ oven for 12 h. After cooling to room temperature, centrifuge, and then wash once with N,N-dimethylformamide and methanol, respectively.

[0133] 2) Take 20 mg of the product obtained in step 1) and 100 mg of NiCl2·6H2O into N,N-dimethylformamide, ultrasonically disperse uniformly, and then magnetically stir in a 85℃ water bath for 2 h to obtain a titanium-based metal organic framework material adsorbed with Ni 2+ .

[0134] 3) The product in step 2) was dispersed in 1 ml of methanol, and 2 mg of sodium borohydride was added to 1 mL of methanol, and was added quickly (3-5 drops / s) under vigorous stirring at room temperature, and the Ni 2+ was reduced to Ni element, thereby obtaining a titanium-based metal organic framework material loaded with Ni nanoparticles;

[0135] 4) 30 mg of the product obtained in step 3) and 600 mg of polyvinylpyrrolidone were dispersed in 15 mL of acetonitrile; 15 mg of 1,3,5-tris(4-aminophenyl)benzene was dissolved in 15 mL of acetonitrile and 2 mL of ethanol, and was added dropwise (20-30 drops / min) to the above solution at a very slow speed, and was stirred magnetically at room temperature for 2 h, and was centrifuged and washed.

[0136] 5) The product obtained in step 4) was dispersed in 15 mL of acetonitrile and 600 mg of polyvinylpyrrolidone was added, and 13 mg of 2,5-dimethoxybenzene-1,4-dicarboxaldehyde was dissolved in 15 mL of acetonitrile, and was added dropwise (20-30 drops / min) to the above solution at a very slow speed, and was stirred magnetically at room temperature for 2 h, and was centrifuged and washed.

[0137] 6) The product obtained in step 5) was dispersed in 15 mL of acetonitrile and 600 mg of polyvinylpyrrolidone was added, and 15 mg of 1,3,5-tris(4-aminophenyl)benzene dissolved in 15 mL of acetonitrile was added dropwise (20-30 drops / min) at a very slow speed, and 13 mg of 2,5-dimethoxybenzene-1,4-dicarboxaldehyde dissolved in 15 mL of acetonitrile was added dropwise (20-30 drops / min) at a very slow speed, and 200 μL of acetic acid was added, and was stirred magnetically at room temperature for 6 h, and was centrifuged and washed, thereby obtaining the final titanium-based nanocomposite for microwave thermal diagnosis and treatment integration.

[0138] The titanium-based nanocomposite for microwave thermal diagnosis and treatment integration obtained in this example has a core-satellite-shell structure, and has microwave response performance and magnetic resonance imaging capability.

[0139] Example 5

[0140] 1) 0.305 mL of titanium isopropoxide, 0.2772 g of 2-amino terephthalic acid, and 2.2 g of benzoic acid were added to 20 mL of N,N-dimethylformamide and 2.2 mL of methanol, and were uniformly dispersed by ultrasonic dispersion, and were placed in a reaction kettle, and were reacted in an oven at 150°C for 12 h. After cooling to room temperature, centrifugation was performed, and N,N-dimethylformamide and methanol were used for washing once, respectively.

[0141] 2) Take 20 mg of the product obtained in step 1) and 100 mg of NiCl2·6H2O and add them to N,N-dimethylformamide, ultrasonically disperse them uniformly, and magnetically stir them at 85°C for 2 h to obtain a Ni-adsorbed titanium-based metal organic framework material; 2+ ;

[0142] 3) Disperse the product in step 2) into 1 ml of methanol, stir vigorously at room temperature, take 2 mg of sodium borohydride and add it to 1 ml of methanol, add it quickly (3-5 drops / s), reduce the Ni 2+ to Ni elementary substance, thereby obtaining a titanium-based metal organic framework material loaded with Ni nanoparticles;

[0143] 4) Take 30 mg of the product obtained in step 3) and 600 mg of polyvinylpyrrolidone and disperse them in 15 ml of acetonitrile; take 6 mg of 1,3,5-tris(4-aminophenyl)benzene and dissolve them in 6 ml of acetonitrile and 0.8 ml of ethanol, add them to the above solution at a very slow speed (20-30 drops / min), magnetically stir them at room temperature for 1 h, and centrifugally wash them.

[0144] 5) Disperse the product obtained in step 4) into 15 ml of acetonitrile and add 600 mg of polyvinylpyrrolidone, take 5.2 mg of 2,5-dimethoxybenzene-1,4-dicarboxaldehyde and dissolve them in 6 ml of acetonitrile, add them to the above solution at a very slow speed (20-30 drops / min), magnetically stir them at room temperature for 1 h, and centrifugally wash them.

[0145] 6) Disperse the product obtained in step 5) into 15 ml of acetonitrile and add 600 mg of polyvinylpyrrolidone, add 6 mg of 1,3,5-tris(4-aminophenyl)benzene dissolved in 6 ml of acetonitrile at a very slow speed (20-30 drops / min), add 5.2 mg of 2,5-dimethoxybenzene-1,4-dicarboxaldehyde dissolved in 6 ml of acetonitrile at a very slow speed (20-30 drops / min), and simultaneously add 100 μl of acetic acid, magnetically stir them at room temperature for 10 h, and centrifugally wash them to obtain the final titanium-based nanocomposite material for microwave thermal diagnosis and treatment integration.

[0146] Transmission electron microscopy characterization proves that the titanium-based nanocomposite material for microwave thermal diagnosis and treatment integration obtained in this embodiment has a core-satellite-shell structure, and it can be confirmed that the nickel nanoparticles are successfully loaded on the titanium-based metal organic framework material and are coated with a covalent organic framework (as shown in Figure 4 ).

[0147] The mass proportion of the titanium-based metal organic framework material in the titanium-based nanocomposite material prepared is 70.5%, the mass proportion of the metal nanoparticles is 1%, and the mass proportion of the covalent organic framework material is 28.5%.

[0148] The titanium-based nanocomposite prepared has microwave thermokinetic performance and magnetic resonance imaging capability.

[0149] Example 6

[0150] 1) Take 0.305 mL titanium isopropoxide, 0.2772 g 2-amino terephthalic acid, and 2.2 g benzoic acid, add them to 20 mL N,N-dimethylformamide and 2.2 mL methanol, ultrasonically disperse them uniformly, and then place them in a reaction kettle, and react in a 150°C oven for 12 h. After cooling to room temperature, centrifugal separation is performed, and then washing is performed once with N,N-dimethylformamide and methanol, respectively.

[0151] 2) Take 20 mg of the product obtained in step 1) and 100 mg NiCl2·6H2O, add them to N,N-dimethylformamide, ultrasonically disperse them uniformly, and then perform magnetic stirring in a 85°C water bath for 2 h, to obtain a titanium-based metal-organic framework material adsorbing Ni 2+ ;

[0152] 3) Disperse the product in step 2) into 1 mL of methanol, and then perform vigorous stirring at room temperature, and then take 2 mg sodium borohydride, add it to 1 mL of methanol, and then add it quickly (3-5 drops / s), to reduce the Ni 2+ to Ni element, thereby obtaining a titanium-based metal-organic framework material loaded with Ni nanoparticles;

[0153] 4) Take 12.5 mg of the product obtained in step 3) and 60 mg polyvinylpyrrolidone, disperse them in 6 mL of acetonitrile; and then take 1 mg p-phenylenediamine, dissolve it in 1 mL of acetonitrile, and then add it dropwise (20-30 drops / min) to the above solution at a very slow speed, and then perform magnetic stirring at room temperature for 0.5 h, and then perform centrifugal separation and washing.

[0154] 5) Disperse the product obtained in step 4) in 6 mL of acetonitrile and add 60 mg of polyvinylpyrrolidone, and then take 1 mg trimesic aldehyde, dissolve it in 1 mL of acetonitrile, and then add it dropwise (20-30 drops / min) to the above solution at a very slow speed, and then perform magnetic stirring at room temperature for 1 h, and then perform centrifugal separation and washing.

[0155] 6) Disperse the product obtained in step 5) in 6 mL of acetonitrile and add 600 mg of polyvinylpyrrolidone, and then add 4 mg of p-phenylenediamine dissolved in 4 mL of acetonitrile dropwise (20-30 drops / min) at a very slow speed, and then add 4 mg of trimesic aldehyde dissolved in 4 mL of acetonitrile dropwise (20-30 drops / min) at a very slow speed, and then add 50 μL of acetic acid, and then perform magnetic stirring at room temperature for 12 h, and then perform centrifugal separation and washing, to obtain the final titanium-based nanocomposite for microwave thermodiagnosis and treatment integration.

[0156] The titanium-based nanocomposite prepared in the embodiment has a core-satellite-shell structure, and can be used for magnetic resonance imaging and microwave treatment of tumors.

[0157] Example 7

[0158] The materials prepared in Examples 1 and 3 were subjected to in vitro microwave heat conversion performance test. The specific experimental method is as follows:

[0159] 1) Sodium chloride was dispersed in deionized water to prepare normal saline (0.9% NaCl);

[0160] 2) The materials prepared in Examples 1 and 3 were added to the normal saline respectively to prepare solutions with concentrations of 0, 0.5, 1 and 2 mg / mL respectively;

[0161] 3) The solutions were irradiated with a microwave of 450 MHz and 1.8 W for 5 min, and the solution concentration was monitored in real time by a near-infrared thermal imager.

[0162] The embodiment proves that the materials prepared in Examples 1 and 3 both have good microwave heat conversion performance (as shown in Figure 5 、 Figure 6 ). The microwave sensitization performance of the titanium-based metal organic framework material prepared has time and concentration dependence. When the concentration is 0.5, 1 and 2 mg / mL, the temperature rise relative to the normal saline control group is 4.4, 7.3 and 8.3 ℃ respectively, and when the concentration is 2 mg / mL, the temperature is above 50 ℃. The microwave sensitization performance of the titanium-based nanocomposite prepared also has time and concentration dependence. When the concentration is 0.5, 1 and 2 mg / mL, the temperature rise relative to the normal saline control group is 8.9, 11.1 and 14.3 ℃ respectively, and when the concentration is 2 mg / mL, the temperature is 60 ℃, the temperature rise relative to the normal saline control group is 14.3 ℃, and the temperature rise is 4-6 ℃ higher than that of the titanium-based metal organic framework material at the same concentration, further improving the microwave heat conversion performance of the material and having better microwave heating effect, which can be used for microwave thermotherapy of tumors (as shown in Figure 7 .

[0163] Example 8

[0164] The materials prepared in Examples 1 and 3 were subjected to in vitro microwave dynamic performance test. The specific experimental method is as follows:

[0165] 1) The materials prepared in Examples 1 and 3 were added to phosphate buffered saline solution (PBS) (PH = 7.2) to prepare solutions with a concentration of 0.5 mg / mL respectively;

[0166] 2) 100 μM hydrogen peroxide was added to the solution;

[0167] 3) Add 10 μM active oxygen detection probe DCFH-DA into the solution;

[0168] 4) Irradiate the solution with 450 MHz, 1.8 W microwave for 5 min;

[0169] 5) Centrifuge after standing in dark for 2 h, and take the supernatant;

[0170] 6) Detect the fluorescence intensity at 520 nm under excitation at 480 nm wavelength.

[0171] This example proves that the materials prepared in Examples 1 and 3 both have good microwave dynamic performance. The fluorescence intensity of the titanium-based metal organic framework material + microwave group is more than 10 times that of the control group and 3 times that of the pure microwave group, which has good microwave dynamic performance. The fluorescence intensity of the titanium-based nanocomposite + microwave group is more than 1.6 times that of the titanium-based metal organic framework material + microwave group, indicating that the coating of the covalent organic framework further improves the ability of the material to produce active oxygen and improves the microwave dynamic performance, which can be used for microwave dynamic treatment of tumors (as shown in Figure 8 ).

[0172] Example 9

[0173] The material prepared in Example 3 was subjected to in vitro magnetic resonance imaging test. The specific experimental method is as follows:

[0174] 1) The material prepared in Example 3 was taken and added into deionized water to prepare a solution with a concentration of 0.1-1.0 mg / mL;

[0175] 2) The solution was placed in a small animal nuclear magnetic resonance imaging instrument for scanning;

[0176] 3) The transverse relaxation time (T1) corresponding to different concentrations was measured respectively;

[0177] 4) The relationship between the material concentration and the relaxation time was fitted through the relationship between 1 / T1 and the concentration of the material prepared in Example 3.

[0178] This example proves that the titanium-based nanocomposite obtained in Example 3 can be used for magnetic resonance imaging. When the concentration increases from 0.1 mg / mL to 1 mg / mL, its 1 / T1 shows a concentration-dependent upward trend, and its linear regression equation is Y = 2.88885C + 3.45528. Therefore, the titanium-based nanocomposite has concentration-dependent magnetic resonance imaging ability (as shown in Figure 9 ).

[0179] Example 10

[0180] In vitro treatment experiments were performed on the materials prepared in Examples 1 and 3. The specific experimental method is as follows:

[0181] 1) Mouse breast cancer cells 4T1 were dispersed in a 6-well plate and incubated for 24 h.

[0182] 2) The materials prepared in Examples 1 and 3 were taken and added to the culture medium (DMEM) to prepare a solution with a concentration of 100 μg / mL.

[0183] 3) The 6 wells were randomly divided into 6 groups, and after adding the corresponding materials (control group, material 1 group, material 3 group, microwave group, material 1 + microwave group, material 3 + microwave group), incubation was continued for 24 h.

[0184] 4) The cells were irradiated with a microwave of 450 MHz, 1.8 W for 5 min.

[0185] 5) Transferred to a 96-well plate and incubated for another 12 h.

[0186] 6) 20 μL thiazolyl blue (MTT) was added to each well and incubated for 4 h.

[0187] 7) The supernatant was removed, and 150 μL dimethyl sulfoxide was added.

[0188] 8) The cell activity was tested with a microplate reader.

[0189] This example demonstrates that the materials prepared in Examples 1 and 3 have a significant inhibitory effect on tumor cells under microwave stimulation. The cell activity of the control group was taken as 100%, and the cell activity of the pure titanium-based metal-organic framework material group and the pure titanium-based nanocomposite material group was more than 85%, indicating that both materials have good biological safety. The cell survival rate of the titanium-based metal-organic framework material + microwave group was 50%, which had a significant killing effect on tumor cells. The cell survival rate of the titanium-based nanocomposite material + microwave group further decreased, and the tumor cell activity was less than 40%, indicating the best tumor treatment effect (as shown in Figure 10 ).

[0190] Example 11

[0191] Cell microwave dynamic performance tests were performed on the materials prepared in Examples 1 and 3. The specific experimental method is as follows:

[0192] 1) Mouse breast cancer cells 4T1 were dispersed in a 6-well plate and incubated for 24 h.

[0193] 2) The materials prepared in Examples 1 and 3 were taken and added to the culture medium (DMEM) to prepare a solution with a concentration of 100 μg / mL.

[0194] 3) 6 wells were randomly divided into 6 groups, and the corresponding materials were added (control group, material 1 group, material 3 group, microwave group, material 1 + microwave group, material 3 + microwave group), and incubated for 4h.

[0195] 4) 100 μM hydrogen peroxide was added to each well, and incubated for 2h.

[0196] 5) The medium was removed, and the reactive oxygen probe (DCFH-DA) was added and incubated for 30 min, then the probe was removed and washed twice.

[0197] 6) The cells were irradiated with a microwave of 450 MHz, 1.8 W for 5 min.

[0198] 7) The intracellular reactive oxygen level was evaluated by flow cytometry characterization.

[0199] This example demonstrates that the materials prepared in Examples 1 and 3 can generate reactive oxygen under microwave stimulation in cells. The amount of reactive oxygen generated is characterized by the average fluorescence intensity determined by flow cytometry. The fluorescence intensity of the titanium-based metal organic framework material + microwave group (i.e., material 1 + microwave group) is about 5000 higher than that of the control group and about 1500 higher than that of the pure microwave group, and has good microwave dynamic performance in cells. The fluorescence intensity of the titanium-based nanocomposite material + microwave group (i.e., material 3 + microwave group) is about 1500 higher than that of the titanium-based metal organic framework material + microwave group, indicating that the coating of the covalent organic framework material further improves the ability of the material to generate reactive oxygen, and demonstrates the good microwave dynamic performance of the titanium-based nanocomposite material in cells (as shown in FIG. 1). Figure 11

[0200] Example 12

[0201] In vivo treatment experiments were performed on the materials prepared in Examples 1 and 3. The specific experimental method is as follows:

[0202] 1) Human breast cancer cells (mda mb 231) were inoculated into Balb / c nude mice, and the mice were fed until the tumor volume reached 250 mm 3 The experiment was started.

[0203] 2) The materials prepared in Examples 1 and 3 were added to physiological saline to prepare a solution with a concentration of 50 mg / kg.

[0204] 3) The mice were randomly divided into 6 groups, with 4 mice in each group.

[0205] 4) On the day of treatment (designated as day 0), different groups of mice were injected with materials (control group, material 1 group, material 3 group, microwave group, material 1 + microwave group, material 3 + microwave group).

[0206] ​4) Six hours after material injection, the cells were irradiated with microwaves at 450 MHz and 1.8 W for 5 minutes.

[0207] 5) Record the mouse's weight and tumor size daily thereafter, and take photos for documentation.

[0208] 6) One month later, the mice were dissected to observe the final treatment effect.

[0209] This embodiment demonstrates that the materials prepared in Examples 1 and 3 under microwave stimulation both exhibit good tumor-suppressing effects. According to... Figure 12 The tumor volume growth curves were as expected: the tumors in the control group grew rapidly; the materials prepared in Examples 1 and 3 had no inhibitory effect on the tumors themselves, and their growth curves were similar to those of the control group; the microwave group showed slow growth in the early stages with a flat curve, followed by rapid growth in the later stages; the tumor volume of the titanium-based metal-organic framework material + microwave group (i.e., material 1 + microwave group) gradually decreased, showing an upward trend after 15 days, but the growth rate was slow; the tumor volume of the titanium-based nanocomposite material + microwave group (i.e., material 3 + microwave group) continued to decrease, and the final tumor volume was smaller than that on day 0. Based on the final tumor volume, the tumor inhibition rate of the titanium-based metal-organic framework material + microwave group was 86%, while the tumor inhibition rate of the titanium-based nanocomposite material + microwave group could reach 97%, exhibiting the best tumor treatment effect (e.g., Figure 12 (As shown).

[0210] The embodiments of the present invention have been described above. However, the present invention is not limited to the above embodiments. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the protection scope of the present invention.

Claims

1. A titanium-based nanocomposite material with properties for tumor microwave hyperthermia, microwave dynamic therapy, and magnetic resonance imaging, wherein, The titanium-based nanocomposite material includes a titanium-based metal-organic framework material, metal nanoparticles, and a covalent organic framework material; the metal nanoparticles are distributed on the surface and / or within the pores of the titanium-based metal-organic framework material to form a titanium-based metal-organic framework material loaded with metal nanoparticles; the covalent organic framework material is coated on the surface of the titanium-based metal-organic framework material loaded with metal nanoparticles. The titanium-based metal-organic framework material is prepared by a titanium source and an organic ligand in the presence of a modifier; the modifier forming the titanium-based metal-organic framework material is selected from at least one of benzoic acid, acetic acid, mercaptoacetic acid and citric acid. The covalent organic framework material is prepared by using covalent organic framework monomers containing amino functional groups and covalent organic framework monomers containing aldehyde functional groups. The metal nanoparticles are selected from at least one of nickel nanoparticles, gadolinium nanoparticles, manganese nanoparticles, and iron nanoparticles. The covalent organic framework material is coated on the surface of the composite to form a shell with a thickness of 2-20 nm; The titanium-based metal-organic framework material has a particle size distribution of 50-400 nm; the metal nanoparticles have a particle size distribution of 1-50 nm. The mass of the metal nanoparticles accounts for 0.5-30% of the total mass of the titanium-based nanocomposite material; the mass of the titanium-based metal-organic framework material accounts for 60-85% of the total mass of the titanium-based nanocomposite material; and the mass of the covalent organic framework material accounts for 10-16.5% of the total mass of the titanium-based nanocomposite material.

2. The titanium-based nanocomposite material according to claim 1, wherein, The titanium source for forming the titanium-based metal-organic framework material is selected from at least one of titanium isopropoxide, titanium tetrachloride, and tetrabutyl titanate; the organic ligand for forming the titanium-based metal-organic framework material is selected from at least one of terephthalic acid, 2-aminoterephthalic acid, 2,5-dihydroxyterephthalic acid, 4-pyridinecarboxylic acid, fumaric acid, and pyromellitic acid.

3. The titanium-based nanocomposite material according to claim 1 or 2, wherein, The molar ratio of the titanium source forming the titanium-based metal-organic framework material to the organic ligand forming the titanium-based metal-organic framework material is 1:0.5-10; the molar ratio of the titanium source forming the titanium-based metal-organic framework material to the modifier forming the titanium-based metal-organic framework material is 1:1-50.

4. The titanium-based nanocomposite material according to claim 1, wherein, The amino-functionalized covalent organic framework monomers forming the covalent organic framework material are selected from at least one of p-phenylenediamine, 1,3,5-tris(4-aminophenyl)benzene, and 2,5-diethoxyterephthalic acid dihydrazide; the aldehyde-functionalized covalent organic framework monomers forming the covalent organic framework material are selected from at least one of benzaldehyde, pyromellitic tricarboxaldehyde, and 2,5-dimethoxybenzene-1,4-dicarboxaldehyde. And / or, the mass ratio of the amino-functionalized covalent organic framework monomer forming the covalent organic framework material to the aldehyde-functionalized covalent organic framework monomer forming the covalent organic framework material is 1:0.5-2.

5. The titanium-based nanocomposite material according to claim 1, wherein, The metal nanoparticles are distributed in a satellite-like manner on the surface and / or within the pores of the titanium-based metal-organic framework material.

6. The titanium-based nanocomposite material according to claim 1, wherein, The particle size distribution of the titanium-based nanocomposite material is 50-500 nm.

7. A method for preparing the titanium-based nanocomposite material according to any one of claims 1-6, the method comprising the following steps: 1) Titanium-based metal-organic framework materials are prepared by dispersing titanium source, organic ligand and regulator in solvent and carrying out solvothermal reaction; 2) The titanium-based metal-organic framework material obtained in step 1) and the metal salt are dispersed in a solvent and mixed to prepare a titanium-based metal-organic framework material that adsorbs metal ions. 3) Mix the titanium-based metal-organic framework material with adsorbed metal ions obtained in step 2) with a reducing agent and react to prepare a titanium-based metal-organic framework material loaded with metal nanoparticles. 4) Mix the titanium-based metal-organic framework material loaded with metal nanoparticles obtained in step 3) with a covalent organic framework monomer containing amino functional groups, and react. 5) Mix the product obtained in step 4) with a covalent organic framework monomer containing an aldehyde functional group and react. 6) The product obtained in step 5), an equal amount of covalent organic framework monomer containing amino functional groups as in step 4), an equal amount of covalent organic framework monomer containing aldehyde functional groups as in step 5), and a catalyst are mixed and reacted to prepare the titanium-based nanocomposite material.

8. The preparation method according to claim 7, wherein, In step 1), the mass concentration of the titanium source is 1-30 mg / mL; the molar ratio of the titanium source to the organic ligand is 1:0.5-10; and the molar ratio of the titanium source to the regulator is 1:1-50.

9. The preparation method according to claim 7, wherein, In step 1), the temperature of the solvothermal reaction is 120-200℃; the time of the solvothermal reaction is 2-48h.

10. The preparation method according to claim 7, wherein, In step 2), the mass ratio of the titanium-based metal-organic framework to the metal salt is 1:0.1-10.

11. The preparation method according to claim 7, wherein, In step 2), the mixing temperature is room temperature, and the mixing time is 1-6 hours.

12. The preparation method according to claim 7, wherein, In step 3), the mass ratio of the reducing agent to the titanium-based metal-organic framework material that adsorbs metal ions is 1:0.1-50.

13. The preparation method according to claim 7, wherein, In step 3), the reducing agent is selected from at least one of sodium borohydride, hydrazine hydrate, and lithium aluminum hydride.

14. The preparation method according to claim 7, wherein, In step 3), the reaction temperature is room temperature, and the reaction time is 10-60 seconds.

15. The preparation method according to claim 7, wherein, Step 3) specifically includes the following steps: The titanium-based metal-organic framework material with adsorbed metal ions obtained in step 2) is dispersed in a solvent to obtain mixed system 1; then the reducing agent is dispersed in the same solvent to obtain mixed system 2; mixed system 2 is added to mixed system 1 at a rate of 3-5 drops / s and stirred at room temperature for 10-60s to prepare titanium-based metal-organic framework material loaded with metal nanoparticles.

16. The preparation method according to claim 7, wherein, In step 4), the mass ratio of the titanium-based metal-organic framework material loaded with metal nanoparticles to the covalent organic framework monomer containing amino functional groups is 5-30:

1.

17. The preparation method according to claim 7, wherein, In step 4), the reaction temperature is room temperature, and the reaction time is 0.5-4 hours.

18. The preparation method according to claim 7, wherein, Step 4) specifically includes the following steps: The titanium-based metal-organic framework material loaded with metal nanoparticles obtained in step 3) is dispersed in a solvent to obtain mixed system 3; then, a covalent organic framework monomer containing amino functional groups is dispersed in the same solvent to obtain mixed system 4; mixed system 4 is added dropwise to mixed system 3 at a rate of 20-30 drops / min, and stirred at room temperature for 0.5-4 hours, the product is collected and washed.

19. The preparation method according to claim 7, wherein, In step 5), the mass ratio of the covalent organic framework monomer containing an amino functional group to the covalent organic framework monomer containing an aldehyde functional group is 1:0.5-2.

20. The preparation method according to claim 7, wherein, In step 5), the reaction temperature is room temperature and the reaction time is 0.5-4 hours.

21. The preparation method according to claim 7, wherein, Step 5) specifically includes the following steps: The product obtained in step 4) is dispersed in a solvent to obtain mixed system 5; then, a covalent organic framework monomer containing an aldehyde functional group is dispersed in the same solvent to obtain mixed system 6; mixed system 6 is added dropwise to mixed system 5 at a rate of 20-30 drops / min, and stirred at room temperature for 0.5-4 hours, the product is collected and washed.

22. The preparation method according to claim 7, wherein, In step 6), the reaction temperature is room temperature, and the reaction time is 2-12 hours.

23. The preparation method according to claim 7, wherein, In step 6), the catalyst is selected from acetic acid.

24. A diagnostic reagent comprising the titanium-based nanocomposite material according to any one of claims 1-6.

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