A gadolinium-based composite material with microwave response heat production and dual-mode imaging effect, and preparation and use thereof

Abstract

The application provides a gadolinium-based composite material with microwave response heat production and double-mode imaging effect, and preparation and application thereof. The gadolinium-based composite material prepared by the application is applied to the fields of microwave hyperthermia and double-mode imaging of tumors, and has good clinical application value. The gadolinium-based composite material synthesized by the application has low cytotoxicity and good cell internalization efficiency, can significantly increase the killing effect of microwaves on tumor sites, and has higher cell internalization efficiency and better microwave heating performance through adjustment of the morphology of the gadolinium-based composite material. In addition, Gd 3+ in the nanomaterial has a nuclear magnetic resonance enhancement effect, and the loaded fluorescent dye has a fluorescence imaging effect, so that the material has double-mode imaging function and can be used as a microwave diagnosis and treatment agent to realize tumor diagnosis and treatment integration.

Description

Technical Field

[0001] This invention relates to the field of sensitizing materials technology, specifically to a gadolinium-based composite material with microwave-responsive heat generation and dual-mode imaging effects, its preparation method, and its applications. Background Technology

[0002] Microwave hyperthermia, as an emerging minimally invasive tumor therapy, boasts advantages such as minimal trauma, rapid patient recovery, and maximum protection of bodily functions, playing an increasingly important role in the treatment of solid tumors. However, current microwave ablation methods still face many challenges, such as insufficient selectivity of microwaves for tumor tissue, the unavoidable damage to surrounding normal tissues while killing tumor cells, and incomplete tumor ablation due to insufficient heating efficiency of existing microwave equipment. There is a strong desire to increase the specific absorption of microwaves within tumor tissue, improve the heating efficiency within the tumor, and enable rapid and uniform heating of the entire tumor tissue to expand the scope of microwave hyperthermia while reducing damage to normal tissues and lowering toxic side effects. Therefore, developing biosafety nanosensitizers with high microwave thermal conversion efficiency, controllable structure, good stability, and easily modifiable surfaces is of great significance for the development of tumor microwave hyperthermia.

[0003] To improve the effectiveness of cancer treatment and alleviate patient suffering, efficient and accurate real-time diagnostic images are essential during treatment. Compared to other imaging techniques with deeper tissue penetration capabilities, magnetic resonance imaging (MRI) offers the advantage of being radiation-free. Due to its deep tissue penetration and high spatial resolution, it has become one of the routine diagnostic tools used in clinical medicine. The signal intensity obtained by MRI depends on the relaxation rate of protons in water molecules, and differences between tissues can be detected by changes in water density, proton relaxation time, or water molecule diffusion rate. However, when the differences between tissues are small, such as when the distinction between a tumor and surrounding tissue is very small, contrast agents that enhance MRI contrast are needed to obtain more effective and clearer diagnostic information. Currently, approximately 50% of clinical MRI examinations require the use of contrast agents to improve image quality. With the help of contrast agents, MRI can provide in-depth details of tumor tissue in real time. However, the delivery efficiency of contrast agents and the absorption efficiency by tumors are low, often requiring excessive amounts of contrast agents in practical applications. Long-term use can increase the risk of cardiovascular and neurotoxic diseases.

[0004] Fluorescence imaging technology uses fluorescent reporter genes or fluorescent dyes for labeling, and the fluorescence generated by the reporter genes, fluorescent proteins, or dyes forms a biological light source in the body. Fluorescence imaging is fast, has low experimental cost, strong labeling ability, and a wide range of fluorescent labels, including animals, cells, microorganisms, antibodies, drugs, and nanomaterials. However, fluorescence imaging also has drawbacks such as limited detection depth, low spatial resolution, and difficulty in accurate in vivo quantification. Summary of the Invention

[0005] Research has found that combining cancer therapy and bioimaging in nanosystems can not only provide accurate tumor location and morphology information before treatment, but also assess the retention and therapeutic effect of diagnostic agents at the tumor site during treatment. Based on this, this invention provides a gadolinium-based composite material with microwave-responsive thermogenic and dual-mode imaging effects, its preparation method, and its applications. The gadolinium-based composite material exhibits low cytotoxicity and good cellular uptake, enabling the combination of microwave thermotherapy for tumors with dual-mode imaging (magnetic resonance imaging and fluorescence imaging) technology, thus integrating diagnostic and therapeutic drugs into one. This will significantly reduce patient suffering and treatment risks, and can be applied simultaneously in the fields of tumor microwave thermotherapy and dual-mode imaging. Furthermore, the nanomaterials prepared by the method exhibit controllable size and morphology.

[0006] The objective of this invention is achieved through the following technical solution:

[0007] A gadolinium-based composite material comprising gadolinium-based nanomaterials, a fluorescent dye, and a surface modifier, wherein the fluorescent dye is encapsulated within the pores of the gadolinium-based nanomaterials to form a composite of the gadolinium-based nanomaterials and the fluorescent dye, and the surface modifier is chemically bonded to the surface of the composite of the gadolinium-based nanomaterials and the fluorescent dye.

[0008] According to an embodiment of the present invention, after the gadolinium-based nanomaterial and the fluorescent dye form a complex through physical interaction, the surface of the complex is then modified, and the surface modification and the complex are bonded by chemical bonds.

[0009] According to an embodiment of the present invention, the fluorescent dye is encapsulated within the pores of the gadolinium-based nanomaterial through a confinement effect to form a composite of the gadolinium-based nanomaterial and the fluorescent dye, that is, the composite is a physical mixture of the gadolinium-based nanomaterial and the fluorescent dye.

[0010] According to an embodiment of the present invention, the gadolinium-based nanomaterial is prepared by a solvothermal reaction of a metal salt containing gadolinium ions and an organic ligand.

[0011] The metal salt containing gadolinium ions is selected from one or two of gadolinium chloride, gadolinium nitrate, and gadolinium bicarbonate.

[0012] The organic ligand is selected from one or two of isophthalic acid, terephthalic acid, fumaric acid, L-tyrosine, L-arginine, and methyl-tetra(4-carboxyphenyl)porphyrin.

[0013] The temperature of the solvothermal reaction is 150-200℃, and the time of the solvothermal reaction is 6-12h.

[0014] According to embodiments of the present invention, the gadolinium-based nanomaterial has at least one morphology selected from rod-shaped, sheet-shaped, and spherical, preferably rod-shaped. When one organic ligand, such as fumaric acid, is selected, the gadolinium-based nanomaterial prepared has a spherical morphology. When two organic ligands, such as methyl-tetra(4-carboxyphenyl)porphyrin and isophthalic acid, are selected, the morphology of the gadolinium-based nanomaterial can be adjusted to be sheet-shaped or rod-shaped by adjusting the ratio of the two ligands. More specifically, when the mass ratio of methyl-tetra(4-carboxyphenyl)porphyrin to isophthalic acid is 1:4, a sheet-shaped material can be obtained; when the mass ratio of methyl-tetra(4-carboxyphenyl)porphyrin to isophthalic acid is 1:6, a rod-shaped material can be obtained.

[0015] The mass ratio of the gadolinium-containing metal salt to the organic ligand is 1:1 to 1:6, for example, 1:1, 1:2, 1:3, 1:4, 1:5 or 1:6.

[0016] According to an embodiment of the present invention, the fluorescent dye is selected from one of indocyanine green (ICG), fluorescein isothiocyanate (FITC), CY5, rhodamine 6G, etc.

[0017] According to an embodiment of the present invention, the mass ratio of the gadolinium-based nanomaterial to the fluorescent dye is 1:1-2:1.

[0018] According to an embodiment of the present invention, the surface modifier is selected from at least one of hyaluronic acid, polyethylene glycol, bovine serum albumin, and dopamine.

[0019] According to an embodiment of the present invention, the mass ratio of the surface modifier to the complex of gadolinium-based nanomaterials and fluorescent dye is 1:5 to 1:15, for example, 1:5, 1:6, 1:7, 1:8, 1:9, 1:10, 1:11, 1:12, 1:13, 1:14 or 1:15.

[0020] According to an embodiment of the present invention, the gadolinium-based composite material has a particle size of 50-1000 nm, preferably 50-400 nm. The gadolinium-based composite material has a narrow particle size distribution and good dispersibility in water and ethanol.

[0021] According to an embodiment of the present invention, the gadolinium-based composite material has nuclear magnetic resonance enhancement properties and fluorescence imaging capabilities.

[0022] According to an embodiment of the present invention, the gadolinium-based composite material has at least one of the following morphologies: rod-shaped, sheet-shaped, and spherical, preferably rod-shaped.

[0023] According to an embodiment of the present invention, the median particle size of the gadolinium-based composite material is 150-900 nm, for example, 150 nm, 200 nm, 250 nm, 300 nm, 350 nm, 400 nm, 450 nm, 500 nm, 550 nm, 600 nm, 650 nm, 700 nm, 750 nm, 800 nm, 850 nm or 900 nm.

[0024] According to an embodiment of the present invention, the potential range of the gadolinium-based composite material is -40 to 50 mV.

[0025] According to an embodiment of the present invention, the gadolinium-based composite material can generate thermal energy under microwaves and can be used for nuclear magnetic resonance imaging and fluorescence imaging.

[0026] The present invention also provides a method for preparing the above-mentioned gadolinium-based composite material, the method comprising the following steps:

[0027] 1) Gadolinium-based nanomaterials are prepared by mixing a metal salt containing gadolinium ions and an organic ligand in an organic solvent and carrying out a solvothermal reaction.

[0028] 2) The obtained gadolinium-based nanomaterials were mixed with fluorescent dyes and stirred in the dark to prepare a composite of gadolinium-based nanomaterials and fluorescent dyes;

[0029] 3) The obtained gadolinium-based nanomaterials and fluorescent dye composites and surface modifiers are mixed and reacted to prepare the gadolinium-based composite material.

[0030] According to an embodiment of the present invention, in step 1), the definition of the metal salt containing gadolinium ions is as described above.

[0031] According to an embodiment of the present invention, in step 1), the organic ligand is defined as described above.

[0032] According to an embodiment of the present invention, in step 1), the mass ratio of the metal salt containing gadolinium ions to the organic ligand is 1:1 to 1:6, for example, 1:1, 1:2, 1:3, 1:4, 1:5 or 1:6.

[0033] According to an embodiment of the present invention, in step 1), the temperature of the solvothermal reaction is 150-200°C, and the time of the solvothermal reaction is 6-12 hours.

[0034] According to an embodiment of the present invention, in step 1), the organic solvent is selected from one or more of N,N-dimethylformamide, dimethyl sulfoxide, acetonitrile, isopropanol, n-hexane, cyclohexane, etc.

[0035] According to an embodiment of the present invention, in step 2), the fluorescent dye is defined as described above.

[0036] According to an embodiment of the present invention, in step 2), the mass ratio of the gadolinium-based nanomaterial to the fluorescent dye is preferably 1:1-2:1.

[0037] According to an embodiment of the present invention, in step 2), the mixture is stirred at room temperature in the dark for 6-24 hours.

[0038] According to an embodiment of the present invention, in step 3), the surface modification is defined as described above.

[0039] According to an embodiment of the present invention, in step 3), the mass ratio of the surface modifier to the complex of gadolinium-based nanomaterials and fluorescent dye is preferably 1:5-1:10, for example, 1:5, 1:6, 1:7, 1:8, 1:9 or 1:10.

[0040] According to an embodiment of the present invention, in step 3), the reaction is carried out at room temperature for 2-6 hours.

[0041] According to an embodiment of the present invention, the method specifically includes the following steps:

[0042] a. Dissolve the metal salt containing gadolinium ions and the organic ligand in an organic solvent, respectively, and sonicate to disperse them evenly to obtain a mixture;

[0043] b. Place the obtained mixture into a reaction vessel and react at 150-200℃ for 6-12 hours;

[0044] c. Centrifuge the product obtained from the reaction and collect the precipitate. Wash the precipitate with anhydrous ethanol to obtain gadolinium-based nanomaterials.

[0045] d. The obtained gadolinium-based nanomaterials and fluorescent dyes were mixed in an ethanol solution at room temperature and stirred in the dark for 6-24 hours. The fluorescent dyes were encapsulated in the pores of the gadolinium-based nanomaterials to form a complex of gadolinium-based nanomaterials and fluorescent dyes.

[0046] e. After ultrasonically dispersing the complex of gadolinium-based nanomaterials and fluorescent dyes in anhydrous ethanol, the surface modifier is dissolved in deionized water and slowly added dropwise to the above dispersion. The mixture is then magnetically stirred at room temperature for 2-6 hours.

[0047] f. After the reaction is complete, centrifuge the resulting solution and wash it with anhydrous ethanol, then collect the precipitate.

[0048] The present invention also provides the application of the above-mentioned gadolinium-based composite material in microwave hyperthermia.

[0049] The present invention also provides the application of the above-mentioned gadolinium-based composite material in microwave hyperthermia and in nuclear magnetic resonance and / or fluorescence imaging.

[0050] According to embodiments of the present invention, the gadolinium-based composite material is used in microwave hyperthermia and magnetic resonance imaging simultaneously, or in microwave hyperthermia and fluorescence imaging simultaneously, or in microwave hyperthermia, magnetic resonance imaging, and fluorescence imaging simultaneously.

[0051] The present invention also provides a formulation comprising the above-mentioned gadolinium-based composite material.

[0052] According to embodiments of the present invention, the preparation is used for microwave hyperthermia of tumors, and can also be used in magnetic resonance and / or fluorescence imaging.

[0053] According to an embodiment of the present invention, the gadolinium-based composite material is dispersed in physiological saline to prepare a concentration range of 1-10 mg / mL. -1 The solution was subjected to continuous microwave irradiation by placing solutions of different concentrations in an external microwave device, and the heating process of the material was monitored in real time.

[0054] The beneficial effects of this invention are:

[0055] This invention provides a gadolinium-based composite material, its preparation method, and its applications. The prepared gadolinium-based composite material is simultaneously applied to microwave hyperthermia and dual-mode imaging of tumors, demonstrating significant clinical application value. The gadolinium-based composite material synthesized in this invention exhibits low cytotoxicity and good cell internalization efficiency, significantly increasing the killing effect of microwaves on tumor sites. Furthermore, by adjusting the morphology of the gadolinium-based composite material, it achieves even higher cell internalization efficiency and better microwave heating performance. In addition, the Gd in the nanomaterial... 3+ This gives it the effect of enhanced nuclear magnetic resonance, and the loaded fluorescent dye gives it the effect of fluorescent imaging. The dual-mode imaging function of this material makes it suitable as a microwave diagnostic agent to achieve integrated tumor diagnosis and treatment. Attached Figure Description

[0056] Figure 1 This is a schematic diagram of the microstructure of the gadolinium-based composite material obtained in Example 1;

[0057] Figure 2 This is a schematic diagram of microwave heating of the gadolinium-based composite material obtained in Example 1;

[0058] Figure 3 This is a comparison of the cell internalization effects of the gadolinium-based composite materials obtained in Example 1 and Example 2. Detailed Implementation

[0059] The present invention will be further described in detail below with reference to specific embodiments. It should be understood that the following embodiments are merely illustrative and explanatory of the present invention and should not be construed as limiting the scope of protection of the present invention. All technologies implemented based on the above content of the present invention are covered within the scope of protection intended by the present invention.

[0060] Unless otherwise specified, the experimental methods used in the following examples are conventional methods; unless otherwise specified, the reagents and materials used in the following examples are commercially available.

[0061] Example 1

[0062] A method for preparing gadolinium-based composite materials with microwave-responsive heat generation and dual-mode imaging effects includes the following steps:

[0063] 1) Weigh 5.08 mg Gd(NO3)3·6H2O, 7.10 mg isophthalic acid and 3.90 mg terephthalene-tetra(4-carboxyphenyl)porphyrin, dissolve them in 90 mL N,N-dimethylformamide, and disperse them evenly by ultrasonication to obtain a mixed solution;

[0064] 2) Place the mixture into a reaction vessel and react at 200°C for 12 hours;

[0065] 3) Centrifuge the solution obtained from the reaction at 12000 r / min for 5 min and collect the precipitate. Wash the precipitate with anhydrous ethanol to obtain gadolinium-based nanomaterials.

[0066] 4) Take 2 mg of the obtained gadolinium-based nanomaterial and 1 mg of indocyanine green (ICG) and mix them in an ethanol solution. Stir the mixture at room temperature in the dark for 12 h to obtain the nanocomposite.

[0067] 5) Dissolve 0.2 mg of hyaluronic acid in 0.2 mL of deionized water, slowly add it dropwise to the above solution, continue magnetic stirring at room temperature for 4 h, and then centrifuge to collect the precipitate GM7IH.

[0068] Transmission electron microscopy (TEM) revealed that the gadolinium-based composite material prepared in this embodiment was rod-shaped, approximately 280 nm long and 80 nm wide. Figure 1 ).

[0069] The gadolinium-based composite material prepared in this embodiment can be used as a microwave diagnostic agent, and when used as a microwave diagnostic agent, it has MR / fluorescence imaging capabilities.

[0070] The nanomaterials prepared in this embodiment were dispersed in physiological saline to prepare concentrations of 1, 3, and 6 mg / mL. -1 Solutions of different concentrations were placed in an external microwave device for continuous microwave irradiation, and the heating process of the materials was monitored in real time. A control group was prepared using an equal volume of physiological saline. The heating results are as follows: Figure 2 As shown, the gadolinium-based microwave therapeutic agent prepared in this embodiment has a good microwave heating effect. After the microwave irradiation begins, the temperature of the material increases with the increase of microwave time, and it exhibits a concentration-dependent effect.

[0071] Example 2

[0072] A method for preparing gadolinium-based composite materials with microwave-responsive heat generation and dual-mode imaging effects includes the following steps:

[0073] 1) Weigh 20.31 mg Gd(NO3)3·6H2O and 5.23 mg fumaric acid, dissolve them in 30 mL N,N-dimethylformamide, and disperse them evenly by ultrasonication to obtain a mixed solution;

[0074] 2) Place the mixture into a reaction vessel and react at 150°C for 12 hours;

[0075] 3) Centrifuge the solution obtained from the reaction at 12000 r / min for 5 min and collect the precipitate. Wash the precipitate with anhydrous ethanol to obtain gadolinium-based nanomaterials.

[0076] 4) Take 2 mg of the obtained gadolinium-based nanomaterial and 2 mg of rhodamine 6G, mix them in an ethanol solution at room temperature and stir for 12 h in the dark to obtain a nanocomposite.

[0077] 5) Dissolve 0.4 mg of polyethylene glycol in 0.2 mL of deionized water, slowly add it dropwise to the above solution, continue magnetic stirring at room temperature for 6 h, and then centrifuge to collect the precipitate GM8IH.

[0078] Transmission electron microscopy (TEM) revealed that the gadolinium-based composite material prepared in this embodiment was spherical with a diameter of approximately 200 nm.

[0079] The gadolinium-based composite material prepared in this embodiment can be used as a microwave diagnostic agent. When used as a microwave diagnostic agent, it has MR / fluorescence imaging capability and microwave response heating capability, and exhibits concentration dependence.

[0080] Example 3

[0081] A method for preparing gadolinium-based composite materials with microwave-responsive heat generation and dual-mode imaging effects includes the following steps:

[0082] 1) Weigh 6.77 mg Gd(NO3)3·6H2O, 12.61 mg isophthalic acid and 11.86 mg methyl-2-tetra(4-carboxyphenyl)porphyrin, dissolve them in 30 mL N,N-dimethylformamide, and disperse them evenly by ultrasonication to obtain a mixed solution;

[0083] 2) Place the mixture into a reaction vessel and react at 150°C for 12 hours;

[0084] 3) Centrifuge the solution obtained from the reaction at 12000 r / min for 5 min and collect the precipitate. Wash the precipitate with anhydrous ethanol to obtain gadolinium-based nanomaterials.

[0085] 4) Take 2 mg of the obtained gadolinium-based nanomaterial and 2 mg of indocyanine green, mix them in an ethanol solution at room temperature and stir for 12 h in the dark to obtain a nanocomposite.

[0086] 5) Dissolve 0.4 mg of hyaluronic acid in 0.2 mL of deionized water, slowly add it dropwise to the above solution, continue magnetic stirring at room temperature for 4 h, and then centrifuge to collect the precipitate.

[0087] Transmission electron microscopy (TEM) revealed that the gadolinium-based composite material prepared in this embodiment was in sheet form, with a length and width of approximately 1000 nm.

[0088] The gadolinium-based composite material prepared in this embodiment can be used as a microwave diagnostic agent. When used as a microwave diagnostic agent, it has MR / fluorescence imaging capability and microwave response heating capability, and exhibits concentration dependence.

[0089] Example 4

[0090] A method for preparing gadolinium-based composite materials with microwave-responsive heat generation and dual-mode imaging effects includes the following steps:

[0091] 1) Weigh 6.77 mg Gd(NO3)3·6H2O, 11.86 mg isophthalic acid and 15.77 mg methyl-2-tetra(4-carboxyphenyl)porphyrin, dissolve them in 30 mL N,N-dimethylformamide, and disperse them evenly by ultrasonication to obtain a mixed solution;

[0092] 2) Place the mixture into a reaction vessel and react at 150°C for 12 hours;

[0093] 3) Centrifuge the solution obtained from the reaction at 12000 r / min for 5 min and collect the precipitate. Wash the precipitate with anhydrous ethanol to obtain gadolinium-based nanomaterials.

[0094] 4) Take 2 mg of the obtained gadolinium-based nanomaterial and 1 mg of rhodamine 6G, mix them in an ethanol solution at room temperature and stir for 12 h in the dark to obtain a nanocomposite.

[0095] 5) Dissolve 0.2 mg of polyethylene glycol in 0.2 mL of deionized water, slowly add it dropwise to the above solution, continue magnetic stirring at room temperature for 6 h, and then centrifuge to collect the precipitate.

[0096] The gadolinium-based composite material prepared in this embodiment can be used as a microwave diagnostic agent. When used as a microwave diagnostic agent, it has MR / fluorescence imaging capability and microwave response heating capability, and exhibits concentration dependence.

[0097] Example 5

[0098] A method for preparing gadolinium-based composite materials with microwave-responsive heat generation and dual-mode imaging effects includes the following steps:

[0099] 1) Weigh 2.26 mg Gd(NO3)3·6H2O, 4.20 mg isophthalic acid and 3.95 mg terephthalene-tetra(4-carboxyphenyl)porphyrin, dissolve them in 30 mL N,N-dimethylformamide, and disperse them evenly by ultrasonication to obtain a mixed solution;

[0100] 2) Place the mixture into a reaction vessel and react at 200°C for 12 hours;

[0101] 3) Centrifuge the solution obtained from the reaction at 12000 r / min for 5 min and collect the precipitate. Wash the precipitate with anhydrous ethanol to obtain gadolinium-based nanomaterials.

[0102] 4) Take 1 mg of the obtained gadolinium-based nanomaterial and 1 mg of indocyanine green, mix them in an ethanol solution at room temperature and stir for 24 h in the dark to obtain a nanocomposite.

[0103] 5) Dissolve 0.2 mg of dopamine in 0.2 mL of deionized water, slowly add it dropwise to the above solution, continue magnetic stirring at room temperature for 6 h, and then centrifuge to collect the precipitate.

[0104] The gadolinium-based composite material prepared in this embodiment can be used as a microwave diagnostic agent. When used as a microwave diagnostic agent, it has MR / fluorescence imaging capability and microwave response heating capability, and exhibits concentration dependence.

[0105] Example 6

[0106] A method for preparing gadolinium-based composite materials with microwave-responsive heat generation and dual-mode imaging effects includes the following steps:

[0107] 1) Weigh 3.71 mg GdCl3·6H2O, 2.21 mg isophthalic acid and 1.22 mg fumaric acid, dissolve them in 30 mL acetonitrile, and disperse them evenly by ultrasonication to obtain a mixed solution;

[0108] 2) Place the mixture into a reaction vessel and react at 200°C for 12 hours;

[0109] 3) Centrifuge the solution obtained from the reaction at 12000 r / min and collect the precipitate. Wash the precipitate with anhydrous ethanol to obtain gadolinium-based nanomaterials.

[0110] 4) Take 2 mg of the obtained gadolinium-based nanomaterial and 2 mg of fluorescein isothiocyanate (FITC), mix them in an ethanol solution at room temperature and stir in the dark for 24 h to obtain the nanocomposite.

[0111] 5) Dissolve 0.8 mg of dopamine in 0.2 mL of deionized water, slowly add it dropwise to the above solution, continue magnetic stirring at room temperature for 6 h, and then centrifuge to collect the precipitate.

[0112] The gadolinium-based composite material prepared in this embodiment can be used as a microwave diagnostic agent. When used as a microwave diagnostic agent, it has MR / fluorescence imaging capability and microwave response heating capability, and exhibits concentration dependence.

[0113] Example 7

[0114] A method for preparing gadolinium-based composite materials with microwave-responsive heat generation and dual-mode imaging effects includes the following steps:

[0115] 1) Weigh 3.71 mg GdCl3·6H2O, 1.22 mg fumaric acid and 8.30 mg of methyl-tetra(4-carboxyphenyl)porphyrin, dissolve them in 30 mL of acetonitrile, and disperse them evenly by ultrasonication to obtain a mixed solution;

[0116] 2) Place the mixture into a reaction vessel and react at 150°C for 9 hours;

[0117] 3) Centrifuge the solution obtained from the reaction at 12000 r / min for 5 min and collect the precipitate. Wash the precipitate with anhydrous ethanol to obtain gadolinium-based nanomaterials.

[0118] 4) Take 2 mg of the obtained gadolinium-based nanomaterial and 1 mg of FITC, mix them in an ethanol solution at room temperature and stir for 12 h in the dark to obtain a nanocomposite.

[0119] 5) Dissolve 0.5 mg of dopamine in 0.1 mL of deionized water, slowly add it dropwise to the above solution, continue magnetic stirring at room temperature for 6 h, and then centrifuge to collect the precipitate.

[0120] The gadolinium-based composite material prepared in this embodiment can be used as a microwave diagnostic agent. When used as a microwave diagnostic agent, it has MR / fluorescence imaging capability and microwave response heating capability, and exhibits concentration dependence.

[0121] Example 8

[0122] A method for preparing gadolinium-based composite materials with microwave-responsive heat generation and dual-mode imaging effects includes the following steps:

[0123] 1) Weigh 3.71 mg GdCl3·6H2O, 1.39 mg fumaric acid and 4.74 mg of 4-tetra(4-carboxyphenyl)porphyrin, dissolve them in 30 mL of N,N-dimethylformamide, and disperse them evenly by ultrasonication to obtain a mixed solution;

[0124] 2) Place the mixture into a reaction vessel and react at 200°C for 12 hours;

[0125] 3) Centrifuge the solution obtained from the reaction at 12000 r / min for 5 min and collect the precipitate. Wash the precipitate with anhydrous ethanol to obtain gadolinium-based nanomaterials.

[0126] 4) Take 2 mg of the obtained gadolinium-based nanomaterial and 1 mg of CY5, mix them in an ethanol solution at room temperature and stir for 12 h in the dark to obtain a nanocomposite.

[0127] 5) Dissolve 0.2 mg of dopamine in 0.2 mL of deionized water, slowly add it dropwise to the above solution, continue magnetic stirring at room temperature for 4 h, and then centrifuge to collect the precipitate.

[0128] The gadolinium-based composite material prepared in this embodiment can be used as a microwave diagnostic agent. When used as a microwave diagnostic agent, it has MR / fluorescence imaging capability and microwave response heating capability, and exhibits concentration dependence.

[0129] Example 9

[0130] A method for preparing gadolinium-based composite materials with microwave-responsive heat generation and dual-mode imaging effects includes the following steps:

[0131] 1) Weigh 4.51 mg Gd(NO3)3·6H2O and 4.20 mg isophthalic acid and dissolve them in 30 mL of n-hexane. After ultrasonic dispersion, a mixed solution is obtained.

[0132] 2) Place the mixture into a reaction vessel and react at 150°C for 10 hours;

[0133] 3) Centrifuge the solution obtained from the reaction at 12000 r / min and collect the precipitate. Wash the precipitate with anhydrous ethanol to obtain gadolinium-based nanomaterials.

[0134] 4) Take 2 mg of the obtained gadolinium-based nanomaterial and 1 mg of ICG, mix them in an ethanol solution at room temperature and stir for 12 h in the dark to obtain a nanocomposite.

[0135] 5) Dissolve 0.5 mg bovine serum albumin in 0.2 mL of deionized water, slowly add it dropwise to the above solution, continue magnetic stirring at room temperature for 6 h, and then centrifuge to collect the precipitate.

[0136] The gadolinium-based composite material prepared in this embodiment can be used as a microwave diagnostic agent. When used as a microwave diagnostic agent, it has MR / fluorescence imaging capability and microwave response heating capability, and exhibits concentration dependence.

[0137] Example 10

[0138] A method for preparing gadolinium-based composite materials with microwave-responsive heat generation and dual-mode imaging effects includes the following steps:

[0139] 1) Weigh 7.42 mg GdCl3·6H2O, 3.98 mg isophthalic acid and 2.08 mg fumaric acid, dissolve them in 30 mL N,N-dimethylformamide, and disperse them evenly by ultrasonication to obtain a mixed solution;

[0140] 2) Place the mixture into a reaction vessel and react at 150°C for 12 hours;

[0141] 3) Centrifuge the solution obtained from the reaction at 12000 r / min and collect the precipitate. Wash the precipitate with anhydrous ethanol to obtain gadolinium-based nanomaterials.

[0142] 4) Take 1 mg of the obtained gadolinium-based nanomaterial and 1 mg of CY5, mix them in an ethanol solution at room temperature and stir for 12 h in the dark to obtain a nanocomposite.

[0143] 5) Dissolve 0.2 mg bovine serum albumin in 0.1 mL of deionized water, slowly add it dropwise to the above solution, continue magnetic stirring at room temperature for 4 h, and then centrifuge to collect the precipitate.

[0144] The gadolinium-based composite material prepared in this embodiment can be used as a microwave diagnostic agent. When used as a microwave diagnostic agent, it has MR / fluorescence imaging capability and microwave response heating capability, and exhibits concentration dependence.

[0145] Performance testing

[0146] The gadolinium-based composite material GM7IH prepared in Example 1 and the gadolinium-based composite material GM8IH prepared in Example 2 were dispersed in culture medium solution, and then co-incubated with L929 cells and H22 cells for 6 hours respectively. The culture medium was then aspirated, the cells were washed with PBS solution, and the cells in the wells were digested and redispersed in PBS solution. The cell uptake of gadolinium-based composite materials was detected by flow cytometry.

[0147] Figure 3 This is a comparison of the cell internalization effects of the gadolinium-based composite materials obtained in Examples 1 and 2. Specifically, it is a single-parameter histogram after processing the flow cytometry measurement results, consisting of a one-dimensional parameter (fluorescence) and cell number (vertical axis), reflecting the number of cells under the same fluorescence intensity. APC-A is the red fluorescence channel, where the right part (APC-A+) represents the proportion of cells with red fluorescence (ICG characteristic light), and the left part represents the proportion of cells without fluorescence. A higher proportion of cells with red fluorescence indicates a higher material internalization rate.

[0148] from Figure 3 As can be seen, compared with ICG alone, the gadolinium-based composite material prepared in this invention has a better cell internalization effect. Among them, the rod-shaped gadolinium-based composite material has a higher cell internalization efficiency than the spherical gadolinium-based composite material.

[0149] The above embodiments are only used to illustrate the technical solutions of the present invention, and are not intended to limit it. Although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art should understand that modifications can still be made to the technical solutions described in the foregoing embodiments, or equivalent substitutions can be made to some of the technical features. Such modifications or substitutions will not cause the essence of the corresponding technical solutions to deviate from the spirit and scope of the technical solutions of the embodiments of the present invention.

[0150] 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 preparation for microwave hyperthermia of tumors, wherein, The formulation includes a gadolinium-based composite material, which comprises gadolinium-based nanomaterials, a fluorescent dye, and a surface modifier. The fluorescent dye is encapsulated within the pores of the gadolinium-based nanomaterials to form a composite of the gadolinium-based nanomaterials and the fluorescent dye. The surface modifier is chemically bonded to the surface of the composite of the gadolinium-based nanomaterials and the fluorescent dye. The gadolinium-based nanomaterials are prepared by a solvothermal reaction of a metal salt containing gadolinium ions and an organic ligand. The organic ligand is selected from one or two of isophthalic acid, terephthalic acid, fumaric acid, L-tyrosine, L-arginine, and methyl-tetra(4-carboxyphenyl)porphyrin; The temperature of the solvothermal reaction is 150-200℃; The gadolinium-based nanomaterial has at least one of the following morphologies: rod-shaped, sheet-shaped, and spherical. The surface modifier is selected from hyaluronic acid; the fluorescent dye is selected from indocyanine green. The median particle size of the gadolinium-based composite material is 150-900 nm.

2. The preparation for microwave hyperthermia of tumors according to claim 1, wherein, The metal salt containing gadolinium ions is selected from one or two of gadolinium chloride, gadolinium nitrate, and gadolinium bicarbonate. And / or, the solvothermal reaction time is 6-12 h; And / or, the mass ratio of the gadolinium-containing metal salt to the organic ligand is 1:1 to 1:

6.

3. The preparation for microwave hyperthermia of tumors according to claim 1, wherein, The mass ratio of the gadolinium-based nanomaterial to the fluorescent dye is 1:1 to 2:

1.

4. The preparation for microwave hyperthermia of tumors according to claim 1, wherein, The mass ratio of the surface modifier to the complex of gadolinium-based nanomaterials and fluorescent dyes is 1:5 to 1:

15.

5. The preparation for microwave hyperthermia of tumors according to claim 1, wherein, The potential range of the gadolinium-based composite material is -40 to 50 mV.

6. The formulation for microwave hyperthermia of tumors according to any one of claims 1-5, wherein the method for preparing the gadolinium-based composite material comprises the following steps: 1) Gadolinium-based nanomaterials are prepared by mixing a metal salt containing gadolinium ions and an organic ligand in an organic solvent and carrying out a solvothermal reaction. 2) The obtained gadolinium-based nanomaterials were mixed with fluorescent dyes and stirred in the dark to prepare a composite of gadolinium-based nanomaterials and fluorescent dyes; 3) The obtained gadolinium-based nanomaterials and fluorescent dye complexes and surface modifiers are mixed and reacted to prepare the gadolinium-based composite material.

7. The preparation for microwave hyperthermia of tumors according to any one of claims 1-5, wherein, The formulation is used in nuclear magnetic resonance and / or fluorescence imaging.

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