Preparation method of multifunctional nanodrug of Fe-based MOF loaded CM-272

Abstract

The application discloses a preparation method of a multifunctional nano drug of Fe-based MOF loaded CM-272, and relates to a preparation method of an anti-cancer nano drug.The application aims at solving the problem that the CM-272 action is limited due to the internal hypoxic environment of a tumor.Method: I, preparing a Fe-based metal organic framework;II, preparing a MIL-53@CM-272 nano drug.The application is used for the preparation of the multifunctional nano drug of the Fe-based MOF loaded CM-272.

Description

Technical Field

[0001] This invention relates to a method for preparing anticancer nanomedicines. Background Technology

[0002] The tumor microenvironment is a crucial area of ​​cancer research. Tumors are abnormally growing tissues that typically exhibit the following characteristics: 1. Rapid cell growth: Tumor cells divide and proliferate uncontrollably, leading to a rapid increase in tumor size; 2. Vigorous metabolism: Cancer cells usually have a high metabolic rate because they require large amounts of energy and nutrients to support rapid growth and division; 3. Lack of a well-developed internal vascular system: Due to the rapid growth of tumors, their internal vascular system cannot keep up with the cells' growth needs, resulting in insufficient blood supply within the tumor; 4. Hypoxic environment: Due to insufficient blood supply, the oxygen supply within the tumor is also insufficient, a condition known as hypoxia; 5. Anaerobic glycolysis: In hypoxic environments, cancer cells often use anaerobic glycolysis to generate energy instead of relying on oxygen respiration. This metabolic pathway produces lactic acid as a byproduct; 6. Acidic microenvironment: Due to lactic acid accumulation, the pH value of the tumor site decreases, creating an acidic environment. This acidic environment promotes the growth of cancer cells while damaging normal cells. These factors collectively create the tumor tissue microenvironment and also increase the invasiveness and metastasis capabilities of cancer cells.

[0003] Recent reports indicate that a novel epigenetic inhibitor, CM-272 (a G9a inhibitor), can effectively inhibit cancer cell proliferation and induce apoptosis. However, the efficacy of CM-272 is significantly limited due to the hypoxic environment within tumors. Summary of the Invention

[0004] This invention aims to address the problem that the function of CM-272 is limited by the hypoxic environment inside tumors, and thus provides a method for preparing a multifunctional nanomedicine of CM-272 supported on Fe-based MOF.

[0005] A method for preparing a multifunctional nanomedicine of Fe-based MOF supported on CM-272, comprising the following steps:

[0006] I. Preparation of Fe-based metal-organic frameworks:

[0007] ① Add 2-aminoterephthalic acid to ethanol and stir for 0.2h to 0.4h in a water bath with magnetic stirring speed of 90rpm to 150rpm and temperature of 30℃ to 50℃ to obtain solution A;

[0008] ② Add FeCl3·6H2O to the ethanol solution and stir for 0.3h to 0.5h in a water bath with magnetic stirring speed of 90rpm~150rpm and temperature of 30℃~50℃ to obtain solution B;

[0009] ③ Mix solution A with solution B and react for 1 to 3 hours in a water bath with magnetic stirring speed of 150 rpm to 200 rpm and temperature of 30℃ to 50℃. Finally, centrifuge, wash and dry to obtain MIL-53.

[0010] II. Preparation of MIL-53@CM-272 nanomedicine:

[0011] MIL-53 and CM-272 were added to dimethyl sulfoxide, mixed by magnetic stirring, and finally washed and dried to obtain a multifunctional nanomedicine of Fe-based MOF loaded with CM-272.

[0012] The beneficial effects of this invention are:

[0013] This invention employs a hydrothermal method to synthesize MIL-53. Subsequently, the drug CM-272 is electron-adsorbed onto the channels of MIL-53 to obtain the final product MIL-53@CM-272, with a CM-272 loading rate of 56.4%. The release rate of CM-272 was 13.1% at pH 7 and 55.6% at pH 6, indicating the acid-responsive disintegration of MIL-53, thereby accelerating the release of CM-272. MIL-53 exhibits a representative IV isotherm, indicating the presence of mesoporous channels. Furthermore, the sample has a specific area of ​​43.97 m². 2 / g, with an average pore size of 4.32nm. The synthesized MIL-53 exhibits a spindle-shaped morphology with uniform size, approximately 100nm.

[0014] This invention effectively prevents Fe from being generated by preparing Fe-based MOFs. 3+ It separates during delivery into the human body and becomes toxic to normal tissues. Iron ions (Fe) 3+ It has vacant orbitals to form coordination catalytic active centers, and can be used as a potent catalase (CAT) catalyst to efficiently catalyze the conversion of H2O2 in tumors into O2 and H2O, effectively alleviating tumor hypoxia and solving the problem that the hypoxic environment inside the tumor limits the function of CM-272.

[0015] This invention provides a highly efficient and controllable drug delivery system. The CM-272 drug, combined with MIL-53 nanomaterials, can be deposited in large quantities at the tumor site through the EPR effect. Simultaneously, while CM-272 is released slowly in normal body fluids, the slightly acidic microenvironment of the tumor site accelerates its release, thus achieving targeted delivery and controlled release of the drug. Ultimately, this results in a significant inhibitory effect on the proliferation and survival of cancer cells.

[0016] This invention can be used in conjunction with sonodynamic therapy. As an oxygen-sensitive treatment method, the efficacy of sonodynamic therapy is often limited by the oxygen concentration at the tumor site. This invention can effectively alleviate the hypoxic microenvironment at the tumor site, improving the efficacy of sonodynamic therapy. Simultaneously, the sound wave energy of sonodynamic therapy enables this invention to more efficiently catalyze the conversion of H2O2 and the generation of reactive oxygen species within the tumor, thereby more effectively killing cancer cells.

[0017] The synthesis process of this invention is relatively simple and uses readily available raw materials, which helps to improve the feasibility and efficiency of the synthesis. The synthesis method of this invention can synthesize the MIL-53@CM-272 complex in high yield, meaning that more product can be produced to meet potential drug delivery needs. The synthesis method is scalable and can be prepared on a large scale as needed, which is important for future clinical applications.

[0018] In summary, this invention utilizes MIL-53 loaded with the CM-272 immunotherapy drug, addressing the limitation of CM-272 efficacy caused by the hypoxic environment within tumors, while simultaneously achieving targeted delivery of nanoparticles and controlled drug release. Therefore, this research provides valuable insights for developing novel tumor treatment strategies and drugs, making a significant contribution to improving patient comfort and enhancing tumor treatment outcomes.

[0019] Instruction manual illustrations

[0020] Figure 1 The image shows a TEM image of MIL-53 prepared in step one of Example 1, with scale bar a at 200 nm, scale bar b at 100 nm, and scale bar c at 50 nm.

[0021] Figure 2 This is a high-angle annular dark-field image of MIL-53 prepared in step one of Example 1;

[0022] Figure 3 The elemental distribution diagram of MIL-53 prepared in step one of Example 1 is shown below. a represents Fe, b represents N, c represents O, and d represents the elemental composite diagram.

[0023] Figure 4The X-ray photoelectron spectrum of MIL-53 prepared in step one of Example 1 is shown in Figure a. The total spectrum is shown in Figure b. The high-resolution Fe 2p XPS spectrum is shown in Figure b.

[0024] Figure 5 A comparison of the zeta potentials of CM-272, MIL-53 prepared in step one of Example 1, and MIL-53@CM-272 prepared in step two of Example 1;

[0025] Figure 6 The N2 adsorption / desorption isotherms and pore size distribution of MIL-53 prepared in step one of Example 1 are shown in Figure a. a is the N2 adsorption / desorption isotherm, and b is the pore size distribution.

[0026] Figure 7 The loading rate and release rate of CM-272 loaded in MIL-53 prepared in step one of Example 1 are shown in Figure a. a) shows the UV absorption spectra of CM-272 dimethyl sulfoxide solutions at different concentrations: 1 (32 μg / mL), 2 (16 μg / mL), 3 (8 μg / mL), 4 (4 μg / mL), and 5 (2 μg / mL); b) shows the standard calibration curve of CM-272 dimethyl sulfoxide solution obtained from a); c) shows the UV absorption spectra of MIL-53 and CM-272 after adding them to dimethyl sulfoxide: 1 (0.2 mg / mL CM-272 dimethyl sulfoxide solution), 2 (20 mg / mL MIL-53 and 5 mg / mL CM-272 added to dimethyl sulfoxide solution in step two of Example 1). The UV absorption spectrum of the supernatant was measured after CM-272 was added to 25 mL of dimethyl sulfoxide; d is the release rate curve of MIL-53@CM-272 prepared in step 2 of Example 1 at different pH values, 1 is pH 6, 2 is pH 7.

[0027] Figure 8 This is a TEM image showing the disintegration of MIL-53@CM-272 prepared in step two of Example 1 under glutathione (GSH) and weakly acidic conditions.

[0028] Figure 9 The XPS and UV-Vis diffuse reflectance spectra of MIL-53@CM-272 prepared in step two of Example 1 are shown. a is the X-ray photoelectron spectrum, and b is the UV-Vis diffuse reflectance spectrum.

[0029] Figure 10 The binding energy curve of MIL-53@CM-272 prepared in step two of Example 1;

[0030] Figure 11 The band structure of MIL-53@CM-272 prepared in step two of Example 1 is related to H2O / ·OH and O2 / ·O2. - A schematic diagram of redox potential;

[0031] Figure 12 It is 1,3-diphenylisobenzofuran (DPBF) and singlet oxygen ( 1 O2) and superoxide anion (·O2) - Interaction diagram;

[0032] Figure 13 The absorbance curves of MIL-53@CM-272 prepared in step two of Example 1 after reacting with 1,3-diphenylisobenzofuran (DPBF) under different ultrasonic irradiation durations are shown. 1 represents 0 min, 2 represents 2 min, 3 represents 4 min, 4 represents 6 min, 5 represents 8 min, and 6 represents 10 min.

[0033] Figure 14 The results of reacting MIL-53@CM-272 prepared in step two of Example 1 with methylene blue under ultrasonic irradiation are shown. 1 represents 0 min, 2 represents 2 min, 3 represents 4 min, 4 represents 6 min, 5 represents 8 min, and 6 represents 10 min.

[0034] Figure 15 The colorimetric reaction of 3,3',5,5'-tetramethylphenyldiaminobiphenyl (TMB) with hydroxyl radicals;

[0035] Figure 16 The UV absorption spectra of MIL-53@CM-272 prepared in step two of Example 1 under ultrasonic irradiation to oxidize TMB are shown below. 1 represents 2 min, 2 represents 4 min, 3 represents 6 min, 4 represents 8 min, and 5 represents 10 min.

[0036] Figure 17 To detect the electron spin resonance (ESR) spectrum of reactive oxygen species generated by MIL-53@CM-272 prepared in step two of Example 1;

[0037] Figure 18 This is a schematic diagram of the reaction between DTNB and GSH.

[0038] Figure 19 The GSH consumption of MIL-53@CM-272 prepared in step two of Example 1 after ultrasonic irradiation treatment is shown in the figures: 1 = 0 min, 2 = 2 min, 3 = 4 min, 4 = 6 min, 5 = 8 min, and 6 = 10 min.

[0039] Figure 20 This is a peroxidase activity test of MIL-53@CM-272 prepared in step two of Example 1. Detailed Implementation

[0040] Specific Implementation Method 1: This implementation method is a method for preparing a multifunctional nanomedicine of Fe-based MOF supported on CM-272, which is carried out according to the following steps:

[0041] I. Preparation of Fe-based metal-organic frameworks:

[0042] ① Add 2-aminoterephthalic acid to ethanol and stir for 0.2h to 0.4h in a water bath with magnetic stirring speed of 90rpm~150rpm and temperature of 30℃~50℃ to obtain solution A;

[0043] ② Add FeCl3·6H2O to the ethanol solution and stir for 0.3h to 0.5h in a water bath with magnetic stirring speed of 90rpm~150rpm and temperature of 30℃~50℃ to obtain solution B;

[0044] ③ Mix solution A with solution B and react for 1 to 3 hours in a water bath with magnetic stirring speed of 150 to 200 rpm and temperature of 30 to 50 ℃. Finally, centrifuge, wash and dry to obtain MIL-53.

[0045] II. Preparation of MIL-53@CM-272 nanomedicine:

[0046] MIL-53 and CM-272 were added to dimethyl sulfoxide, mixed by magnetic stirring, and finally washed and dried to obtain a multifunctional nanomedicine of Fe-based MOF loaded with CM-272.

[0047] Iron ions (Fe) 3+ Fe is an effective catalase catalyst (CAT) that efficiently catalyzes the conversion of H2O2 within tumors into O2 and H2O, thereby alleviating tumor hypoxia and enhancing the efficacy of certain O2-dependent treatments (such as photodynamic and sonodynamic therapy). 3+ As a catalyst, it is of great significance to overcome the hypoxic microenvironment at tumor sites and enhance the efficacy of oxygen-dependent therapies. However, free Fe... 3+ It has toxic effects on normal tissues and cells; therefore, constructing a suitable Fe... 3+ Vectors are crucial for the effective treatment of tumors.

[0048] Reactive oxygen species (ROS) kill cancer cells through various mechanisms. ROS mainly include singlet oxygen. 1 O2, hydroxyl radical ·OH and superoxide anion ·O2 -These substances can directly damage cellular proteins, lipids, and DNA, leading to cellular structural and functional disorders. Cancer cells are often under oxidative stress, impairing their antioxidant mechanisms and making them more susceptible to attack by reactive oxygen species (ROS). High concentrations of ROS exacerbate oxidative stress, leading to cell damage and death. Furthermore, ROS readily induce DNA damage, including single-strand and double-strand breaks, which helps inhibit the growth and division of cancer cells. ROS can also interfere with multiple cellular signaling pathways, including those for cell proliferation and survival, thereby inhibiting cancer cell growth.

[0049] The beneficial effects of this embodiment are:

[0050] This method employs a hydrothermal synthesis of MIL-53. Subsequently, the drug CM-272 was electron-adsorbed onto the channels of MIL-53, yielding the final product MIL-53@CM-272, with a CM-272 loading of 56.4%. The release rate of CM-272 was 13.1% at pH 7 and 55.6% at pH 6, indicating acid-responsive disintegration of MIL-53, thereby accelerating the release of CM-272. MIL-53 exhibited a representative IV isotherm, indicating the presence of mesoporous channels. Furthermore, the sample had a specific area of ​​43.97 m². 2 / g, with an average pore size of 4.32nm. The synthesized MIL-53 exhibits a spindle-shaped morphology with uniform size, approximately 100nm.

[0051] This method effectively prevents Fe from being generated by preparing Fe-based MOFs. 3+ It separates during delivery into the human body and becomes toxic to normal tissues. Iron ions (Fe) 3+ It has vacant orbitals to form coordination catalytic active centers, and can be used as a potent catalase (CAT) catalyst to efficiently catalyze the conversion of H2O2 in tumors into O2 and H2O, effectively alleviating tumor hypoxia and solving the problem that the hypoxic environment inside the tumor limits the function of CM-272.

[0052] This method provides a highly efficient and controllable drug delivery system. The CM-272 drug, combined with MIL-53 nanomaterials, can be deposited in large quantities at the tumor site through the EPR effect. Simultaneously, while CM-272 is released slowly in normal body fluids, the slightly acidic microenvironment at the tumor site accelerates its release, thus achieving targeted delivery and controlled release of the drug. Ultimately, this results in a significant inhibitory effect on the proliferation and survival of cancer cells.

[0053] This method can be used in conjunction with sonodynamic therapy. As an oxygen-sensitive treatment method, the efficacy of sonodynamic therapy is often limited by the oxygen concentration at the tumor site. This method can effectively alleviate the hypoxic microenvironment at the tumor site, improving the efficacy of sonodynamic therapy. Simultaneously, the sound wave energy of sonodynamic therapy enables this invention to more efficiently catalyze the conversion of H2O2 and the generation of reactive oxygen species within the tumor, thereby more effectively killing cancer cells.

[0054] The synthesis process described in this embodiment is relatively simple, utilizing readily available raw materials, which contributes to improved feasibility and efficiency. This method enables the synthesis of the MIL-53@CM-272 complex in high yield, meaning that larger quantities can be produced to meet potential drug delivery needs. The synthesis method is scalable and can be scaled up as needed, which is crucial for future clinical applications.

[0055] In summary, this method utilizes MIL-53 loaded with CM-272 immunotherapy, addressing the limitation of CM-272 efficacy caused by the hypoxic environment within tumors, while simultaneously achieving targeted delivery of nanoparticles and controlled drug release. Therefore, this research provides valuable insights for developing novel tumor treatment strategies and drugs, making a significant contribution to improving patient comfort and enhancing tumor treatment outcomes.

[0056] Specific Implementation Method Two: This implementation method differs from Specific Implementation Method One in that the molar ratio of 2-aminoterephthalic acid to ethanol in step one ① is 1 mmol:(20-30) mL. Everything else is the same as in Specific Implementation Method One.

[0057] Specific Implementation Method Three: This implementation method differs from Specific Implementation Method One or Two in that the molar ratio of FeCl3·6H2O to the volume ratio of ethanol in step one, ②, is 1 mmol:(5-10) mL. Everything else is the same as in Specific Implementation Method One or Two.

[0058] Specific Implementation Method Four: This implementation method differs from Specific Implementation Methods One to Three in that the volume ratio of solution A to solution B in step one ③ is 1:(0.1~0.5). Everything else is the same as in Specific Implementation Methods One to Three.

[0059] Specific Implementation Method Five: This implementation method differs from Specific Implementation Methods One to Four in that the centrifugation described in step one ③ is specifically carried out at a centrifugation speed of 10,000 rpm to 16,000 rpm for 10 to 30 minutes. Everything else is the same as in Specific Implementation Methods One to Four.

[0060] Specific Implementation Method Six: This implementation method differs from Specific Implementation Methods One to Five in that the mass ratio of MIL-53 to CM-272 in step two is 1:(0.2 to 0.4). Everything else is the same as in Specific Implementation Methods One to Five.

[0061] Specific Implementation Method Seven: This implementation method differs from Specific Implementation Methods One to Six in that the total mass ratio of MIL-53 and CM-272 to the volume ratio of dimethyl sulfoxide in step two is 1 mg:(0.6-1.5) mL. Everything else is the same as in Specific Implementation Methods One to Six.

[0062] Specific Implementation Method Eight: This implementation method differs from Specific Implementation Methods One to Seven in that the magnetic stirring and mixing described in step two is specifically carried out in a water bath at a magnetic stirring speed of 150 rpm to 300 rpm and a temperature of 30°C to 50°C for 24 to 48 hours. Everything else is the same as in Specific Implementation Methods One to Seven.

[0063] Specific Implementation Method Nine: This implementation method differs from Specific Implementation Methods One to Eight in that: the washing described in step one ③ specifically involves washing with ethanol 3 to 5 times; the washing described in step two specifically involves washing with deionized water 3 to 5 times. Everything else is the same as in Specific Implementation Methods One to Eight.

[0064] Specific Implementation Method Ten: This implementation method differs from Specific Implementation Methods One to Nine in that the drying described in Step One ③ and Step Two is specifically carried out overnight in a vacuum drying oven at a temperature of 30℃~50℃. Everything else is the same as in Specific Implementation Methods One to Nine.

[0065] The beneficial effects of the present invention are verified using the following embodiments:

[0066] Example 1:

[0067] A method for preparing a multifunctional nanomedicine of Fe-based MOF supported on CM-272, comprising the following steps:

[0068] I. Preparation of Fe-based metal-organic frameworks:

[0069] ① Add 0.4 mmol of 2-aminoterephthalic acid (2-NH2BDC) to 10 mL of ethanol, and stir for 0.2 h in a water bath at 30 °C with magnetic stirring at 90 rpm to obtain solution A;

[0070] ② Add 0.4 mmol FeCl3·6H2O to 3 mL of ethanol solution, and stir for 0.3 h in a water bath with magnetic stirring speed of 90 rpm and temperature of 30℃ to obtain solution B;

[0071] ③ Mix solution A with solution B and react for 2 hours in a water bath with magnetic stirring speed of 160 rpm and temperature of 30℃. Finally, centrifuge, wash and dry to obtain MIL-53.

[0072] The volume ratio of solution A to solution B is 1:0.3;

[0073] II. Preparation of MIL-53@CM-272 nanomedicine:

[0074] 20 mg MIL-53 and 5 mg CM-272 were added to 25 mL of dimethyl sulfoxide, mixed by magnetic stirring, and finally washed and dried to obtain a multifunctional nanomedicine of Fe-based MOF loaded with CM-272, named MIL-53@CM-272.

[0075] The centrifugation described in step 1③ specifically refers to centrifugation at a speed of 14000 rpm for 20 minutes;

[0076] The magnetic stirring and mixing described in step two is specifically carried out in a water bath at a magnetic stirring speed of 160 rpm and a temperature of 30°C for 24 hours.

[0077] The washing described in step 1, ③ specifically involves washing three times with ethanol; the washing described in step 2 specifically involves washing three times with deionized water.

[0078] The drying process described in Step 1③ and Step 2 specifically involves drying overnight in a vacuum drying oven at a temperature of 30°C.

[0079] Figure 1 The image shows a TEM image of MIL-53 prepared in step one of Example 1. The scale bar is 200 nm in a, 100 nm in b, and 50 nm in c. As can be seen from the image, the prepared MIL-53 is spindle-shaped with uniform morphology and a size of about 100 nm. At this size, it can be deposited at the tumor site through the EPR effect, enhancing the permeability and retention effect, which is suitable for the absorption and permeation of MIL-53 in tumors.

[0080] Figure 2 The image shows a high-angle annular dark-field image of MIL-53 prepared in step one of Example 1; as can be seen from the image, MIL-53 was successfully synthesized.

[0081] Figure 3 The image shows the elemental distribution of MIL-53 prepared in step one of Example 1. a represents Fe, b represents N, c represents O, and d is an elemental composite diagram; it illustrates the elemental distribution within the Fe-based MOF structure. Figure 3 and Figure 2 Together, they proved the successful synthesis of MIL-53 nanoparticles.

[0082] Figure 4 The X-ray photoelectron spectrum of MIL-53 prepared in step one of Example 1 is shown in a. a is the total spectrum, and b is the high-resolution Fe 2p XPS spectrum. From a, it can be seen that Fe... 2p Peak, O at 531.95 eV 1s1 / 2 Peak and N at 399.47 eV 1s1 / 2 The peak and Figure b also confirm the successful synthesis of MIL-53.

[0083] Figure 5 A comparison of the zeta potentials of CM-272, MIL-53 prepared in step one of Example 1, and MIL-53@CM-272 prepared in step two of Example 1 is shown in the figure. As can be seen from the figure, the surface zeta potential of MIL-53 decreased from 24.5mV to 6.3mV after loading CM-272, which proves that the drug CM-272 was successfully loaded.

[0084] Figure 6 The figures show the N2 adsorption / desorption isotherms and pore size distribution of MIL-53 prepared in step one of Example 1. Figure a shows the N2 adsorption / desorption isotherm, and figure b shows the pore size distribution. As can be seen from the figures, MIL-53 exhibits a typical IV isotherm, indicating the presence of mesoporous channels. The final measured comparative area of ​​the sample is 43.97 m². 2 / g, with an average pore size of 4.32nm.

[0085] Figure 7 The loading rate and release rate of CM-272 loaded in MIL-53 prepared in step one of Example 1 are shown in Figure a. a) shows the UV absorption spectra of CM-272 dimethyl sulfoxide solutions at different concentrations: 1 (32 μg / mL), 2 (16 μg / mL), 3 (8 μg / mL), 4 (4 μg / mL), and 5 (2 μg / mL); b) shows the standard calibration curve of CM-272 dimethyl sulfoxide solution obtained from a); c) shows the UV absorption spectra of MIL-53 and CM-272 after adding them to dimethyl sulfoxide: 1 (0.2 mg / mL CM-272 dimethyl sulfoxide solution), 2 (20 mg / mL MIL-53 and 5 mg / mL CM-272 added to dimethyl sulfoxide solution in step two of Example 1). The UV absorption spectrum of the supernatant was measured after CM-272 was added to 25 mL of dimethyl sulfoxide; d is the release rate curve of CM-272 prepared by step two of Example 1 at different pH values, 1 is pH 6, 2 is pH 7; ac indicates that the loading rate of CM-272 in the final product MIL-53@CM-272 is 56.4%, and d shows that the release rate of CM-272 is 13.1% at pH 7 and 55.6% at pH 6.

[0086] Figure 8The image shows a TEM image of the disintegration of MIL-53@CM-272 prepared in step two of Example 1 under glutathione (GSH) and weak acidic conditions. It shows that the morphology of MIL-53 changed under GSH and weak acidic conditions at pH 6.5, and the original spindle shape showed obvious disintegration, indicating that MIL-53 has good degradability.

[0087] Figure 9 The XPS and UV-Vis diffuse reflectance spectra of MIL-53@CM-272 prepared in step two of Example 1 are shown. a is the X-ray photoelectron spectrum and b is the UV-Vis diffuse reflectance spectrum. By calculating the slope of the Tacu plot, the narrow bandgap of MIL-53@CM-272 was finally determined to be 2.72 eV.

[0088] Figure 10 The binding energy curve of MIL-53@CM-272 prepared in step two of Example 1 is shown; (ahν) 2 The band gap is obtained by plotting the photon energy (hν), and the maximum valence band value (E) is calculated. VB The conduction band value is 2.25 eV, and the maximum conduction band value is (E CB The value is 0.47 eV.

[0089] Figure 11 The band structure of MIL-53@CM-272 prepared in step two of Example 1 is related to H2O / ·OH and O2 / ·O2. - A schematic diagram of redox potentials; showing the band structure of MIL-53 and H2O / ·OH and O2 / ·O2. - The redox potential of MIL-53, the maximum valence band value (E) VB The redox potential (2.25 eV) is greater than that of H₂O / ·OH (2.01 eV). Therefore, the holes generated in the valence band of MIL-53 can oxidize H₂O to ·OH. Similarly, electrons in the conduction band (CB) contribute to the formation of singlet oxygen (·OH). 1 O2) and superoxide anion (·O2) - ).

[0090] Figure 12 It is 1,3-diphenylisobenzofuran (DPBF) and singlet oxygen ( 1 O2) and superoxide anion (·O2) - Schematic diagram of interactions; singlet oxygen ( 1 O2) and superoxide anion (·O2) - Both can oxidize DPBF, altering its molecular structure and causing it to change from yellow to colorless. Therefore, the change in the absorption characteristics of DPBF before and after the reaction can be used to detect and measure singlet oxygen. 1O2) and superoxide anion (·O2) - The generation of ).

[0091] Two mL of the MIL-53@CM-272 dimethyl sulfoxide solution (50 μg / mL) prepared in step two of Example 1 was mixed with one mL of a dimethyl sulfoxide solution of 1,3-diphenylisobenzofuran (DPBF) (0.1 mg / mL), and then subjected to different ultrasonic irradiations (1 W / cm²). 2 Absorbance curves were tested at 1.0 MHz and 50% duty cycle. Figure 13 The absorbance curves of MIL-53@CM-272 prepared in step two of Example 1 after reacting with 1,3-diphenylisobenzofuran (DPBF) under different ultrasonic irradiation durations are shown below. 1 represents 0 min, 2 represents 2 min, 3 represents 4 min, 4 represents 6 min, 5 represents 8 min, and 6 represents 10 min. The curves show that the absorbance of 1,3-diphenylisobenzofuran (DPBF) decreases significantly with increasing ultrasonic irradiation time, directly proving that singlet oxygen (… 1 O2) and superoxide anion (·O2) - The generation of ).

[0092] 2 mL of the MIL-53@CM-272 aqueous solution (50 μg / mL) prepared in step two of Example 1 was mixed with 1 mL of methylene blue (MB) aqueous solution (0.1 mg / mL), and the mixture was subjected to ultrasonic irradiation (1 W / cm²). 2 Under ultrasonic irradiation at 1.0 MHz and 50% duty cycle for 0 min, 2 min, 4 min, 6 min, 8 min and 10 min, the generation of hydroxyl radicals was detected by monitoring the degradation of methylene blue; Figure 14 The results of the reaction between MIL-53@CM-272 prepared in step two of Example 1 and methylene blue under ultrasonic irradiation are shown in the figure. 1 represents 0 min, 2 represents 2 min, 3 represents 4 min, 4 represents 6 min, 5 represents 8 min, and 6 represents 10 min. It can be clearly observed in the figure that the absorbance of methylene blue at 660 nm decreases significantly with the extension of ultrasonic irradiation time, which indicates the generation of hydroxyl radicals (·OH) under ultrasonic action.

[0093] Figure 15This describes the colorimetric reaction of 3,3',5,5'-tetramethylphenyldiaminobiphenyl (TMB) with hydroxyl radicals; TMB is a colorless organic molecule. When TMB reacts with hydroxyl radicals (·OH), the hydroxyl radical captures a hydrogen atom from the TMB molecule, forming a TMB hydroxyl radical. This hydroxyl radical further reacts with other TMB molecules, leading to the oxidation of TMB. The product of TMB oxidation is blue, a color that can be measured using an optical spectrophotometer. Therefore, by measuring the change in absorbance of the TMB solution, the amount of hydroxyl radicals produced can be determined.

[0094] 2 mL of the MIL-53@CM-272 aqueous solution (50 μg / mL) prepared in step two of Example 1 was mixed with 1 mL of LTMB ethanol solution (3.2 mM), and the mixture was subjected to ultrasonic irradiation (1 W / cm²). 2 The ultraviolet absorption spectra of oxidized TMB were measured after ultrasonic irradiation for 2 min, 4 min, 6 min, 8 min and 10 min at 1.0 MHz and 50% duty cycle. Figure 16 The image shows the UV absorption spectrum of TMB oxidized by MIL-53@CM-272 prepared in step two of Example 1 under ultrasonic irradiation. 1 represents 2 min, 2 represents 4 min, 3 represents 6 min, 4 represents 8 min, and 5 represents 10 min. In the figure, the characteristic peaks of oxidized TMB at 375 nm and 675 nm gradually increase, which confirms the generation of hydroxyl radicals (·OH) under ultrasonic irradiation.

[0095] In order to detect 1 O2, ·OH and ·O2 - The ESR signal was measured by taking 1 mL of the MIL-53@CM-272 aqueous solution (100 μg / mL) prepared in step two of Example 1 and mixing it with TEMP (0.1 M, 50 μL). 1 O2 detection), DMPO (0.02M, 50μL, ·OH detection), BMPO (0.02M, 50μL, ·O2 detection), - (Detection) Mixed, under US irradiation (1W / cm) 2 After 5 minutes (1.0MHz and 50% duty cycle), the ROS signal was measured using a Bruker E500. Figure 17 To detect the electron spin resonance (ESR) spectrum of reactive oxygen species generated by MIL-53@CM-272 prepared in step two of Example 1; the ESR spectrum shows the singlet oxygen, superoxide anion, and hydroxyl radical detected under ultrasonic irradiation, with characteristic peaks corresponding to 1 O2 (1:1:1), ·OH (1:2:2:1) and ·O2 - (2:2:1:2:1:2) demonstrates the ROS generation induced by ultrasound irradiation in MIL-53@CM-272.

[0096] Figure 18 This is a schematic diagram of the reaction between DTNB and GSH; the reaction between 5,5'-dithiobenzoic acid (DTNB) and glutathione (GSH) is a common biochemical reaction used to detect the presence and activity of GSH. This reaction changes the color of DTNB by reducing the thioacryl group, resulting in a change in the absorption peak; DTNB changes from yellow to the blue TNB observed at 412 nm.

[0097] Mix 2 mL of the MIL-53@CM-272 aqueous solution prepared in step two of Example 1 with 1 mL of glutathione (GSH) (10 mM), then add 20 μL of DTNB (2 mM), and then place the mixture in a 1 W / cm solution. 2 The change in absorbance at 412 m was measured using a UV-Vis spectrophotometer under ultrasonic irradiation at 1.0 MHz and 50% duty cycle. Figure 19 The GSH consumption of MIL-53@CM-272 prepared in step two of Example 1 after ultrasonic irradiation treatment is shown in the figures: 1 = 0 min, 2 = 2 min, 3 = 4 min, 4 = 6 min, 5 = 8 min, and 6 = 10 min. The absorbance change at 412 nm was detected by spectrophotometry. It was found that the curve at 412 nm decreased significantly with the extension of reaction time, indicating the consumption of glutathione (GSH).

[0098] 10 mL of the MIL-53@CM-272 aqueous solution (100 μg / mL) prepared in step two of Example 1 was mixed with 10 mL of H2O2 (0.1 mM) in a beaker. The beaker was then sealed with wax, and dissolved oxygen was measured using an AR8406 dissolved oxygen meter with and without US irradiation (1 W / cm²). 2 The ability to generate O2 under conditions of 1.0 MHz and 50% duty cycle was divided into two groups: ultrasonic irradiation and no ultrasonic irradiation. Figure 20 The peroxidase activity of MIL-53@CM-272 prepared in step two of Example 1 was tested. It can be clearly seen that the O2 generated under 30 min of ultrasonic irradiation was 10.6 mg / L, which is significantly higher than 8.5 mg / L without ultrasonic treatment.

Claims

1. A method for preparing a multifunctional nanomedicine of Fe-based MOF supported on CM-272, characterized in that... It is done in the following steps: I. Preparation of Fe-based metal-organic frameworks: ① Add 2-aminoterephthalic acid to ethanol and stir for 0.2h to 0.4h in a water bath with magnetic stirring speed of 90rpm to 150rpm and temperature of 30℃ to 50℃ to obtain solution A; ② Add FeCl3·6H2O to the ethanol solution and stir for 0.3h to 0.5h in a water bath with magnetic stirring speed of 90rpm~150rpm and temperature of 30℃~50℃ to obtain solution B; ③ Mix solution A with solution B and react for 1 to 3 hours in a water bath with magnetic stirring speed of 150 to 200 rpm and temperature of 30 to 50 ℃. Finally, centrifuge, wash and dry to obtain MIL-53. II. Preparation of MIL-53@CM-272 nanomedicine: MIL-53 and CM-272 were added to dimethyl sulfoxide, mixed by magnetic stirring, and finally washed and dried to obtain a multifunctional nanomedicine of Fe-based MOF loaded with CM-272.

2. The method for preparing a multifunctional nanomedicine of Fe-based MOF supported on CM-272 according to claim 1, characterized in that... The molar ratio of 2-aminoterephthalic acid to ethanol in step 1① is 1 mmol:(20-30) mL.

3. The method for preparing a multifunctional nanomedicine of Fe-based MOF supported on CM-272 according to claim 1, characterized in that... The molar ratio of FeCl3·6H2O to ethanol mentioned in step 1② is 1 mmol: (5~10) mL.

4. The method for preparing a multifunctional nanomedicine of Fe-based MOF supported on CM-272 according to claim 1, characterized in that... The volume ratio of solution A to solution B in step 1③ is 1:(0.1~0.5).

5. The method for preparing a multifunctional nanomedicine of Fe-based MOF supported on CM-272 according to claim 1, characterized in that... The centrifugation described in step 1③ specifically refers to centrifugation at a speed of 10,000 rpm to 16,000 rpm for 10 to 30 minutes.

6. The method for preparing a multifunctional nanomedicine of Fe-based MOF supported on CM-272 according to claim 1, characterized in that... The mass ratio of MIL-53 to CM-272 mentioned in step two is 1:(0.2 to 0.4).

7. The method for preparing a multifunctional nanomedicine of Fe-based MOF supported on CM-272 according to claim 1, characterized in that... The total mass of MIL-53 and CM-272 mentioned in step two is in the volume ratio of dimethyl sulfoxide to 1 mg: (0.6-1.5) mL.

8. The method for preparing a multifunctional nanomedicine of Fe-based MOF supported on CM-272 according to claim 1, characterized in that... The magnetic stirring and mixing described in step two is specifically carried out in a water bath with a magnetic stirring speed of 150 rpm to 300 rpm and a temperature of 30°C to 50°C for 24 to 48 hours.

9. The method for preparing a multifunctional nanomedicine of Fe-based MOF supported on CM-272 according to claim 1, characterized in that... The washing described in step 1, ③ specifically involves washing with ethanol 3 to 5 times; the washing described in step 2 specifically involves washing with deionized water 3 to 5 times.

10. The method for preparing a multifunctional nanomedicine of Fe-based MOF supported on CM-272 according to claim 1, characterized in that... The drying process described in Step 1, Step 2 specifically involves drying overnight in a vacuum drying oven at a temperature of 30℃ to 50℃.

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