Application of deferasirox in preparation of medicine for preventing ventricular remodeling after acute myocardial infarction

Abstract

The invention discloses application of deferasirox in preparation of a medicine for preventing ventricular remodeling after acute myocardial infarction, and relates to the technical field of medicines, in particular to application of deferasirox in preparation of a medicine for preventing ventricular remodeling after acute myocardial infarction. The medicine plays a role in inhibiting ferroptosis and inflammatory reaction of myocardial cells so as to prevent ventricular remodeling after acute myocardial infarction; deerasirox plays an iron chelation role and an anti-oxidative stress injury role in the medicine so as to inhibit ferroptosis and inflammatory response of myocardial cells; the inhibition effect of the medicine on myocardial cell ferroptosis is that GPX4 protein is up-regulated; the dosage of deferasirox in the medicine is 30 mg / Kg every day; the deferasirox can prevent ventricular remodeling after myocardial infarction of mice by inhibiting ferroptosis and inflammatory response, including improvement of cardiac function indexes and reduction of myocardial infarction area, and a new idea and a new scheme are provided for clinical prevention of ventricular remodeling after acute myocardial infarction.

Description

Technical Field

[0001] The present invention relates to the field of medical technology, and in particular to application of deferasirox in the preparation of a medicament for preventing ventricular remodeling after acute myocardial infarction. Background Art

[0002] Acute myocardial infarction (AMI) is a cardiovascular disease that severely impacts both life expectancy and quality of life. AMI is characterized by rapid onset, numerous complications, severe illness, and high mortality, resulting from acute obstruction and interruption of coronary blood flow, leading to localized ischemic necrosis of the corresponding myocardium. Epidemiological surveys indicate that the overall mortality rate among AMI patients in China continues to rise, posing a significant health risk to the Chinese public. While the widespread use and standardization of percutaneous coronary intervention (PCI) and the establishment of numerous chest pain centers have significantly reduced the mortality rate in the acute phase of AMI in recent years, the prevalence of heart failure caused by post-AMI ventricular remodeling has not decreased. Ventricular remodeling, driven by myocardial cell death after AMI, is a key determinant of heart failure and significantly impacts the life expectancy and quality of life of AMI patients.

[0003] Currently, most drugs that delay ventricular remodeling work by inhibiting or interacting with the renin-angiotensin-aldosterone system (RAAS). These drugs include angiotensin-converting enzyme inhibitors (ACEIs), angiotensin II receptor blockers (ARBs), and aldosterone antagonists, as well as β-adrenergic receptor antagonists, sodium-glucose cotransporter 2 (SGLT2) inhibitors, and angiotensin receptor neprilysin inhibitors (ARNIs). Long-term combined therapy can improve clinical symptoms, hemodynamic status, and ventricular remodeling. However, ACEIs and ARBs increase plasma renin activity, only temporarily lower plasma aldosterone levels, and fail to block mineralocorticoid receptors, resulting in unsatisfactory clinical results. The incidence of ventricular remodeling remains 25% to 30% in STEMI patients receiving comprehensive treatment. In high-risk myocardial infarction patients with decreased left ventricular function, the combination of conventional ACEIs / ARBs and β-blockers fails to improve ventricular remodeling, leading to progressive worsening of heart failure and even more adverse events. In addition, some drugs that inhibit ventricular remodeling still have certain limitations for patients with renal insufficiency, hypotension, etc.

[0004] Ferroptosis is a unique, newly discovered process of programmed cell death. In recent years, a growing body of research has demonstrated that ferroptosis, as a link between redox biology and cellular biological function, has a potential role in a wide range of diseases, including heart disease, degenerative diseases, retinal diseases, tumors, diabetes, autoimmune diseases, and infectious diseases. It differs significantly from apoptosis, necrosis, and autophagy in morphology, biochemistry, and genetics. Its key characteristics are iron dependency and lipid peroxidation, ultimately leading to the accumulation of large amounts of reactive oxygen species, which in turn cause cell death and damage. Ferroptosis is a complex process, and its mechanisms are not fully understood. Existing research suggests that the mechanisms of ferroptosis primarily involve abnormal iron metabolism, lipid peroxidation, and an imbalance in oxidative-antioxidant homeostasis. These mechanisms are intertwined and mutually influential, collectively regulating the onset and progression of ferroptosis. Ferroptosis is an oxidative cell death induced by small molecules and is iron-dependent, characterized by the following features: The cell death process is accompanied by the accumulation of large amounts of iron ions and lipid peroxidation. Within the cell's microstructure, mitochondria appear smaller than normal, with wrinkled mitochondrial membranes, reduced or absent mitochondrial cristae, and fragmented outer membranes. Electron microscopy reveals reduced mitochondrial size and increased double membrane density. Ferroptosis is caused by an imbalance in the generation and degradation of reactive oxygen species (ROS) in intracellular lipids. Ferroptosis inducers act directly or indirectly on glutathione peroxidase (GPXs) through various pathways, including those regulating iron homeostasis, the RAS pathway, and the cystine transport pathway, leading to a decrease in cellular antioxidant capacity and the accumulation of ROS. The generated ROS directly damage various subcellular organelles and / or disrupt normal cellular structure, ultimately leading to rapid oxidative cell death. A variety of substances and external conditions can trigger ferroptosis. For example, the small molecule erastin inhibits the cystine-glutamate exchanger on the plasma membrane, reducing cellular cystine availability and hindering the synthesis of glutathione, a substrate for GPX4. This in turn triggers the accumulation of membrane lipid ROS and ferroptosis. A small molecule, RSL3, acts as a GPX4 inhibitor and can also induce ferroptosis. Furthermore, nuclear factor erythroid 2-related factor 2 (Nrf2) increases SLC7A11 levels and transcriptionally induces GPX4 expression, thereby reducing ROS. Consequently, overexpression of Nrf2 can inhibit ferroptosis, while Kelch-like ECH-associated protein 1 (Keap1), which binds to and negatively regulates Nrf2, can reverse this process and exert its ferroptotic effects. Nrf2 can also induce ferroptosis by upregulating Hmox1, which then degrades heme and releases free iron.

[0005] Iron chelators have been clinically used to treat iron overload and hold promise as a therapeutic strategy for preventing ferroptosis. Deferoxamine (DFO) is a clinically approved iron chelator with a high affinity for binding Fe3+. In an isolated mouse heart model of I / R injury, DFO administration exerted cardioprotective effects by inhibiting cytosolic ROS production. Dexazolidine (DXZ), another iron chelator, also exerted cardioprotective effects by inhibiting ferroptosis in an in vivo mouse model of I / R injury. Iron chelators, used to inhibit ferroptosis, restored function in both in vitro and in vivo experiments and exerted overall cardioprotective effects against I / R injury. Furthermore, by inhibiting ferroptosis, inflammation was suppressed, limiting the extent of left ventricular remodeling after I / R injury. Therefore, inhibiting ferroptosis may prevent left ventricular remodeling after myocardial infarction by reducing cardiomyocyte death and inflammatory responses, and may represent a potential therapeutic target for the prevention of subsequent heart failure.

[0006] Deferasirox is an orally effective iron chelator primarily used to treat iron overload caused by chronic transfusion-dependent diseases associated with blood transfusions, such as thalassemia and myelodysplastic syndrome. Compared with traditional iron chelators (such as deferoxamine), deferasirox has high oral bioavailability and myocardial targeting. Clinical studies have shown that deferasirox can effectively reduce iron deposition in organs such as the liver and heart, reducing complications associated with iron overload. In addition to its iron chelation properties, deferasirox also possesses multiple pharmacological properties, including antifungal, anti-cellular, anti-malarial, anti-oxidative stress, and anti-cytotoxicity-induced apoptosis. During acute myocardial infarction (AMI), myocardial cells undergo severe oxidative stress, and deferasirox mitigates this damage through its antioxidant properties, thereby slowing the progression of ventricular remodeling. Deferasirox also resists cytotoxicity-induced apoptosis, which helps reduce the number of myocardial cell deaths after AMI and further protects myocardial function. Although deferasirox's primary effect is not anti-inflammatory, its antifungal and anti-cell proliferation properties indirectly affect the inflammatory response after AMI. However, the inflammatory response plays a key role in ventricular remodeling, and deferasirox mitigates ventricular remodeling by affecting the inflammatory response. Its iron chelation and antioxidant effects, through the inhibition of ferroptosis, make deferasirox important in preventing ventricular remodeling after myocardial infarction. The present invention provides a method for preparing a drug for preventing ventricular remodeling after acute myocardial infarction. Summary of the Invention

[0007] In view of this, the present invention addresses the deficiencies in the prior art, and its main purpose is to provide a use of deferasirox in the preparation of a drug for preventing ventricular remodeling after acute myocardial infarction. Deferasirox can prevent ventricular remodeling after myocardial infarction in mice by inhibiting ferroptosis and inhibiting inflammatory response, including improving cardiac function indicators and reducing myocardial infarction area, providing new ideas and new solutions for the clinical prevention of ventricular remodeling after acute myocardial infarction.

[0008] To achieve the above object, the present invention adopts the following technical solutions:

[0009] Use of deferasirox in the preparation of drugs for preventing ventricular remodeling after acute myocardial infarction.

[0010] As a preferred embodiment: the drug exerts its effect by inhibiting myocardial cell ferroptosis and inflammatory response, thereby preventing ventricular remodeling after acute myocardial infarction; deferasirox exerts iron chelation and anti-oxidative stress damage effects in the drug, thereby inhibiting myocardial cell ferroptosis and inflammatory response.

[0011] As a preferred embodiment, the drug achieves its effect of inhibiting myocardial cell ferroptosis by upregulating the relative expression level of GPX4 protein.

[0012] As a preferred embodiment, the drug is administered 6 hours after the occurrence of acute myocardial infarction.

[0013] As a preferred solution: the drug administration cycle is from the 1st day to the 14th day after the occurrence of acute myocardial infarction, the administration frequency is once a day, and the dosage of deferasirox in the drug is 20-30 mg / K per day.

[0014] As a preferred embodiment, the deferasirox is used to prepare a drug for preventing ventricular remodeling after acute myocardial infarction, and the dosage of the deferasirox in the drug is 30 mg / kg per day.

[0015] As a preferred embodiment, the administration mode of the drug is selected from oral administration and injection administration.

[0016] As a preferred solution: the Deferasirox is obtained by the following steps: S1, obtaining a crude Deferasirox product; S2, obtaining Deferasirox.

[0017] As a preferred embodiment, the specific steps for obtaining the crude Deferasirox are as follows:

[0018] S11, reacting a salicylic acid derivative monomer and cyanuric chloride under stirring and condensation reflux conditions;

[0019] S12, gradually increasing the temperature of the reaction system from 60°C to 120°C, and continuing the reaction;

[0020] S13, stopping the reaction, and after the reaction product is cooled into a solid block, adding DMF to dissolve the product, using deionized water to precipitate, and filtering to obtain a powder;

[0021] S14. Wash the powder with deionized water for multiple times, and then freeze-dry to obtain crude deferasirox.

[0022] As a preferred embodiment, the specific steps for obtaining the deferasirox are as follows:

[0023] S21, dissolving the crude deferasirox and the stabilizer in DMSO respectively to prepare solutions;

[0024] S22, mixing the deferasirox solution and the stabilizer solution to obtain an oil phase, and adding the oil phase to water with stirring to obtain a solution;

[0025] S23. The solution is transferred into an ultrafiltration tube with a molecular weight cut-off greater than 100 kDa, centrifuged, and washed to obtain deferasirox.

[0026] Compared with the prior art, the present invention has obvious advantages and beneficial effects. Specifically, it can be seen from the above technical scheme that the deferasirox of the present application can prevent ventricular remodeling after myocardial infarction in mice by inhibiting ferroptosis and inhibiting inflammatory response, including improving cardiac function indicators and reducing myocardial infarction area. The degree of cardiac fibrosis in mice is also significantly reduced, providing new ideas and new solutions for the clinical prevention of ventricular remodeling after acute myocardial infarction; deferasirox inhibits ferroptosis by targeted chelation of free iron ions, reduces myocardial cell death and inflammatory infiltration, and significantly reduces the degree of cardiac fibrosis in mice.

[0027] To more clearly illustrate the structural features and effects of the present invention, it is described in detail below with reference to the accompanying drawings and specific embodiments. BRIEF DESCRIPTION OF THE DRAWINGS

[0028] Figure 1 A roadmap for the use of delarotux of the present invention in preventing ventricular remodeling after acute myocardial infarction;

[0029] Figure 2 A technical roadmap for the administration of delarotix according to the present invention;

[0030] Figure 3 Representative M-mode echocardiogram images of the blank group, sham operation group, model group, and different doses of deferasirox in mice 28 days after myocardial infarction in the examples of the present invention;

[0031] Figure 4 Schematic diagram of left ventricular ejection fraction, a cardiac function index, before, 14 days after, and 28 days after myocardial infarction in each group of mice in an embodiment of the present invention;

[0032] Figure 5 Schematic diagram of the short-axis shortening rate of cardiac function indicators before, 14 days after, and 28 days after myocardial infarction in each group of mice in the examples of the present invention;

[0033] Figure 6 This is a diagram showing the QPCR results of each group of mice in the examples of the present invention;

[0034] Figure 7This is a WB analysis chart of each group of mice in the examples of the present invention;

[0035] Figure 8 Schematic diagram of the ELISA test results of myocardial tissue in an embodiment of the present invention;

[0036] Figure 9 Schematic diagram of the analysis of serum BNP results at different points in each group in the examples of the present invention;

[0037] Figure 10 This is an analysis chart of the serological CRP results of each group at different points in the embodiment of the present invention;

[0038] Figure 11 This is a diagram showing the TTC staining results of each group of mice on the 28th day of TTC in the examples of the present invention;

[0039] Figure 12 This is a representative cardiac MRI image of each group of mice 28 days after surgery in the examples of the present invention;

[0040] Figure 13 Representative transmission electron micrographs of the embodiments of the present invention. DETAILED DESCRIPTION

[0041] The present invention Figures 1 to 13 As shown, the use of deferasirox in the preparation of a drug for preventing ventricular remodeling after acute myocardial infarction.

[0042] The drug works by inhibiting myocardial cell ferroptosis and inflammatory response, thereby preventing ventricular remodeling after acute myocardial infarction; deferasirox exerts iron chelation and anti-oxidative stress damage effects in the drug, thereby inhibiting myocardial cell ferroptosis and inflammatory response.

[0043] The drug inhibits myocardial cell ferroptosis by upregulating the relative expression of GPX4 protein.

[0044] The drug should be administered starting 6 hours after the onset of acute myocardial infarction.

[0045] The drug's administration cycle is from the 1st day to the 14th day after acute myocardial infarction, with a dosing frequency of once a day. The dosage of deferasirox in the drug is 20-30 mg / K per day.

[0046] The deferasirox is used for preparing a medicine for preventing ventricular remodeling after acute myocardial infarction. The dosage of the deferasirox in the medicine is 30 mg / kg per day.

[0047] The administration mode of the drug is selected from oral administration and injection administration.

[0048] The deferasirox is obtained by the following steps: S1, obtaining a crude product of deferasirox; S2, obtaining deferasirox.

[0049] The specific steps for obtaining the crude product of Deferasirox are as follows:

[0050] S11, reacting a salicylic acid derivative monomer (2-hydroxybenzoic acid-4-trifluoromethylaniline Schiff base) with cyanuric chloride under stirring and reflux conditions at 60° C. for 1 h;

[0051] S12, increasing the temperature of the reaction system from 60°C to 120°C at a rate of 10°C per 15 minutes, and continuing the reaction for 4 hours;

[0052] S13, stopping the reaction, and after the reaction product is cooled into a solid block, adding DMF to dissolve the product, using deionized water to precipitate, and filtering to obtain a powder;

[0053] S14. Wash the powder with deionized water for multiple times, and then freeze-dry to obtain crude deferasirox.

[0054] The specific steps for obtaining Deferasirox are:

[0055] S21, dissolving the crude deferasirox and the stabilizer DSPE-PEG2000 in DMSO to prepare a 20 mg / mL solution;

[0056] S22, uniformly mixing the deferasirox solution and the DSPE-PEG2000 solution to obtain an oil phase, adding the oil phase dropwise to water under stirring at 1600 rpm, and continuing stirring for 1 min before stopping stirring;

[0057] S23. Transfer the solution to an ultrafiltration tube with a molecular weight cutoff greater than 100 kDa, centrifuge at 2000 rpm for 8-10 min to remove the organic solvent, and add ultrapure water for washing. Repeat this process twice to obtain a refined product of Deferasirox.

[0058] Unless otherwise specified, the test materials used in the following examples can be purchased through conventional commercial channels.

[0059] Example: Use of Deferasirox in the preparation of a drug for preventing ventricular remodeling after acute myocardial infarction: Deferasirox prevents the occurrence of ventricular remodeling after acute myocardial infarction in mice.

[0060] (1) Preparation of acute myocardial infarction model in mice

[0061] All mice were housed individually in an SPF animal facility for one week at a room temperature of (24±2)°C, with a 12-hour day / night cycle and free access to food and water. After one week of adaptive feeding, an AMI model was established. Following ligation of the left anterior descending coronary artery, myocardial pallor and decreased wall motion were observed in the anterior wall of the left ventricle, along with elevated ST segments on the electrocardiogram (ECG).

[0062] (2) Method of administration

[0063] The drug was administered by gavage and the rats were returned to their cages after administration. The therapeutic dose was 30 mg / kg and was administered every day after surgery for 14 days.

[0064] (3) Improve the echocardiography process

[0065] ① Mouse anesthesia and hair removal: Anesthetize the mouse. After complete anesthesia, use hair removal cream to remove the hair from the chest to the upper abdomen. Place the mouse in a supine position on the ultrasound operation panel.

[0066] ② Use tape to fix the mouse's limbs to the ultrasound electrode sheet, and apply coupling agent to the skin of its limbs and heart area to avoid bubbles.

[0067] ③ Perform detection using an echocardiography system with a 30-MHz imaging sensor. When detecting the long axis of the heart, point the ultrasound probe notch toward the mouse's head, rotate it counterclockwise about 45°, and adjust the probe to fit the mouse's skin.

[0068] ④ Turn on the B-mode ultrasound mode and adjust the operation panel according to the image until the B-mode echocardiogram of the left ventricular long axis can be clearly observed.

[0069] ⑤Then turn on the M-mode ultrasound mode, adjust the sampling line to the correct position, and press the M-mode ultrasound mode again to obtain the M-mode echocardiogram of the left ventricular long axis.

[0070] When detecting the short axis of the heart, rotate the probe notch 90° clockwise and adjust the ultrasound operation panel according to the screen image until a clear circular cardiogram with two papillary muscles is displayed.

[0071] ⑥ Repeat the above steps (3)-(6) to obtain B-mode and M-mode echocardiograms.

[0072] ⑦Throughput: left ventricular ejection fraction (EF%), left ventricular fractional shortening (FS%), left ventricular end-diastolic diameter (LVID.d), and left ventricular end-systolic diameter (LVID.s).

[0073] (4) MRI

[0074] Scanning was performed using a 9.4T MRI. Mice were anesthetized with isoflurane mixed with high-concentration pure oxygen (99.5%, 1 L / min). After anesthesia took effect, the mice were placed on the scanning bed and maintained in the prone position under continuous anesthesia. After positioning scanning, a fast low-angle excitation cine sequence (FLASH-cine) was scanned. The scan included vertical long-axis and horizontal long-axis views of the left ventricle, as well as short-axis views of the heart. Scanning parameters included echo time (TE) 2.5ms, repetition time (TR) 40ms, flip angle (FS) 35, imaging field of view (FOV) 60cm×60cm, matrix: 256×256, imaging thickness 1mm, interslice spacing: 0, and number of excitations 20. After scanning, cardiac MRI short-axis cine images were imported into the analysis software CVI42short3D module, and the end-diastolic and end-systolic myocardial endomyocytes and epicardials of each layer of the left ventricle were manually traced. Import long-axis two-chamber / four-chamber and short-axis cine images into the tissue tracking module and manually trace the end-diastolic endocardium and epicardium of each layer of the left ventricle, marking the superior and inferior poles of the ventricular septum. Include the left ventricular papillary muscles in the blood pool and the left ventricular outflow tract in the ventricular volume.

[0075] (5) Tissue embedding and sectioning

[0076] ① Fix fresh tissue in fixative for at least 24 hours. Remove the tissue from the fixative and trim the desired area with a scalpel in a fume hood. Place the trimmed tissue in an embedding frame, mark the frame, and place the frame containing the tissue in a dehydration box.

[0077] ② Dehydration and wax dipping: Place the dehydration box in the hanging basket and dehydrate in a dehydrator using a gradient of alcohol: 75% alcohol for 4 hours, 85% alcohol for 2 hours, 90% alcohol for 2 hours, 95% alcohol for 1 hour, absolute ethanol I for 30 minutes, absolute ethanol II for 30 minutes, benzene for 5-10 minutes, xylene I for 5-10 minutes, xylene II for 5-10 minutes, paraffin wax I at 65° for 1 hour, paraffin wax II at 65° for 1 hour, paraffin wax III at 65° for 1 hour.

[0078] ③ Embedding: Remove the tissue from the embedding frame and place the melted wax into the frame. Before the wax solidifies, place the tissue into the frame according to the embedding surface requirements. Cool in a -20°C freezer. Once the wax solidifies, remove the wax block from the embedding frame and trim it.

[0079] ④ Sectioning: Place the trimmed wax block on a paraffin microtome and slice to 4 μm thickness. Float the slices on a 40°C warm water slide to flatten the tissue. Remove the tissue with a glass slide and bake in a 60°C oven. Once the wax is melted, remove the slices and store at room temperature until ready for use.

[0080] (6) HE staining

[0081] ① Paraffin section deparaffinization to water: put the section into xylene I 20 min-xylene II 20 min-anhydrous ethanol I 10 min-anhydrous ethanol II 10 min-95% alcohol 5 min-90% alcohol 5 min-80% alcohol 5 min-70% alcohol 5 min-distilled water.

[0082] ② Hematoxylin staining of cell nucleus: put the section into hematoxylin dye for 3-8 min, wash with tap water, differentiate with 1% hydrochloric acid alcohol for several seconds, wash with tap water, return to blue with 0.6% ammonia water, and wash with running water.

[0083] ③ Eosin staining of cytoplasm: put the section into eosin dye for 1-3 min.

[0084] ④ Dehydration and mounting: put the section into 95% alcohol I 5 min-95% alcohol II 5 min-anhydrous ethanol I 5 min-anhydrous ethanol II 5 min-xylene I 5 min-xylene II 5 min for dehydration and transparency, take the section out of xylene, dry slightly, and mount with neutral balsam.

[0085] ⑤ Microscope examination, image collection and analysis.

[0086] (7) TTC staining

[0087] ① Sample pretreatment: immediately after the death of the animal subjected to myocardial ischemia-reperfusion experiment, take the ischemic infarct tissue, wrap it with OCT embedding agent in a clean culture dish, and freeze it at -20°C for 30-60 min. If the frozen section cannot be prepared in time, it needs to be stored at -80°C. Before staining, slightly thaw the tissue at room temperature, and cut it into a section with a sharp knife, with a thickness of 2-3 mm, and ensure that the section is neat.

[0088] ② TTC staining: preheat the TTC dye at 37°C for 30 min, immerse the tissue in the dye at 37°C for 30 min in a dark water bath (every 10 min, gently shake the dye to make the tissue appear dark red, recover the dye, add fixing solution to terminate the staining, and store in the dark.

[0089] (8) Immunofluorescence

[0090] ① According to the amount of section, add an appropriate amount of acid / alkali antigen repair solution, heat and boil with high fire, then put the washed tissue section into a pressure cooker, cover the pot cover and check the safety raft, after boiling, adjust to low fire to maintain boiling and spray for 33 seconds, and turn off the electromagnetic stove switch.

[0091] ② After 33 seconds, move the pressure cooker to tap water for cooling.

[0092] ③ After the antigen repair solution in the pressure cooker is completely cooled, open the pressure cooker and rinse the tissue section with distilled water for 2 minutes.

[0093] ④ Soak in 3% H2O2 for 20 minutes, rinse with distilled water 3 times, and rinse with PBS buffer 4 times, each time for 2 minutes.

[0094] ⑤ Incubate with BSA for 30 minutes (Cat. No. 2306001, Manufacturer: SOlario)

[0095] ⑥ Add the antibody dropwise. Also add the same antibody to the positive control slide, using a sufficient amount to fully cover the tissue. Add PBS buffer to the negative control slide. Note: The slides should be moist before adding the antibody; avoid drying them out. However, avoid excessive moisture, which can dilute the antibody.

[0096] ⑦ Place the slices in a wet incubation box, cover it with the lid, and then place the incubation box in a -4°C refrigerator for overnight incubation.

[0097] ⑧Take out the slices, rewarm for 30 minutes, put them back into the slide rack, and wash them 5 times with PBS, 2 minutes each time.

[0098] ⑨ Remove the slices, add secondary antibody to the specimen to be tested, and incubate at room temperature for 60 minutes. The reagent should fully cover the tissue.

[0099] ⑩ Remove the sections, insert them back into the slide rack, and wash them 6 times with PBS for 2 minutes each. Stain with API staining reagent (ready-to-use) for 5 minutes, and wash them 3 times with PBS for 2 minutes each.

[0100] The sections were placed in PBS to stop color development and then sealed with an anti-fluorescence quencher.

[0101] (9) Immunohistochemistry

[0102] ① Antigen retrieval: After heating, microwave the slides in a microwave oven at P14 for 3 minutes and 30 seconds. During this process, prevent excessive evaporation of the buffer and do not allow the slides to dry out. After cooling naturally, wash the slides three times in PBS (pH 7.4) with shaking for 5 minutes each time.

[0103] ② Block endogenous peroxidase: Place the sections in 3% hydrogen peroxide solution and incubate at room temperature in the dark for 10 minutes. Place the slides in PBS (PH7.4) and wash them three times with shaking, each time for 5 minutes.

[0104] ③ Add primary antibody: Add a certain proportion of prepared primary antibody on the slice, place the slice flat in a humidified box and incubate at room temperature for 1 hour.

[0105] ④ Add secondary antibody: Wash the slides three times in PBS (pH 7.4) with shaking, 5 minutes each time. After the sections are slightly dried, add the secondary antibody (HRP-labeled) of the same species as the primary antibody in the histochemistry kit to the circle, covering the tissue and incubating at room temperature for 30 minutes.

[0106] ⑤Immune color development: the slide was shaken in PBS (PH 7.4) for 3 times, 5 min each time. After the slice was slightly shaken dry, fresh prepared immune color development solution was added in the circle, incubated for 3-5 min, and the positive was brown. The slice was washed by tap water to stop the color development.

[0107] ⑥Re-stain the cell nucleus: stained by hematoxylin for about 3 min, washed by tap water, differentiated by hematoxylin differentiation solution for several seconds, washed by tap water, and returned to blue by hematoxylin return blue solution, and washed by running water.

[0108] ⑦Mounting: after dehydration by gradient alcohol, the slice was mounted by gum, and naturally air-dried.

[0109] ⑧Microscopy: the result was judged under white light microscope.

[0110] (10) Transmission electron microscope:

[0111] ①Material taking and fixing: when the cell grew to about 80% of 100 mm culture dish, the culture medium was discarded, and electron microscope fixing solution was added for 2-4 h at 4°C. The cell was collected in a centrifuge tube, centrifuged at 800 rpm for 5 min, the supernatant was discarded, and 0.1M phosphate buffer (PH 7.4) was washed for 3 times, 15 min each time.

[0112] ②Osmium fixation: fixed by 1% osmium acid for 2 h, and then washed by 0.1M phosphate buffer (PH 7.4) for 3 times, 15 min each time.

[0113] ③Dehydration: the cell was dehydrated by 30%, 50%, 70%, 80%, 85%, 90%, and 100% (twice) alcohol gradient, 20 min each time.

[0114] ④Permeation: the permeation agent was acetone: epoxy resin (2:1), acetone: epoxy resin (1:1), and epoxy resin, 10 h each time in a 37°C incubator.

[0115] ⑤Embedding: the permeated sample was placed in an embedding plate, and embedding agent epoxy resin was added, and polymerized for 48 h in a 60°C incubator, and placed at room temperature for about 20 days.

[0116] ⑥Slicing: the ultrathin slicer was used to slice 80-100 nm.

[0117] ⑦Double staining: uranium-lead double staining (2% acetic acid uranium saturated alcohol solution, citric acid lead), room temperature staining for 15 min, the slice was dried at room temperature, and observed.

[0118] (11) Q-PCR experiment

[0119] (I) RNA extraction

[0120] 1. Tissue: Pre-chill a mortar with liquid nitrogen and mince an appropriate amount of tissue in liquid nitrogen. Place the minced tissue into a 1.5 mL centrifuge tube pre-added with 150 μL of Buffer RZ, which is free of nuclease. Grind thoroughly with a homogenizer. Add 850 μL of Buffer RZ and vortex thoroughly to mix.

[0121] 2. Place the homogenized sample at 15-30°C for 5 minutes to completely separate the nucleic acid-protein complex;

[0122] Centrifuge at 3.4°C, 12,000 rpm for 5 min, remove the supernatant, and transfer it to a new RNase-free centrifuge tube;

[0123] 4. Add 200 μL of chloroform, cover the tube, shake vigorously for 15 seconds, and let it stand at room temperature for 3 minutes;

[0124] Centrifuge at 5.4°C, 12,000 rpm for 10 minutes. The sample will separate into three layers: a yellow organic phase, an intermediate layer, and a colorless aqueous phase. The RNA is primarily in the aqueous phase, which accounts for approximately 50% of the volume of Buffer RZ. Transfer the aqueous phase to a fresh RNase-free tube and proceed to the next step.

[0125] 6. Slowly add 0.5 times the volume of anhydrous ethanol and mix thoroughly. Transfer the resulting solution and precipitate to adsorption column CR3. Centrifuge at 12,000 rpm for 30 seconds at 4°C. Discard the waste liquid from the collection tube. If the entire solution and mixture cannot be added to adsorption column CR3 in one go, transfer them in two batches.

[0126] 7. Add 500 μL of deproteinization solution RD to the adsorption column CR3 (please check whether ethanol has been added before use). Centrifuge at 12,000 rpm (13,400 × g) at 4°C for 30 seconds. Discard the waste liquid and place CR3 in a collection tube.

[0127] 8. Add 500 μL of Buffer RW to the adsorption column CR3, let it stand at room temperature for 2 minutes, centrifuge at 12,000 rpm for 30 seconds at 4°C, and discard the waste liquid;

[0128] 9. Repeat step 8;

[0129] 10. Place the adsorption column in a 2 mL collection tube and centrifuge at 12,000 rpm (13,400 × g) at 4°C for 2 min to remove any residual liquid.

[0130] 11. Open the lid at room temperature, place flat on paper, and let dry for 5 minutes;

[0131] 12. Transfer the adsorption column CR3 to a new RNase-free 1.5 mL centrifuge tube, add 50 μL RNase-free H2O, incubate at room temperature for 2 minutes, and centrifuge at 12,000 rpm for 2 minutes at 4°C.

[0132] 13. The volume of elution buffer should not be less than 30 μL. A smaller volume will affect the recovery efficiency. RNA should be stored at -80°C to prevent degradation.

[0133] Take 2.5 μL for quantification using an enzyme-labeled instrument.

[0134] (2) Reverse transcription (synthesis of the first strand of cDNA)

[0135] (3) Real-time fluorescence quantitative PCR reaction

[0136] (12) WB experiment

[0137] (1) Cell protein extraction

[0138] Pre-chill RIPA protein extraction reagent and add protease (phosphatase) inhibitors. Add 0.1M PMSF stock solution before protein extraction to a final PMSF concentration of 1mM. Add 1×106 cells to 100μL of lysis buffer (weigh the tissue weight and add 0.2mL of lysis buffer for 20mg of tissue). Cryo-grind and continue lysis on ice for 20 minutes. Centrifuge at 4°C, 13,000 rpm, for 10 minutes (can be repeated multiple times). After centrifugation, remove the supernatant and store in aliquots until assayed.

[0139] (2) BCA protein quantification

[0140] Prepare BCA working solution A:B in a 50:1 ratio and dilute each extracted BSA standard. Dilute the sample with PBS. Mix well and incubate at 37°C for 30 minutes or at room temperature for 60 minutes. Read the OD value on a microplate reader using a 570 nm filter. Adjust the protein concentration using RIPA to a final concentration of 4-8 mg / mL. Add 5x reducing sample buffer and denature by boiling for 5 minutes.

[0141] (3) WB experiment

[0142] 1. Prepare 6-12% separation gel and 5% stacking gel according to the molecular weight of the target protein.

[0143] 2. Sample volume of protein sample to be tested: 5-10 μL (30 μg)

[0144] 3. Electrophoresis conditions: Constant voltage of stacking gel 80V, about 30min; constant voltage of separation gel 130V, and the stopping time of electrophoresis is determined by pre-stained protein marker.

[0145] 4. Wet transfer method. General transfer conditions: 300mA constant current; 0.45μm pore size NC membrane; transfer time 1h. Slight adjustments may be made (30-90min) depending on molecular weight.

[0146] 5. Blocking: Immerse the membrane completely in 3% BSA-TBST and shake gently at room temperature for 60 minutes.

[0147] 6. Primary antibody incubation: Dilute the primary antibody with antibody diluent (1:1000, or refer to the instructions), incubate at room temperature for 10 minutes, and then incubate at 4°C overnight.

[0148] 7. The next day, remove the membrane from the 4°C incubation and incubate at room temperature for 30 minutes. Wash the membrane five times with TBST, each time for 5 minutes.

[0149] 8. Secondary Antibody Incubation: Dilute the secondary antibody (goat anti-rabbit / mouse IgG (H+L) HRP) at 1:10,000 in 5% skim milk powder-TBST and gently shake at room temperature for 40 minutes. Wash the membrane: Wash the membrane five times with TBST for 5 minutes each.

[0150] 9. After adding ECL to the membrane, react for 2-5 minutes, place it in a developer for development and exposure. Depending on the difference in antibodies, the development interval is set to: 0.5s-5min (exposure time is adjusted with different light intensities). Develop 1-10 images continuously and save the images for backup for data analysis.

[0151] (4) Data processing

[0152] After development, the integrated optical density (IOD) values ​​of the bands were read and the data were analyzed using Image J software.

[0153] (13) ELISA test

[0154] Directly frozen myocardial tissue was used for mitochondrial membrane potential (MMP) measurement (JC-1 staining kit), iron level measurement, lipid ROS detection, lipid peroxidation detection, and glutathione level detection. That is, 162 samples of 3 samples per group for each indicator (54 serum samples after successful modeling + 54 serum samples on Day 42 + 54 serum samples on Day 48) were subjected to ELISA detection: relative content of cytokines IgE, IL-10, and TGF-β1 was determined, with two replicates for each sample; ELISA detection of 54 bronchoalveolar lavage fluids: relative content of cytokines IL-4 and TNF-α was determined, with two replicates for each sample.

[0155] Preventive effect of deferasirox on ventricular remodeling after acute myocardial infarction in mice

[0156] Deferasirox can significantly inhibit ventricular remodeling after myocardial infarction in mice. Figure 1 、 Figure 2As shown, in order to explore whether deferasirox can prevent ventricular remodeling after acute myocardial infarction in mice, this study performed permanent ligation of the left anterior descending coronary artery on mice. After surgery, the mice were randomly divided into 5 groups: blank control group, sham operation group, model group, deferasirox 2 mg / kg group, and deferasirox 3 mg / kg group, with 15 mice in each group. After surgery, each group was given saline and different doses of deferasirox by gavage every day for 14 days. The cardiac function was tested on the 0th, 14th, and 28th days after surgery. It was found that on the 28th day after surgery, the cardiac function of the different doses of deferasirox groups was improved to varying degrees compared with the model group.

[0157] Analyze cardiac ultrasound:

[0158] At 0 weeks, there was no significant difference in EF% among the groups; at 2 weeks, compared with the low-dose deferasirox group, the EF% of the model group and the high-dose deferasirox group increased significantly (P < 0.05); at 4 weeks, there was no significant difference in EF% among the groups; Figure 4 shown.

[0159] At 0 weeks, compared with the low-dose deferasirox group, the FS% of the model group and the high-dose deferasirox group increased significantly (P < 0.05); at 2 weeks, there was no significant difference in EF% among the groups; at 4 weeks, there was no significant difference in FS% among the groups; Figure 5 shown.

[0160] Analytical qPCR:

[0161] Compared with the blank control group, the relative expression of SLC7A11 in the sham operation group, model group and high-dose deferasirox group decreased, but there was no significant difference; the relative expression of SLC7A11 in the low-dose deferasirox group decreased without significant change; compared with the model group, the relative expression of SLC7A11 in the low-dose deferasirox group increased, but there was no significant difference; Figure 6 shown.

[0162] Compared with the blank control group, the relative expression of p62 in the sham-operated group increased, but there was no significant difference; the relative expression of p62 in the model group, low-dose deferasirox group, and high-dose deferasirox group decreased, but there was no significant difference; compared with the model group, the relative expression of p62 in the sham-operated group increased significantly (P < 0.05), and the relative expression of p62 in the low-dose deferasirox group and high-dose deferasirox group increased, but there was no significant difference;

[0163] Compared with the blank control group, the relative expression of NRF2 in the sham operation group increased, but there was no significant difference; the relative expression of NRF2 in the model group, the low-dose deferasirox group and the high-dose deferasirox group decreased, but there was no significant difference; compared with the model group, the relative expression of NRF2 in the low-dose deferasirox group and the high-dose deferasirox group decreased, but there was no significant difference;

[0164] Compared with the blank control group, the relative expression of Keap1 in the sham operation group increased, but there was no significant difference; the relative expression of Keap1 in the model group, the low-dose deferasirox group and the high-dose deferasirox group decreased, but there was no significant difference; compared with the model group, the relative expression of Keap1 in the sham operation group was significantly increased (P<0.05), and the relative expression of Keap1 in the low-dose deferasirox group and the high-dose deferasirox group increased, but there was no significant difference;

[0165] Compared with the blank control group, the relative expression of GPX4 protein in the sham operation group increased, but there was no significant difference; the relative expression of GPX4 protein in the high-dose deferasirox group did not change significantly, and the relative expression of GPX4 protein in the model group and the low-dose deferasirox group decreased, but there was no significant difference; compared with the model group, the relative expression of GPX4 protein in the low-dose deferasirox group and the high-dose deferasirox group increased, but there was no significant difference;

[0166] Compared with the blank control group, the relative expression of Ferriitin in the sham operation group increased, but there was no significant difference; the relative expression of Ferriitin in the model group, the low-dose deferasirox group and the high-dose deferasirox group decreased, but there was no significant difference; compared with the model group, the relative expression of Ferriitin in the low-dose deferasirox group and the high-dose deferasirox group increased, but there was no significant difference;

[0167] WB analysis: compared with the blank control group, the relative expression of SLC7A11 in the sham operation group, the model group, the low-dose deferasirox group and the high-dose deferasirox group decreased, but there was no significant difference; compared with the model group, the relative expression of SLC7A11 in the low-dose deferasirox group did not change significantly; as Figure 7 shown.

[0168] Compared with the blank control group, the relative expression of p62 in the sham operation group, the model group, the low-dose deferasirox group and the high-dose deferasirox group decreased, but there was no significant difference; compared with the model group, the relative expression of p62 in the low-dose deferasirox group and the high-dose deferasirox group decreased, but there was no significant difference;

[0169] Compared with the blank control group, the relative expression levels of NFR2 in the sham operation group, model group, low-dose deferasirox group, and high-dose deferasirox group were decreased, but there was no significant difference; compared with the model group, the relative expression levels of NFR2 in the low-dose deferasirox group and high-dose deferasirox group did not change significantly;

[0170] Compared with the blank control group, the relative expression levels of Keap1 in the sham operation group, model group, low-dose deferasirox group, and high-dose deferasirox group were decreased, but there was no significant difference. Compared with the model group, the relative expression level of Keap1 in the low-dose deferasirox group was decreased, while the relative expression level of Keap1 in the high-dose deferasirox group was increased, but there was no significant difference.

[0171] Compared with the blank control group, the relative expression levels of GPX4 protein in the sham operation group, model group, low-dose deferasirox group, and high-dose deferasirox group were decreased, but there was no significant difference. Compared with the model group, the relative expression level of GPX4 protein in the low-dose deferasirox group was decreased, while the relative expression level of GPX4 protein in the high-dose deferasirox group was increased, but there was no significant difference.

[0172] Compared with the blank control group, the relative expression levels of Ferritin in the sham operation group, model group, low-dose deferasirox group and high-dose deferasirox group were decreased, but there was no significant difference; compared with the model group, the relative expression levels of Ferritin in the low-dose deferasirox group and high-dose deferasirox group were increased, but there was no significant difference.

[0173] Analyze ELISA test results:

[0174] Compared with the blank control group, the myocardial GSH concentrations in the sham operation group, model group, low-dose deferasirox group, and high-dose deferasirox group were decreased, but there was no significant difference (P>0.05); compared with the model group, the myocardial GSH concentrations in the low-dose deferasirox group and high-dose deferasirox group were increased, among which the increase in the low-dose deferasirox group was greater, but there was no significant difference (P>0.05).

[0175] Compared with the blank control group, the ROS fluorescence intensity of myocardial tissue in the sham operation group increased, but there was no significant difference (P>0.05); compared with the blank control group, the ROS fluorescence intensity of myocardial tissue in the model group increased significantly (P<0.01); compared with the blank control group, the ROS fluorescence intensity of myocardial tissue in the low-dose deferasirox group and the high-dose deferasirox group increased significantly (P<0.05); compared with the model group, the ROS fluorescence intensity of myocardial tissue in the low-dose deferasirox group and the high-dose deferasirox group decreased, among which the decrease in the low-dose deferasirox group was greater, but there was no significant difference (P>0.05); Figure 8 shown.

[0176] The BNP and CRP serological indexes were analyzed sequentially.

[0177] On D1, compared with the blank control group, the serum BNP concentration of the model group and the high-dose deferasirox group was significantly increased (P<0.0001), and there was no significant difference in the serum BNP concentration of the sham operation group and the low-dose deferasirox group; compared with the model group, the serum BNP concentration of the low-dose deferasirox group was significantly decreased (P<0.0001); there was no significant difference between the remaining groups;

[0178] On D3, compared with the blank control group, the serum BNP concentration of the model group was significantly increased (P<0.001), the serum BNP concentration of the low-dose deferasirox group was significantly increased (P<0.0001), the serum BNP concentration of the high-dose deferasirox group was significantly increased (P<0.01), and there was no significant difference in the serum BNP concentration of the sham operation group; compared with the model group, there was no significant difference in each group;

[0179] On D7, compared with the blank control group, the serum BNP concentration of the sham operation group was significantly decreased (P<0.05), the serum BNP concentration of the model group was significantly decreased (P<0.01), the serum BNP concentration of the low-dose deferasirox group was significantly decreased (P<0.001), and the serum BNP concentration of the high-dose deferasirox group was significantly decreased (P<0.0001); compared with the model group, there was no significant difference in each group;

[0180] On D14, compared with the blank control group, the serum BNP concentration of the model group was significantly decreased (P<0.05), and the serum BNP concentration of the high-dose deferasirox group was significantly decreased (P<0.05); there was no significant difference in the serum BNP concentration of the sham operation group and the low-dose deferasirox group; compared with the model group, there was no significant difference in each group;

[0181] On D21, compared with the blank control group, there was no significant difference in each group; compared with the model group, there was no significant difference in each group;

[0182] On D28, compared with the blank control group, the serum BNP concentration of the model group was significantly increased (P<0.001), and there was no significant difference in the serum BNP concentration of the sham operation group, the high-dose deferasirox group and the low-dose deferasirox group; compared with the model group, the serum BNP concentration of the high-dose deferasirox group was significantly decreased (P<0.001); as shown in the figure. Figure 9

[0183] ​On D1, compared with the blank control group, the serum CRP concentrations in the model group and the high-dose deferasirox group increased significantly (P < 0.0001), while there was no significant difference in the serum CRP concentrations between the sham-operated group and the low-dose deferasirox group. Compared with the model group, the serum CRP concentration in the low-dose deferasirox group decreased significantly (P < 0.0001). There were no significant differences among the other groups.

[0184] On D3, compared with the blank control group, the serum CRP concentration in the model group increased significantly (P < 0.001), the serum CRP concentration in the low-dose deferasirox group increased significantly (P < 0.0001), and the serum CRP concentration in the high-dose deferasirox group increased significantly (P < 0.01). There was no significant difference in the serum CRP concentration in the sham operation group; compared with the model group, there was no significant difference in the serum CRP concentration in all groups.

[0185] On D7, compared with the blank control group, the serum CRP concentration in the sham-operated group decreased significantly (P < 0.05), the serum CRP concentration in the model group decreased significantly (P < 0.01), the serum CRP concentration in the low-dose deferasirox group decreased significantly (P < 0.001), and the serum CRP concentration in the high-dose deferasirox group decreased significantly (P < 0.0001). Compared with the model group, there were no significant differences among the groups.

[0186] On D14, compared with the blank control group, the serum CRP concentration in the model group decreased significantly (P < 0.05), and the serum CRP concentration in the high-dose deferasirox group decreased significantly (P < 0.05). There was no significant difference in serum CRP concentration between the sham operation group and the low-dose deferasirox group; compared with the model group, there was no significant difference in serum CRP concentration between the groups.

[0187] On D21, compared with the blank control group, there was no significant difference in each group; compared with the model group, there was no significant difference in each group;

[0188] On D28, compared with the blank control group, the serum CRP concentration in the model group increased significantly (P < 0.001), and there was no significant difference in the serum CRP concentration among the sham group, high-dose deferasirox group and low-dose deferasirox group; compared with the model group, the serum CRP concentration in the high-dose deferasirox group decreased significantly (P < 0.001); Figure 10 shown.

[0189] Analyze TTC staining results:

[0190] On D28, there was no significant difference in the sham operation group compared with the blank control group; the myocardial infarction area ratio of the model group, low-dose deferasirox group and high-dose deferasirox group increased significantly (P < 0.001); compared with the model group, the myocardial infarction area ratio of the blank control group and the sham operation group decreased significantly (P < 0.001); Figure 11 shown.

[0191] Figures 3 to 10 Center: EF (Ejection Fraction) is left ventricular ejection fraction; FS is fractional shortening. VS,d: left ventricular anterior wall diastolic thickness. IVS,s: left ventricular anterior wall systolic thickness. LVPW,d: left ventricular posterior wall diastolic thickness. LVPW,s: left ventricular posterior wall systolic thickness. LVID,d: left ventricular end-systolic internal diameter. LVID,s: left ventricular end-diastolic internal diameter.

[0192] Analysis of cardiac MRI results:

[0193] At D28, the degree of ventricular remodeling in the high-dose and low-dose deferasirox groups was significantly lower than that in the model group.

[0194] The design focus of the present invention is that the present invention's deferasirox can prevent ventricular remodeling after myocardial infarction in mice by inhibiting ferroptosis and inhibiting inflammatory response, including improving cardiac function indicators and reducing myocardial infarction area. The degree of cardiac fibrosis in mice is also significantly reduced, providing new ideas and new solutions for the clinical prevention of ventricular remodeling after acute myocardial infarction; deferasirox inhibits ferroptosis by targeted chelation of free iron ions, reduces myocardial cell death and inflammatory infiltration, and significantly reduces the degree of cardiac fibrosis in mice.

[0195] The above description is merely a preferred embodiment of the present invention and does not limit the technical scope of the present invention. Therefore, any minor modifications, equivalent changes and modifications made to the above embodiments based on the technical essence of the present invention are still within the scope of the technical solution of the present invention.

Claims

1. Application of deferasirox in the preparation of drugs for preventing ventricular remodeling after acute myocardial infarction.

2. The use according to claim 1, characterized in that: The drug exerts its effect by inhibiting myocardial cell ferroptosis and inflammatory response, thereby preventing ventricular remodeling after acute myocardial infarction; deferasirox exerts iron chelation and anti-oxidative stress damage effects in the drug, thereby inhibiting myocardial cell ferroptosis and inflammatory response.

3. The use according to claim 2, characterized in that: The drug achieves its effect of inhibiting myocardial cell ferroptosis by upregulating the relative expression level of GPX4 protein.

4. The use according to claim 1, characterized in that: The drug is administered 6 hours after the occurrence of acute myocardial infarction.

5. The use according to claim 1, characterized in that: The drug administration cycle is from the 1st day to the 14th day after the occurrence of acute myocardial infarction, the administration frequency is once a day, and the dosage of deferasirox in the drug is 20-30 mg / K per day.

6. The use according to claim 5, characterized in that: The deferasirox is used to prepare a medicine for preventing ventricular remodeling after acute myocardial infarction. The dosage of the deferasirox in the medicine is 30 mg / kg per day.

7. The use according to claim 1, characterized in that The administration mode of the drug is selected from oral administration and injection administration.

8. The use according to claim 1, characterized in that The Deferasirox is obtained by the following steps: S1, obtaining a crude Deferasirox product; S2, obtaining Deferasirox.

9. The use according to claim 8, characterized in that The specific steps for obtaining the crude Deferasirox are as follows: S11, reacting a salicylic acid derivative monomer and cyanuric chloride under stirring and condensation reflux conditions; S12, gradually increasing the temperature of the reaction system from 60°C to 120°C, and continuing the reaction; S13, stopping the reaction, and after the reaction product is cooled into a solid block, adding DMF to dissolve the product, using deionized water to precipitate, and filtering to obtain a powder; S14. Wash the powder with deionized water for multiple times, and then freeze-dry to obtain crude deferasirox.

10. The use according to claim 9, characterized in that The specific steps of obtaining the deferasirox are: S21, dissolving the crude deferasirox and the stabilizer in DMSO respectively to prepare solutions; S22, mixing the deferasirox solution and the stabilizer solution to obtain an oil phase, and adding the oil phase to water with stirring to obtain a solution; S23. The solution is transferred to an ultrafiltration tube with a molecular weight cutoff greater than 100 kDa, centrifuged, and washed to obtain deferasirox.

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