Small molecule compound and application of preparation thereof in treating acute kidney injury

Abstract

The invention relates to a small molecule compound AO and application thereof in preparation of a medicine for treating kidney injury. The compound AO has the effect of effectively relieving acute kidney injury, can reduce the expression of a kidney injury index KIM-1, is used for recovering kidney functions, and has no obvious toxic or side effect. Therefore, the AO compound has a wide clinical application prospect as an acute kidney injury therapeutic agent.

Description

Technical Field

[0001] The present invention relates to the field of drugs, and particularly to a drug for treating acute kidney injury. Background Art

[0002] Acute kidney injury (AKI) is a clinical syndrome characterized by a sudden decline in kidney function within a short period (hours to days), which can lead to the retention of urea and other nitrogenous wastes, as well as disorders in body fluid and electrolyte regulation. It is usually induced by various factors such as ischemia-reperfusion injury, sepsis, nephrotoxic drugs, and contrast agents. When kidney injury persists without improvement or is severe, it can cause incomplete repair and progress to fibrosis, leading to the development of chronic kidney disease in patients and, in severe cases, end-stage renal disease, which is also a major cause of patient death.

[0003] Global epidemiological statistics show that the incidence of AKI in adult inpatients is 21.6%, while in patients in the intensive care unit (ICU), the incidence of AKI can be as high as 30% - 60%. Approximately 13.3 million people develop AKI each year, and approximately 1.7 million people die from acute kidney injury and its complications. The high incidence, high mortality, and poor prognosis of AKI, along with the lack of effective treatment methods, have made AKI an urgent health problem to be solved globally. Therefore, it is of great significance to find kidney protection drugs that can reduce kidney injury and prevent the progression to end-stage renal disease. Summary of the Invention

[0004] The purpose of the present invention is to overcome the deficiencies in the prior art and provide a small molecule compound for treating acute kidney injury.

[0005] Based on the above findings, the technical solutions for solving the above technical problems proposed by the present invention are as follows.

[0006] The first aspect of the present invention provides a small molecule compound for treating acute kidney injury, and the structure of the compound is shown in formula (I):

[0007]

[0008] Preferably, the acute kidney injury is acute kidney injury caused by renal ischemia-reperfusion or acute kidney injury caused by cisplatin.

[0009] The second aspect of the present invention provides a pharmaceutical composition for treating acute kidney injury, and the pharmaceutical composition contains the compound of formula (I) and pharmaceutically acceptable excipients.

[0010] Preferably, the acute kidney injury is acute kidney injury caused by renal ischemia-reperfusion or acute kidney injury caused by cisplatin.

[0011] Preferably, the pharmaceutical composition further comprises other drugs for treating acute kidney injury.

[0012] Preferably, the other drugs for treating acute kidney injury are selected from one or more of furosemide, bumetanide, and aliskiren.

[0013] Preferably, the pharmaceutically acceptable excipients are selected from one or more of fillers, disintegrants, binders, lubricants, solvents, emulsifiers, cosolvents, solubilizers, preservatives, pH regulators, osmotic pressure regulators, surfactants, coating materials, antioxidants, flavoring agents, and coloring agents.

[0014] Preferably, the dosage form of the pharmaceutical composition is tablets, capsules, granules, pills, oral liquids, injections, patches, gels, or ointments.

[0015] Preferably, the concentration of the compound of formula (I) in the pharmaceutical composition is 8 - 64 μM. More preferably, the concentration of the compound of formula (I) in the pharmaceutical composition is 16 - 64 μM. Further preferably, the concentration of the compound of formula (I) in the pharmaceutical composition is 32 - 64 μM. Further preferably, the concentration of the compound of formula (I) in the pharmaceutical composition is 64 μM.

[0016] The third aspect of the present invention provides the use of the above-mentioned compound of formula (I) or the above-mentioned pharmaceutical composition in the preparation of a drug for treating acute kidney injury.

[0017] Preferably, the acute kidney injury is acute kidney injury caused by renal ischemia-reperfusion or acute kidney injury caused by cisplatin.

[0018] Preferably, the concentration of the compound of formula (I) in the drug is 8 - 64 μM. More preferably, the concentration of the compound of formula (I) in the drug is 16 - 64 μM. Further preferably, the concentration of the compound of formula (I) in the drug is 32 - 64 μM. Further preferably, the concentration of the compound of formula (I) in the drug is 64 μM.

[0019] Preferably, the single-dose administration amount of the compound of formula (I) is 10 - 50 mg / kg body weight. More preferably, the single-dose administration amount of the compound of formula (I) is 25 - 50 mg / kg body weight. Further preferably, the single-dose administration amount of the compound of formula (I) is 50 mg / kg body weight.

[0020] The fourth aspect of the present invention provides the use of the compound of formula (I) or the above-mentioned pharmaceutical composition in the preparation of a medicament for treating renal tubular epithelial cell injury.

[0021] Preferably, the renal tubular epithelial cells are human renal tubular epithelial cells HK2.

[0022] Preferably, the renal tubular epithelial cell injury is renal tubular epithelial cell injury caused by hypoxia-reoxygenation or cisplatin-induced renal tubular epithelial cell injury.

[0023] Preferably, the concentration of the compound of formula (I) in the medicament is 8 - 64 μM. More preferably, the concentration of the compound of formula (I) in the medicament is 16 - 64 μM. Further preferably, the concentration of the compound of formula (I) in the medicament is 32 - 64 μM. Further preferably, the concentration of the compound of formula (I) in the medicament is 64 μM.

[0024] Preferably, the single-dose administration amount of the compound of formula (I) is 10 - 50 mg / kg body weight. More preferably, the single-dose administration amount of the compound of formula (I) is 25 - 50 mg / kg body weight. Further preferably, the single-dose administration amount of the compound of formula (I) is 50 mg / kg body weight.

[0025] The fifth aspect of the present invention provides the use of the compound of formula (I) or the above-mentioned pharmaceutical composition in the preparation of a medicament for treating inflammatory diseases.

[0026] Preferably, the inflammatory disease is related to one or more inflammatory factors selected from TNF-α, IL-6, and MCP-1.

[0027] Preferably, the inflammatory disease is simultaneously related to the inflammatory-related factors TNF-α, IL-6, and MCP-1.

[0028] Preferably, the compound of formula (I) or the pharmaceutical composition simultaneously reduces the expression levels of the inflammatory-related factors TNF-α, IL-6, and MCP-1.

[0029] Preferably, the concentration of the compound of formula (I) in the medicament is 8 - 64 μM. More preferably, the concentration of the compound of formula (I) in the medicament is 16 - 64 μM. Further preferably, the concentration of the compound of formula (I) in the medicament is 32 - 64 μM. Further preferably, the concentration of the compound of formula (I) in the medicament is 64 μM.

[0030] Preferably, the single-dose of the compound of formula (I) is 10 - 50 mg / kg body weight. More preferably, the single-dose of the compound of formula (I) is 25 - 50 mg / kg body weight. Even more preferably, the single-dose of the compound of formula (I) is 50 mg / kg body weight.

[0031] Advantages of the present invention:

[0032] Surprisingly, it was found that the AO compound of the present invention can reduce the expression levels of kidney injury molecule-1 (KIM-1) and inflammation-related factors TNF-α, IL-6, and MCP-1 in animal models of acute kidney injury and renal tubular epithelial cells, improve the histopathology of the kidney tissue in animals with renal tubular injury, effectively alleviate acute kidney injury, can be used for the protection of renal function, and has no obvious toxic and side effects. Therefore, the AO compound of the present invention has broad clinical application prospects as a therapeutic agent for acute kidney injury. Brief Description of the Drawings

[0033] Figure 1 It is a CCK8 graph showing the protective effect of compound AO on cisplatin-induced renal tubular epithelial cell injury;

[0034] Figure 2 It is a Western Blot graph showing the protective effect of compound AO on hypoxia-reoxygenation-induced renal tubular epithelial cell injury;

[0035] Figure 3 It is a Real-time PCR graph showing the protective effect of compound AO on hypoxia-reoxygenation-induced renal tubular epithelial cell injury;

[0036] Figure 4 It is a Western Blot graph showing the effect of compound AO on cisplatin-induced renal tubular epithelial cell injury and the protein level of inflammatory protein p-P65;

[0037] Figure 5 It is a Real-time PCR graph showing the mRNA levels of inflammation-related factors TNF-α and IL-6 induced by cisplatin;

[0038] Figure 6 It is the values of serum creatinine and blood urea nitrogen in a mouse model of acute kidney injury induced by renal ischemia-reperfusion after treatment with different concentrations of compound AO;

[0039] Figure 7 It is a Western Blot graph showing the protein level of kidney injury molecule-1 (KIM-1) in renal injury induced by renal ischemia-reperfusion after treatment with different concentrations of compound AO;

[0040] Figure 8It is a Real-time PCR graph of the mRNA levels of kidney injury factor KIM-1 and inflammation-related factors MCP-1, TNF-α, and IL-6 caused by renal ischemia-reperfusion with different concentrations of compound AO;

[0041] Figure 9 It is a glycogen staining graph of the pathological structural changes of the mouse kidney caused by renal ischemia-reperfusion with different concentrations of AO compound;

[0042] Figure 10 It is the values of serum creatinine and blood urea nitrogen in the acute kidney injury model of mice caused by cisplatin with compound AO;

[0043] Figure 11 It is a Real-time PCR graph of the mRNA levels of kidney injury factor KIM-1 and inflammation-related factors MCP-1, TNF-α, and IL-6 caused by cisplatin with compound AO;

[0044] Figure 12 It is a glycogen staining graph of the pathological structural changes of the mouse kidney caused by cisplatin with compound AO;

[0045] Figure 13 It is the mass spectrometry spectrum of the AO compound of the present invention. Specific Embodiments

[0046] The present invention will be further described in detail below in conjunction with experimental examples and the accompanying drawings, but the embodiments of the present invention are not limited thereto. Unless otherwise specified, the definitions of the significance symbols of P values in the experimental examples and the accompanying drawings of this application are as follows: *P<0.05, **P<0.01, ***P<0.001, #P<0.05, ##P<0.01, P<0.001.

[0047] The compound of formula (I) used in the experimental examples of the present invention was purchased from Shanghai TargetMol Biotechnology Co., Ltd., with the product number AO-022 / 43513826, abbreviated as "AO compound" in the experimental examples of the specification, and its structural confirmation spectrum is as Figure 13 shown.

[0048] Experimental Example 1: Effect of AO Compound on the Viability of Cisplatin-Induced Renal Tubular Epithelial Cells

[0049] 1. Experimental Method

[0050] Human renal tubular epithelial cells (HK2) were seeded in 96-well plates at a seeding density of approximately 4,000 cells / well. After the cells adhered, they were starved with serum-free medium for 12 hours. The first column was set as the control group, and the second column was the cisplatin model group without adding drugs. The remaining columns were sequentially added with the prepared AO (the concentrations of AO were 0.5, 1, 2, 4, 8, 16, 32, and 64 μM) according to the concentration gradient from left to right, so that the volume of each well system was 100 μL. After 12 hours, except for the control group in the first column, cisplatin with a final concentration of 20 μM was added to stimulate the other wells, and after continued culture for 24 hours, 10 μL of CCK8 solution was added to each well. After the CCK8 solution acted for 2 hours, the OD values of each well were measured at 450 nm with an enzyme-linked immunosorbent assay (ELISA) reader, and the results were recorded. The cell viability was plotted against the dose. Cell viability = (OD value of experimental group cells - OD value of blank group cells) / (OD value of control group cells - OD value of blank group cells) × 100%.

[0051] 2. Experimental results

[0052] The results of the CCK8 experiment were as Figure 1 shown. Under cisplatin stimulation, the viability of HK2 cells decreased significantly. After treatment with the AO compound, the relative survival rate of the cells increased significantly, especially at a concentration of 64 μM ( Figure 1 ). The above experimental results indicate that the AO compound has a good protective effect on renal tubular epithelial cells stimulated by cisplatin.

[0053] Experimental example 2. Protective effect of the AO compound on hypoxia-reoxygenation-induced injury and inflammatory response of renal tubular epithelial cells

[0054] 1. Experimental method

[0055] 1.1 Western Blot

[0056] HK2 cells in the logarithmic growth phase were seeded in 6-well plates at a seeding density of approximately 1.0×10 5 cells / ml and were divided into a normal group (NC), a simple AO group (AO 64 μM), a hypoxia-reoxygenation model group (HR), a low-dose AO group (HR + AO 16 μM), a medium-dose AO group (HR + AO 32 μM), and a high-dose AO group (HR + AO 64 μM). After incubation for 24 hours, they were treated with different concentrations of the AO compound for 12 hours. The cells were placed in a hypoxia incubator (1% O2, 5% CO2) for 12 hours, and then the cells were placed in a normal incubator for reoxygenation and continued to be cultured for 6 hours. The cells were washed three times with PBS, collected, and total protein was extracted. The protein expression of kidney injury molecule-1 (KIM-1) was detected by Western Blot. Each group repeated the experiment 3 times.

[0057] 1.2, Real-time PCR

[0058] HK2 cells were seeded in 12-well plates and divided into a normal group (NC), a simple AO group (AO 64 μM), a hypoxia-reoxygenation model group (HR), a low-dose AO group (HR+AO 16 μM), a medium-dose AO group (HR+AO 32 μM), and a high-dose AO group (HR+AO 64 μM). The seeding density was approximately 0.5×10 5 cells / well. After incubation for 24 hours, different concentrations of AO compounds were added and the cells were treated for 12 hours. Then the cells were placed in a hypoxia incubator (1% O2, 5% CO2) for 12 hours, and then placed in a normal incubator for reoxygenation and continued to be cultured for 3 hours. The cells were washed three times with PBS, collected, and RNA was extracted, reverse-transcribed, and amplified. Each group repeated the experiment 3 times.

[0059] 2. Experimental results

[0060] The results of Western blot are as Figure 2 shown (using β-actin as an internal reference). After treatment with AO compounds, the protein expression level of kidney injury molecule KIM-1 in HK2 cells treated with hypoxia-reoxygenation was significantly inhibited, and the effect was most obvious in the high-dose group. The results of Real-time PCR are as Figure 3 shown in Figures 3A and 3B. After hypoxia-reoxygenation treatment, the mRNA levels of kidney injury molecule KIM-1 and inflammatory factor MCP-1 in the model group were significantly up-regulated. This phenomenon was improved after treatment with AO compounds, and the effect was most obvious in the high-dose group, suggesting that AO compounds can inhibit cell damage and inflammatory responses caused by hypoxia-reoxygenation.

[0061] Experimental Example 3. Inhibitory effect of AO compounds on cisplatin-induced cell damage and inflammatory responses in vitro

[0062] 1. Experimental method

[0063] 1.1. Western Blot

[0064] HK2 cells in the logarithmic growth phase were seeded in 6-well plates at a seeding density of approximately 1.0×10 5 cells / ml and divided into a normal group (NC), a simple AO group (64 μM), a model group (cisplatin 20 μM), and a model+AO group (cisplatin 20 μM+AO 64 μM). After incubation for 24 hours and starvation with serum-free medium for 12 hours, cisplatin and AO compounds were added respectively. The cells were cultured for another 24 hours. The cells were washed three times with PBS, collected, and total protein was extracted. The expression level of kidney injury factor KIM-1 and the phosphorylation level of P65 in the inflammatory signaling NF-κB pathway were detected by Western Blot. Each group repeated the experiment 3 times.

[0065] 1.2, Real-time PCR

[0066] HK2 cells were seeded in 12-well plates and divided into a normal group (NC), a simple AO group (64 μM), a model group (cisplatin 20 μM), and a model + AO group (cisplatin 20 μM + AO 64 μM). The seeding density was approximately 0.5×10 5 cells / well. After incubation for 24 hours and starvation with serum-free medium for 12 hours, cisplatin and AO compounds were added respectively. Cultivation was continued for 24 hours. The cells were washed three times with PBS, collected, and RNA was extracted, reverse-transcribed, and amplified. Each group repeated the experiment 3 times.

[0067] 2. Experimental results

[0068] The results of Western Blot are as shown in Figure 4 Figures 4A and 4B. The AO compound significantly down-regulated the expression of kidney injury molecule-1 (KIM-1) in cisplatin-induced HK2 cells and the phosphorylation level of p65 in the inflammatory signaling NF-κB pathway. The results of Real-time PCR are as shown in Figure 5 Figures 5A and 5B. The AO compound significantly down-regulated the mRNA levels of inflammation-related factors TNF-α and IL-6 induced by cisplatin. In summary, the AO compound can significantly inhibit cisplatin-induced cell damage and inflammatory responses.

[0069] Experimental Example 4. Effects of AO compound on serum creatinine and blood urea nitrogen in an acute kidney injury model induced by renal ischemia-reperfusion in vivo

[0070] 1. Experimental method

[0071] 6- to 8-week-old C57BL / 6 mice were adaptively cultured for 1-2 days. The experiments were divided into a sham operation group (saline), a renal ischemia-reperfusion group (IRI), an AO low-dose group (IRI + AO 12.5 mg / kg), an AO medium-dose group (IRI + AO 25 mg / kg), and an AO high-dose group (IRI + AO 50 mg / kg), with 6 mice in each group. The AKI mouse model induced by renal ischemia-reperfusion was established by anesthetizing the mice with intraperitoneal injection of 4% pentobarbital sodium, removing the hair on the back surgical site with a hair clipper, bluntly separating the bilateral renal pedicles on a thermostatic plate, simultaneously clamping the bilateral renal pedicles with non-invasive micro arterial clips, and removing the bilateral micro arterial clips after 40 minutes. Subsequently, the muscle layer and skin layer were sutured respectively. Before the renal ischemia-reperfusion surgery, low, medium, and high doses of the AO compound were injected for drug intervention. Serum samples and kidney tissues were collected under anesthesia 1 day after the surgery, and the contents of creatinine (SCr) and blood urea nitrogen (BUN) in the serum of the animal model were detected according to the instructions of the creatinine and blood urea nitrogen assay kits (purchased from Nanjing Jiancheng Bioengineering Institute).

[0072] The method for creatinine determination is as follows:

[0073]

[0074] Creatinine content (μmol / L) = [(Measured A2 - K * Measured A1) - (Blank A2 - K * Blank A1)] / [(Standard A2 - K * Standard A1) - (Blank A2 - K * Blank)] * Standard concentration (442 μmol / L)

[0075] Note: Dilution factor K = (Sample volume + Volume of Enzyme solution A) / (Sample volume + Volume of Enzyme solution A + Volume of Enzyme solution B) = 186 / 246

[0076] The method for urea nitrogen test is as follows:

[0077]

[0078] Urea nitrogen content (mmol / L) = (Measured OD value - Blank measured value) / (Standard OD value - Blank OD value) * Standard concentration (10 mmol / L) * Dilution factor before sample test

[0079] 2. Experimental results

[0080] In the mouse model group induced by renal ischemia - reperfusion to cause acute kidney injury, the blood creatinine and blood urea nitrogen contents increased significantly, and renal function deteriorated. However, AO compounds at different concentrations effectively reduced the blood creatinine level and blood urea nitrogen level in the model group (see Figure 6 Figures A and 6B), and the down - regulation effect was the best in the high - dose group. The above experimental results further confirmed that AO compounds can restore renal function and have a kidney - protecting effect on acute kidney injury.

[0081] Experimental Example 5. Effects of AO compounds on kidney injury factor proteins and inflammatory responses in the in - vivo model of acute kidney injury induced by renal ischemia - reperfusion

[0082] 1. Experimental methods

[0083] 1.1 Western Blot method

[0084] Weigh the renal tissue, add protein lysate according to the mass - volume ratio of 14 μl / mg, grind it thoroughly, transfer it to an EP tube, and lyse it on a shaker at 4°C for 30 min. The whole process is carried out on ice. For subsequent steps, refer to the Western Blot method in the cell experiment of Experimental Example 1.

[0085] 1.2 Real - time PCR method

[0086] Weigh the renal tissue, add Trizol lysis buffer at a mass-to-volume ratio of 600 μl / 15 mg, grind thoroughly, transfer it to an EP tube, and lyse it at 4 °C for 30 min. The whole process is carried out on ice. For subsequent steps, refer to the cell experiment Real-time PCR method in Experimental Example 1.

[0087] 2. Experimental Results

[0088] The results of Western Blot showed that the level of kidney injury molecule KIM-1 was significantly increased in the acute kidney injury model, while the AO compound could significantly reduce the expression of KIM-1 protein (see Figure 7 ). In addition, as shown in Figure 8 A-8D for the results of Real-time PCR, after renal ischemia-reperfusion treatment, the mRNA levels of kidney injury molecule KIM-1 and inflammatory factors MCP-1, TNF-α, and IL-6 in the model group were significantly upregulated. This phenomenon was improved after treatment with the AO compound, and the high-dose group had the most obvious effect. The above experimental results suggest that the AO compound can alleviate kidney injury caused by ischemia.

[0089] Experimental Example 6. Effect of AO Compound on the Kidney Tissue Pathology of Mice in an In Vivo Model of Acute Kidney Injury Induced by Renal Ischemia-Reperfusion

[0090] 1. Experimental Method (Glycogen Staining Method)

[0091] Embed the taken mouse renal tissue in paraffin, and then section the paraffin. After placing the sections in an oven at 65 °C for 2 hours, successively place them in xylene and gradient ethanol for dewaxing. Drop the periodic acid staining solution in the glycogen staining kit on the tissue and stain for 15 min. After the staining is completed, wash three times with PBS solution, then use the Schiff staining solution in the glycogen staining kit, stain for 30 min, wash three times with PBS solution, stain with hematoxylin for 5 min, then place the sections in a container and rinse with running water for 5 min. Dehydrate successively in ethanol solutions with increasing concentrations. Place the sections in a fume hood and air dry them completely at room temperature. After drying, use a plastic pipette to suck a small amount of viscous neutral resin, drop it on the tissue and cover it with a coverslip, exhaust the air bubbles, and air dry it again. Observe the PAS staining situation under a microscope, collect appropriate images and analyze them.

[0092] 2. Experimental Results

[0093] The results are as shown in Figure 9As shown, the level of renal tubular injury in the renal ischemia-reperfusion group of mice increased significantly, manifested as renal tubular dilation and rupture, while the level of renal tubular injury in the AO compound-treated group of mice decreased significantly. Compared with the model group, the situation of renal tubular rupture in the AO compound-treated group decreased, and the tubular dilation was improved, indicating that the AO compound had an obvious improvement effect on the renal tissue pathology of mice with renal ischemia-reperfusion.

[0094] Experimental Example 7. Effects of AO Compound on Serum Creatinine and Blood Urea Nitrogen in an In Vivo Model of Cisplatin-Induced Acute Kidney Injury

[0095] 1. Experimental Method

[0096] C57BL / 6 mice aged 6 - 8 weeks were adaptively cultured for 1 - 2 days. The experiment was divided into a normal control group (saline), a model group (cisplatin 20 mg / kg), and an AO group (cisplatin 20 mg / kg + AO 50 mg / kg), with 6 mice in each group. The mice were intraperitoneally injected with 20 mg / kg cisplatin to establish an acute kidney injury model and were injected with the AO compound for drug intervention. After 3 days, serum samples and kidney tissues were collected under anesthesia, and the contents of creatinine and blood urea nitrogen in the serum of the animal model were detected according to the instructions of the creatinine and blood urea nitrogen kits (purchased from Nanjing Jiancheng Bioengineering Institute).

[0097] The method for creatinine determination is as follows:

[0098]

[0099] Creatinine content (μmol / L) = [(Measured A2 - K * Measured A1) - (Blank A2 - K * Blank A1)] / [(Standard A2 - K * Standard A1) - (Blank A2 - K * Blank)] * Standard concentration (442 μmol / L)

[0100] Note: Dilution factor K = (Sample volume + Volume of Enzyme Solution A) / (Sample volume + Volume of Enzyme Solution A + Volume of Enzyme Solution B) = 186 / 246

[0101] The method for blood urea nitrogen test is as follows:

[0102]

[0103]

[0104] Blood urea nitrogen content (mmol / L) = (Measured OD value - Blank measured value) / (Standard OD value - Blank OD value) * Standard concentration (10 mmol / L) * Dilution factor before sample testing

[0105] 2. Experimental Results

[0106] The experimental results showed that the levels of serum creatinine and blood urea nitrogen in the cisplatin-induced acute kidney injury model group were significantly increased, and renal function deteriorated. However, after intervention with AO compounds, the levels of serum creatinine and blood urea nitrogen in the model group could be effectively reduced (see Figure 10 Figures A and 10B). The above experimental results further confirmed that AO compounds had a renal protective effect on acute kidney injury.

[0107] Experimental Example 8. Effects of AO compounds on kidney injury factors and inflammatory factors in the in vivo cisplatin-induced acute kidney injury model

[0108] 1. Experimental method (Real-time PCR method)

[0109] Weigh the kidney tissue, add Trizol lysate at a mass-volume ratio of 600 μL / 15 mg, fully grind it, and then transfer it to an EP tube. Lyse it at 4 °C for 30 min, and operate on ice throughout the process. For subsequent steps, refer to the Real-time PCR method in the cell experiment of Test Example 1.

[0110] 2. Experimental results

[0111] The results of the Real-time PCR experiment were as shown in Figure 11 Figures A-11D. After cisplatin treatment, the mRNA levels of the kidney injury molecule KIM-1 and the inflammatory factors MCP-1, TNF-α, and IL-6 in the model group were significantly up-regulated. This phenomenon was improved after treatment with AO compounds, suggesting that AO compounds could alleviate kidney injury caused by drug toxicity.

[0112] Experimental Example 9. Effects of AO compounds on the pathology of mouse kidney tissue in the in vivo cisplatin-induced acute kidney injury model

[0113] 1. Experimental method (glycogen staining method)

[0114] Embed the taken mouse kidney tissue in paraffin, and then section the paraffin. After placing the sections in an oven at 65 °C for 2 hours, sequentially place them in xylene and gradient ethanol for dewaxing. Drop the periodic acid staining solution in the glycogen staining kit on the tissue and stain for 15 min. After the staining is completed, wash it three times with PBS solution, and then use the Schiff staining solution in the glycogen staining kit. After staining for 30 min, wash it three times with PBS solution. After hematoxylin staining for 5 min, place the sections in a container and rinse them with running water for 5 min. Dehydrate them sequentially in ethanol solutions with increasing concentrations. Place the sections in a fume hood and air-dry them completely at room temperature. After air-drying, use a plastic pipette to suck a small amount of viscous neutral resin, drop it on the tissue, cover it with a coverslip, exhaust the bubbles, and air-dry it again. Observe the PAS staining under a microscope, collect appropriate images, and perform analysis.

[0115] 2. Experimental results

[0116] The results were as Figure 12 shown. The level of renal tubular injury in the cisplatin group of mice increased significantly, manifested as a large number of ruptured renal tubules and glycogen deposition in the tubules. However, after treatment with the AO compound, the level of renal tubular injury in mice decreased significantly. Compared with the cisplatin model group, the number of ruptured renal tubules in the AO compound treatment group decreased significantly, and the glycogen deposition improved, indicating that the AO compound has an obvious improvement effect on the renal tissue pathology caused by cisplatin in mice.

[0117] The above experimental examples are preferred embodiments of the present invention. However, the embodiments of the present invention are not limited by the above experimental examples. Any other changes, modifications, substitutions, combinations, and simplifications made without departing from the spirit and principle of the present invention shall be equivalent replacement methods and are all included in the protection scope of the present invention.

Claims

1. A compound for treating acute kidney injury, characterized in that, The structure of the said compound is shown in formula (I):

2. A pharmaceutical composition for treating acute kidney injury, characterized in that, The said pharmaceutical composition comprises the compound of formula (I) as claimed in claim 1 and a pharmaceutically acceptable excipient.

3. The pharmaceutical composition according to claim 2, wherein, The said pharmaceutical composition further comprises other drugs for treating acute kidney injury.

4. The pharmaceutical composition according to claim 3, characterized in that, The said other drugs for treating acute kidney injury are selected from one or more of furosemide, bumetanide, and aliskiren.

5. The pharmaceutical composition according to claim 2, wherein The dosage form of the said pharmaceutical composition is tablet, capsule, granule, pill, oral liquid, injection, patch, gel or ointment.

6. Use of the compound of formula (I) as claimed in claim 1 or the pharmaceutical composition as claimed in any one of claims 2 - 5 in the preparation of a drug for treating acute kidney injury.

7. The application according to claim 6, characterized in that, The said acute kidney injury is acute kidney injury caused by renal ischemia - reperfusion or acute kidney injury caused by cisplatin.

8. Use of the compound of formula (I) as claimed in claim 1 or the pharmaceutical composition as claimed in any one of claims 2 - 5 in the preparation of a drug for treating renal tubular epithelial cell injury.

9. The application according to claim 8, characterized in that, The said renal tubular epithelial cell injury is renal tubular epithelial cell injury caused by hypoxia - reoxygenation or renal tubular epithelial cell injury induced by cisplatin.

10. Use of the compound of formula (I) according to claim 1 or the pharmaceutical composition according to any one of claims 2 - 5 in the preparation of a medicament for treating inflammatory diseases, characterized in that, The said inflammatory disease is related to one or more inflammatory factors selected from TNF - α, IL - 6, and MCP - 1.

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