Application of tazemetostat in the preparation of medicine for treating acute kidney injury

Abstract

The present invention discloses the use of tazemetostat in the preparation of drugs for treating acute kidney injury, and belongs to the field of biomedicine technology. Specifically, it also relates to the use of tazemetostat in the preparation of drugs for reducing the mRNA expression levels of lipocalin Ngal, kidney injury factor KIM1, chemokine MCP-1, inflammatory factors IL-1β, IL-6, and TNF-α, and the use of tazemetostat in the preparation of drugs for reducing the levels of creatinine and urea nitrogen in serum. Beneficial effect: The present invention proposes a new drug use of tazemetostat in the treatment of acute kidney injury. It effectively reduces the inflammatory response and renal damage level caused in an in vitro model of acute kidney injury; it can also reduce the expression levels of inflammatory factors IL-6 and chemokine MCP-1 in the serum of an in vivo animal model of renal injury, improve the kidney tissue pathology of animals with acute kidney injury, inhibit the protein expression of KIM1 in kidney tissue, reduce the levels of creatinine and urea nitrogen in kidney tissue serum, and has a potential therapeutic effect on acute kidney injury.

Description

Technical Field

[0001] The present invention relates to the technical field of biomedicine, and in particular to application of tazemetostat in preparing a medicament for treating acute kidney injury. Background Art

[0002] Acute kidney injury (AKI) is a common clinical condition characterized by a sudden (within 1-7 days) and sustained (>24 hours) decline in renal function, defined as an increase in serum creatinine (SCr) of at least 0.5 mg / dL. Symptoms include azotemia, changes in water, electrolyte, and acid-base balance, and systemic symptoms, which may be accompanied by oliguria (<400 ml / 24 hours or 17 ml / hour) or anuria (<100 ml / 24 hours). Depending on the location and etiology, AKI can be categorized as prerenal, renal, and postrenal. The main causes of AKI include ischemia / reperfusion (I / R), sepsis, and toxic injury. Treatment primarily involves addressing the underlying cause, maintaining a stable internal environment, providing nutritional support, managing complications, and administering blood purification therapy. While most patients who survive AKI recover from renal failure, 5% do not and require maintenance renal replacement therapy, a proportion that can reach as high as 16% in elderly patients. Approximately 5% of patients will experience recovery in renal function but will gradually develop chronic renal impairment, manifested by persistent hypertension with or without proteinuria, despite recovery of Scr levels. This may be related to compensatory glomerular hypertrophy and secondary focal segmental glomerulosclerosis. Furthermore, the mortality rate of acute kidney injury is high in elderly patients, those with sepsis, multiple organ dysfunction syndrome, and those following cardiac surgery. A growing number of epidemiological studies have shown that the mortality rate of AKI patients is as high as 60%-80%, but there are currently no effective treatments or therapeutic strategies. Therefore, it is of great significance to identify renal protective drugs that can reduce tissue damage, promote renal repair, and prevent the development of chronic fibrosis.

[0003] Tazemetostat is an inhibitor of the methyltransferase EZH2 and some EZH2 gain-of-function mutations and is used to treat adults and pediatric patients 16 years and older with metastatic / locally advanced epithelioid sarcoma who are not candidates for complete resection. However, the use of tazemetostat for the prevention and treatment of acute kidney injury has not been reported. Summary of the Invention

[0004] The technical problem to be solved by the present invention is how to provide a new pharmaceutical use of tazemetostat.

[0005] The present invention solves the above technical problems through the following technical means:

[0006] The present invention proposes the use of tazemetostat in any one or more of the following situations:

[0007] (1) Application in the preparation of drugs for treating acute kidney injury;

[0008] (2) Use in the preparation of a drug for reducing the expression level of lipocalin Ngal;

[0009] (3) Application in the preparation of drugs for reducing the mRNA expression levels of kidney injury factor KIM1, chemokine MCP-1, inflammatory factors IL-1β, IL-6, and TNF-α;

[0010] (4) Application in the preparation of drugs for improving the pathological characteristics of renal tissue;

[0011] (5) Use in the preparation of drugs for lowering serum creatinine and urea nitrogen levels;

[0012] The structural formula of tazemetostat is

[0013]

[0014] Preferably, the concentration of tazemetostat is 0.1-100 μM.

[0015] Preferably, the concentration of tazemetostat is 2.5-100 μM.

[0016] Preferably, the concentration of tazemetostat is 12.5-50 μM.

[0017] Preferably, the concentration of tazemetostat is 10 μM.

[0018] Preferably, the concentration of tazemetostat is 3-13 mg / kg.

[0019] Preferably, in (1)-(5), the drug further comprises a pharmaceutically acceptable carrier.

[0020] Preferably, the pharmaceutically acceptable carrier is selected from one or more of a diluent, an excipient, a filler, a binder, a wetting agent, a disintegrant, an absorption promoter, a surfactant, an adsorption carrier and a lubricant.

[0021] Preferably, in (1)-(5), the drug is prepared into a pharmaceutically acceptable dosage form.

[0022] Preferably, the dosage form is tablet, pill, paste, capsule, oral solution or granule.

[0023] The beneficial effects of the present invention are:

[0024] The present invention proposes a new use of tazemetostat for preparing a drug for treating acute kidney injury. The drug has been shown to reduce the expression and secretion levels of intracellular inflammatory factors such as TNF-α, IL-1β, and IL-6, the chemokine MCP-1, neutrophil gelatinase-associated lipocalin Ngal, and the kidney injury factor KIM1. It also reduces the mRNA expression levels of the inflammatory factor IL-6, the chemokine MCP-1, the neutrophil gelatinase-associated lipocalin Ngal, and the kidney injury factor KIM1 in animal models of acute kidney injury, improves kidney tissue pathology in animals with acute kidney injury, inhibits the protein expression of the kidney injury factor KIM1 in kidney tissue, and reduces the levels of creatinine and urea nitrogen in kidney serum. This drug has a potential therapeutic effect on acute kidney injury and has good application prospects. BRIEF DESCRIPTION OF THE DRAWINGS

[0025] Figure 1 This is a comparison of the effects of different concentrations of Tazemetostat on the viability of renal tubular epithelial HK2 cells induced by cisplatin in an in vitro experiment in Example 1 of the present invention;

[0026] Figure 2 This is a graph showing the effect of Tazemetostat on the mRNA expression levels of neutrophil gelatinase-associated lipocalin, chemokines, and inflammatory factors in cisplatin-induced HK2 cells measured by real-time PCR in Example 2 of the present invention;

[0027] Figure 3 This is a staining image of the kidney injury factor KIM1 in the cisplatin-induced acute kidney injury model treated with Tazemetostat in vitro using the IF method in Example 2 of the present invention;

[0028] Figure 4 This is a graph showing the effect of Tazemetostat on the mRNA expression levels of neutrophil gelatinase-associated lipocalin, kidney injury factor, chemokines, and inflammatory factors in HK2 cells induced by LPS, as determined by real-time PCR in Example 3 of the present invention;

[0029] Figure 5 This is a staining image of the kidney injury factor KIM1 in the LPS-induced acute kidney injury model treated with Tazemetostat in vitro using the IF method in Example 3 of the present invention;

[0030] Figure 6 This is a graph showing the effect of Tazemetostat on the mRNA expression levels of renal injury factors, chemokines, and inflammatory factors in HK2 cells induced by H / R, as determined by real-time PCR in Example 4 of the present invention;

[0031] Figure 7This is a staining image of the kidney injury factor KIM1 in the H / R-induced acute kidney injury model treated with Tazemetostat in vitro using the IF method in Example 4 of the present invention;

[0032] Figure 8 This is a graph showing the effect of Tazemetostat on serum creatinine (a) and urea nitrogen (b) levels in the serum of CIS model mice in Example 7 of the present invention;

[0033] Figure 9 This is a graph showing the effect of Tazemetostat on serum creatinine (a) and urea nitrogen (b) levels in the serum of I / R model mice in Example 7 of the present invention;

[0034] Figure 10 This is a graph showing the effect of Tazemetostat on the mRNA expression levels of renal injury factors and inflammatory factors in a cisplatin mouse model determined by real-time PCR in Example 5 of the present invention;

[0035] Figure 11 This is the result of HE staining of kidney tissue in a cisplatin mouse model treated with Tazemetostat in Example 5 of the present invention;

[0036] Figure 12 The figure shows the results of kidney tissue staining using a glycogen (PAS) staining kit in Example 5 of the present invention for a cisplatin mouse model and a cisplatin mouse model treated with Tazemetostat;

[0037] Figure 13 This is a diagram showing the results of F4 / 80 staining of kidney tissue in a CIS mouse model after treatment with Tazemetostat using the immunohistochemical staining method in Example 5 of the present invention;

[0038] Figure 14 This is the result of immunofluorescence staining of KIM1 in kidney tissue of a CIS mouse model treated with Tazemetostat in Example 5 of the present invention;

[0039] Figure 15 This is a graph showing the effect of Tazemetostat on the mRNA expression levels of neutrophil gelatinase-associated lipocalin Ngal, kidney injury factor KIM1, and chemokine MCP-1 in an I / R-induced acute kidney injury model measured by real-time PCR in Example 6 of the present invention;

[0040] Figure 16 This is the result of HE staining of kidney tissue in an I / R mouse model after treatment with Tazemetostat in Example 6 of the present invention;

[0041] Figure 17 The figure shows the results of kidney tissue staining using a glycogen (PAS) staining kit in Example 6 of the present invention for an I / R mouse model and an I / R mouse model treated with Tazemetostat;

[0042] Figure 18 This is the result of F4 / 80 staining of kidney tissue in an I / R mouse model after treatment with Tazemetostat using the immunohistochemical staining method in Example 9 of the present invention;

[0043] Figure 19 This is a graph showing the results of KIM1 staining of kidney tissue in an I / R mouse model after Tazemetostat treatment using the immunofluorescence staining method in Example 8 of the present invention;

[0044] In the figure, *( # ) indicates significant difference at P<0.05 level, **( ## ) represents extremely significant difference at the P<0.01 level, ***( ### ) indicates that the difference is extremely significant at the P<0.001 level, ****( #### ) represents a significant difference at the P<0.001 level, * represents a significant difference compared with the control group, # Compared with the model group. DETAILED DESCRIPTION

[0045] To make the objectives, technical solutions, and advantages of the embodiments of the present invention more clear, the technical solutions in the embodiments of the present invention will be clearly and completely described below in conjunction with the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making creative efforts shall fall within the scope of protection of the present invention.

[0046] Unless otherwise specified, the test materials and reagents used in the following examples can be obtained from commercial sources.

[0047] If no specific techniques or conditions are specified in the examples, they can be carried out according to the techniques or conditions described in the literature in the field or according to the product instructions.

[0048] The compound Tazemetostat used in the following examples has the structure

[0049]

[0050] Purchased from Taoshu Biotechnology Co., Ltd.

[0051] Example 1: Study on the effect of different concentrations of Tazemetostat on the viability of human renal tubular epithelial cells (HK2 cells) in vitro

[0052] An in vitro cisplatin cell model was constructed using HK2 cells. HK2 cells were cultured with a dedicated HK2 cell culture medium containing different concentrations of Tazemetostat + CIS, and the CCK8 method was used to detect cell proliferation to determine cell viability. A dedicated HK2 cell culture medium without Tazemetostat + Cisplatin was used as a normal control group (NC), and a dedicated HK2 cell culture medium without Tazemetostat but containing Cisplatin was used as a damage control (denoted as CIS). The concentration of Cisplatin was 20 μM, and the concentration of Tazemetostat was 2.5, 5, 10, 20, 40, 80, 160, and 320 μM. The test results are shown in Figure 3. Figure 1 shown.

[0053] from Figure 1 As shown in the results, tazemetostat can mitigate cisplatin-induced renal tubular epithelial cell damage in vitro. Within the 2.5-10 μM concentration range, the mitigation effect is dose-dependent, with increased tazemetostat concentration leading to less renal tubular epithelial cell damage. CCK8 assay results indicate that tazemetostat significantly mitigates the growth inhibitory effect of cisplatin on HK2 cells within the 2.5-10 μM range.

[0054] Example 2: Study on the effect of Tazemetostat on cisplatin (Cis)-induced cell inflammation and cell death response in vitro

[0055] HK2 cells were cultured in different experimental groups of culture media, which were grouped as follows:

[0056] The normal control group (NC), Cis model group (Cis 20 μM), and Cis model + Tazemetostat high-dose group (Cis 20 μM + Tazemetostat 10 μM) were respectively recorded as NC, A, and B groups.

[0057] After culturing for 24 h, HK2 cells were collected, RNA was extracted from each group, and the expression levels of neutrophil gelatinase-associated lipocalin Ngal, chemokine MCP-1, and inflammatory factor TNF-α mRNA in the supernatant were determined by real-time fluorescence quantitative PCR (Real-time PCR). Figure 2The results showed that Tazemetostat significantly reduced the expression levels of Ngal, MCP-1, and TNF-α mRNA in cells induced by cisplatin in vitro.

[0058] The IF method was used to stain the kidney injury factor KIM1 in the supernatant of HK2 cells cultured in the NC normal control group (NC), Cis model group (Cis20μM) (group B), and Cis model + Tazemetostat high-dose group (Cis20μM+Tazemetostat10μM) (group C). Figure 3 As shown, consistent with the above results, Tazemetostat significantly reduced the level of cisplatin-induced renal injury in HK2 cells in vivo.

[0059] Example 3: Study on the effect of Tazemetostat on LPS (lipopolysaccharide)-induced cell inflammation and damage response in vitro

[0060] HK2 cells were cultured in different experimental groups of culture media, which were grouped as follows:

[0061] The normal control group (NC), LPS model group (LPS 1 μg / ml), and LPS model + Tazemetostat high-dose group (LPS 1 μg / ml + Tazemetostat 10 μM) were respectively recorded as NC, C, and D groups.

[0062] After culturing for 24 h, HK2 cells were collected, RNA was extracted from each group, and the expression levels of renal injury factor KIM1, neutrophil gelatinase-associated lipocalin Ngal, chemokine MCP-1, and inflammatory factor IL-1β mRNA in the supernatant were determined by real-time fluorescence quantitative PCR (RT-PCR). Figure 4 The results showed that Tazemetostat significantly reduced the expression levels of Ngal, KIM1, MCP-1, and IL-1β mRNA in LPS-induced cells in vitro.

[0063] The IF method was used to stain the kidney injury factor KIM1 in the supernatant of HK2 cells cultured in the NC normal control group (NC), LPS model group (LPS 1 μg / ml) (i.e., group C), and LPS model + Tazemetostat high-dose group (LPS 1 μg / ml + Tazemetostat 100 μM) (i.e., group D). Figure 5 As shown, consistent with the above results, Tazemetostat significantly reduced the level of LPS-induced renal injury in HK2 cells in vivo.

[0064] Example 4: Study on the effect of Tazemetostat on H / R (hypoxia / reoxygenation)-induced cell inflammation and damage response in vitro

[0065] HK2 cells were cultured in different experimental groups of culture media, which were grouped as follows:

[0066] The normal control group (NC), H / R model group, H / R model + Tazemetostat low-dose group (H / R + Tazemetostat 2.5 μM), H / R model + Tazemetostat medium-dose group (H / R + Tazemetostat 5 μM), and H / R model + Tazemetostat high-dose group (H / R + Tazemetostat 10 μM) were recorded as NC, E, F, G, and H groups.

[0067] The H / R model specifically used 0.5% low-glucose medium for 12 hours of hypoxia followed by replacement with 2% normal medium for 6 hours of reoxygenation.

[0068] The supernatant of HK2 cells was collected, RNA was extracted from the supernatant, and the expression levels of renal injury factor KIM1, chemokine MCP-1, inflammatory factors IL-1β, IL-6, and TNF-α mRNA in the supernatant were determined by real-time fluorescence quantitative PCR (Real-time PCR). Figure 6 As shown in the figure, Tazemetostat significantly reduced the expression levels of KIM1, MCP-1, IL-1β, IL-6, and TNF-α mRNA in cells induced by the H / R model in vitro, and the best reduction effect was achieved when the Tazemetostat concentration was 10 μM, i.e., in the H group.

[0069] The IF method was used to stain the kidney injury factor KIM1 in the supernatant of HK2 cells cultured in the normal control group (NC), H / R model group (i.e., E group), and H / R model + Tazemetostat high-dose group (i.e., H group). Figure 7 As shown in the figure, it can be seen that Tazemetostat significantly inhibited the expression of kidney injury factor KIM1 protein in HK2 cells in the H / R model and reduced the level of H / R-induced kidney injury in vivo.

[0070] Example 5: Study on the effect of Tazemetostat on the pathological structure, damage factors and inflammatory factor levels in the kidney tissue of mice in the cisplatin (CIS) model

[0071] Cisplatin administration method for mice: once by gavage 6 hours before surgery, twice a day after surgery, with a time difference of 12 hours between each administration. Three days after Cis administration, the mice were sacrificed and kidney tissues were obtained.

[0072] To establish the Cis model, mice were randomly divided into three groups: the Wt group, the Cis group, and the Cis + Tazemetostat (13 mg / kg) group (denoted as Wt, a, and b groups, with six mice in each group). The Wt group received a saline injection, the Cis group received a single injection of cisplatin, and the Cis + Tazemetostat group received a cisplatin injection and the corresponding concentration of Tazemetostat.

[0073] After sacrificing the mice, the kidney tissues were taken for embedding and slicing, protein extraction, and RNA extraction. At the same time, blood was taken from the mice's eyeballs for later serum index determination.

[0074] Real-time PCR was used to determine the effect of Tazemetostat on the mRNA expression levels of renal injury factors and inflammatory factors in the Cis mouse model. Figure 10 As shown;

[0075] HE staining was used to stain the kidney tissues of the Cis mouse model and mice treated with Tazemetostat. Figure 11 As shown;

[0076] The results of kidney tissue staining using the glycogen (PAS) staining kit for the simple Cis mouse model and the mouse model treated with Tazemetostat are as follows: Figure 12 As shown;

[0077] The results of F4 / 80 staining of kidney tissues of CIS mouse models treated with Tazemetostat were analyzed by immunohistochemical staining. Figure 13 As shown;

[0078] The immunofluorescence staining method was used to stain the KIM1 in the kidney tissue of the CIS mouse model after Tazemetostat treatment. Figure 14 shown.

[0079] Example 6: Study on the effect of Tazemetostat on the pathological structure, injury factors and inflammatory factor levels in the kidney tissue of mice in a myocardial ischemia-reperfusion injury (I / R) mouse model

[0080] Dosing method for I / R mice: administer the drug once by gavage 12 hours before surgery and once by gavage 12 hours after surgery, for a total of two doses. Sacrifice the mice 12 hours after the last gavage to obtain kidney tissue.

[0081] Establishment of I / R model: Mice were randomly divided into sham group, I / R group, I / R + tazemetostat (3.25 mg / kg), I / R + tazemetostat (6.5 mg / kg), and I / R + tazemetostat (13 mg / kg) groups (denoted as sham, c, d, e, and f groups), with 6 mice in each group. Among them, mice in the sham group underwent sham surgery, mice in the I / R group underwent ischemia-reperfusion surgery with hemostatic clamps for 40 minutes, and mice in the I / R + tazemetostat group underwent surgery and received the corresponding concentration of tazemetostat for drug treatment.

[0082] After sacrificing the mice, the kidney tissues were taken for embedding and slicing, protein extraction, and RNA extraction. At the same time, blood was taken from the mice's eyeballs for later serum index determination.

[0083] Real-time PCR was used to determine the effect of Tazemetostat on the mRNA expression levels of neutrophil gelatinase-associated lipocalin, renal injury factor, and chemokine in the I / R mouse model. Figure 15 As shown;

[0084] HE staining was used to stain the kidney tissues of I / R mouse models and mice treated with Tazemetostat. Figure 16 As shown;

[0085] The results of kidney tissue staining using a glycogen (PAS) staining kit in simple I / R mouse models and Tazemetostat-treated mouse models are as follows: Figure 17 As shown;

[0086] The results of F4 / 80 staining of kidney tissue in I / R mouse model after Tazemetostat treatment were analyzed by immunohistochemical staining. Figure 18 As shown;

[0087] The immunofluorescence staining method was used to stain the kidney tissue of the I / R mouse model after Tazemetostat treatment with KIM1. Figure 19 shown.

[0088] Example 7: Study on the effect of Tazemetostat on serum creatinine and urea nitrogen in CIS model and I / R model mice

[0089] ① The creatinine level (CRE) in the serum of renal tissue of I / R model and CIS model mice was measured. The specific operation process of the measurement was as follows: according to Table 1, the corresponding amount of sample, standard (concentration of 442 μmol / L), double distilled water, and enzyme solution A were added to the test tube of each experimental group. After incubation at 37°C for 5 minutes, the absorbance A1 was measured at a wavelength of 546 nm. After the measurement, the same amount of enzyme solution B was added to the above test tubes, incubated at 37°C for 5 minutes, and the absorbance A2 was measured at a wavelength of 546 nm.

[0090] The formula for calculating creatinine content is as follows:

[0091] Creatinine content (μmol / L) = [(Assay A2 - K * Assay A1) - (Blank A2 - K * Blank A1)] / [(Standard A2 - K * Standard A1) - (Blank A2 - K * Blank A1)] * Standard concentration (442 μmol / L)

[0092] Wherein, the dilution factor K=(sample amount+volume of enzyme solution A) / (sample amount+volume of enzyme solution A+volume of enzyme solution B)=186 / 246.

[0093] The results of determination of creatinine levels (CRE) in renal tissue and serum of I / R model and Cis model mice were as follows: Figure 8 (a) Figure 9 As shown in (a), Tazemetostat significantly reduced the creatinine level in the kidney tissue and serum of mice in the I / R and Cis models, with the highest reduction effect in the I / R model when the concentration of Tazemetostat was 13 mg / kg.

[0094] Table 1: Components added to the test tubes of each experimental group and their contents

[0095]

[0096] ② The urea nitrogen (BUN) level in the serum of the renal tissue of mice in the I / R model and CIS model was measured. The specific operation process of the measurement was as follows: according to Table 2, the corresponding amount of components were added to the test tubes of each experimental group, mixed, and placed in a 37°C water bath for 10 minutes. Then, phenol color developer and alkaline sodium hypochlorite were added in sequence and placed in a 37°C water bath for 10 minutes; the OD value of each tube was measured at a wavelength of 640 nm, a light path of 1 cm, and double distilled water was used to adjust the zero value.

[0097]

Note

[0098] The calculation formula of urea nitrogen content is as follows:

[0099] Urea nitrogen content (mmol / L) = [(measurement A - blank A) - (standard A - blank A)] * standard concentration (10mmol / L) * sample dilution factor

[0100] The results of the determination of urea nitrogen (BUN) levels in the serum of renal tissues of I / R model and CIS model mice were as follows: Figure 8 As shown in Figures (b) and 9(b), it can be seen that Tazemetostat significantly reduced the level of urea nitrogen in the serum of renal tissue of I / R model mice and CIS model mice, and the best reduction effect was achieved when the concentration of Tazemetostat was 10 mg / kg.

[0101] Table 2: Components added to the test tubes of each experimental group and their contents

[0102]

[0103] ③ Real-time PCR was used to measure the mRNA expression levels of renal injury factors, inflammatory factors, and chemokines in renal tissue. Figure 15 As shown in the figure, it can be seen that the mRNA expression levels of neutrophil gelatinase-associated lipocalin Ngal, kidney injury factor KIM1, and chemokine MCP-1 in the kidney tissue of mice in the I / R group were significantly increased, making the mice infected;

[0104] The above-mentioned renal injury factors, inflammatory factors and chemokines in the I / R+Tazemetostat group of mice were significantly reduced, indicating that Tazemetostat can significantly inhibit renal injury and inflammation in the I / R model, thereby improving renal function damage in infected mice and reducing the levels of creatinine and urea nitrogen in the serum of the mice's kidney tissue. Figure 10 It can be seen that the mRNA expression levels of the kidney injury factor KIM1 and the inflammatory factor IL-6 in the kidney tissue of mice in the CIS group were significantly increased, consistent with the I / R model. The relevant inflammatory indicators of mice in the CIS+Tazemetostat group were significantly reduced, indicating that Tazemetostat can significantly inhibit the expression of the above factors, thereby improving the renal function damage of CIS model mice, and then reducing the levels of creatinine and urea nitrogen in the serum of the mouse kidney tissue.

[0105] Example 7: Study on the effect of Tazemetostat on renal tissue pathology in CIS model and I / R model mice

[0106] Example 5 After establishing a CIS model in mice for 24 hours, the mice were anesthetized and kidney tissues were collected. The kidney tissues of the mice were stained using a hematoxylin and eosin (H&E) staining kit and a glycogen (PAS) staining kit. The staining results are shown in FIG. Figure 11 、 12 As shown; and observing the renal tubular damage of CIS model mice, it can be seen that the level of renal tubular damage in CIS group mice was significantly increased, mainly manifested as renal tubular dilation and glycogen deposition, while the level of renal tubular damage in CIS+Tazemetostat group mice was significantly reduced, indicating that Tazemetostat has a significant improvement effect on the renal tissue pathology of cisplatin mice.

[0107] Example 6 I / R model and CIS model were established in mice. 24 hours later, the mice were anesthetized and kidney tissues were collected. Tissue samples were stained with hematoxylin and eosin (H&E) staining kit and glycogen (PAS) staining kit to observe the renal tubular damage in the model mice. The results are as follows: Figure 16 、 17 As shown in the results, the level of renal tubular injury in mice in the I / R group was significantly increased, manifested by dilation of renal tubules and cytoplasmic vacuoles, while the level of renal tubular injury in mice in the I / R+Tazemetostat group was significantly reduced, indicating that Tazemetostat has a significant improvement effect on renal tissue pathology in mice with ischemia-reperfusion.

[0108] Example 8: Study on the effect of Tazemetostat on renal tissue damage in CIS model and I / R model mice in vivo

[0109] In Example 5 and Example 6, CIS and I / R models were established in mice, and the mice were anesthetized and kidney tissues were collected. KIM1 fluorescence staining was performed on the kidneys to observe the effect on the renal injury of the mice. Figure 14 The renal tubular injury of CIS model mice was observed. The level of renal tubular injury in CIS group mice was significantly increased, while the level of renal tubular injury in CIS+Tazemetostat group mice was significantly reduced, indicating that Tazemetostat can improve acute kidney injury in cisplatin-induced mice. Figure 19 As shown, Tazemetostat can improve renal injury in mice induced by ischemia-reperfusion acute kidney injury.

[0110] Example 9: Study on the effect of Tazemetostat on renal tissue inflammation in CIS model and I / R model mice in vivo

[0111] After establishing CIS and I / R models in mice in Example 5 and Example 6, the mice were anesthetized and kidney tissues were collected. The kidneys were stained with F4 / 80 histochemically to observe the effect on acute kidney injury inflammation in mice. The staining results of the CIS model are shown in Figure 5. Figure 13 and 18As shown in the results, the renal inflammation level of mice in the model group was significantly increased, indicating that the model was successfully constructed. The renal inflammation level of mice in the CIS(I / R)+Tazemetostat group was significantly reduced, indicating that Tazemetostat can reduce the number of infiltrating macrophages in the renal tissue of CIS and ischemia-reperfusion mice and has a significant anti-inflammatory effect.

[0112] In summary, experiments have shown that tazemetostat plays an important role in acute kidney injury and has a significant protective effect on kidney damage. Therefore, tazemetostat is expected to become a key drug for the prevention and treatment of acute kidney injury.

[0113] The above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit the same. Although the present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art should understand that they can still modify the technical solutions described in the aforementioned embodiments, or make equivalent replacements for some of the technical features therein. However, these modifications or replacements do not deviate the essence of the corresponding technical solutions from the spirit and scope of the technical solutions of the various embodiments of the present invention.

Claims

1. Use of tazemetostat in the preparation of a medicament for treating acute kidney injury, characterized in that: The structural formula of tazemetostat is 。 2. The use according to claim 1, characterized in that Tazemetostat treats acute kidney injury by reducing the expression level of lipocalin Ngal.

3. The use according to claim 1, characterized in that Tazemetostat treats acute kidney injury by reducing the mRNA expression levels of kidney injury factor KIM1, chemokine MCP-1, and inflammatory factors IL-1β, IL-6, and TNF-α.

4. The use according to claim 1, characterized in that The concentration of tazemetostat is 0.1-100 μM.

5. The use according to claim 4, characterized in that The concentration of tazemetostat is 2.5-100 μM.

6. The use according to claim 4, characterized in that The concentration of tazemetostat is 12.5-50 μM.

7. The use according to claim 1, characterized in that The drug further includes a pharmaceutically acceptable carrier.

8. The use according to claim 7, characterized in that The pharmaceutically acceptable carrier is selected from one or more of a diluent, a filler, a binder, a wetting agent, a disintegrant, an absorption promoter, a surfactant, and a lubricant.

9. The use according to claim 1, characterized in that The drug is prepared into a pharmaceutically acceptable dosage form.

10. The use according to claim 9, characterized in that The dosage form is tablet, pill, paste, capsule, oral liquid or granule.

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