Application of tazeostat in preparation of medicine for treating acute kidney injury

Abstract

The invention discloses application of tazeostat in preparation of a medicine for treating acute kidney injury, and belongs to the technical field of biological medicine. The invention specifically relates to application of the tazeostat in preparation of drugs for reducing mRNA expression levels of lipocalin Ngal, a kidney injury factor KIM1, a chemotactic factor MCP-1, inflammatory factors IL-1beta, IL-6 and TNF-alpha, and application of the tazeostat in preparation of drugs for reducing levels of creatinine and urea nitrogen in serum. The invention has the beneficial effect that the invention provides a new medicinal application of the tazeostat in treating the acute kidney injury. The inflammatory response and the renal injury level caused in an acute renal injury in-vitro model are effectively reduced; in addition, the pharmaceutical composition can also reduce the expression level of inflammatory factors IL-6 and chemotactic factors MCP-1 in serum of a kidney injury in-vivo animal model, improve kidney tissue pathology of acute kidney injury animals, inhibit protein expression of KIM1 in kidney tissues and reduce the levels of creatinine and urea nitrogen in serum of the kidney tissues, and has a potential treatment 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 medicine for treating acute kidney injury. Background Art

[0002] Acute kidney injury (AKI) is a common clinical disease, which refers to 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, manifested by azotemia, 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). According to the location of the lesion and the cause, acute kidney injury can be divided into three categories: prerenal, renal, and postrenal. The main causes of acute kidney injury are ischemia / reperfusion (I / R), sepsis, and toxic injury. Treatment mainly includes removing the cause, maintaining a stable internal environment, nutritional support, dealing with complications, and blood purification therapy. Most patients who survive acute kidney injury can restore their renal function to normal, but 5% of patients cannot restore their renal function and require maintenance renal replacement therapy, and the proportion can be as high as 16% in elderly patients. About 5% of patients will gradually develop chronic renal damage even though their renal function has recovered. This is manifested by persistent hypertension with or without proteinuria, although Scr has returned to normal levels. This may be related to compensatory glomerular hypertrophy and secondary focal segmental glomerulosclerosis. In addition, the mortality rate of acute kidney injury is high in elderly patients, patients with sepsis, multiple organ dysfunction syndrome, and after cardiac surgery. More and more epidemiological studies have shown that the mortality rate of AKI patients is as high as 60%-80%, but there is currently no effective treatment method or treatment strategy. Therefore, it is of great significance to find kidney-protective drugs that can reduce tissue damage, promote kidney repair, and prevent the occurrence of chronic fibrosis.

[0003] Tazemetostat is an inhibitor of the methyltransferase EZH2 and some EZH2 gain-of-function mutations, used to treat adult and pediatric patients 16 years and older with metastatic / locally advanced epithelioid sarcoma who are not amenable to 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 drug 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 tazepam 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 kidney tissue;

[0011] (5) Use in the preparation of drugs for reducing 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 tazestat for preparing a drug for treating acute kidney injury, and proves that it can reduce the expression and secretion levels of intracellular inflammatory factors TNF-α, IL-1β, IL-6, chemokine MCP-1, neutrophil gelatinase-associated lipocalin Ngal, and kidney injury factor KIM1; it can also reduce the expression levels of inflammatory factors IL-6, chemokine MCP-1mRNA, neutrophil gelatinase-associated lipocalin Ngal, and kidney injury factor KIM1mRNA in acute kidney injury animal models, improve kidney tissue pathology in acute kidney injury animals, inhibit kidney injury factor KIM1 protein expression in kidney tissue, reduce creatinine and urea nitrogen levels in kidney tissue serum, and has a potential therapeutic effect on acute kidney injury. It has good application prospects. BRIEF DESCRIPTION OF THE DRAWINGS

[0025] Figure 1 This is a comparison diagram 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, renal injury factor, chemokine and inflammatory factor in HK2 cells induced by LPS, measured 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, measured by Real-time PCR in Example 4 of the present invention;

[0031] Figure 7This is a staining diagram 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] Fig. 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] Fig.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] Fig.11 This is a diagram showing the result of HE staining of kidney tissue in a cisplatin mouse model treated with Tazemetostat in Example 5 of the present invention;

[0036] Fig.12 The figure is the result of kidney tissue staining of the cisplatin mouse model and the cisplatin mouse model treated with Tazemetostat using a glycogen (PAS) staining kit in Example 5 of the present invention;

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

[0038] Fig.14 This is a diagram showing the results of KIM1 staining of kidney tissue of a CIS mouse model treated with Tazemetostat using an immunofluorescence staining method in Example 5 of the present invention;

[0039] Fig.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 determined by real-time PCR in Example 6 of the present invention;

[0040] Fig.16 This is a diagram showing the results of renal tissue staining of an I / R mouse model after Tazemetostat treatment using the HE staining method in Example 6 of the present invention;

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

[0042] Fig.18 This is a diagram showing the results of F4 / 80 staining of kidney tissue of an I / R mouse model treated with Tazemetostat using the immunohistochemical staining method in Example 9 of the present invention;

[0043] Fig.19 This is a diagram showing the results of KIM1 staining of kidney tissue of an I / R mouse model treated with Tazemetostat using an 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 P<0.01 level, ***( ### ) indicates that the difference is extremely significant at the P<0.001 level, ****( #### ) indicates that the difference is extremely significant at the P<0.001 level, and * indicates that compared with the control group, # Compared with the model group. DETAILED DESCRIPTION

[0045] In order to make the purpose, technical solution and advantages of the embodiments of the present invention clearer, the technical solution in the embodiments of the present invention will be clearly and completely described in combination with the embodiments of the present invention. Obviously, the described embodiments are 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 creative work are 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 art 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 special culture medium containing different concentrations of Tazemetostat+CIS, and the CCK8 method was used to detect cell proliferation to determine cell viability. A special culture medium for HK2 cells without Tazemetostat+Cisplatin was used as a normal control group (NC), and a special culture medium for HK2 cells 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 2. Figure 1 shown.

[0053] from Figure 1 It can be seen that Tazemetostat can reduce the damage of renal tubular epithelial cells caused by cisplatin in vitro experiments, and within the concentration range of 2.5-10μM, as the concentration of Tazemetostat increases, the mitigation effect is dose-dependent, and the damage to renal tubular epithelial cells decreases. The CCK8 results show that Tazemetostat can significantly reduce the growth inhibitory effect of cisplatin on HK2 cells within 2.5-10μM.

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

[0055] HK2 cells were cultured in culture media of different experimental groups, and the culture media 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 mRNA expression levels of neutrophil gelatinase-associated lipocalin Ngal, chemokine MCP-1, and inflammatory factor TNF-α in the supernatant were determined by real-time fluorescence quantitative PCR (Real-time PCR). Figure 2As shown in the results, 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), the Cis model group (Cis20μM) (group B), and the Cis model + Tazemetostat high-dose group (Cis20μM+Tazemetostat10μM) (group C). The results are shown in 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 culture media of different experimental groups, and the culture media 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 referred to as NC, C, and D groups.

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

[0065] HK2 cells were cultured in culture media of different experimental groups, and the culture media 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 2% normal medium for reoxygenation for 6 hours.

[0068] The supernatant of HK2 cells was collected, RNA was extracted from the supernatant, and the mRNA expression levels of renal injury factor KIM1, chemokine MCP-1, inflammatory factors IL-1β, IL-6, and TNF-α in the supernatant were determined by real-time fluorescence quantitative PCR (Real-time PCR). Figure 6 As shown in the figure, it can be seen that Tazemetostat significantly reduced the expression levels of mRNA of KIM1, MCP-1, IL-1β, IL-6, and TNF-α in cells induced by the H / R model in vitro, and the reduction effect was best when the concentration of Tazemetostat was 10 μM, that is, 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., group E), and H / R model + Tazemetostat high-dose group (i.e., group H). 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, injury 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 each time. Three days after Cis administration, the mice were sacrificed to obtain kidney tissue.

[0072] Construction of Cis model: Mice were randomly divided into Wt group, Cis group, and Cis+Tazemetostat (13 mg / kg) group, denoted as Wt, a, and b groups, with 6 mice in each group. Among them, mice in the Wt group were injected with normal saline, mice in the Cis group were injected with cisplatin once, and mice in the Cis+Tazemetostat group were injected with cisplatin and given the corresponding concentration of Tazemetostat for drug treatment.

[0073] After sacrificing the mice, 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 determination of serum indicators.

[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. Fig.10 As shown;

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

[0076] The results of kidney tissue staining of the simple Cis mouse model and the mouse model treated with Tazemetostat using a glycogen (PAS) staining kit are as follows: Fig.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. Fig.13 As shown;

[0078] Immunofluorescence staining was used to stain KIM1 in the kidney tissue of CIS mouse model after Tazemetostat treatment. Fig.14 shown.

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

[0080] Dosing method for I / R mice: one gavage administration 12 hours before surgery and one gavage administration 12 hours after surgery, a total of two gavage administrations, and sacrifice the mice 12 hours after the last gavage administration to obtain kidney tissue.

[0081] Construction of I / R model: The mice were randomly divided into Sham group, I / R group, I / R+Tazemetostat (3.25 mg / kg), I / R+Tazemetostat (6.5 mg / kg), I / R+Tazemetostat (13 mg / kg) group, denoted as Sham, c, d, e, f groups, 6 mice in each group. Among them, the mice in Sham group only underwent sham surgery, the mice in I / R group underwent ischemia-reperfusion surgery, and the hemostatic clamp was used for ischemia for 40 minutes, and the mice in I / R+Tazemetostat group underwent surgery and were given the corresponding concentration of Tazemetostat for drug treatment.

[0082] After sacrificing the mice, 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 determination of serum indicators.

[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. Fig.15 As shown;

[0084] HE staining was used to stain the kidney tissues of I / R mice and mice treated with Tazemetostat. Fig.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: Fig.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. Fig.18 As shown;

[0087] The immunofluorescence staining method was used to stain the kidney tissue KIM1 of the I / R mouse model after Tazemetostat treatment. Fig.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 samples, standard (concentration of 442 μmol / L), double distilled water, and enzyme solution A were added to the test tubes of each test group, incubated at 37°C for 5 minutes, and the absorbance A1 was measured at a wavelength of 546 nm; after the measurement, equal amounts of enzyme solution B were 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 calculation formula of 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 (442μmol / 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) Fig. 9 As shown in (a), Tazemetostat significantly reduced the creatinine level in the serum of renal tissues of mice in the I / R and Cis models, and the best reduction effect was achieved when the Tazemetostat concentration was 13 mg / kg in the I / R model.

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

[0095]

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

[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 (b) and 9(b), it can be seen that Tazemetostat significantly reduced the urea nitrogen level in the serum of kidney 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 test group and their contents

[0102]

[0103] ③ Real-time quantitative PCR (Real-time PCR) was used to measure the mRNA expression levels of renal injury factors, inflammatory factors, and chemokines in renal tissue. The results of the I / R model are as follows Fig.15 As shown in the figure, it can be seen that the mRNA expression levels of neutrophil gelatinase-associated lipocalin Ngal, renal 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 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 mouse kidney tissue. Fig.10 It can be seen that the mRNA expression levels of kidney injury factor KIM1 and inflammatory factor IL-6 in the kidney tissue of mice in the CIS group were significantly increased, which is 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 the 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. Fig.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 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 After establishing I / R model and CIS model in mice for 24 hours, the mice were anesthetized and kidney tissues were collected. The 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: Fig.16 , 17 As shown, 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 injury in CIS model and I / R model mice in vivo

[0109] After establishing CIS and I / R models in mice in Example 5 and Example 6, the mice were anesthetized and kidney tissues were collected. KIM1 fluorescence staining was performed on the kidneys to observe the effects on kidney damage in mice. The staining results Fig.14 The renal tubular injury of CIS model mice was observed. The renal tubular injury level of CIS group mice was significantly increased, while that of CIS+Tazemetostat group mice was significantly reduced, indicating that Tazemetostat can improve acute kidney injury in cisplatin-induced mice. Fig.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 histochemical staining to observe the effects on acute renal injury inflammation in mice. The staining results of the CIS model are shown in Fig.13 and 18As shown, the renal inflammation level of mice in the model group was significantly increased, indicating that the model was successfully constructed, and the renal inflammation level of mice in the CIS(I / R)+Tazemetostat group was significantly reduced, indicating that Tazemetostat can reduce the number of infiltrated 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 can play 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 the technical solutions described in the aforementioned embodiments may still be modified, or some of the technical features may be replaced by equivalents. 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 embodiments of the present invention.

Claims

1. The use of tazemetostat in any one or more of the following situations: (1) Application in the preparation of drugs for treating acute kidney injury; (2) Use in the preparation of a drug for reducing the expression level of lipocalin Ngal; (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; (4) Application in the preparation of drugs for improving the pathological characteristics of kidney tissue; (5) Use in the preparation of drugs for reducing serum creatinine and urea nitrogen levels; The structural formula of tazemetostat is 2. The use according to claim 1, characterized in that: The concentration of tazemetostat is 0.1-100 μM.

3. The use according to claim 2, characterized in that: The concentration of tazemetostat is 2.5-100 μM.

4. The use according to claim 2, characterized in that: The concentration of tazemetostat is 12.5-50 μM.

5. The use according to claim 2, characterized in that: The concentration of tazemetostat was 10 μM.

6. The use according to claim 1, characterized in that: The concentration of tazemetostat is 3-13 mg / kg.

7. The use according to claim 1, characterized in that: In (1)-(5), the drug further comprises 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, an excipient, a filler, a binder, a wetting agent, a disintegrant, an absorption promoter, a surfactant, an adsorption carrier and a lubricant.

9. The use according to claim 1, characterized in that: In (1)-(5), 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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