Use of vps34 inhibitors in the preparation of a medicament for preventing anthracycline-induced cardiotoxicity

Abstract

The present application relates to the application of VPS34 inhibitor in the preparation of the drug for preventing and treating anthracycline-induced cardiotoxicity. The present application creatively finds that VPS34 inhibitor represented by SAR405 has obvious prevention and treatment effect on anthracycline-induced cardiotoxicity (AIC), and the present application verifies the role of autophagy in AIC mouse model based on cardiomyocyte strain and gene knockout mouse as the research object, and proves that early inhibition of autophagy by Atg7 gene knockout can reduce AIC, and VPS34 inhibitor can inhibit Vps34 kinase activity by interacting with the ATP binding domain of Vps34, thereby interfering with the occurrence of autophagy, which provides a new strategy for preventing and treating anthracycline-induced cardiotoxicity (AIC).

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of biological medicine, and relates to a new strategy for preventing and treating anthracycline-induced cardiotoxicity, in particular to application of a VPS34 inhibitor in preparation of a medicine for preventing and treating anthracycline-induced cardiotoxicity. BACKGROUND

[0002] Anthracyclines are a class of chemotherapy drugs containing anthracene rings, including doxorubicin (doxorubicin), epirubicin (epirubicin), pirarubicin, daunorubicin, aclarubicin, idarubicin, etc. Anthracyclines play an important role in the chemotherapy of tumors, and have the advantages of broad-spectrum and high efficiency. The main mechanism of action is to directly insert between DNA base pairs, interfere with the transcription process, prevent the formation of mRNA, and inhibit the proliferation of tumor cells. It inhibits both DNA and RNA synthesis, so it has effects on all stages of the cell cycle and is a cell cycle non-specific drug. However, the dose-dependent cardiotoxicity of anthracyclines limits their clinical application and increases the pain and burden of patients. Myocardial cell death is the main pathophysiological cause of cardiotoxicity. Taking doxorubicin (DOX) as an example, the mechanisms of DOX-induced myocardial injury include oxidative stress, apoptosis and autophagy disorder.

[0003] One of the ways to prevent and treat anthracycline cardiotoxicity is to find a myocardial protective agent targeting the cardiotoxicity mechanism of anthracyclines. Existing studies have used several drugs to prevent the cardiotoxicity of anthracyclines, but the effect is very small. Dexrazoxane (DEX) is approved by the US Food and Drug Administration for clinical prevention and treatment of doxorubicin cardiotoxicity. Dexrazoxane has a relatively certain effect, but it is expensive and can only be dissolved in lactic acid solution and slowly injected intravenously. In addition, dexrazoxane can exacerbate the inhibition of doxorubicin on bone marrow, and some patients cannot tolerate it, so it is still necessary to develop a myocardial protective agent with a certain effect, convenient administration and a reasonable price.

[0004] Dexrazoxane is a ring-opening chelating agent, which has been used to achieve cardioprotection against AIC due to its possible interaction with Top2B. However, dexrazoxane can achieve certain clinical efficacy in short-term application, but there is great uncertainty in terms of long-term efficacy and prevention of long-term cardiotoxicity adverse events. Moreover, dexrazoxane can only be administered by injection, and it is relatively expensive, and its adverse reactions cannot be ignored. Therefore, it is of great clinical application value to find a safe and effective anthracycline protective agent. SUMMARY

[0005] In view of the deficiencies of the prior art, the purpose of the present application is to provide a new strategy for preventing and treating anthracycline-induced cardiotoxicity, and specifically to provide application of a VPS34 inhibitor in preparation of a medicine for preventing and treating anthracycline-induced cardiotoxicity.

[0006] To achieve the object of the present application, the following technical solutions are adopted:

[0007] In a first aspect, the present application provides a use of a VPS34 inhibitor in the preparation of a medicament for preventing and treating anthracycline-induced cardiotoxicity (AIC).

[0008] The present application creatively discovers that VPS34 inhibitors represented by SAR405 have obvious preventive and therapeutic effects on anthracycline-induced cardiotoxicity (AIC), and based on cardiomyocyte strains and gene knockout mice as research objects, the present application verifies the role of autophagy in AIC mouse models, and proves that early inhibition of autophagy through Atg7 gene knockout can reduce AIC, and VPS34 inhibitors inhibit Vps34 kinase activity by interacting with the ATP binding domain of Vps34, thereby interfering with the occurrence of autophagy, which provides a new strategy for preventing and treating anthracycline-induced cardiotoxicity (AIC).

[0009] Preferably, the anthracycline includes doxorubicin or a pharmaceutically acceptable salt thereof, epirubicin, pirarubicin, daunorubicin, aclarubicin, idarubicin, or a pharmaceutically acceptable salt of the above compounds.

[0010] Preferably, the VPS34 inhibitor includes SAR405.

[0011] SAR405 is an ATP-competitive Vps34 selective inhibitor, and the chemical formula of SAR405 is:

[0012]

[0013] The present application creatively discovers that SAR405 has obvious preventive and therapeutic effects on anthracycline-induced cardiotoxicity (AIC), and can improve the ejection fraction (EF) and fractional shortening (FS) of AIC model mice, reduce the number of myocardial cell vacuoles and myocardial fiber diameter and the percentage of myocardial fibrosis of mice, and provide effective protection against AIC in vivo, thereby reducing myocardial toxicity caused by anthracyclines. The present application repositions the drug function of SAR405, further expands its new application in preventing and treating doxorubicin cardiotoxicity drugs, and provides potential treatment ideas and reference basis for clinical use.

[0014] Preferably, the medicament further contains a pharmaceutically acceptable excipient.

[0015] Preferably, the pharmaceutically acceptable excipient includes any one or a combination of at least two of a carrier, an excipient, a filler, a binder, a wetting agent, a disintegrant, an emulsifying agent, a cosolvent, a solubilizer, an osmotic pressure regulator, a surfactant, a coating material, a coloring agent, a pH regulator, an antioxidant, a bacteriostatic agent, or a buffer.

[0016] Preferably, the dosage form of the drug is any one of pharmaceutically acceptable dosage forms, such as tablets, powders, suspensions, granules, capsules, solutions, enemas, emulsions, etc.

[0017] In a second aspect, the present application provides a use of a VPS34 inhibitor in the preparation of a synergist for an anthracycline-induced cardiotoxicity-preventing drug.

[0018] The present application has found that the AIC-preventing drug represented by dexrazoxane (DEX) alone is not sufficient to completely reverse the damage to myocardial cells, and the preventing effect is limited, but the VPS34 inhibitor can promote the efficacy of the AIC-preventing drug, and greatly improve the effect of reversing the damage to myocardial cells.

[0019] Preferably, the anthracycline-induced cardiotoxicity-preventing drug comprises dexrazoxane.

[0020] Preferably, the anthracycline comprises doxorubicin or a pharmaceutically acceptable salt thereof.

[0021] Preferably, the VPS34 inhibitor comprises SAR405.

[0022] In a third aspect, the present application provides a combined drug composition for preventing and treating anthracycline-induced cardiotoxicity, which comprises a VPS34 inhibitor and dexrazoxane.

[0023] The present application also creatively combines the VPS34 inhibitor represented by SAR405 and dexrazoxane as a drug for preventing and treating anthracycline-induced cardiotoxicity. The combination of the two not only can reduce the dosage of the VPS34 inhibitor or dexrazoxane, improve the safety of drug use, but also has a more significant effect on preventing and treating anthracycline-induced cardiotoxicity than the single VPS34 inhibitor or single dexrazoxane, and plays a synergistic promoting effect. The present application has proved that the combined drug composition can improve the ejection fraction (EF) and fractional shortening (FS) of AIC model mice, reduce the number of myocardial cell vacuoles, myocardial fiber diameter and myocardial fibrosis percentage of mice, and provide more effective protection for AIC in vivo. The present application provides an effective drug combination strategy for the prevention and treatment of AIC, which has very significant meaning.

[0024] Preferably, the VPS34 inhibitor comprises SAR405.

[0025] Preferably, the combined drug composition is a single compound preparation or a combination of two separate preparations.

[0026] Preferably, the combined drug composition is a combination of two separate preparations, and the two separate preparations are administered simultaneously or sequentially.

[0027] The combination drug composition can be in the form of a single complex preparation or a combination of two separate preparations. When it is a combination of two separate preparations, it can be administered simultaneously, or administered in a cross or sequential manner.

[0028] Preferably, the preparation is in any one of the pharmaceutically acceptable dosage forms, such as tablets, powders, suspensions, granules, capsules, solutions, enemas, emulsions, etc.

[0029] Preferably, the combination drug composition further comprises a pharmaceutically acceptable excipient.

[0030] Preferably, the pharmaceutically acceptable excipient comprises any one or a combination of at least two of carriers, excipients, fillers, binders, wetting agents, disintegrants, emulsifiers, co-solvents, solubilizers, osmotic pressure regulators, surfactants, coating materials, coloring agents, pH regulators, antioxidants, bacteriostatic agents, or buffers.

[0031] Compared with the prior art, the present application has the following beneficial effects:

[0032] The present application creatively discovers that the VPS34 inhibitor represented by SAR405 has a significant preventive and therapeutic effect on anthracycline-induced cardiotoxicity (AIC). The present application also creatively combines the VPS34 inhibitor represented by SAR405 and dexrazoxane as a drug for preventing and treating anthracycline-induced cardiotoxicity. The combination of the two not only reduces the dosage of the VPS34 inhibitor or dexrazoxane, improves the safety of drug use, but also has a more significant effect on preventing and treating anthracycline-induced cardiotoxicity than a single VPS34 inhibitor or a single dexrazoxane, and has a synergistic effect. BRIEF DESCRIPTION OF DRAWINGS

[0033] Figure 1 is a schematic diagram of the operation process of Example 1;

[0034] Figure 2 is a representative echocardiogram of each group of mice in Example 1;

[0035] Figure 3 is a statistical analysis result graph of the ejection fraction (EF) and fractional shortening (FS) of each group of mice in Example 1;

[0036] Figure 4 is a statistical analysis result graph of hematoxylin-eosin (HE) staining and WGA staining of myocardial cell vacuoles and myocardial fiber diameters in Example 1;

[0037] Figure 5This is a graph showing the statistical results of cell viability of AC16 and H9c2 cardiomyocytes in Example 1;

[0038] Figure 6 This is an immunoblot image and quantitative statistical results of LC3II protein expression levels after DOX treatment of HL-1 cells in Example 2;

[0039] Figure 7 This is an immunoblot image and quantitative statistical results of Atg7 protein expression levels after DOX treatment of HL-1 cells in Example 2;

[0040] Figure 8 This is an immunoblot image and quantitative statistical results of P62 protein expression levels after DOX treatment of HL-1 cells in Example 2;

[0041] Figure 9 This is a laser confocal image of Atg7 staining in cells after DOX treatment in Example 2;

[0042] Figure 10 This is a laser confocal image of LC3 staining in cells treated with DOX in Example 2;

[0043] Figure 11 This is a schematic diagram of the operation process of Example 3;

[0044] Figure 12 These are representative echocardiograms of mice from each group in Example 3;

[0045] Figure 13 This is a graph showing the statistical analysis results of the ejection fraction (EF) and shortening fraction (FS) of mice in each group in Example 3;

[0046] Figure 14 The image shows the HE staining pattern (left) and the statistical analysis of myocardial cell vacuoles (right) in Example 3.

[0047] Figure 15 The image shows the WGA staining pattern (left) and the statistical analysis of myocardial fiber diameter (right) from Example 3.

[0048] Figure 16 The image shows the LC3 staining plot (left) and the statistical analysis plot of LC3 AOD (right) in Example 3.

[0049] Figure 17 These are transmission electron micrographs of heart samples from the WT and AKO groups of mice in Example 3;

[0050] Figure 18 This is a schematic diagram of the operation process of Example 4;

[0051] Figure 19 These are representative echocardiograms of mice from each group in Example 4;

[0052] Figure 20 Figure 1 is a graph of the statistical analysis of the ejection fraction (EF) and fractional shortening (FS) of each group of mice in Example 4;

[0053] Figure 21 Figure 2 is a graph of HE staining (left) and statistical analysis of myocardial cell vacuoles (right) in Example 4;

[0054] Figure 22 Figure 3 is a graph of WGA staining (left) and statistical analysis of myocardial fiber diameter (right) in Example 4;

[0055] Figure 23 Figure 4 is a graph of LC3 staining (left) and statistical analysis of LC3 AOD (right) in Example 4;

[0056] Figure 24 Figure 5 is a graph of transmission electron microscopy of heart samples of WT and AKO group mice in Example 4. DETAILED DESCRIPTION

[0057] The technical solutions of the present application will be further described below through specific embodiments. Those skilled in the art should understand that the embodiments are only to help understand the present application and should not be regarded as specific limitations of the present application.

[0058] The dexrazoxane (DEX) used in the following examples is a product purchased from MedChemExpress with model number HY-76201-50mg, doxorubicin (DOX) is a product purchased from MedChemExpress with model number HY-15142-50mg, SAR405 is a product purchased from Glpbio with model number GC11471-10mg, and tamoxifen is a product purchased from Sigma-Aldrich with model number T5648-1g (the concentration in the test is calculated based on the actual active ingredient in the drug).

[0059] The AC16 myocardial cells used in the following examples are derived from ATCC, the H9c2 myocardial cells are donated by the laboratory of York University in Canada, the HL-1 myocardial cells are donated by the laboratory of Qi Xufeng of Jinan University, and the C57 mice are derived from Jackson Laboratory in the United States, male, 9 to 10 weeks old.

[0060] Example 1

[0061] Reversal effect of single drug dexrazoxane (DEX) on doxorubicin (DOX)-induced myocardial cell damage:

[0062] (1) Animal test:

[0063] (1.1) C57 mice were used as animal models and divided into 3 groups, 3 mice in each group, normal control group (CON group), DOX treatment group (DOX group), and DEX pretreatment + DOX treatment group (DEX + DOX group).

[0064] (1.2) The experimental operation process is shown in Figure 1 , and the DEX + DOX group mice were pretreated with DEX (30 mg / kg) by intraperitoneal injection on the 1st, 7th, 14th and 28th days of the experiment, 30 min later, 5 mg / kg of DOX or normal saline was injected intravenously to the CON group and DOX group mice, and on the 42nd day of the experiment, the heart ultrasound of each group of mice was detected, and the ventricular size and function of the mice after anesthesia were examined by echocardiography (Vevo 2100, MS400C scan head), as shown in Figure 2 . The M-mode image of the left ventricle was obtained at the papillary muscle level. The left ventricular internal diameter at end diastole (LVIDd) and end systole (LVID) was measured according to the M-mode recording. The fractional shortening was calculated as (LVIDd-LVID) / LVIDd (percentage). Six representative systolic cycles were selected for analysis, and the ejection fraction (EF) and fractional shortening (FS) of each mouse were calculated, and the statistical results are shown in Figure 3 . As can be seen from the figure, compared with the DOX group, the EF and FS of the DEX + DOX group mice were significantly increased, indicating that DEX had obvious protective effect on doxorubicin (DOX) induced cardiac injury, but the EF and FS were still significantly lower than those of the control group mice, indicating that DEX alone could not completely reverse the DOX induced cardiotoxicity injury.

[0065] (1.3) The hearts of mice in each group were collected, fixed, sectioned, and stained with hematoxylin-eosin (HE) and WGA, and the number of myocardial cell vacuoles and myocardial fiber diameter of each mouse were counted, and the results are shown in Figure 4 . As can be seen from the figure, compared with the DOX group, the number of myocardial cell vacuoles and myocardial fiber diameter of the DEX + DOX group mice were significantly reduced, indicating that DEX had obvious protective effect on doxorubicin (DOX) induced cardiac injury, but the number of myocardial cell vacuoles and myocardial fiber diameter were still higher than those of the control group mice, indicating that DEX alone could not completely reverse the DOX induced cardiotoxicity injury.

[0066] (2) Cell test:

[0067] (2.1) AC16 myocardial cells and H9c2 myocardial cells were respectively inoculated in 96-well plates at a density of 5×10 4The HL-1 cells were inoculated in 96-well plates at a cell inoculation density of 1 x 10

[0068] (2.2) The DEX+DOX group was added with DEX (20 μM) for pretreatment, and after mixing, the 96-well plate was placed in a cell culture box for 30 min, and then DOX (0.5 μM) or normal saline was added to treat the cells for 48 h; then 10 μL of CCK8 working solution was added to each well, mixed, and then incubated in the dark for 3 h in the incubator, and then the absorbance at 450 nm was detected by an enzyme-labeled instrument, and the cell survival rate of each group was calculated. The results are shown in Figure 5 The results show that DEX has a significant protective effect on doxorubicin (DOX)-induced cardiac injury, but is not enough to completely reverse DOX-induced myocardial cell injury.

[0069] Example 2

[0070] This experiment explores whether autophagy is activated in the process of DOX-induced myocardial cell toxicity:

[0071] (1) The HL-1 myocardial cells were inoculated in 5 cm plates at a cell inoculation density of 1 x 10 7 mL, and DOX (0.5 μM) or normal saline (control group) was added to treat the cells for 48 h, and the protein expression of LC3-II, Atg7 and P62 in the HL-1 myocardial cells was detected by Western blot at 0 h, 12 h, 24 h and 48 h, respectively, and the results are shown in Figure 6 (LC3-II), Figure 7 (Atg7), Figure 8 (P62), respectively. As can be seen from the figures, after DOX treatment, the expression of LC3-II and Atg7 gradually increased, while the expression of P62 gradually decreased, indicating that autophagy was activated in the process of DOX-induced myocardial cell toxicity.

[0072] (2) Further detection of the expression of Atg7 and LC3-II fluorescent proteins by laser confocal detection, as shown in Figure 9 (Atg7) and Figure 10 (LC3-II), respectively. As can be seen from the figures, the expression of LC3-II and Atg7 gradually increased, indicating that autophagy was activated in the process of DOX-induced myocardial cell toxicity.

[0073] Example 3

[0074] This experiment explores whether inhibition of autophagy can reduce DOX-induced cardiac dysfunction in vivo:

[0075] Autophagy related genes (Atg) control the extension of autophagic membranes and the formation of autophagosomes by coupling complex formation in the first stage of autophagy. Among them, Atg7 is a key inducer of autophagy. In order to determine whether autophagy plays a protective or harmful role in DOX-induced cardiotoxicity, we established a model of AIC chronic mice with myocardial cell-specific deletion of Atg7 (AKO mice) by injecting Atg7 flox / flox The mice were administered tamoxifen at 40 mg / kg once a week for 3 weeks to induce Atg7 gene-specific deletion in myocardial cells, and the body weight gain of the mice was monitored during the period.

[0076] (1) C57 mice and AKO mice described above were used as animal models, each divided into 2 groups, 4 in each group, and administered with DOX and normal saline respectively.

[0077] (2) The experimental operation process is shown in Figure 11 5 mg / kg of DOX or normal saline was intravenously injected on the 1st, 7th, 14th and 28th day of the experiment, and on the 42nd day of the experiment, the heart ultrasound of each group of mice was detected, and the ventricular size and function of the mice after anesthesia were examined by echocardiography (Vevo 2100, MS400C scan head), as shown in Figure 12 The left ventricular internal diameter at end diastole (LVIDd) and end systole (LVID) was measured according to M-mode recording. Six representative systolic cycles were selected for analysis, and the ejection fraction (EF) and fractional shortening (FS) of each mouse were calculated, and the statistical results are shown in Figure 13 As can be seen from the figure, the echocardiogram of DOX-treated WT mice showed a decrease in EF and fractional shortening FS, while the EF and FS of DOX-treated AKO mice were higher than those of the WT group, indicating that myocardial cell-specific knockout of Atg7 gene in mice can reduce the effect of DOX on myocardial ejection function.

[0078] (3) The hearts of each group of mice were collected, fixed, sectioned, and stained with hematoxylin-eosin (HE) and WGA, and the number of myocardial cell vacuoles of each mouse was counted, and the results are shown in Figure 14 As can be seen from the figure, the number of myocardial cell vacuoles in DOX-treated AKO mice was reduced compared with that in the DOX-treated WT group, indicating that myocardial cell-specific knockout of Atg7 gene in mice can significantly reduce the vacuolization of myocardial cells caused by DOX. The diameter of myocardial fibers was counted, and the results are shown in Figure 15As shown in the figure, the myocardial cell fiber diameter of the DOX-treated AKO mice is smaller than that of the DOX-treated WT group, indicating that the specific knockout of the Atg7 gene in the myocardial cells of the mice can weaken DOX-induced myocardial hypertrophy.

[0079] The hearts of the mice in each group were collected, fixed, sectioned, and subjected to LC3 immunohistochemical staining, and the results are shown in Figure 16 As shown in the figure, the expression amount of LC3 in the myocardial cells of the DOX-treated AKO mice is less than that of the DOX-treated WT group, indicating that the specific knockout of the Atg7 gene in the myocardial cells of the mice weakens the autophagy of the myocardial cells caused by DOX.

[0080] (4) Freshly excised myocardial tissue was quickly cut into 1 mm cubes and fixed in 2.5% glutaraldehyde overnight, then post-fixed with 1% osmium tetroxide, dehydrated through a graded ethanol series and embedded in epoxy resin. Ultrathin sections (70 nm) were collected and double-stained with uranyl acetate and lead citrate. Transmission electron microscopy (TEM) using a Philips TECNA110 electron microscope was used to observe the mitochondrial state and autophagic vacuoles of the tissue, as shown in Figure 17 As shown in the figure, the autophagy-related structure of the myocardial cells of the DOX-treated AKO mice is less than that of the DOX-treated WT group, indicating that the specific knockout of the Atg7 gene in the myocardial cells of the mice can weaken the autophagy of the myocardial cells caused by DOX.

[0081] Example 4

[0082] This experiment explores the protective effect of single-drug SAR405 or combined drug composition with DEX on DOX-induced cardiotoxicity:

[0083] (1) C57 mice were used as animal models and divided into 4 groups, 4 in each group, namely normal control group (CON group), DOX treatment group (DOX group), SAR405 pretreatment + DOX treatment group (SAR405 + DOX group), and SAR405 combined with DEX pretreatment + DOX treatment group (SAR405 + DEX + DOX group).

[0084] (2) The test operation process is as shown in Figure 18As shown, on the 1st, 7th, 14th and 28th day of the experiment, the mice were pretreated with SAR405 (10 mg / kg, gavage) or pretreated with SAR405 (10 mg / kg, gavage) combined with DEX (40 mg / kg, intraperitoneal injection) for 30 min, and then intravenously injected with 5 mg / kg of DOX or normal saline for the CON group and the DOX group. On the 42nd day of the experiment, the heart function of each group of mice was detected by echocardiography (Vevo 2100, MS400C scan head), and the ventricular size and function of the mice after depilation and anesthesia were examined by echocardiography (Vevo 2100, MS400C scan head), as shown in Figure 19 The M-mode image of the left ventricle was obtained at the papillary muscle level. The left ventricular internal diameter at end diastole (LVIDd) and end systole (LVID) was measured according to the M-mode recording. The fractional shortening was calculated as (LVIDd-LVID) / LVIDd (percentage). Six representative systolic cycles were selected for analysis, and the ejection fraction (EF) and fractional shortening (FS) of each mouse were calculated, and the statistical results are shown in Figure 20 As can be seen from the figure, compared with the DOX group, the EF and FS of the SAR405+DOX group of mice were significantly increased, indicating that SAR405 had a significant protective effect on doxorubicin (DOX)-induced cardiac injury, but the EF and FS were still significantly lower than those of the control group of mice, indicating that the use of SAR405 alone could not completely reverse the DOX-induced cardiotoxicity. But the combination of SAR405 and DEX can completely protect the heart function, which is manifested as the percentage of EF and FS increasing to the same level as the normal control group.

[0085] (3) The hearts of the mice in each group were collected, fixed, sectioned, and stained with hematoxylin-eosin (HE) and WGA, and the number of myocardial cell vacuoles of each mouse was counted, and the results are shown in Figure 21 As can be seen from the figure, the increase in the number of myocardial cell vacuoles induced by DOX can be partially and completely reversed by the use of SAR405 alone or the combination of SAR405 and DEX, respectively. The diameter of the myocardial fibers was counted, and the results are shown in Figure 22 As can be seen from the figure, the increase in the diameter of the myocardial fibers induced by DOX can be partially and completely reversed by the use of SAR405 alone or the combination of SAR405 and DEX, respectively.

[0086] The hearts of the mice in each group were collected, fixed, sectioned, and stained with LC3 immunohistochemistry, and the results are shown in Figure 23 As can be seen from the figure, in the DOX-treated heart tissue, LC3 punctate cell accumulation indicates that autophagy is activated, and compared with the SAR405-only group, the autophagy degree of the SAR405 and DEX combined group is reduced, which is comparable to the control group.

[0087] (4) Freshly resected myocardial tissue was rapidly cut into 1 mm cubes and fixed overnight in 2.5% glutaraldehyde, followed by post-fixation with 1% osmium tetroxide, dehydrated through graded ethanol series and embedded in epoxy resin. Ultrathin sections (70 nm) were collected and double stained with uranyl acetate and lead citrate. Mitochondrial status and autophagic vacuoles of the tissue were observed using transmission electron microscopy (TEM) on a Philips TECNA110 electron microscope, as shown in Figure 24 As shown in the figure, autophagy-related structures in the combination of SAR405 and DEX group were significantly reduced compared with the single use of SAR405 group, indicating that the combination can more effectively reduce autophagy.

[0088] The applicant states that the application of the VPS34 inhibitor of the present application in the preparation of a drug for preventing and treating anthracycline-induced cardiotoxicity is illustrated by the above examples, but the present application is not limited to the above examples, that is, it does not mean that the present application must rely on the above examples to be implemented. Those skilled in the art should understand that any improvement on the present application, equivalent replacement of each raw material of the product of the present application, addition of auxiliary ingredients, selection of specific modes, etc. all fall within the protection scope and disclosure scope of the present application.

[0089] The preferred embodiments of the present application are described in detail above, but the present application is not limited to the specific details in the above embodiments, and within the technical concept scope of the present application, various simple modifications can be made to the technical solutions of the present application, and these simple modifications all belong to the protection scope of the present application.

[0090] In addition, it should be noted that each specific technical feature described in the above specific embodiments can be combined in any appropriate manner without contradiction, and in order to avoid unnecessary repetition, the present application will not further describe various possible combination manners.

Claims

1. Use of VPS34 inhibitor SAR405 for the preparation of a medicament for preventing or treating anthracycline-induced cardiotoxicity. The anthracycline is doxorubicin or a pharmaceutically acceptable salt thereof.

2. Use according to claim 1, characterized in that, The medicament further comprises a pharmaceutically acceptable excipient.

3. Use according to claim 1, characterized in that, The medicament is in any pharmaceutically acceptable dosage form.

4. Use of a combination of VPS34 inhibitor SAR405 and dexrazoxane for the preparation of a medicament for preventing or treating anthracycline-induced cardiotoxicity. The anthracycline is doxorubicin or a pharmaceutically acceptable salt thereof.

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