A pharmaceutical composition for preventing or treating ventricular arrhythmia after myocardial infarction

Abstract

The application discloses a kind of for preventing or treating post myocardial infarction ventricular arrhythmia pharmaceutical composition, the pharmaceutical composition is by everolimus, ethmolam and fluoxetine, belongs to the field of medicine technology.Immunoblotting proves that everolimus, ethmolam and fluoxetine can inhibit fatty acid oxidation protein.In vitro and in vitro cardiac electrophysiology experiment proves that the pharmaceutical composition can effectively reduce post myocardial infarction ventricular arrhythmia susceptibility.TTC staining proves that the pharmaceutical composition can effectively reduce post myocardial infarction myocardial damage area.Based on the above composition in the regulation of post myocardial infarction ventricular arrhythmia susceptibility, it can be further developed in the clinical application of the disease.

Description

Technical Field

[0001] This invention belongs to the field of medicine and relates to a pharmaceutical composition for the prevention or treatment of ventricular arrhythmias after myocardial infarction. Background Technology

[0002] Cardiovascular disease is the leading cause of death worldwide, and related diseases, particularly myocardial ischemia, have particularly poor clinical outcomes. Ischemic myocardium can lead to serious complications such as ventricular arrhythmias and acute heart failure. Therefore, developing novel drugs or combination strategies that can effectively prevent and treat ventricular arrhythmias after myocardial infarction is of great clinical significance for improving the long-term prognosis of these patients.

[0003] Everolimus is an important targeted therapy and immunosuppressant, belonging to the mTOR inhibitor class. Currently, its clinical application is mainly for cancer treatment. In this study, we found that everolimus can inhibit the fatty acid oxidation-related gene Fitm1; however, it remains unclear whether everolimus can be used to prepare drugs for the prevention or treatment of ventricular arrhythmias after myocardial infarction.

[0004] Etomoxir is a mitochondrial carnitine palmitoyltransferase 1 (CPT-1) inhibitor, belonging to the fatty acid β-oxidation inhibitors. Currently, the efficacy and feasibility of this compound in the prevention or treatment of ventricular arrhythmias after myocardial infarction remain unclear and require further research confirmation.

[0005] Fluoxetine is a classic selective serotonin reuptake inhibitor (SSRI) antidepressant. Previous studies have shown that fluoxetine is an inhibitor of VDAC1 (voltage-dependent anion channel 1), a protein located on the outer mitochondrial membrane involved in fatty acid transport. Whether fluoxetine can be used to develop drugs for the prevention or treatment of ventricular arrhythmias following myocardial infarction remains unclear.

[0006] Based on currently available scientific and technological information, it is not possible to predict whether the combination therapy consisting of everolimus, ethmoxel, and fluoxetine is suitable for the prevention or treatment of ventricular arrhythmias after myocardial infarction, nor is it possible to determine whether this drug combination can produce a synergistic effect. Summary of the Invention

[0007] To address the aforementioned clinical needs, this invention aims to provide a pharmaceutical composition for the prevention or treatment of ventricular arrhythmias following myocardial infarction. The pharmaceutical composition comprises everolimus, ethmoxel, and fluoxetine, wherein the mass ratio of everolimus, ethmoxel, and fluoxetine is 1:1:1.

[0008] Furthermore, the pharmaceutical composition also includes pharmaceutically acceptable excipients and / or carriers.

[0009] Furthermore, the pharmaceutical composition is an oral or parenteral dosage form, and also includes a pharmaceutically acceptable carrier or excipient.

[0010] Furthermore, the oral dosage form is a tablet, capsule, oral liquid, syrup, granule, drop pill, orally disintegrating tablet or sustained-release tablet; the parenteral dosage form is an aqueous injection, lyophilized powder injection, sterile powder injection or infusion.

[0011] Specifically, the drug described in this invention can reduce the incidence and duration of ventricular arrhythmias after ischemia-reperfusion. Simultaneously, the drug described in this invention can reduce the increase in myocardial infarction area after ischemia-reperfusion.

[0012] Furthermore, the combined use of everolimus, ethmoxel, and fluoxetine in the drug composition can synergistically increase the efficacy of preventing, alleviating, and treating ventricular arrhythmias after myocardial infarction.

[0013] To solve the above-mentioned technical problems, the technical solution of the present invention is as follows:

[0014] In this invention, unless otherwise stated, everolimus includes everolimus compounds (drugs), free forms, pharmaceutically acceptable salts, esters, solvates, hydrates, isomers, prodrugs, deuterated compounds, or isotope-labeled forms.

[0015] In this invention, unless otherwise stated, ethmoxel includes ethmoxel compounds (drugs), free forms, pharmaceutically acceptable salts, cocrystallizations, solvates, hydrates, isomers (including stereoisomers and tautomers), or prodrugs.

[0016] In this invention, unless otherwise stated, the fluoxetine includes fluoxetine compounds (drugs), free forms, pharmaceutically acceptable salts, cocrystallizations, solvates, hydrates, isomers (including stereoisomers and tautomers), or prodrugs.

[0017] Optionally, everolimus is an everolimus compound. Optionally, ethmoxel is an ethmoxel compound. Optionally, fluoxetine is a fluoxetine compound.

[0018] Optionally, in a specific embodiment, the pharmaceutically acceptable salt may be in a salt form conventionally used in the art. The salt may be selected from one or more of the following compounds: acetate, hydrochloride, hydrobromide, nitrate, sulfate, phosphate, benzoate, fumarate, maleate, succinate, tartrate, citrate, oxalate, glyoxylate, aspartate, hydrogen tartrate, 2,5-dihydroxybenzoate, methanesulfonate, ethanesulfonate, benzenesulfonate, lauryl sulfonate, hydroquinone sulfonate, p-toluenesulfonate, or a salt formed with a carboxylic acid (e.g., formic acid, acetic acid, propionic acid).

[0019] Generally, the structural formula of everolimus compounds is shown in Formula I, and its molecular formula is C1. 53 H 83 NO 14 Its molecular weight is 958.23.

[0020] Generally, the structural formula of everolimus compounds is shown in Formula I, and its molecular formula is C1. 11 H 23 N3O5 has a molecular weight of 277.32.

[0021] Formula I

[0022] Generally, the structural formula of ethmoxan compounds is shown in Formula II, and their molecular formula is C. 11 H 23 N3O5 has a molecular weight of 277.32.

[0023] Formula II

[0024] Generally, the structural formula of fluoxetine compounds is shown in Formula III, and its molecular formula is C. 17 H 18 F3NO has a molecular weight of 309.33.

[0025] Formula III

[0026] Preferably, the concentration of everolimus in the pharmaceutical composition of the present invention is 1-1000 μM.

[0027] Preferably, the concentration of ethmoxanil in the pharmaceutical composition of the present invention is 1-1000 μM.

[0028] Preferably, the concentration of fluoxetine in the pharmaceutical composition of the present invention is 1-1000 μM.

[0029] The pharmaceutical composition described in this invention can be prepared into any pharmaceutically acceptable dosage form using conventional pharmaceutical techniques. Optional dosage forms include, but are not limited to: tablets, capsules (including granules and powders), oral liquids (such as suspensions and decoctions), injections (including ready-to-use or temporarily prepared solutions), topical preparations (such as ointments, creams, and gels), suppositories, sprays / aerosols, and novel delivery systems (such as nanoparticles, microspheres, and implants). To prepare the above dosage forms, various applicable excipients known in the art can be used, such as: solvents, excipients, disintegrants, binders, lubricants, coating agents, stabilizers, preservatives, thickeners, emulsifiers, surfactants, flavoring agents, and colorants. The specific excipients chosen depend on the specific requirements of the target dosage form and route of administration. The composition is preferably presented in a single dosage form, such as tablets, capsules, or oral liquids, for ease of administration. Its preparation can employ conventional processes such as mixing, granulation, emulsification, and freeze-drying. The content of the active ingredient in the final composition is typically from 0.05% to 90% (by weight), with a preferred range of 0.1% to 50%.

[0030] The pharmaceutical excipients used in the pharmaceutical compositions of this invention can be selected from the following commonly used substances: water, starch, dextrin, various sugars (such as sucrose, lactose, glucose, mannitol), cellulose and its derivatives (such as microcrystalline cellulose, sodium carboxymethyl cellulose, hydroxypropyl cellulose), inorganic salts (such as calcium carbonate, calcium hydrogen phosphate, magnesium oxide), binders (such as gelatin paste, polyvinylpyrrolidone), disintegrants (such as croscarmellose sodium, croscarmellose), lubricants (such as magnesium stearate, talc, polyethylene glycol), and thickeners (such as sodium alginate, hydroxypropyl methyl cellulose), etc. The specific excipients can be combined according to the dosage form and process requirements.

[0031] For adult patients, this dose can be administered once daily or divided into multiple doses as needed. Administration routes include injection and non-injection. It should be noted that the actual dosing regimen needs to be individualized and optimized based on factors such as the specific type of disease being treated, the patient's age, weight, and the severity of clinical symptoms.

[0032] This invention provides a method for preventing, alleviating, and treating ventricular arrhythmias following myocardial infarction. The pharmaceutical composition comprises everolimus, ethmoxel, and fluoxetine. The combination of these three drugs achieves a synergistic effect. Attached Figure Description

[0033] Figure 1 The expression levels of Fitm1 and Cpt1b proteins in the ventricles of each mouse were shown.

[0034] Figure 2 The results of electrophysiological testing of the mouse heart are shown.

[0035] Figure 3 The results of electrophysiological testing of the mouse heart are shown.

[0036] Figure 4 The results of TTC staining of the myocardium in each group are shown. Detailed Implementation

[0037] The invention is described in more detail below to aid in understanding it.

[0038] Unless otherwise explicitly stated in the embodiments, all experimental procedures described in this section are performed using conventional methods in the art. Any specific experimental techniques or operating conditions not described in detail should be understood as being performed based on common knowledge in the relevant technical field, published literature methods, or the official product instructions for the reagents used.

[0039] Example 1: A pharmaceutical composition for detecting the inhibitory effects of everolimus on Fitm1 protein, ethmoxel on Cpt1b, and fluoxetine on VDAC1 by immunoblotting.

[0040] 1. Establishment of the MI model (myocardial infarction model):

[0041] This part of the experiment used 120 C57 mice, half male and half female, with a weight range of 200-220 grams (average 210 grams). All mice were housed in a barrier-compliant animal facility with environmental conditions set as follows: room temperature 20-23℃, relative humidity 60-70%, and a 12-hour light / 12-hour dark circadian rhythm. After acclimatization, all mice were randomly assigned to the following twelve groups, with 10 mice in each group:

[0042] The first part consists of: sham-operated group (each mouse was given 0.3 ml DMSO daily), myocardial infarction group (each mouse was given 0.3 ml DMSO daily), everolimus group (each mouse was given 20 mg / kg), and everolimus + myocardial infarction group (each mouse was given 20 mg / kg).

[0043] The second part consisted of: sham-operated group (each mouse was given 0.3 ml DMSO daily), myocardial infarction group (each mouse was given 0.3 ml DMSO daily), ethmoxel group (30 mg / kg), and ethmoxel + myocardial infarction group (30 mg / kg).

[0044] The third group consisted of: sham-operated group (0.3 ml DMSO per mouse per day), myocardial infarction group (0.3 ml DMSO per mouse per day), fluoxetine group (15 mg / kg), and fluoxetine + myocardial infarction group (15 mg / kg). All groups received intraperitoneal injections once daily, starting 7 days before the modeling surgery; and were given a second injection 30 minutes before the modeling surgery.

[0045] To establish a myocardial infarction injury model, this study followed the method reported by Jo et al. (2020) in the *International Journal of Molecular Sciences*, employing ligation of the left anterior descending coronary artery. The specific procedure was as follows: Anesthetized rats were fixed in a supine position, connected to a ventilator, and their electrocardiograms were continuously monitored. The pleural cavity was opened at the 3rd and 4th intercostal spaces on the left side of the sternum, and the pericardium was cut to fully expose the heart. A needle was inserted approximately 2 mm below the lower border of the left atrial appendage and exited beside the pulmonary artery conus, with a needle depth of approximately 1 mm. The left anterior descending coronary artery was ligated using 6-0 sutures along with a pre-placed inflatable balloon, and then the pleural cavity was closed. After ligation, the balloon was inflated. If the electrocardiogram showed a typical upward-convex ST segment elevation, the myocardial ischemia model was considered successfully established, and ischemia was maintained for 30 minutes. The balloon was then aspirated to restore coronary blood flow, and reperfusion was performed for 3 hours. Reperfusion was considered successful if the electrocardiogram showed that the previously elevated ST segment had decreased by more than 50%. Rats in the sham-operated group underwent the same surgical procedure, including thoracotomy and suture threading, but without coronary artery ligation.

[0046] 2. Immunoblotting detection method:

[0047] The methodology for detecting Fitm1, Cpt1b, and VDAC1 protein expression in C57 mouse myocardial tissue purchased from Cyagen Biosciences Co., Ltd. (Suzhou) using Western blotting is as follows: First, C57 mice were sacrificed according to the experimental design (e.g., genotype or treatment group), and target tissues (e.g., skeletal muscle, liver) were rapidly collected and flash-frozen in liquid nitrogen. Tissue samples were thoroughly homogenized and lysed at low temperature using lysis buffer (containing protease inhibitors and phosphatase inhibitors), and the supernatant was collected by centrifugation to obtain total protein. Protein concentration was accurately quantified using the BCA method, and the samples were denatured by boiling with 5× loading buffer to prepare loading samples. Subsequently, an equal volume of total protein (usually 20-50 µg) was loaded onto a 10%-12% SDS-PAGE gel for electrophoretic separation and then transferred to a PVDF membrane using wet transfer. After transfer, the membrane was blocked with 5% skim milk at room temperature for 1 hour. Then, it was incubated with primary and secondary antibodies sequentially: specific primary antibodies against Fitm1, Cpt1b, and VDAC1 (dilution ratio recommended according to the instructions, usually 1:1000) were incubated overnight at 4°C. After washing with TBST, the membrane was incubated with the corresponding species' HRP-labeled secondary antibody (1:5000-1:10000) at room temperature for 1 hour. Finally, the membrane was developed using ECL chemiluminescent developing solution on an imaging system, with GAPDH or β-actin as internal controls. The grayscale values ​​of the target bands were analyzed using software such as ImageJ, and the ratio of the target protein to the internal control was calculated for statistical comparison.

[0048] 3. Statistical Analysis

[0049] All statistical analyses in this study were performed using GraphPad Prism 8.0 software (GraphPad Software, Inc., USA). Data are expressed as mean ± standard error. The strategy for comparisons between groups was as follows: when the data were confirmed to be normally distributed and homogeneous in variance by the Shapiro-Wilk test, an unpaired t-test was used; otherwise, the Mann-Whitney U test was used. One-way ANOVA was used for comparisons among multiple groups, and Tukey's method was used for post-hoc tests. Categorical variables were described as frequencies (percentages), and chi-square tests were used for comparisons between groups. Furthermore, univariate and multivariate logistic regression models were used to analyze clinical relevance. A p-value less than 0.05 was used as the criterion for statistical significance.

[0050] like Figure 1 As shown in Figures AB, compared with the sham-operated group, the expression level of Fitm1 protein in the ventricles of C57 mice was significantly reduced in the sham-operated + everolimus group. Compared with the simple myocardial ischemia group, everolimus also significantly inhibited the expression level of Fitm1 protein in the ventricles of C57 mice; Figure 1As shown in CD, compared with the sham-operated group, the expression level of Cpt1b protein in the ventricles of C57 mice in the sham-operated + ethmoxel group was significantly reduced. Compared with the simple myocardial ischemia group, ethmoxel also significantly inhibited the expression level of Cpt1b protein in the ventricles of C57 mice; Figure 1 As shown in Figure EF, compared with the sham-operated group, the expression level of VDAC1 protein in the ventricles of C57 mice was significantly reduced in the sham-operated + fluoxetine group. Compared with the myocardial ischemia-only group, fluoxetine also significantly inhibited the expression level of VDAC1 protein in the ventricles of C57 mice.

[0051] Example 2: The preventive effect of a pharmaceutical composition of everolimus, ethmoxel, and fluoxetine on ventricular arrhythmias after myocardial infarction (MI).

[0052] Main reagents: Everolimus, Etoxol, and Fluoxetine were purchased from MCE (China) Co., Ltd.

[0053] 1. Establishment of the MI model:

[0054] This part of the experiment used 60 C57 mice, half male and half female, with a weight range of 200-220 grams (average 210 grams). All mice were housed in a barrier-compliant animal room with environmental conditions set as follows: room temperature 20-23℃, relative humidity 60-70%, and a circadian rhythm of 12 hours light / 12 hours dark. After acclimatization, all mice were randomly assigned to the following six groups, 10 mice per group: sham-operated group (0.3 ml DMSO per mouse per day), model group (0.3 ml DMSO per mouse per day), everolimus group (20 mg / kg), esmoxel group (30 mg / kg), fluoxetine group (15 mg / kg), and everolimus + esmoxel + fluoxetine combined administration group (all three drugs at a dose of 10 mg / kg). All groups received intraperitoneal injections once daily, starting 7 days before the modeling surgery; and a second injection 30 minutes before the modeling surgery.

[0055] To establish a myocardial infarction model, this study followed the method reported by Jo et al. (2020) in the *International Journal of Molecular Sciences*, employing ligation of the left anterior descending coronary artery. The specific procedure was as follows: Anesthetized rats were fixed in a supine position, connected to a ventilator, and their electrocardiograms were continuously monitored. The thoracic cavity was opened at the 3rd and 4th intercostal spaces on the left side of the sternum, and the pericardium was cut to fully expose the heart. A needle was inserted approximately 2 mm below the lower border of the left atrial appendage and exited beside the pulmonary artery conus, with a depth of approximately 1 mm. The left anterior descending coronary artery was ligated using 6-0 sutures along with a pre-placed inflatable balloon, and then the thoracic cavity was closed. After ligation, the balloon was inflated. If the electrocardiogram showed a typical upward-arching ST segment elevation, the myocardial ischemia model was considered successfully established. Rats in the sham-operated group underwent the same surgical procedure, including thoracotomy and suture threading, but without coronary artery ligation.

[0056] 2. Evaluation Methods

[0057] On the second day after myocardial infarction surgery, C57 mice were weighed and anesthetized by intraperitoneal injection of sodium pentobarbital at a dose of 50 mg / kg. After the animals were stabilized under anesthesia, surface electrocardiogram electrodes were placed at specific locations on their chests and connected to an electrocardiograph for real-time monitoring. Basic cardiac electrical activity was recorded, with a focus on observing and identifying abnormal cardiac rhythms such as ventricular tachycardia or ventricular fibrillation. Finally, the number of animals exhibiting the above-mentioned ventricular arrhythmias in each group was counted, and their incidence was calculated.

[0058] 3. Statistical Analysis

[0059] All statistical analyses in this study were performed using GraphPad Prism 8.0 software (GraphPad Software, Inc., USA). Data are expressed as mean ± standard error. The strategy for comparisons between groups was as follows: when the data were confirmed to be normally distributed and homogeneous in variance by the Shapiro-Wilk test, an unpaired t-test was used; otherwise, the Mann-Whitney U test was used. One-way ANOVA was used for comparisons among multiple groups, and Tukey's method was used for post-hoc tests. Categorical variables were described as frequencies (percentages), and chi-square tests were used for comparisons between groups. Furthermore, univariate and multivariate logistic regression models were used to analyze clinical relevance. A p-value less than 0.05 was used as the criterion for statistical significance.

[0060] Results of electrophysiological testing of mouse heart are as follows Figure 2 As shown. Figure 2 In the diagram, A represents a schematic representation of each group of electrocardiograms. For example... Figure 2As shown in Figure BC, compared with the control group, the incidence of ventricular arrhythmias such as ventricular tachycardia and ventricular fibrillation was significantly increased in the MI group. The everolimus group, semaphore group, fluoxetine group, and the everolimus + semaphore + fluoxetine combination group all significantly reduced the incidence of ventricular arrhythmias following myocardial infarction. This indicates that everolimus, semaphore, and fluoxetine can, individually or in combination, reduce the incidence and duration of ventricular arrhythmias after myocardial infarction, thereby preventing ventricular arrhythmias after myocardial infarction (p†<0.05 vs solvent + sham operation group, p**<0.01 vs solvent + myocardial ischemia group, p***<0.001 vs solvent + myocardial ischemia group).

[0061] Example 3: The therapeutic effect of a pharmaceutical composition of everolimus, ethmoxel, and fluoxetine on ventricular arrhythmias following myocardial infarction (MI).

[0062] Main reagents: Everolimus, Etimoxel, and Fluoxetine were all purchased from MCE Company.

[0063] 1. MI model establishment and arrhythmia index detection:

[0064] This part of the experiment used 60 C57 mice of similar weight (230±10 g), half male and half female, housed in a barrier environment with controlled temperature and humidity (20-23℃, 60-70% humidity) and a 12-hour light cycle. After acclimatization, the mice were randomly divided into 6 groups (n=10 / group).

[0065] First, C57 mice were anesthetized and heparinized. The heart was then rapidly removed via thoracotomy and placed in a 4°C cold perfusion solution. The aortic arch was immediately connected to a constant-temperature (37°C), constant-pressure Langendorff perfusion system, and retrograde perfusion was performed using a modified Krebs-Henseleit buffer with continuous oxygenation (95% O2 / 5% CO2). After the heart resumed beating and stabilized for approximately 20 minutes, a reversible ischemia model was created by encircling the left anterior descending coronary artery with a 6-0 or 7-0 surgical suture approximately 2 mm below the lower border of the left atrial appendage, with both ends of the suture passed through a small silicone tube. Coronary blood flow was then completely blocked by tightening the suture and fixing the silicone tube. Significant ST-segment elevation was observed on the electrocardiogram during ischemia. Programmed electrical stimulation electrodes placed at the apex of the right ventricle were applied using a specific stimulation protocol to quantitatively assess the arrhythmia induction threshold. Drugs were administered via the Langendorff perfusion system during this experiment. The groups were as follows: sham surgery group, everolimus group (3 mmol / ml), esmoxel (2 mmol / ml) or fluoxetine (1.5 mmol / ml); and a triple therapy group was set up, which was given everolimus, esmoxel and fluoxetine (1 mmol / ml each).

[0066] 2. Statistical Analysis

[0067] Statistical analysis was performed using GraphPad Prism 8.0 software. Continuous data were expressed as mean ± SEM, and categorical data as n (%). For comparisons of continuous data between two groups, after confirming normality via the Shapiro-Wilk test, unpaired t-tests were used for those meeting the criteria; otherwise, the Mann-Whitney U test was used. One-way ANOVA was used for comparisons among multiple groups, and Tukey's method was used for post-hoc pairwise comparisons. Chi-square tests were used for comparisons among categorical data. Logistic regression (univariate and multivariate) was used to analyze clinical relevance. p < 0.05 was considered statistically significant.

[0068] Results of electrophysiological testing of mouse heart are as follows Figure 3 As shown. Figure 3 In the diagram, A represents a schematic representation of each group of electrocardiograms. For example... Figure 3 As shown in Figure BC, compared with the control group, the incidence of ventricular arrhythmias such as ventricular tachycardia and ventricular fibrillation was significantly increased in the myocardial ischemia group. The everolimus group, semaphore group, fluoxetine group, and the everolimus + semaphore + fluoxetine combination group all significantly reduced the incidence of ventricular arrhythmias after myocardial infarction. This indicates that everolimus, semaphore, and fluoxetine can, individually or in combination, reduce the incidence and duration of ventricular arrhythmias after myocardial infarction, thereby treating ventricular arrhythmias after myocardial infarction (p†<0.05 vs solvent + sham operation group, p††<0.01 vs solvent + sham operation group, p**<0.01 vs solvent + myocardial ischemia group, p***<0.001 vs solvent + myocardial ischemia group).

[0069] Example 4: A pharmaceutical composition of everolimus, ethmoxel, and fluoxetine reduced the severity of myocardial infarction.

[0070] The extent of myocardial infarction is directly related to the risk of ventricular arrhythmias; an increased infarct area significantly increases the probability of arrhythmia. Therefore, based on the aforementioned model construction and treatment intervention, this embodiment used TTC staining to quantitatively assess the myocardial infarction area in four groups of mice. The specific experimental steps are as follows:

[0071] TTC staining method:

[0072] Mice were euthanized by cervical dislocation, and their hearts were quickly removed. The myocardial tissue was placed in a specialized slide mold and continuously sliced ​​into 2 mm thick sections using a scalpel. The obtained heart sections were then immersed in prepared TTC staining solution and incubated at 37°C for 30 minutes to ensure thorough staining. After staining, the sections were gently washed with PBS buffer for 3 to 5 minutes, and finally, images were acquired from the stained sections.

[0073] The TTC staining results of the myocardium in each group are as follows: Figure 4 As shown. Figure 4 In the diagram, A represents a schematic representation of TTC staining in the myocardium of each group. The results are as follows: Figure 4 The BC analysis showed that, compared with the sham-operated group, the myocardial infarction area in the MI group mice was significantly larger, confirming that myocardial infarction led to significant cardiomyocyte death. The myocardial infarction area in the everolimus group, semaphore group, fluoxetine group, and the everolimus + semaphore + fluoxetine combination group was significantly smaller than that in the model group, indicating that everolimus, semaphore, and fluoxetine can alleviate myocardial infarction individually or in combination. Therefore, it can be inferred that everolimus, semaphore, and fluoxetine can reduce the risk of ventricular arrhythmias after myocardial infarction by reducing the infarct area (p*<0.05 vs solvent + myocardial ischemia group, p**<0.01 vs solvent + myocardial ischemia group, p***<0.001 vs solvent + myocardial ischemia group, p****<0.0001 vs solvent + myocardial ischemia group).

Claims

1. A pharmaceutical composition for the prevention or treatment of ventricular arrhythmias following myocardial infarction, wherein the active ingredients of the pharmaceutical composition consist of everolimus, ethmoxel, and fluoxetine, wherein the mass ratio of everolimus, ethmoxel, and fluoxetine is 1:1:

1.

2. The pharmaceutical composition according to claim 1, characterized in that, The pharmaceutical composition also includes pharmaceutically acceptable excipients.

3. The pharmaceutical composition according to claim 1, characterized in that, The pharmaceutical composition is in the form of an oral or parenteral dosage form and also includes a pharmaceutically acceptable carrier.

4. The pharmaceutical composition according to claim 1, characterized in that, The pharmaceutical composition is in the form of an oral or parenteral dosage form and also includes pharmaceutically acceptable excipients.

5. The pharmaceutical composition according to claim 3 or 4, characterized in that, The oral dosage form is tablet, capsule, oral liquid, syrup, granule or drop pill; the parenteral dosage form is water injection, lyophilized powder injection or infusion.

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