Compositions for the prevention and treatment of heart disease caused by pulmonary hypertension, and functional preparations containing the same.

Abstract

The present invention provides a composition for the prevention and treatment of heart disease caused by pulmonary hypertension, and a functional formulation containing the same. The composition comprises pimobendan, enalapril, torsemide, and spironolactone, and is effective in the prevention and treatment of heart disease and pulmonary hypertension in animals.

Description

Technical Field

[0001] The present invention relates to a composition for preventing heart diseases and treating the heart, and a functional preparation containing the same. More specifically, the present invention relates to a composition for preventing heart diseases such as heart failure caused by pulmonary hypertension and treating the heart, and a functional preparation for animals containing the same, which are targeted at animals such as dogs and cats rather than humans.

Background Art

[0002] The heart plays a role in receiving blood from veins and continuously circulating blood throughout the body via arteries. In order for the heart to pump blood, it needs to contract, and the myocardium composed of muscle causes such a contraction effect. In this regard, when there is a disorder in the myocardial cells themselves due to an increase in ventricular load caused by hypertension or valvular heart disease, myocardial infarction, myocarditis or cardiomyopathy, etc., it becomes impossible to send a sufficient amount of blood to the organs of the whole body, the cardiac output decreases, and a myocardial hypertrophy phenomenon is induced. The heart is an organ in which differentiation has been completely completed in terms of embryology, and further cell proliferation is impossible. Therefore, when it becomes necessary to increase the cardiac function, the only countermeasure is to increase the size of existing myocardial cells to enhance the contractility of the myocardium, and such a physiological phenomenon is called myocardial hypertrophy. If such myocardial hypertrophy persists for a certain period, there is a very high possibility of progressing to heart failure.

Summary of the Invention

Problems to be Solved by the Invention

[0003] One of the causes of heart failure is pulmonary hypertension. Blood is passed from the right heart to the lungs via the pulmonary artery to remove carbon dioxide in the blood and supply oxygen. Here, a state in which the blood pressure of the pulmonary artery rises to a high level is called pulmonary hypertension, and in such a case, it may become difficult to send blood from the right heart to the pulmonary artery. As a result, the right ventricle of the heart may hypertrophy and cause right heart failure.

[0004] The problem that this invention aims to solve is to provide a composition effective for the prevention and treatment of heart diseases such as heart failure, as well as a functional formulation containing the same.

[0005] Furthermore, the problem that the present invention aims to solve is to provide a composition effective in preventing and treating heart diseases such as heart failure and pulmonary hypertension, which may be a cause of such diseases, as well as a functional preparation containing the same.

[0006] The technical problems of the present invention are not limited to those described above, and other technical problems not mentioned will be clearly understood by those skilled in the art from the following description. [Means for solving the problem]

[0007] A composition and functional formulation for the prevention and treatment of heart disease and pulmonary hypertension in animals according to one embodiment for solving the aforementioned problems comprises pimobendan, enalapril, torsemide, and spironolactone.

[0008] The formulation may contain 0.5 mg of pimobendan, 1 mg of enalapril, 0.2 mg of torsemide, and 2 mg of spironolactone.

[0009] Based on the weight of the subject, the single dose may be 0.25 mg / kg of pimobendan, 0.5 mg / kg of enalapril, 0.1 mg / kg of torsemide, and 1 mg / kg of spironolactone, and may be administered twice daily.

[0010] The subject may be a dog or a cat.

[0011] Specific details of other embodiments are included in the detailed description. [Effects of the Invention]

[0012] The composition according to the examples of the present invention comprises pimobendan, enalapril, torsemide, and spironolactone, and is effective in the prevention and treatment of heart disease and pulmonary hypertension in animals.

[0013] The composition of the present invention is not particularly limited to a specific formulation form and can be manufactured in various formulation forms and used as an additive in various formulations.

[0014] The compositions according to the embodiments of the present invention are effective against pet animals such as dogs and cats, and can be manufactured as treats and food for pet animals.

[0015] The effects of the present invention are not limited to those exemplified above, and a wider range of effects are included herein. [Brief explanation of the drawing]

[0016] [Figure 1] This graph shows the clinical test results for Example 1. [Figure 2] This graph shows the clinical test results for Example 1. [Figure 3] This graph shows the clinical test results for Example 1. [Figure 4] This graph shows the clinical test results for Example 1. [Figure 5] This graph shows the clinical test results for Example 1. [Figure 6] This graph shows the evaluation results of the blood index test in Example 1. [Figure 7] This graph shows the evaluation results of the blood index test in Example 1. [Figure 8] This graph shows the evaluation results of the blood index test in Example 1. [Figure 9] This graph shows the evaluation results of the serum biochemical indicator test in Example 1. [Figure 10] This graph shows the evaluation results of the serum biochemical indicator test in Example 1. [Figure 11] This graph shows the evaluation results of the serum biochemical indicator test in Example 1. [Figure 12] This graph shows the evaluation results of the serum biochemical indicator test in Example 1. [Figure 13]It is a graph showing the evaluation results of the serum biochemical index test in Example 1. [Figure 14] It is a graph showing the evaluation results of the serum biochemical index test in Example 1. [Figure 15] It is a graph showing the evaluation results of the serum electrolyte index in Example 1. [Figure 16] It is a graph showing the evaluation results of the serum electrolyte index in Example 1. [Figure 17] It is a graph showing the evaluation results of the serum electrolyte index in Example 1. [Figure 18] It is a graph showing the evaluation results of the serum electrolyte index in Example 1. [Figure 19] It is a graph showing the biomarker test results in Example 1. [Figure 20] It is a graph showing the biomarker test results in Example 1. [Figure 21] It is a graph showing the biomarker test results in Example 1. [Figure 22] It is a graph showing the evaluation results of the chest radiograph in Example 1. [Figure 23] It is a graph showing the evaluation results of the chest radiograph in Example 1. [Figure 24] It is a graph showing the evaluation results of the chest radiograph in Example 1. [Figure 25] It is a graph showing the evaluation results of the echocardiogram in Example 1. [Figure 26] It is a graph showing the evaluation results of the echocardiogram in Example 1. [Figure 27] It is a graph showing the evaluation results of the echocardiogram in Example 1. [Figure 28] It is a graph showing the evaluation results of the echocardiogram in Example 1. [Figure 29] It is a graph showing the evaluation results of the echocardiogram in Example 1. [Figure 30] It is a graph showing the evaluation results of the echocardiogram in Example 1. [Figure 31] It is a graph showing the evaluation results of the echocardiogram in Example 1. [Figure 32]This graph shows the evaluation results of the echocardiogram in Example 1. [Figure 33] This graph shows the evaluation results of the ISACHC stage in Example 1. [Figure 34] This graph shows the clinical test results for Example 2. [Figure 35] This graph shows the clinical test results for Example 2. [Figure 36] This graph shows the clinical test results for Example 2. [Figure 37] This graph shows the clinical test results for Example 2. [Figure 38] This graph shows the clinical test results for Example 2. [Figure 39] This graph shows the evaluation results of chest radiation in Example 2. [Figure 40] This graph shows the evaluation results of chest radiation in Example 2. [Figure 41] This graph shows the evaluation results of chest radiation in Example 2. [Figure 42] This graph shows the evaluation results of the echocardiogram in Example 2. [Figure 43] This graph shows the evaluation results of the echocardiogram in Example 2. [Figure 44] This graph shows the evaluation results of the echocardiogram in Example 2. [Figure 45] This graph shows the evaluation results of the echocardiogram in Example 2. [Figure 46] This graph shows the evaluation results for the severity of pulmonary hypertension in Example 2. [Figure 47] This graph shows the evaluation results of the ISACHC stage in Example 2. [Modes for carrying out the invention]

[0017] The advantages and features of the present invention, as well as methods for achieving them, will become clearer with reference to the examples described below in detail, along with the accompanying drawings. However, the present invention is not limited to the examples disclosed below and can be realized in a variety of different forms, and the examples are merely provided to complete the disclosure of the present invention and to fully inform those who are ordinary skill in the art to which the present invention pertains, and the present invention is defined solely by the scope of the claims.

[0018] Furthermore, "and / or" means that each item listed and all combinations of one or more items are included. A numerical range indicated using "or" means a numerical range that includes the values ​​listed before and after it as the lower and upper limits, respectively. "Approximately" or "roughly" means a value or numerical range that is within 20% of the value or numerical range listed after it.

[0019] The following describes embodiments of the present invention.

[0020] One embodiment of the present invention provides a composition for the prevention and treatment of heart disease, comprising pimobendan, enalapril, torsemide, and spironolactone. The composition of the present invention is effective for the prevention and treatment of heart disease in animals such as dogs and cats, rather than in humans. In particular, the composition of the present invention may be effective in the prevention and treatment of heart diseases that involve changes in the size of the heart, such as heart failure, and pulmonary hypertension, which is one of its causes.

[0021] In this application, "heart failure" refers to any systolic disorder or heart disease. Generally, clinical symptoms appear as a result of changes in the cellular and molecular components of the heart, and changes in the mediators that drive homeostasis. Typically, heart failure is associated with an increase in heart size and deterioration of cardiac function.

[0022] Pimobendan is a muscle contractile vasodilator compound that exhibits calcium sensitization and several phosphodiesterase type III inhibitory effects. Calcium sensitizers achieve positive muscle contraction not by increasing calcium influx into cardiomyocytes, but by sensitizing contractile proteins to cytosolic calcium and altering the binding of calcium to troponin-C. Achieving positive muscle contraction through calcium sensitization prevents some of the side effects of cytosolic calcium overload. Increased cytosolic calcium levels are associated with an increased tendency towards arrhythmias and sudden death. Clinical trials of long-term oral administration of pimobendan in dogs with heart failure demonstrated improved exercise tolerance and quality of life without significant survival-related side effects.

[0023] Pimobendan is an inodilator that possesses both positive myocardial force-increasing and vasodilatory effects. These effects, in addition to the PDEi-3 effect, exert a cardiotonic effect by increasing the calcium sensitivity of the contractile machinery within myocardial cells; therefore, it is also called a calcium sensitizer. Because this drug exerts a cardiotonic effect without increasing the oxygen demand of the myocardium, it enhances myocardial contractility without exacerbating myocardial ischemic changes in heart failure, and unlike digitalis, it carries virtually no risk of arrhythmia. Pimobendan is prescribed for chronic treatment in all patients with heart failure due to CMVI or DCM. The efficacy of pimobendan is highly beneficial in the treatment of clinically symptomatic heart failure patients, demonstrating outstanding effects in improving clinical symptoms and extending survival (PITCH study 2000, QUEST study 2008). The therapeutic effects of pimobendan have been shown to be even better in canine CMVI and DCM patients. The EPIC study (2018) investigated whether administering pimobendan to patients with subclinical (asymptomatic) CMVI resulted in improvements in clinical and radiographic findings, and whether there were differences in disease progression. In this study, the pimobendan group showed a decrease in radiographic LVIDd and LA / Ao at day 35 compared to the placebo group.

[0024] Enalapril is a medication used to treat hypertension, diabetic nephropathy, and heart failure. In cases of heart disease, it is commonly used in combination with diuretics such as furosemide. It can be administered orally or intravenously. When taken orally, the effects usually appear within one hour and typically last up to one day. Enalapril is classified as an ACE inhibitor, and common side effects include headache, fatigue, and cough.

[0025] Angiotensin-converting enzyme inhibitors (ACEi), such as enalapril, induce vasodilation by inhibiting the formation of angiotensin II, a vasoconstrictive factor produced by the action of renin secreted by the kidneys. Clinically, ACEi are primarily used to treat heart failure and hypertension and proteinuria caused by renal disease. The efficacy of several ACEi (enalapril, benazapril, imidapril, captopril) has been evaluated in patients with clinical symptoms (COVE study-1995; Live study-1998; BENCH trial-1999). These studies demonstrated that administering ACEi to patients with heart failure due to DCM and CMVI delayed the progression of heart failure, improved clinical symptoms, and extended survival.

[0026] Torsemide is a diuretic used to treat fluid overload caused by heart failure, kidney disease, liver disease, and hypertension. Torsemide is generally not preferred for treating hypertension itself and, like sulfonamides, acts as a loop diuretic by inhibiting sodium reabsorption in the kidneys. Common side effects of torsemide include headache, increased urination, diarrhea, cough, and dizziness, while other side effects include hearing loss and hypokalemia.

[0027] Diuretics are the most commonly and effectively used medications in the treatment of heart failure. Diuretics reduce venous return by decreasing circulating blood volume, ultimately lowering the blood flow into the ventricles (preload). Diuretics mainly used in small animal clinical practice are broadly classified into (1) loop diuretics, (2) thiazide diuretics, and (3) potassium-sparing diuretics. Of these, loop diuretics such as torsemide have the strongest diuretic effect. Furosemide has traditionally been the most widely used drug in the veterinary field, but it exhibits a ceiling effect (a phenomenon in which the drug's effect weakens after a certain period of time), and its bioavailability when administered orally is lower than that of torsemide. In a study investigating the pharmacokinetic properties of torsemide in dogs, torsemide showed almost no ceiling effect and demonstrated sustained efficacy even with long-term administration. Furthermore, in a recent randomized controlled trial (RCT) comparing the two drugs conducted in France, torsemide demonstrated superior efficacy. While torsemide is typically prescribed for dogs that do not respond to furosemide, its high bioavailability, long half-life in the blood, and minimal ceiling effect make it advantageous to administer torsemide as a substitute for furosemide in managing heart failure patients without increasing the dose of diuretics.

[0028] Spironolactone is an anti-aldosterone agent, potassium-sparing diuretic, and spiro compound. It exerts a diuretic effect by inhibiting the binding of aldosterone to its receptors in the body, thereby retaining potassium ions. It is mainly used to treat heart failure, ascites due to cirrhosis, and hypertension, and can also be used as an adjunctive treatment for hypokalemia.

[0029] Unlike other diuretics, potassium-sparing diuretics such as spironolactone do not directly act on sodium transport but rather antagonize the action of aldosterone in the collecting duct. This blocks the reabsorption of sodium (water) and induces diuresis. However, since potassium and hydrogen ion excretion does not occur at this site, it does not alter blood potassium or hydrogen concentrations. For this reason, this class of diuretics is called potassium-sparing diuretics. However, because the amount of sodium absorbed at this site accounts for a very small proportion of the total sodium reabsorption, the diuretic effect is extremely weak (~10% of the efficacy of loop diuretics). Therefore, it is not used alone and is often used in combination with thiazide diuretics or loop diuretics (to prevent hypokalemia). Studies on whether spironolactone slows the progression of heart failure were conducted in patients with quasi-clinical MMVD, but the effect was not clearly demonstrated. However, compared to the placebo group, the increase in cardiac size on echocardiography and cardiac biomarker concentrations was much smaller in the spironolactone group.

[0030] According to one embodiment, the composition of the present application may contain 5 parts by weight of pimobendan, 10 parts by weight of enalapril, 2 parts by weight of torsemide, and 20 parts by weight of spironolactone. The composition of the present application may contain pimobendan, enalapril, torsemide, and spironolactone in amounts effective for the prevention and treatment of heart disease, and may contain each component in the amount required for a single dose, taking into account the body weight of the target group to which the composition of the present application is administered, such as dogs and cats. According to one embodiment, the composition may contain 0.5 mg of pimobendan, 1 mg of enalapril, 0.2 mg of torsemide, and 2 mg of spironolactone. Tablets produced by compressing the composition may contain 0.5 mg of pimobendan, 1 mg of enalapril, 0.2 mg of torsemide, and 2 mg of spironolactone per tablet. The total weight of a single compressed tablet is not limited but may be set considering the patient's type, ability to take medication, and the absorption rate of the tablet's components. For example, a tablet produced by compressing the composition may have a weight of 100 mg per tablet and may contain 0.5 mg of pimobendan, 1 mg of enalapril, 0.2 mg of torsemide, and 2 mg of spironolactone per tablet. For a more detailed explanation, please refer to the experimental examples described later.

[0031] Compositions for the prevention and treatment of heart disease according to the embodiments of the present invention can be manufactured in various oral or parenteral dosage forms. When formulated, they are prepared using commonly used fillers, bulking agents, binders, wetting agents, disintegrants, surfactants, and other diluents or excipients. Solid formulations for oral administration include tablets, pills, powders, granules, capsules, etc. These solid formulations are prepared by mixing one or more compounds with at least one or more excipients, such as starch, calcium carbonate, sucrose or lactose, gelatin, etc. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Liquid formulations for oral administration include suspensions, oral solutions, emulsions, syrups, etc. In addition to commonly used simple diluents such as water and liquid paraffin, various excipients, such as wetting agents, sweeteners, fragrances, and preservatives, may be included. Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized agents, and suppositories. Non-aqueous solvents and suspending agents may include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate. Suppository bases may include witepsol, macrogol, tween 61, cocoa butter, lauric acid butter, and glycerol gelatin.

[0032] According to one embodiment, a composition for the prevention and treatment of heart disease can be formulated by including excipients. For example, a composition for the prevention and treatment of heart disease can be manufactured as a tablet containing oils and fats such as chicken oil and tocopherol acetate. By coating the surface of the tablet with oils and fats and tocopherol acetate, the oils and fats are not absorbed into the inner part of the tablet, thus preventing the tablet from crumbling or becoming damp.

[0033] The compositions for the prevention and treatment of heart disease according to the embodiments of the present invention can be manufactured as various functional foods. For example, they may be various foods such as chewing gum, caramel products, candies, frozen desserts, and confectionery; beverages such as soft drinks, mineral water, and alcoholic beverages; and health functional foods such as vitamins and minerals. Furthermore, they can be formulated as oral preparations for pets, specifically tablets, capsules, liquids, gels, pastes, oral sprays, buccal tablets, powders, and chewable treats or animal feed for pets, but are not limited thereto.

[0034] The compositions for the prevention and treatment of heart disease according to the embodiments of the present invention can be manufactured as food on their own or used in combination with other food ingredients, and can be used appropriately according to conventional methods. The amount of active ingredients mixed can be appropriately determined depending on the purpose of use.

[0035] The functional food of the present invention may contain various flavorings or natural carbohydrates as additional ingredients. The aforementioned natural carbohydrates include monosaccharides such as glucose and fructose, and sugar alcohols such as sorbitol and erythritol. As sweeteners, natural sweeteners such as thaumatin and stevia extract, or synthetic sweeteners such as saccharin and aspartame can be used. The ratio of the natural carbohydrates is preferably selected in the range of 0.01 to 0.04 parts by weight, more preferably about 0.02 to 0.03 parts by weight, per 100 parts by weight of the health functional food of the present invention.

[0036] In addition to the above, the functional food of the present invention may contain various nutrients, vitamins, electrolytes, flavorings, colorings, pectin acid and its salts, alginic acid and its salts, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohol, carbonation agents used in carbonated beverages, etc. Furthermore, the functional food of the present invention may contain fruit pulp for the production of natural fruit juices, fruit juice beverages, and vegetable beverages. These components can be used individually or in combination. The ratio of these additives is not important, but it is generally selected in the range of 0.01 to 0.1 parts by weight per 100 parts by weight of the functional food of the present invention.

[0037] The present invention will be described in more detail below using examples. This is for illustrative purposes only and does not limit the scope of the present invention.

[0038] <Example 1> Manufacture of a composition for the prevention and treatment of heart disease A liver treatment composition was prepared by mixing the components shown in Table 1 below. The units in Table 1 below are mg. Tablets containing 100 mg each were prepared by compressing the composition having the following composition.

[0039] [Table 1]

[0040] <Experimental Example 1> Confirmation of the preventive and therapeutic effects of heart disease To confirm the preventive and therapeutic effects of the composition of Example 1 on heart disease, the following experiment was conducted.

[0041] <Experimental Method> Experiments were conducted to evaluate the efficacy and stability of the composition of Example 1 by selecting a test group to receive the composition and a placebo group to receive a placebo. The test group and placebo group were selected from 30 dogs diagnosed with heart failure of ISACHC stage III or higher. The 30 test animals were small breed dogs with the same sex composition (15 females and 15 males).

[0042] The selected test animals were classified for heart failure status based on direct examination by a veterinary cardiologist through clinical examination, chest radiography, and echocardiography, as well as the ACVIM guidelines for pulmonary hypertension. Dogs with other organ diseases were excluded during clinical examination.

[0043] The test drug was administered orally twice a day using the tablets prepared in Example 1, with 1 / 2 tablet per kg of body weight of the test animal administered each time. Administration was carried out at fixed times, 12 hours apart in the morning and evening, regardless of whether the animal had eaten, and the administration period was 56 days.

[0044] The plan for this experiment involved administering drugs to dogs that had developed heart failure due to various causes, including valvular heart disease. ISACHC stage III heart failure is a severe state of heart failure, characterized by clear clinical symptoms of cardiac disease such as pulmonary edema and syncope. Among these dogs, those with pulmonary hypertension as a complication of cardiac disease had their pulmonary hypertension indicators recorded separately, and the improvement effect of drug administration was also examined.

[0045] The intervals between drug administration tests were set as follows: before administration (DAY 0), 7 days after administration (DAY 7), 14 days after administration (DAY 14), 28 days after administration (DAY 28), and 56 days after administration (DAY 56).

[0046] The specific observation items and examination methods are as follows:

[0047] Observation items 1) CBC: Hematological indicator (RBC index and WBC index) 2) Serum biochemistry tests: Renal function indicators (BUN, creatinine), liver function indicators (ALT, ALP), total protein (TP, albumin), electrolyte indicators (Na, K, Cl, P, etc.) 3) Evaluation of cardiac and lung fields in radiographic images: VHS, VLAS, lung field pattern 4) Echocardiography: Measurement of LA / Ao, LVIDdn, MVE, E / E', TR velocity, PR velocity, RVOT ET / AT, and RVOT AT. 5) Blood symmetric dimethyl arginine (SDMA) concentration: Evaluation of improvement in renal function values 6) Blood N-terminal pro-brain natriuretic peptide (NT-proBNP) concentration: Evaluation of improvement in cardiac function values 7) Serum canine pancreatic lipase immunoactivity (cPL) concentration: Evaluation of improvement in pancreatic function 8) Clinical laboratory tests were evaluated separately for exercise intolerance (1-4; 1 being best, 4 being worst), appetite (1-4), respiratory effort (1-4), cough (1-4), and syncope (1-4).

[0048] The observation and testing method involved performing clinical tests on the following dates: before administration (Day 0), 7 days after administration (Day 7), 14 days after administration (Day 14), 28 days after administration (Day 28), and 56 days after administration (Day 56). The veterinarian and guardian each recorded the results. For the observation items, including radiography, echocardiography, and biomarker tests, the animals were brought to designated animal hospitals where the tests were performed and recorded.

[0049] The evaluation criteria for the effectiveness of each clinical test item on the test results were as follows:

[0050] 1) Criteria for evaluating effectiveness - Improvement in clinical symptoms: Increased vitality, decreased frequency of coughing, increased appetite, reduced respiratory effort, decreased frequency of fainting spells. - Improvement of CBC and serum biochemical indicators - Improvement of cardiac size and lung field infiltration on chest radiographs. - Improvement of cardiac examination indicators on echocardiography - Improvement findings in cardiac biomarker testing 2) Method for evaluating the effects - Improvement in clinical symptoms: Evaluated based on records from guardians and veterinarians' examination records. - Improvement of CBC and serum biochemical indicators: Evaluation of liver, kidney, pancreas, and electrolyte parameters. - Alleviation of pulmonary hypertension on chest radiography: Determined by evaluation of VHS (heart size), VLAS (left atrium size), and lung field pattern (infiltration findings in the lung fields). - Improvement of cardiac examination indicators on echocardiography: Determined by evaluation of LA / Ao (left atrial-to-aortic diameter ratio), LVIDdn (left ventricular end-diastolic diameter), MVE (mitral valve filling velocity), E / E' (ratio of mitral valve filling velocity to tissue Doppler velocity), TR velocity (tricuspid regurgitation velocity), PR velocity (pulmonary artery regurgitation velocity), RVOT ET / AT (ratio of right ventricular outflow tract ejection time to acceleration time), and RVOT AT (ratio of right ventricular outflow tract acceleration time). - Improvement findings in cardiac biomarker tests: SDMA concentration, which indicates renal perfusion status, is measured to evaluate the degree of improvement in the renal circulation and suppression of renal remodeling, and NT-proBNP concentration is measured to evaluate whether there is a decrease in right ventricular pressure due to pulmonary hypertension. 3) Methods of statistical analysis -The normality of each test index was tested using the Kolmogorov-Smirnov test.

[0051] - Differences in test indicators between groups were compared using the Mann-Whitney U test, and the Wilcoxson signed-rank test was used for comparisons before and after administration.

[0052] A P<0.05 value was considered statistically significant.

[0053] <Experimental Result 1> 1. Clinical Tests The clinical test results for Example 1, including evaluations of exercise intolerance, appetite, syncope, respiratory effort, and cough, are shown in Table 2 and Figures 1 to 5 below.

[0054] [Table 2]

[0055] In Table 1 above, the numerical values ​​1 to 4 in the clinical evaluation results were evaluated based on the following conditions, respectively.

[0056] - Exercise intolerance: 1. None at all, 2. Shows signs of fatigue after long walks, 3. Seems to have difficulty walking during walks, 4. Refuses to go for walks. - Appetite: 1. Very good, 2. Good, 3. Picky eater (only wants to eat tasty food), 4. Refuses food -Respiratory effort: 1. None at all, 2. Hyperventilation occurs after playing, 3. Hyperventilation occurs within 30 minutes even during rest, 4. Hyperventilation occurs during sleep -Syncope: 1. None at all, 2. Syncope once or less per day after exercise or excitement, 3. Syncope once or less per day regardless of exercise or excitement, 4. Syncope once or more per day regardless of exercise or excitement - Cough: 1. None at all, 2. Coughs when excited or after drinking water, 3. Coughs 10 times or less per day, regardless of excitement or drinking water, 4. Coughs so severe that they prevent sleep. Figures 1 to 5 are graphs showing the clinical test results for Example 1. Figures 1 to 5 show the evaluation results for exercise intolerance, appetite, syncope, respiratory effort, and cough for the test group and the placebo group, respectively, as clinical test results.

[0057] Referring to Figures 1 through 5, in the exercise intolerance assessment, the affected dogs showed significant improvement in exercise intolerance from the first week of administration (DAY 7) onward, and no longer exhibited any particular fatigue in their daily lives. From day 14 onward, the vast majority of the affected dogs did not show any symptoms of exercise intolerance in their daily lives.

[0058] In appetite assessments, the affected dogs showed a significant improvement in appetite from the first week of administration (DAY 7) onward, and did not refuse food in their daily lives. From day 14 onward, the majority of the affected dogs showed a strong appetite in their daily lives.

[0059] In respiratory effort assessments, the affected dogs showed significant improvement in symptoms of labored breathing from the first week after administration (DAY 7), and their daily lives were not affected. From the 14th day after administration (DAY 14) onward, the vast majority of affected dogs did not show any particular symptoms of labored breathing in their daily lives.

[0060] In the cough evaluation, the affected dogs showed significant improvement in cough symptoms from the first week of administration (DAY 7) onward, and their daily lives were not affected. From the 14th day of administration (DAY 14) onward, the vast majority of affected dogs did not exhibit cough symptoms unless stimulated in their daily lives.

[0061] In the syncope evaluation, the affected dogs showed almost no symptoms of syncope after the first week of administration (DAY 7), and very few dogs were observed to reappear with syncope symptoms during the subsequent administration period.

[0062] 2. Blood indicator tests In the blood index tests of Example 1, anemia and polycythemia (occurring as a reaction to cyanosis) were evaluated during the administration period by assessing red blood cell count (RBC) and hematocrit (PCV). In addition, the white blood cell count (WBC) was evaluated to check for the presence or absence of inflammation. The evaluation results of the blood index tests are shown in Table 3 and Figures 6 to 8 below.

[0063] [Table 3]

[0064] Figures 6 to 8 are graphs showing the evaluation results of blood index tests in Example 1. Figures 6 to 8 show the evaluation results of red blood cell count (RBC), hematocrit (PCV), and white blood cell count (WBC), respectively, during the administration period.

[0065] Referring to Table 3 and Figures 6 through 8, no significant changes were observed in the percentage of blood cells when comparing the period before and after administration.

[0066] 3. Testing of serum biochemical indicators In Example 1, during the administration period, biochemical indicators for renal function (BUN, creatinine), biochemical indicators for liver function (ALT, ALP), and indicators for serum protein (TP, albumin) were evaluated as part of the serum biochemical indicator testing. The evaluation results of the serum biochemical indicator tests are shown in Table 4 and Figures 9 to 14 below.

[0067] [Table 4]

[0068] Figures 9 to 14 are graphs showing the evaluation results of serum biochemical indicator tests in Example 1. Figures 9 to 14 show the evaluation results for BUN, creatinine, ALP, ALT, TP, and Alb, respectively.

[0069] Referring to Table 4 and Figures 9 to 14, while elevated renal function indicators are commonly reported with excessive administration of cardiac medications (e.g., diuretics), no significant changes in renal function indicators were observed when comparing the BUN evaluation results shown in Figure 9 and the Creatinine evaluation results shown in Figure 10 with the administration of Example 1 containing the composition of the present invention.

[0070] Elevated liver enzyme levels are widely reported in dogs with right heart failure (pulmonary hypertension). These elevated liver enzyme levels are known to subside as heart failure symptoms improve. As shown in Figure 11 for ALP and Figure 12 for ALT, this experiment also confirmed a gradual improvement (decrease) in liver enzyme indicators before and after administration.

[0071] Total protein and albumin tend to increase with excessive administration of diuretics included in cardiac medications (in cases of severe dehydration), but tend to decrease in cases of severe heart failure symptoms such as impaired liver function and fluid retention. As shown in the TP evaluation results in Figure 13 and the Alb evaluation results in Figure 14, these serum protein indicators tended to gradually increase after administration in this experiment. However, no significant increase suggestive of dehydration was observed; only a gradual upward trend associated with the suppression of fluid retention and liver treatment was confirmed.

[0072] 4. Testing of serum electrolyte indicators As part of the serum electrolyte index testing in Example 1, changes in the concentrations of serum electrolytes Na, K, Cl, and P were evaluated during the administration period. The results are shown in Table 5 and Figures 15 to 18 below.

[0073] [Table 5]

[0074] Figures 15 to 18 are graphs showing the evaluation results of serum electrolyte indicators in Example 1. Figures 15 to 18 show the changes in blood concentrations of Na, Cl, K, and P as serum electrolytes, respectively.

[0075] Due to the nature of cardiac drugs, electrolyte imbalances can occur as a common side effect in association with cardiac drug administration. Among the components of this study drug, torsemide exerts its pharmacological effect by acting on the loop of Henle in the kidneys to remove sodium, leading to hyponatremia, hypokalemia, and hypochloremia. On the other hand, drugs such as spironolactone and enalapril act on the collecting ducts to retain potassium, potentially causing hyperkalemia and hypernatremia. Therefore, such electrolyte imbalances can be prevented by appropriately combining these drugs.

[0076] Referring to Table 5 and Figures 15 to 18, in this experiment as well, no statistically significant changes in sodium, potassium, and chlorine levels were observed in relation to administration when comparing pre- and post-administration results. Phosphorus is an important electrolyte whose levels increase due to impaired excretion and nephrogenic hyperparathyroidism when 85% of renal function is lost. Since overdose of cardiac drugs impairs renal function, monitoring phosphorus levels is very important in dogs receiving cardiac drugs. In this experiment, no changes in phosphorus levels were observed before and after administration, and no dogs exhibiting clinically significant hyperphosphatemia were observed throughout the administration period.

[0077] 5. Testing of biomarker indicators In Example 1, changes in the renal marker (SDMA), pancreatitis marker (cPL), and heart failure marker (NT-proBNP) were examined as biomarker indicators. The main biomarker findings in dogs with heart failure due to pulmonary hypertension are elevated renal marker (SDMA), elevated pancreatitis marker (cPL), and elevated heart failure marker (NT-proBNP).

[0078] SDMA stands for Symmetric dimethylarginine and is the most widely used prognostic marker for renal failure in current veterinary clinical practice. Dogs with renal failure stage 1 are classified as having a SDMA level of 18 or less, stage 2 dogs as 18-35, stage 3 dogs as 36-54, and end-stage dogs as having a SDMA level of 54 or more.

[0079] cPL stands for canine pancreatic lipase and is the most widely used prognostic marker for pancreatitis in dogs in current veterinary practice. A value of 200 or less is considered normal, 200-400 indicates borderline pancreatitis, and 400 or more indicates pancreatitis.

[0080] NT-proBNP stands for N-terminal pro-B-type natriuretic peptide and is currently the most widely used prognostic marker for heart failure in dogs in veterinary clinical practice. Levels below 900 are considered normal, levels between 900 and 1800 indicate borderline heart failure, and levels above 1800 indicate heart failure.

[0081] In this evaluation, changes in biomarker indicators were examined, and the results are shown in Table 6 and Figures 19 to 21 below.

[0082] [Table 6]

[0083] Figures 19 to 21 are graphs showing the biomarker test results for Example 1. Figures 19 to 21 show the changes in SDMA, NT-proBNP, and cPL as biomarkers, respectively.

[0084] Referring to Table 6 and Figures 19 to 21, NT-proBNP, a marker for heart failure, gradually decreased after administration and showed a tendency to decline in stages as the administration period progressed. In particular, the average NT-proBNP concentration 56 days after administration decreased to one-third of the level before administration. No significant changes were observed in the values ​​of SDMA, a renal marker, and cPL, a pancreatic marker, before and after administration. It is presumed that there were no significant changes in the values ​​of renal and pancreatic markers before and after administration because dogs with systemic diseases were excluded during the selection process of the dogs that participated in the study.

[0085] 6. Evaluation of chest radiography In Example 1, the chest radiography evaluation included VHS (a measure of cardiac size), VLAS (a measure of left atrium size), and the infiltration pattern of the lung fields.

[0086] VHS, an abbreviation for Vertebral heart scale, is a method for quantitatively evaluating heart size using radiographic images.

[0087] VLAS, an abbreviation for Vertebral left atrial score, is an index for evaluating the size of the left atrium, and is a method for evaluating the size of the heart on chest radiographs.

[0088] Lung infiltration patterns are a method for evaluating how clearly the lung lobes are visualized on radiographic images, and are primarily used to check for the presence or absence of pulmonary edema or inflammation.

[0089] The evaluation results of the chest radiation are shown in Table 7 and Figures 22 to 24 below.

[0090] [Table 7]

[0091] Figures 22 to 24 are graphs showing the evaluation results of chest radiation in Example 1. Figures 22 to 24 show the evaluation results of VHS, VLAS, and lung field pattern, respectively, as chest radiation evaluation results.

[0092] Referring to Table 7 and Figures 22 to 24, the VHS, an index for evaluating cardiac size, began to decrease from day 7 of administration and continued to decrease until day 56. Similarly, the VLAS, an index for left atrial size, also began to decrease from day 7 of administration and continued to decrease until day 56. Regarding lung field infiltration, the majority of affected dogs showed improvement to a normal lung lobe pattern at the examination on day 7 of administration, and no further dogs were observed to show lung field infiltration at the examination on day 14 of administration.

[0093] 7. Echocardiographic evaluation In Example 1, echocardiographic evaluation was performed to assess left heart function by examining LA / Ao, LVIDDn, MVE, and MV E / E' using echocardiography. In addition, right heart function was assessed by examining TR velocity, PR velocity, RVOT ET / AT, and RVOT AT using echocardiography.

[0094] LA / Ao is an abbreviation representing the left atrial-to-aortic diameter ratio and is an indicator used to assess the degree of left atrial dilation. Dogs with ISACHC I heart failure typically show a value of 1.4-1.8, those with ISACHC II heart failure 1.8-2.0, and those with ISACHC III heart failure 2.0 or greater. The normal range is 1.2-1.4, and a value of 1.2 or less indicates a sub-normal size, mainly caused by dehydration.

[0095] LVIDdN is an index that evaluates the end-diastolic diameter of the left ventricle regardless of the dog's body weight, and generally, a value of 20 or higher is considered to indicate an increased end-diastolic diameter.

[0096] MVE (Mitt Valve Elevation) indicates the velocity of mitral valve blood flow during the filling phase and is generally higher with increasing severity of heart failure. Dogs with ISACHC I heart failure typically show values ​​of 0.8–1.0 m / s, ISACHC II dogs 1.0–1.2 m / s, and ISACHC III dogs 1.2 m / s or higher. The normal value is 0.8 m / s or less.

[0097] MV E / E' is the ratio of blood flow velocity in the mitral valve to tissue Doppler velocity of the mitral annulus, and is an indicator used to predict left atrial pressure. A value of 9 or higher indicates a left atrial pressure of 20 mmHg, and a value of 12 or higher indicates a left atrial pressure of 30 mmHg or higher. Normal left atrial pressure should not exceed 20 mmHg, and an increase in this value indicates an increase in left atrial pressure.

[0098] TR velocity is a method for measuring the regurgitation velocity of tricuspid valve blood flow (in dogs with pulmonary hypertension, increased resistance in the right heart system and pulmonary blood vessels causes tricuspid valve blood flow to flow in the opposite direction to its normal direction during cardiac systole). The faster the regurgitation velocity, the more severe the pulmonary hypertension. Pulmonary hypertension is diagnosed if the velocity is 2.8 m / s or higher.

[0099] PR velocity is a method for measuring the regurgitation velocity of pulmonary artery blood flow (in dogs with pulmonary hypertension, increased resistance in the right heart system and pulmonary blood vessels causes pulmonary artery blood to flow in the opposite direction to its normal direction during cardiac systole). The faster the regurgitation velocity, the more severe the pulmonary hypertension. A velocity of 2.2 m / s or higher indicates pulmonary hypertension.

[0100] RVOT ET / AT is a measure of pulmonary artery blood flow, which is largely composed of acceleration time (AT), ejection time (ET), and deceleration time (DT). As pulmonary vascular resistance increases, acceleration time shortens and ejection time lengthens. AT / ET is an echocardiographic index that assesses the severity of pulmonary hypertension by comparing these two times. The normal value is 0.42 or higher, and in dogs with severe pulmonary hypertension, the value drops significantly to 0.30 or lower. In other words, a lower value indicates more severe pulmonary hypertension.

[0101] RVOT AT is a measure of pulmonary artery blood flow, consisting of acceleration time (AT), ejection time (ET), and deceleration time (DT). Increased pulmonary vascular resistance shortens the acceleration time. This index is an echocardiographic indicator that can assess the severity of pulmonary hypertension. A lower value indicates a shorter acceleration time and a more severe pulmonary hypertension.

[0102] The evaluation results of this echocardiogram are shown in Table 8 and Figures 25 to 32 below.

[0103] [Table 8]

[0104] Figures 25 to 32 are graphs showing the echocardiographic evaluation results for Example 1. Figures 25 to 32 show the evaluation results for LA / Ao, LVIDDn, MVE, MV E / E', TR velocity, PR velocity, RVOT ET / AT, and RVOT AT, respectively, as echocardiographic evaluation results. Referring to Table 8 and Figures 25 to 32, it was confirmed that in this experiment as well, these left heart system indicators improved significantly in evaluations from day 7 (DAY 7) onwards, and this improvement effect persisted until day 56 (DAY 56). Since the circulatory system of dogs and cats is a closed circulatory system, diseases that occur on the left or right side of the heart will eventually transfer pressure to the opposite side of the heart. Therefore, in many dogs with left heart failure, right heart failure develops over time, and a typical cause of this phenomenon is pulmonary hypertension. In this experiment as well, pulmonary hypertension was observed in the majority of the affected dog group, and it was confirmed that the laboratory indicators related to pulmonary hypertension improved significantly when the administration of the test drug was started to these dogs. Furthermore, it was confirmed that these improvement effects persisted until day 56 of administration.

[0105] 8. Evaluation of heart failure indicators As an indicator of heart failure in Example 1, the ISACHC stage was evaluated. The ISACHC stage evaluation method is a classification index for dogs with heart failure proposed by the International Society of Small Animal Cardiology, and is divided into five stages: 1. ISACHC IA: Asymptomatic dogs without cardiac hypertrophy, 2. ISACHC IB: Asymptomatic dogs with cardiac hypertrophy, 3. ISACHC II: Dogs showing mild to moderate heart failure, 4. ISACHC IIIA: Dogs showing severe heart failure with pulmonary edema, 5. ISACHC IIIB: Dogs showing end-stage heart failure with recurrent pulmonary edema.

[0106] The evaluation results for the ISACHC stage are shown in Table 9 and Figure 33 below.

[0107] [Table 9]

[0108] Figure 33 is a graph showing the evaluation results of the ISACHC stage in Example 1.

[0109] Referring to Table 9 and Figure 33 above, in this study, the majority of affected dogs were in ISACHC IIIA and IIIB condition, but significant improvement was observed from day 7 of administration, and this improvement persisted until day 56 of administration.

[0110] 9. Conclusions based on experimental results Based on the evaluation results of Example 1, it was revealed that the composition for the prevention and treatment of heart disease according to one example contains the cardiotonic component pimobendan, the vasodilator enalapril, and the diuretic components torsemide and spironolactone, and is effective in preventing and treating heart disease in animals such as dogs and cats.

[0111] A composition for the prevention and treatment of heart disease in animals according to one embodiment of the present invention showed improvement in cardiac function in clinical indicators for heart failure, demonstrating therapeutic effects against heart failure in clinical trials. In particular, it was confirmed that the composition of the present invention also simultaneously improved pulmonary hypertension, a complication of heart failure. No adverse effects related to the kidneys, liver, and electrolytes associated with drug administration were observed during the administration period. The stage of heart failure was also confirmed to improve gradually and sustainably after administration.

[0112] <Example 2> Manufacture of a composition for the prevention and treatment of heart disease To more specifically confirm the efficacy of the composition of Example 1 against pulmonary hypertension, various comparative examples were prepared and evaluated in a manner similar to that described above.

[0113] The compositions shown in Table 10 below were mixed to produce Production Example 1, a composition for liver treatment. The units in Table 10 below are mg. Tablets with a composition of 100 mg each were produced by compressing the composition having the following composition.

[0114] Furthermore, comparative examples 1 to 3, which are compositions for liver treatment, were prepared by mixing the compositions shown in Table 11 below. The units in Table 11 below are mg. Tablets with a composition having the following composition were produced by compressing them to produce tablets with a weight of 100 mg per tablet.

[0115] [Table 10]

[0116] [Table 11]

[0117] <Experiment Example 2> Confirmation of the preventive and therapeutic effects of heart disease To confirm the preventive and therapeutic effects of the composition of Manufacturing Example 1 and the compositions of Comparative Examples 1 to 3 on heart disease, the following experiments were conducted.

[0118] <Experimental Method> Experiments were conducted to evaluate the efficacy and stability of the compositions of Production Example 1 and Comparative Examples 1 to 3, selecting test groups to receive the compositions and placebo groups to receive the compositions. The test and placebo groups were selected from 40 dogs diagnosed with heart failure of ISACHC stage III or higher. The 40 dogs were small breeds and had the same sex composition (20 females and 20 males).

[0119] The selected test animals were directly examined by a veterinary cardiologist through clinical examination, chest radiography, and echocardiography, and their heart failure status was classified based on the ACVIM guidelines for pulmonary hypertension. Dogs with other organ diseases were excluded during the clinical examination.

[0120] The test drug was administered orally twice a day using tablets of Production Example 1, which were manufactured in Example 2, with each dose being 1 / 2 tablet per kg of body weight of the test animal. Administration was carried out at fixed times, 12 hours apart in the morning and evening, regardless of whether the animals had eaten, and the administration period was 30 days.

[0121] The tablets of Comparative Examples 1 to 3, manufactured in Example 2, were similarly administered orally twice a day. However, for Comparative Example 1, one tablet was administered per 5 kg of body weight of the test animals, and for Comparative Examples 2 and 3, half a tablet was administered per 1 kg of body weight of the test animals. Administration was carried out at fixed times, with 12-hour intervals in the morning and evening, regardless of whether the animals had eaten, and the administration period was 30 days.

[0122] The plan for this experiment involved administering drugs to dogs that had developed heart failure due to various causes, including valvular heart disease. ISACHC stage III heart failure is a severe state of heart failure, characterized by clear clinical symptoms of cardiac disease such as pulmonary edema and syncope. Among these dogs, those with pulmonary hypertension as a complication of cardiac disease had their pulmonary hypertension indicators recorded separately, and the improvement effect of drug administration was also examined. If pulmonary edema or clinical symptoms worsened during the administration period, the drug was discontinued and a rescue drug (diuretic or cardiac stimulant) was administered instead. Indices were evaluated only for individuals whose administration was not changed until the end of the experiment, and the average values ​​were compared.

[0123] The intervals for testing following drug administration were set as before administration (DAY 0) and 30 days after administration (DAY 30).

[0124] The specific observation items and examination methods are as follows:

[0125] Observation items 1) Evaluation of cardiac and lung fields in radiographic images: VHS, VLAS, lung field pattern 2) Echocardiography: Measurement of LA / Ao, LVIDdn, MVE, E / E', TR velocity, PR velocity, RVOT ET / AT, and RVOT AT. 3) Assessment of pulmonary hypertension: None (0), mild (1), moderate (2), severe (3) 4) Heart failure stages: ISACHC1A(1), ISACHC1B(2), ISACHC2(3), ISACHC3A(4), ISACHC3B(5) 5) Clinical laboratory tests were conducted separately for exercise intolerance (1-4; 1 being best, 4 being worst), appetite (1-4), respiratory effort (1-4), cough (1-4), and syncope (1-4).

[0126] The observation and testing method involved performing clinical tests before administration (Day 0) and 30 days after administration (Day 30), with the veterinarian and guardian each recording the results. The observation items, including radiography, echocardiography, and biomarker tests, were performed and recorded at designated animal hospitals.

[0127] The evaluation criteria for the effectiveness of each clinical test item on the test results were as follows:

[0128] 1) Criteria for evaluating effectiveness - Improvement in clinical symptoms: Increased vitality, decreased frequency of coughing, increased appetite, reduced respiratory effort, decreased frequency of fainting spells. - Improvement of cardiac size and lung field infiltration on chest radiographs. - Improvement of cardiac examination indicators on echocardiography 2) Method for evaluating the effects - Improvement in clinical symptoms: Evaluated based on records from guardians and veterinarians' examination records. - Improvement of CBC and serum biochemical indicators: Evaluation of liver, kidney, pancreas, and electrolyte parameters. - Alleviation of pulmonary hypertension on chest radiography: Determined by evaluation of VHS (heart size), VLAS (left atrium size), and lung field pattern (infiltration findings in the lung fields). - Improvements in cardiac examination indicators on echocardiography: LA / Ao (left atrial-to-aortic diameter ratio), LVIDdn (left ventricular end-diastolic diameter), MVE (mitral valve filling velocity), E / E' (ratio of mitral valve filling velocity to tissue Doppler velocity), - Decreased grade of pulmonary hypertension: Determined by evaluation of TR velocity (tricuspid valve regurgitation velocity), PR velocity (pulmonary artery regurgitation velocity), RVOT ET / AT (ratio of right ventricular outflow tract ejection time to acceleration time), and RVOT AT (ratio of right ventricular outflow tract acceleration time). - Improvement in heart failure stage: Reduction in heart failure stage according to the classification system of the International Society of Small Animal Cardiology (ISACHC). 3) Methods of statistical analysis -The normality of each test index was tested using the Kolmogorov-Smirnov test.

[0129] - Differences in test indicators between groups were compared using the Mann-Whitney U test, and the Wilcoxson signed-rank test was used for comparisons before and after administration.

[0130] A P<0.05 value was considered statistically significant.

[0131] <Experimental Results 2> 1. Clinical Tests The clinical test results for Example 2, including evaluations of exercise intolerance, appetite, syncope, respiratory effort, and cough, are shown in Table 12 and Figures 34 to 38 below.

[0132] [Table 12]

[0133] In Table 12 above, the numerical values ​​1 to 4 in the clinical evaluation results were evaluated based on the following conditions, respectively.

[0134] - Exercise intolerance: 1. None at all, 2. Shows signs of fatigue after long walks, 3. Seems to have difficulty walking during walks, 4. Refuses to go for walks. - Appetite: 1. Very good, 2. Good, 3. Picky eater (only wants to eat tasty food), 4. Refuses food -Respiratory effort: 1. None at all, 2. Hyperventilation occurs after playing, 3. Hyperventilation occurs within 30 minutes even during rest, 4. Hyperventilation occurs during sleep - Cough: 1. None at all, 2. Coughs when excited or after drinking water, 3. Coughs 10 times or less per day, regardless of excitement or drinking water, 4. Coughs so severe that they prevent sleep. -Syncope: 1. None at all, 2. Syncope once or less per day after exercise or excitement, 3. Syncope once or less per day regardless of exercise or excitement, 4. Syncope once or more per day regardless of exercise or excitement Figures 34 to 38 are graphs showing the clinical test results for Example 2. Figures 34 to 38 show the evaluation results for exercise intolerance, appetite, respiratory effort, cough, and syncope for the test group and the placebo group, respectively, as clinical test results.

[0135] Referring to Figures 34 to 38, in the exercise intolerance evaluation, the groups administered with Production Example 1 or Comparative Example 3 showed significant improvement in exercise intolerance symptoms at the end of the study (DAY 30), while the groups administered with Comparative Example 1 and Comparative Example 2 did not show significant improvement. This improvement in symptoms was most pronounced in the group administered with Production Example 1.

[0136] In appetite assessment, the groups administered with Production Example 1 or Comparative Example 3 showed a significant improvement in appetite at the end of the study (DAY 30), while the groups administered with Comparative Example 1 and Comparative Example 2 did not show significant improvement. This improvement in symptoms was most pronounced in the group administered with Production Example 1.

[0137] In the respiratory effort assessment, the groups administered with Production Example 1 or Comparative Example 3 showed significant improvement in symptoms of labored breathing at the end of the study (DAY 30), while the group administered with Comparative Example 2 showed only partial improvement, and the group administered with Comparative Example 1 showed no significant improvement in symptoms. This improvement in symptoms was most pronounced in the groups administered with Production Example 1 and Comparative Example 3, and the degree of improvement was almost the same.

[0138] In the cough evaluation, the groups administered with Production Example 1 or Comparative Example 3 showed significant improvement in cough symptoms at the end of the study (DAY 30), while the group administered with Comparative Example 2 showed only partial improvement, and the group administered with Comparative Example 1 did not show significant improvement in symptoms. This symptom improvement effect was most pronounced in the group administered with Production Example 1.

[0139] In the syncope evaluation, improvement in syncope symptoms was observed in most drug administration groups, but this effect was most pronounced in the group administered with Production Example 1.

[0140] As mentioned above, 1. According to the clinical test results, the composition of Production Example 1 shows improved effects in evaluations of exercise intolerance, appetite, respiratory effort, cough, and syncope compared to the compositions of Comparative Examples 1 to 3.

[0141] 2. Evaluation of chest radiography For the chest radiography evaluation in Example 2, we assessed the VHS (vein size scale), the VLAS (left atrium size scale), and the lung field pattern.

[0142] VHS, an abbreviation for Vertebral heart scale, is a method for quantitatively evaluating heart size using radiographic images.

[0143] VLAS, an abbreviation for Vertebral left atrial score, is an index for evaluating the size of the left atrium, and is a method for evaluating the size of the heart on chest radiographs.

[0144] Lung infiltration patterns are a method for evaluating how clearly the lung lobes are visualized on radiographic images, and are primarily used to check for the presence or absence of pulmonary edema or inflammation.

[0145] The evaluation results of the chest radiography are shown in Table 13 and Figures 39 to 41 below.

[0146] [Table 13]

[0147] Figures 39 to 41 are graphs showing the evaluation results of chest radiation in Example 2. Figures 39 to 41 show the evaluation results of VHS, VLAS, and Lung fielded pattern (PE), respectively, as chest radiation evaluation results.

[0148] As heart failure progresses, the heart enlarges and the left atrium expands. Excessive left atrium expansion causes dilation of pulmonary blood vessels, leading to fluid infiltration into the lung fields and pulmonary edema, which results in symptoms such as cough and labored breathing.

[0149] The VHS evaluation results in Figure 39 quantitatively evaluate the size of the heart on radiographic images. Referring to Table 13 and Figure 39, the VHS, an evaluation index for heart size, showed no significant change in heart size at the end of the study (DAY 30) in the administration groups of Comparative Example 1 and Comparative Example 2, whereas a significant decrease in heart size was observed at the end of the study (DAY 30) compared to before administration in the administration groups of Manufacturing Example 1 and Comparative Example 3.

[0150] The VLAS evaluation results in Figure 40 assess the size of the left atrial region on a chest radiograph. Referring to Table 13 and Figure 40, the VLAS, an evaluation index for left atrial size, showed no significant change in left atrial size at the end of the study (DAY 30) in the treatment groups of Comparative Example 1 and Comparative Example 2, whereas in the treatment groups of Manufacturing Example 1 and Comparative Example 3, a significant decrease in left atrial size was observed at the end of the study (DAY 30) compared to before administration.

[0151] The evaluation results for lung field infiltration (PE) in Figure 41 are a method for evaluating how clearly the lung lobes are visualized on radiographic images, and are mainly used to check for the presence or absence of pulmonary edema or inflammation. Referring to Table 13 and Figure 41, the lung field infiltration findings did not significantly improve in the treatment groups of Comparative Example 1 and Comparative Example 2 at the end of the study (DAY 30). In particular, it was confirmed that the lung field infiltration worsened further in the treatment group of Comparative Example 1 compared to before administration. The lung field infiltration findings were significantly improved in the treatment groups of Manufacturing Example 1 and Comparative Example 3 at the end of the study (DAY 30) compared to before administration.

[0152] 3. Echocardiographic evaluation In Example 2, echocardiographic evaluation was performed to assess left heart function by examining LA / Ao, LVIDDn, MVE, and MV E / E' using echocardiography. In addition, right heart function was assessed by examining TR velocity, PR velocity, RVOT ET / AT, and RVOT AT using echocardiography.

[0153] LA / Ao is an abbreviation representing the left atrial-to-aortic diameter ratio and is an indicator used to assess the degree of left atrial dilation. Dogs with ISACHC I heart failure typically show a value of 1.4-1.8, those with ISACHC II heart failure 1.8-2.0, and those with ISACHC III heart failure 2.0 or greater. The normal range is 1.2-1.4, and a value of 1.2 or less indicates a sub-normal size, mainly caused by dehydration.

[0154] LVIDdN is an index that evaluates the end-diastolic diameter of the left ventricle regardless of the dog's body weight, and generally, a value of 20 or higher is considered to indicate an increased end-diastolic diameter.

[0155] MVE (Mitt Valve Elevation) indicates the velocity of mitral valve blood flow during the filling phase and is generally higher with increasing severity of heart failure. Dogs with ISACHC I heart failure typically show values ​​of 0.8–1.0 m / s, ISACHC II dogs 1.0–1.2 m / s, and ISACHC III dogs 1.2 m / s or higher. The normal value is 0.8 m / s or less.

[0156] MV E / E' is the ratio of blood flow velocity in the mitral valve to tissue Doppler velocity of the mitral annulus, and is an indicator used to predict left atrial pressure. A value of 9 or higher indicates a left atrial pressure of 20 mmHg, and a value of 12 or higher indicates a left atrial pressure of 30 mmHg or higher. Normal left atrial pressure should not exceed 20 mmHg, and an increase in this value indicates an increase in left atrial pressure.

[0157] The results of the echocardiogram evaluation are shown in Table 14 and Figures 42 to 45 below.

[0158] [Table 14]

[0159] Figures 42 to 45 are graphs showing the echocardiographic evaluation results for Example 2. Figures 42 to 45 show the evaluation results for LA / Ao, LVIDDn, MVE, and MV E / E', respectively, as echocardiographic evaluation results. As heart failure progresses, the heart enlarges and the left atrium dilates. Therefore, the main diagnostic indicators used with echocardiography are LA / Ao, which evaluates the degree of left atrial dilation; LVIDDn, which evaluates the degree of left ventricular dilation; and MVE and MV E / E', which evaluate the increase in left atrial pressure.

[0160] Referring to Table 13 and Figures 42 to 45, in the LA / Ao evaluation, which assesses the degree of left atrial dilation, no significant change in left atrial size was observed at the end of the study (DAY 30) in the administration groups of Comparative Example 1 and Comparative Example 2, whereas a significant decrease in left atrial size was observed at the end of the study (DAY 30) in the administration groups of Manufacturing Example 1 and Comparative Example 3 compared to before administration.

[0161] In the evaluation of LVIDDn, which assesses the degree of left ventricular dilation, no significant changes in left ventricular size were observed in the treatment groups of Comparative Example 1 and Comparative Example 2 at the end of the study (DAY 30), whereas a significant decrease in left ventricular size was observed in the treatment groups of Manufacturing Example 1 and Comparative Example 3 at the end of the study (DAY 30) compared to before administration.

[0162] In the MVE and MV E / E' evaluations, which assess the increase in left atrial pressure, no significant changes in mitral valve blood flow velocity or left atrial pressure were observed in the treatment groups of Comparative Example 1 and Comparative Example 2 at the end of the study (DAY 30). In contrast, in the treatment groups of Manufacturing Example 1 and Comparative Example 3, a significant decrease in mitral valve blood flow velocity and a decrease in left atrial pressure were observed at the end of the study (DAY 30) compared to before administration. In particular, the decrease in left atrial pressure was observed most significantly in the treatment group of Manufacturing Example 1.

[0163] 4. Evaluation of pulmonary hypertension indicators In Example 2, to evaluate pulmonary hypertension and heart failure indicators, TR velocity, PR velocity, RVOT ET / AT, and RVOT AT were examined using echocardiography to assess right heart function.

[0164] TR velocity is a method for measuring the regurgitation velocity of tricuspid valve blood flow (in dogs with pulmonary hypertension, increased resistance in the right heart system and pulmonary blood vessels causes tricuspid valve blood flow to flow in the opposite direction to its normal direction during cardiac systole). The faster the regurgitation velocity, the more severe the pulmonary hypertension. Pulmonary hypertension is diagnosed if the velocity is 2.8 m / s or higher.

[0165] PR velocity is a method for measuring the regurgitation velocity of pulmonary artery blood flow (in dogs with pulmonary hypertension, increased resistance in the right heart system and pulmonary blood vessels causes pulmonary artery blood to flow in the opposite direction to its normal direction during cardiac systole). The faster the regurgitation velocity, the more severe the pulmonary hypertension. A velocity of 2.2 m / s or higher indicates pulmonary hypertension.

[0166] RVOT ET / AT is a measure of pulmonary artery blood flow, which is largely composed of acceleration time (AT), ejection time (ET), and deceleration time (DT). As pulmonary vascular resistance increases, acceleration time shortens and ejection time lengthens. AT / ET is an echocardiographic index that assesses the severity of pulmonary hypertension by comparing these two times. The normal value is 0.42 or higher, and in dogs with severe pulmonary hypertension, the value drops significantly to 0.30 or lower. In other words, a lower value indicates a more severe state of pulmonary hypertension.

[0167] RVOT AT is a measure of pulmonary artery blood flow, consisting of acceleration time (AT), ejection time (ET), and deceleration time (DT). Increased pulmonary vascular resistance leads to a shorter acceleration time. This index is an echocardiographic indicator that can assess the severity of pulmonary hypertension. A lower value indicates a shorter acceleration time and a more severe pulmonary hypertension.

[0168] The echocardiographic evaluation results are shown in Table 15 and Figure 46 below. The evaluation results in Table 15 and Figure 46 below comprehensively evaluate the TR velocity, PR velocity, RVOT ET / AT, and RVOT AT results to assess the severity of pulmonary hypertension.

[0169] [Table 15]

[0170] Figure 46 is a graph showing the evaluation results of the severity of pulmonary hypertension in Example 2. Figure 46 shows the changes in the severity of pulmonary hypertension evaluated based on the evaluation results of TR velocity, PR velocity, RVOT ET / AT, and RVOT AT. Referring to Table 15 and Figure 46, the administration group of Comparative Example 1 actually worsened pulmonary hypertension compared to before administration, and no significant change was observed in the administration group of Comparative Example 2 before and after administration. A significant decrease in the severity of pulmonary hypertension was observed in the administration groups of Manufacturing Example 1 and Comparative Example 3 compared to before administration. In particular, a clear decrease in the severity of pulmonary hypertension was observed in the administration group of Manufacturing Example 1 before and after administration.

[0171] 5. Evaluation of heart failure indicators As an indicator of heart failure in Example 2, the ISACHC stage was evaluated. The ISACHC stage evaluation method is a classification index for dogs with heart failure proposed by the International Society of Small Animal Cardiology, and is classified into five stages: 1. ISACHCIA: Asymptomatic group of dogs without cardiac hypertrophy. 2. A group of asymptomatic dogs with ISACHCIB cardiac hypertrophy. 3. ISACHCII: A group of dogs exhibiting mild to moderate heart failure. 4. ISACHC IIIA: A group of dogs exhibiting severe heart failure with pulmonary edema. 5. ISACHC IIIB: A group of dogs exhibiting end-stage heart failure with recurrent pulmonary edema.

[0172] The evaluation results for the ISACHC stage are shown in Table 16 and Figure 47 below.

[0173] [Table 16]

[0174] Figure 47 is a graph showing the evaluation results of the ISACHC stage in Example 2.

[0175] Referring to Table 16 and Figure 47 above, in Comparative Example 1, the treatment group actually experienced a worsening of heart failure compared to before administration, while in Comparative Example 2, no significant change was observed before and after administration. In the treatment groups of Manufacturing Example 1 and Comparative Example 3, an improvement in heart failure was observed compared to before administration, and the improvement in heart failure was most pronounced in the treatment group of Manufacturing Example 1.

[0176] 6. Exam completion date In this experiment, if pulmonary edema or clinical symptoms worsened during the administration period, administration was discontinued and a rescue drug (diuretic or cardiac stimulant) was administered instead. The number of individuals who received a rescue drug as a substitute during the administration period was compared in the administration groups of Production Example 1 and Comparative Examples 1 to 3.

[0177] In Comparative Example 1, out of 10 affected dogs, only 2 reached the end of the final test (DAY 30). Of these, 3 required rescue medication due to worsening clinical symptoms or pulmonary edema within one week of starting administration, 3 required it within 2 weeks, and 2 required it within 3 weeks.

[0178] In Comparative Example 2, out of a total of 10 affected dogs, only 4 reached the end of the final test (DAY 30). Of these, 2 dogs required rescue medication due to worsening clinical symptoms or the development of pulmonary edema within one week of the start of administration, 2 dogs discontinued the study drug after 2 weeks, and 2 dogs discontinued rescue medication after 3 weeks.

[0179] In Comparative Example 3, out of 10 affected dogs, 9 reached the end of the final trial (DAY 30). Of these, one dog required rescue medication after 3 weeks due to worsening clinical symptoms or the development of pulmonary edema.

[0180] In contrast, in the group administered with Production Example 1, all 10 affected dogs reached the end of the final trial (DAY 30).

[0181] 7. Conclusions based on experimental results The evaluation results of Example 2 confirmed that the composition for the prevention and treatment of heart disease according to one example contains the cardiotonic component pimobendan, the vasodilator enalapril, and the diuretic components torsemide and spironolactone, and is effective in preventing and treating heart disease in animals such as dogs and cats.

[0182] In this study, comparing the number of individuals maintained without rescue medication until the end of the study (DAY 30), it was found that when a composition from one example, such as Manufacturing Example 1, was administered, severe heart failure (pulmonary hypertension) could be managed more effectively when other types of drugs were added compared to the administration of the inotropic agent pimobendan alone. Furthermore, compositions like Comparative Example 2, which did not include a diuretic, were less effective in managing symptoms. Regarding diuretics, it was found that the composition from Manufacturing Example 1, which combined two types of diuretics, prevented symptom exacerbation and was more advantageous in survival than when used alone.

[0183] In Comparative Example 1, where pimobendan was administered alone, a slight improvement in clinical symptoms was observed in dogs with severe heart failure (pulmonary hypertension). However, in the majority of affected dogs, this improvement was short-lived and temporary, and ultimately, symptoms tended to worsen. Regarding clinical symptoms related to heart failure, the combination formulations with the addition of enalapril (Production Example 1, Comparative Examples 2 and 3) showed better improvement, and the improvement was most pronounced in Production Example 1 and Comparative Example 3, which were combination formulations with diuretics. Among these, Production Example 1, which combined four drugs, showed the best improvement in clinical indicators. Furthermore, in evaluations of chest radiography and echocardiography, particularly regarding heart failure and pulmonary hypertension, the composition of Production Example 1 was confirmed to show superior effects compared to the compositions of the other comparative examples.

[0184] One embodiment of the composition comprises pimobendan, enalapril, torsemide, and spironolactone, and is effective in treating heart failure and pulmonary hypertension in animals. Furthermore, the composition, which includes drugs with different mechanisms of action, has the advantage of being even easier to manage in patients.

[0185] Although embodiments of the present invention have been described above with reference to the attached drawings, any person with ordinary skill in the art to which the present invention pertains will understand that the present invention can be implemented in other specific forms without altering its technical idea or essential features. Therefore, the above embodiments are merely illustrative in all respects and should not be interpreted as limiting.

Claims

1. Pimobendan; Enalapril; Torsemid; and A composition containing spironolactone for the prevention and treatment of heart disease and pulmonary hypertension in animals.

2. The composition for the prevention and treatment of heart disease and pulmonary hypertension in animals according to claim 1, comprising 0.5 mg of pimobendan, 1 mg of enalapril, 0.2 mg of torsemide, and 2 mg of spironolactone.

3. The composition for the prevention and treatment of heart disease and pulmonary hypertension in animals according to claim 1, wherein the single dose is 0.25 mg / kg of pimobendan, 0.5 mg / kg of enalapril, 0.1 mg / kg of torsemide, and 1 mg / kg of spironolactone, based on the weight of the subject, and is administered twice daily.

4. The composition for the prevention and treatment of heart disease and pulmonary hypertension in animals according to claim 3, wherein the subject is a dog or a cat.

5. Pimobendan; Enalapril; Torsemid; and A functional formulation containing spironolactone for the prevention and treatment of heart disease and pulmonary hypertension in animals.

6. A functional preparation for the prevention and treatment of heart disease and pulmonary hypertension in animals according to claim 5, comprising 0.5 mg of pimobendan, 1 mg of enalapril, 0.2 mg of torsemide, and 2 mg of spironolactone.

7. A functional preparation for the prevention and treatment of heart disease and pulmonary hypertension in animals according to claim 5, wherein the single dose is 0.25 mg / kg of pimobendan, 0.5 mg / kg of enalapril, 0.1 mg / kg of torsemide, and 1 mg / kg of spironolactone, based on the weight of the subject, and is administered twice daily.

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