Pharmaceutical composition for preventing or treating ischemic heart disease comprising aspartic acid

Abstract

One aspect provides a pharmaceutical composition for preventing or treating ischemic heart disease, the composition comprising aspartic acid or a pharmaceutically acceptable salt thereof. The pharmaceutical composition significantly reduces the volume of infarcted areas within the myocardium, thereby exhibiting an excellent effect of preventing or treating ischemic heart disease. In addition, by further comprising ornithine or a pharmaceutically acceptable salt thereof, the composition induces a greater reduction in the volume of infarcted areas within the myocardium compared to administering aspartic acid or a pharmaceutically acceptable salt thereof alone, thereby exhibiting a superior effect of preventing or treating ischemic heart disease.

Description

Pharmaceutical composition for preventing or treating ischemic heart disease containing aspartic acid

[0001] It relates to a pharmaceutical composition for preventing or treating ischemic heart disease containing aspartic acid.

[0002]

[0003] Ischemia is a condition in which blood supply to an organ, tissue, or area is reduced due to the constriction or occlusion of blood vessels. Ischemic heart disease (IHD) occurs when blood flow to the heart is blocked, resulting in a significant insufficient oxygen supply to the myocardium compared to its needs. Even if reperfusion occurs after ischemia, cardiac cells are damaged, ultimately leading to irreversible myocardial damage, i.e., cell and tissue necrosis.

[0004] In the early stages when the degree of damage is reversible, reperfusion therapy such as percutaneous coronary intervention, coronary artery bypass grafting, or drug therapy using thrombolytics can be used. However, it is known that reperfusion injury such as recurrent myocardial infarction, decline in cardiac function, and arrhythmia occur at a high rate even after such reperfusion therapy. Ischemic heart disease caused by myocardial cell damage and decline in cardiac function during ischemia / reperfusion has a high morbidity and mortality rate and is difficult to completely cure, so intensive basic and clinical research has been conducted for the past 50 years.

[0005] To date, active research has been conducted on mechanisms, the development of therapeutics targeting novel target sites, and the development of surgical procedures. However, technologies capable of protecting cardiomyocytes from ischemia / reperfusion have not yet been clinically commercialized. Therefore, there is a pressing need for treatments for ischemic heart disease that can slow the progression of ischemic cardiomyocyte damage and mitigate reperfusion damage.

[0006] Blood vessels are the conduits that circulate blood between the heart and each organ and tissue in the body. If any of these blood vessels become narrowed or bleed, preventing proper oxygen and nutrient supply to tissues, cells can suffer serious damage.

[0007] Currently, vascular endothelial growth factor (VEGF) and other agents are used as treatments for ischemic diseases. Nitric oxide (NO) is a well-known regulator of endothelial function and plays a crucial role in endothelial vasorelaxation. NO also upregulates VEGF (Vascular Endothelial Growth Factor), a key stimulator of angiogenesis. However, clinically applicable treatments for ischemic diseases, which involve numerous interrelated mechanisms, remain lacking.

[0008] Accordingly, the inventors of the present invention intend to provide a pharmaceutical composition for preventing or treating ischemic heart disease, including aspartic acid, in order to develop a treatment agent for ischemic heart disease that can slow the progression of ischemic-induced cardiomyocyte damage and alleviate reperfusion damage.

[0009]

[0010] One aspect is to provide a pharmaceutical composition for preventing or treating ischemic heart disease, comprising aspartic acid or a pharmaceutically acceptable salt thereof.

[0011] Another aspect is to provide a health functional food for preventing or improving ischemic heart disease, which contains aspartic acid or a food-related acceptable salt thereof.

[0012] Another aspect provides a method for preventing or treating ischemic heart disease, comprising administering aspartic acid or a pharmaceutically acceptable salt thereof to a subject in need thereof.

[0013] Another aspect provides the use of aspartic acid or a pharmaceutically acceptable salt thereof for the manufacture of a medicament for the prevention or treatment of ischemic heart disease.

[0014]

[0015] One aspect provides a pharmaceutical composition for preventing or treating ischemic heart disease, comprising aspartic acid or a pharmaceutically acceptable salt thereof.

[0016] According to one aspect, the aspartic acid may be L-aspartic acid, D-aspartic acid, or a mixture thereof, specifically, it may be L-aspartic acid, and more specifically, it may be a compound represented by the following chemical formula 1:

[0017] [Chemical Formula 1]

[0018] .

[0019] In one aspect, the aspartic acid or a pharmaceutically acceptable salt thereof may be derived from natural sources or synthesized using known organic synthesis methods. Furthermore, the aspartic acid or a pharmaceutically acceptable salt thereof may be a non-protein compound, a peptide, an extract of plant-derived tissue or cells, or a product obtained by culturing microorganisms (e.g., bacteria or fungi, and particularly yeast).

[0020] The above aspartic acid or a pharmaceutically acceptable salt thereof reduces the volume of the infarcted area in the myocardium, increases perfusion, increases the density of CD31-positive capillary lumen, and promotes angiogenesis by overexpressing VEGF, thereby exhibiting an excellent effect in preventing or treating ischemic heart disease.

[0021] More specifically, the aspartic acid or a pharmaceutically acceptable salt thereof promotes the conversion of citrulline or a pharmaceutically acceptable salt thereof into arginine or a pharmaceutically acceptable salt thereof within the administered subject (see Fig. 2). The converted arginine produces nitric oxide (NO) together with endothelial nitric oxide synthase (eNOS), and the nitric oxide upregulates the expression of VEGF, thereby exhibiting an excellent angiogenic effect and an excellent preventive or therapeutic effect on ischemic heart disease.

[0022] In one aspect, the composition may further comprise ornithine or a pharmaceutically acceptable salt thereof.

[0023] According to one aspect, the ornithine may be L-ornithine, D-ornithine or a mixture thereof, specifically, L-ornithine, and more specifically, a compound represented by the following chemical formula 2:

[0024] [Chemical Formula 2]

[0025] .

[0026] In one aspect, the ornithine or a pharmaceutically acceptable salt thereof may be derived from natural sources or may be synthesized using known organic synthesis methods. Furthermore, the ornithine or a pharmaceutically acceptable salt thereof may be a non-protein compound, a peptide, an extract of plant-derived tissue or cells, or a product obtained by culturing microorganisms (e.g., bacteria or fungi, and particularly yeast).

[0027] When the above composition further comprises ornithine or a pharmaceutically acceptable salt thereof, the volume of the infarcted area in the myocardium is further reduced, angiogenesis is further promoted, and thereby a better effect in preventing or treating ischemic heart disease is exhibited.

[0028] More specifically, the ornithine or its pharmaceutically acceptable salt is converted into citrulline or its pharmaceutically acceptable salt within the mitochondria. As a result, the aspartic acid or its pharmaceutically acceptable salt contained in the composition produces more arginine, ultimately exhibiting a superior angiogenic effect and an excellent preventive or therapeutic effect against ischemic heart disease.

[0029] The term "salt" means a salt prepared using a specific compound and a relatively non-toxic acid or base, depending on the nature of the compound.

[0030] Additionally, the “salt” in the present specification may be a “pharmaceutically acceptable salt.”

[0031] The term "pharmaceutically acceptable" means that a compound exhibits the property of being non-toxic to cells or humans when exposed to it under certain conditions.

[0032] The term "pharmaceutically acceptable salt" refers to a salt prepared using a compound according to one aspect and a relatively non-toxic acid or base. When the compound contains a relatively acidic functional group, a base addition salt can be obtained by contacting the neutral form of such compound with a sufficient amount of a base, either in a pure solution or in a suitable inert solvent. Pharmaceutically acceptable base addition salts include salts of sodium, potassium, calcium, ammonium, organic amines, or magnesium, or similar salts. When the compound contains a relatively basic functional group, an acid addition salt can be obtained by contacting the neutral form of such compound with a sufficient amount of an acid, either in a pure solution or in a suitable inert solvent. Pharmaceutically acceptable acid addition salts include salts of inorganic acids such as hydrochloric acid, hydrobromic acid, nitric acid, carbonic acid, hydrogen carbonate ion, phosphoric acid, monohydrogen phosphate ion, dihydrogen phosphate ion, sulfuric acid, hydrogen sulfate ion, hydroiodic acid or phosphorous acid, and salts of organic acids such as acetic acid, propionic acid, isobutyric acid, maleic acid, malonic acid, benzoic acid, succinic acid, suberic acid, fumaric acid, lactic acid, mandelic acid, phthalic acid, benzenesulfonic acid, p-tolylsulfonic acid, citric acid, tartaric acid and methanesulfonic acid, and further include salts of amino acids (e.g., arginine) and salts of organic acids such as glucuronic acid.

[0033] The above pharmaceutically acceptable salts can be synthesized by conventional chemical methods from parent compounds containing acidic or basic moieties. Typically, these salts are prepared by reacting the free acid or base form of these compounds with a stoichiometrically appropriate amount of base or acid in water, an organic solvent, or a mixture of the two. Non-aqueous media such as ether, ethyl acetate, ethanol, isopropanol, or acetonitrile are generally preferred.

[0034] In one aspect, the composition may reduce the volume of an infarcted area within the myocardium.

[0035] In one example, in a rat model of myocardial infarction induced by LAD I / R surgery, it was confirmed that the experimental group injected intraperitoneally with LOLA (333 mg / kg) showed a significantly superior reduction in the volume of the infarcted area in the myocardium compared to the control group injected intraperitoneally with only saline (1.7 mL / kg) (21.8±4.04% vs. 7.1±3.44%). Accordingly, it was confirmed that when a pharmaceutical composition containing aspartic acid is administered, a significantly superior effect in preventing or treating ischemic heart disease can be obtained (see Example 1).

[0036] In one aspect, the composition may reduce cardiac cell death. Specifically, the reduction in cardiac cell death may be at least one selected from the group consisting of a reduction in dead cardiac cells and an inhibition of cardiac cell death, and more specifically, both a reduction in dead cardiac cells and an inhibition of cardiac cell death.

[0037] In addition, in one embodiment, in a rat model in which myocardial infarction was induced by LAD I / R surgery, a significantly superior reduction in apoptotic cells was observed in the experimental group that received intraperitoneal injection of LOLA (333 mg / kg) compared to the control group that received intraperitoneal injection of only saline (1.7 mL / kg). Accordingly, it was confirmed that tissue death caused by ischemic heart disease can be reduced when a pharmaceutical composition containing aspartic acid is administered (see Example 2).

[0038] Additionally, in one aspect, the composition may increase at least one selected from the group consisting of perfusion, CD31-positive capillary lumen density, and nitric oxide, specifically, may increase at least two selected from the group consisting of perfusion, CD31-positive capillary lumen density, and nitric oxide, and more specifically, may increase all of perfusion, CD31-positive capillary lumen density, and nitric oxide.

[0039] In one example, in a rat model of induced ischemic disease, it was confirmed that the experimental group injected intraperitoneally with LOLA (333 mg / kg) showed significantly better perfusion recovery rate (62.97 ± 11.02 vs 72.76 ± 18.66), increased CD31-positive lumen density (157.25 ± 34.49 vs 352.25 ± 44.3), and increased VEGF expression (0.76 ± 0.12 vs vs 1.00 ± 0.19) than the control group injected intraperitoneally with only saline (1.7 mL / kg). Through the above results, it was confirmed that a pharmaceutical composition containing aspartic acid increases angiogenesis (see Examples 3 to 5).

[0040] In addition, in one aspect, the composition may be one that promotes the expression of one or more proteins selected from the group consisting of VEGF (Vascular Endothelial Growth Factor), VEGFR (Vascular Endothelial Growth Factor Receptor), FGF (Fibroblast Growth Factor), FGFR (Fibroblast Growth Factor Receptor), CD31 and endothelial nitric oxide synthase (eNOS), and specifically, one or more proteins selected from the group consisting of VEGFR (Vascular Endothelial Growth Factor Receptor), FGF (Fibroblast Growth Factor), FGFR (Fibroblast Growth Factor Receptor) and endothelial nitric oxide synthase (eNOS), VEGF (Vascular Endothelial Growth Factor) and CD31, and more specifically, VEGF (Vascular Endothelial Growth Factor), VEGFR (Vascular It may enhance the expression of all of the endothelial growth factor receptors (EGFR), fibroblast growth factor receptor (FGF), fibroblast growth factor receptor (FGFR), CD31, and endothelial nitric oxide synthase (eNOS).

[0041] In addition, in one aspect, the composition may promote the expression of at least one selected from the group consisting of a gene encoding VEGF (Vascular Endothelial Growth Factor), a gene encoding VEGFR (Vascular Endothelial Growth Factor Receptor), a gene encoding FGF (Fibroblast Growth Factor), a gene encoding FGFR (Fibroblast Growth Factor Receptor), a gene encoding CD31, and a gene encoding endothelial nitric oxide synthase (eNOS), and specifically, at least one selected from the group consisting of a gene encoding VEGFR (Vascular Endothelial Growth Factor Receptor), a gene encoding FGF (Fibroblast Growth Factor), a gene encoding FGFR (Fibroblast Growth Factor Receptor), and a gene encoding endothelial nitric oxide synthase (eNOS), VEGF (Vascular Endothelial Growth Factor) It may be to promote the expression of a gene encoding a growth factor (Growth Factor) and a gene encoding CD31, and more specifically, a gene encoding a vascular endothelial growth factor (VEGF), a gene encoding a vascular endothelial growth factor receptor (VEGFR), a gene encoding a fibroblast growth factor (FGF), a gene encoding a fibroblast growth factor receptor (FGFR),It may be by enhancing the expression of both the gene encoding CD31 and the gene encoding endothelial nitric oxide synthase (eNOS).

[0042] In addition, in one aspect, the ischemic heart disease may be one or more diseases selected from the group consisting of coronary artery disease, stable angina, unstable angina, variant angina, myocardial infarction, heart attack, heart failure, arteriosclerosis, ischemic heart failure, and cardiomyopathy.

[0043] The above term “prevention” may mean any act of inhibiting or delaying ischemic heart disease in a subject by administering a pharmaceutical composition according to one aspect.

[0044] The above term “treatment” may mean any action that improves or beneficially changes the symptoms of ischemic heart disease in a subject by administering a pharmaceutical composition according to one aspect.

[0045] The term "administration" refers to introducing a given substance into a subject in an appropriate manner, and "subject" refers to any living organism, including humans, rats, mice, and livestock, that may have ischemic heart disease. A specific example may be a mammal, including humans.

[0046] Additionally, the pharmaceutical composition may be provided as a pharmaceutical composition containing only the active ingredient, or including one or more pharmaceutically acceptable carriers, excipients or diluents.

[0047] Specifically, the carrier may be, for example, a colloidal suspension, a powder, a saline solution, a lipid, a liposome, microspheres, or nano-spheres. These may be complexed or associated with a carrier vehicle and may be transported in vivo using carrier systems known in the art, such as lipids, liposomes, microparticles, gold, nanoparticles, polymers, condensation agents, polysaccharides, polyamino acids, dendrimers, saponins, adsorption enhancing substances, or fatty acids.

[0048] When the above pharmaceutical composition is formulated, it can be prepared using diluents or excipients such as lubricants, sweeteners, flavoring agents, emulsifiers, suspending agents, preservatives, fillers, bulking agents, binders, wetting agents, disintegrating agents, and surfactants that are commonly used. Solid preparations for oral administration may include tablets, pills, powders, granules, capsules, etc., and such solid preparations can be prepared by mixing at least one excipient, such as starch, calcium carbonate, sucrose or lactose, gelatin, etc., with the above composition. In addition to simple excipients, lubricants such as magnesium stearate and talc can also be used. Liquid preparations for oral administration include suspensions, solutions, emulsions, and syrups. In addition to commonly used simple diluents such as water and liquid paraffin, they may contain various excipients such as wetting agents, sweeteners, fragrances, and preservatives. Preparations for parenteral administration may include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, and suppositories. Non-aqueous solvents and suspensions 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, cacao butter, laurin butter, and glycerogelatin. When manufacturing in the form of eye drops, known diluents or excipients may be used.

[0049] In one aspect, the pharmaceutical composition may further comprise an agent for treating ischemic heart disease in addition to aspartic acid or a pharmaceutically acceptable salt thereof and / or ornithine or a pharmaceutically acceptable salt thereof.

[0050] The above pharmaceutical composition may be provided in combination with another ischemic heart disease treatment agent, and the other ischemic heart disease treatment agent may be a conventionally known ischemic heart disease treatment agent or a newly developed ischemic heart disease treatment agent.

[0051] When the above pharmaceutical composition further includes another ischemic heart disease treatment agent, it is important to mix the agent in an amount that can achieve the maximum effect with the minimum amount without causing side effects, and this can be easily determined by a person skilled in the art.

[0052] Furthermore, in one aspect, the pharmaceutical composition may be administered alone or in combination with another ischemic heart disease treatment agent. Specifically, the pharmaceutical composition may be administered in combination with a known composition having an ischemic heart disease preventive or therapeutic effect or with another ischemic heart disease treatment agent, and may be administered simultaneously, separately, or sequentially, and may be administered singly or in multiple doses. Taking all of the above factors into consideration, it is important to administer an amount that achieves maximum effect with the minimum amount without causing side effects, and this can be readily determined by those skilled in the art.

[0053] The above pharmaceutical composition can be administered orally or parenterally, and when administered parenterally, the method of injection can be selected from external application to the skin or intraperitoneal injection, intrarectal injection, subcutaneous injection, intravenous injection, intramuscular injection, intraarterial injection, intramedullary injection, intracardiac injection, intrathecal injection, percutaneous injection, intranasal injection, intraenteric injection, local injection, sublingual injection, rectal injection, or intrathoracic injection.

[0054] The above pharmaceutical composition is administered in a pharmaceutically effective amount. The term "pharmaceutically effective amount" means an amount sufficient to treat a disease with a reasonable benefit / risk ratio applicable to medical treatment. The effective dosage level may be determined based on the type and severity of the patient's disease, the activity and sensitivity of the drug, the time of administration, the route of administration and excretion rate, the duration of treatment, concomitant medications, and other factors well known in the medical field. The above administration may be administered once a day or in several divided doses. For example, it may be administered every other day or once a week.

[0055] In one aspect, the pharmaceutical composition comprising the aspartic acid or a pharmaceutically acceptable salt thereof may be administered at a dose of 0.1 mg / kg to 1000 mg / kg. Specifically, the pharmaceutical composition comprising the aspartic acid or a pharmaceutically acceptable salt thereof is 0.1 mg / kg to 1000 mg / kg, 0.1 mg / kg to 800 mg / kg, 0.1 mg / kg to 500 mg / kg, 0.1 mg / kg to 300 mg / kg, 0.1 mg / kg to 100 mg / kg, 0.1 mg / kg to 10 mg / kg, 10 mg / kg to 1000 mg / kg, 10 mg / kg to 800 mg / kg, 10 mg / kg to 500 mg / kg, 10 mg / kg to 300 mg / kg, 10 mg / kg to 100 mg / kg, 100 mg / kg to 1000 mg / kg, 100 mg / kg to 800 mg / kg, 100 mg / kg to 500 mg / kg, It may be administered at 100 mg / kg to 300 mg / kg, 300 mg / kg to 1000 mg / kg, 300 mg / kg to 800 mg / kg, 300 mg / kg to 500 mg / kg, 500 mg / kg to 1000 mg / kg, 500 mg / kg to 800 mg / kg, or 800 mg / kg to 1000 mg / kg.

[0056] In addition, in one aspect, the pharmaceutical composition comprising the aspartic acid or a pharmaceutically acceptable salt thereof; and ornithine or a pharmaceutically acceptable salt thereof may be administered at a dose of 0.1 mg / kg to 1000 mg / kg. Specifically, the aspartic acid or a pharmaceutically acceptable salt thereof; And a pharmaceutical composition comprising ornithine or a pharmaceutically acceptable salt thereof, wherein the pharmaceutical composition comprises 0.1 mg / kg to 1000 mg / kg, 0.1 mg / kg to 800 mg / kg, 0.1 mg / kg to 500 mg / kg, 0.1 mg / kg to 300 mg / kg, 0.1 mg / kg to 100 mg / kg, 0.1 mg / kg to 10 mg / kg, 10 mg / kg to 1000 mg / kg, 10 mg / kg to 800 mg / kg, 10 mg / kg to 500 mg / kg, 10 mg / kg to 300 mg / kg, 10 mg / kg to 100 mg / kg, 100 mg / kg to 1000 mg / kg, 100 mg / kg to 800 mg / kg, 100 mg / kg to 500 mg / kg, It may be administered at 100 mg / kg to 300 mg / kg, 300 mg / kg to 1000 mg / kg, 300 mg / kg to 800 mg / kg, 300 mg / kg to 500 mg / kg, 500 mg / kg to 1000 mg / kg, 500 mg / kg to 800 mg / kg, or 800 mg / kg to 1000 mg / kg.

[0057] According to one aspect, a pharmaceutical composition significantly reduces the volume of an infarcted area in the myocardium, increases perfusion, increases the density of CD31-positive capillary lumen, and promotes angiogenesis by overexpressing VEGF by including aspartic acid or a pharmaceutically acceptable salt thereof, and when the pharmaceutical composition further includes ornithine or a pharmaceutically acceptable salt thereof, it exhibits a superior effect of reducing the volume of an infarcted area in the myocardium and angiogenesis compared to administration of aspartic acid alone, and therefore, the pharmaceutical composition can be effectively utilized as an agent for preventing, improving, or treating ischemic heart disease.

[0058]

[0059] Another aspect provides a health functional food for preventing or improving ischemic heart disease, comprising aspartic acid or a food-wise acceptable salt thereof.

[0060] The above “aspartic acid”, “salt”, “ischemic heart disease”, “prevention”, etc. may be within the aforementioned range.

[0061] The above aspartic acid or a food-based acceptable salt thereof reduces the volume of the infarcted area in the myocardium, increases perfusion, increases the density of CD31-positive capillary lumen, and promotes angiogenesis by overexpressing VEGF, thereby exhibiting an excellent effect in preventing or improving ischemic heart disease.

[0062] More specifically, the aspartic acid or a food-acceptable salt thereof promotes the conversion of citrulline or a food-acceptable salt thereof into arginine or a food-acceptable salt thereof within the administered subject. The converted arginine produces nitric oxide (NO) together with endothelial nitric oxide synthase (eNOS), and the nitric oxide upregulates the expression of VEGF, thereby exhibiting an excellent angiogenic effect and an excellent preventive or ameliorating effect on ischemic heart disease.

[0063] In one aspect, the health functional food may further contain ornithine or a food-wise acceptable salt thereof.

[0064] When the above health functional food further contains ornithine or a food-related acceptable salt thereof, it further reduces the volume of the infarcted area in the myocardium, promotes angiogenesis, and thereby exhibits a better effect in preventing or improving ischemic heart disease.

[0065] More specifically, the above-mentioned ornithine or a food-acceptable salt thereof is converted into citrulline or a food-acceptable salt thereof within the mitochondria. As a result, the aspartic acid or a food-acceptable salt thereof contained in the health functional food produces more arginine, ultimately demonstrating a superior angiogenic effect and an excellent preventive or ameliorating effect against ischemic heart disease.

[0066] The term "improvement" can refer to any action that at least reduces a parameter associated with the condition being treated, such as the severity of symptoms. In this case, the health functional food can be used to prevent or improve ischemic heart disease, either before or after the onset of the disease, or simultaneously with or separately from a treatment medication.

[0067] In the above health functional food, the active ingredient can be added directly to the food or used in combination with other foods or food ingredients, and can be used appropriately according to conventional methods. The amount of the active ingredient mixed can be appropriately determined depending on the intended use (prevention or improvement). Generally, when manufacturing a food or beverage, the health functional food can be added in an amount of about 15% by weight or less, more specifically about 10% by weight or less, based on the raw material. However, in the case of long-term intake for the purpose of health and hygiene or health control, the amount may be below the above range.

[0068] The above health functional food may be formulated into one selected from the group consisting of tablets, pills, powders, granules, powders, capsules, and liquid formulations, further comprising one or more of a carrier, diluent, excipient, and additive. Foods to which compounds according to one aspect may be added include various foods, powders, granules, tablets, capsules, syrups, beverages, gum, tea, vitamin complexes, and health functional foods.

[0069] Specific examples of the carrier, excipient, diluent and additive may include at least one selected from the group consisting of lactose, dextrose, sucrose, sorbitol, mannitol, erythritol, starch, acacia gum, calcium phosphate, alginate, gelatin, calcium phosphate, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, polyvinylpyrrolidone, methylcellulose, water, sugar syrup, methylcellulose, methyl hydroxy benzoate, propyl hydroxy benzoate, talc, magnesium stearate and mineral oil.

[0070] In addition to containing the above-mentioned effective ingredient, the above-mentioned health functional food may contain other ingredients as essential ingredients without special restrictions. For example, it may contain various flavoring agents or natural carbohydrates as additional ingredients, as in conventional beverages. Examples of the above-mentioned natural carbohydrates may include conventional sugars such as monosaccharides, such as glucose, fructose, etc.; disaccharides, such as maltose, sucrose, etc.; and polysaccharides, such as dextrin, cyclodextrin, etc.; and sugar alcohols, such as xylitol, sorbitol, and erythritol. As flavoring agents other than those described above, natural flavoring agents (thaumatin, stevia extracts (e.g., rebaudioside A, glycyrrhizin, etc.)) and synthetic flavoring agents (saccharin, aspartame, etc.) may be advantageously used. The proportion of the above-mentioned natural carbohydrates may be appropriately determined by a person skilled in the art.

[0071] In addition to the above, health functional foods according to the aspect may contain various nutrients, vitamins, minerals (electrolytes), flavoring agents such as synthetic flavoring agents and natural flavoring agents, coloring agents and thickening agents (cheese, chocolate, etc.), pectic acid and its salts, alginic acid and its salts, organic acids, protective colloid thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohol, carbonating agents used in carbonated beverages, etc. These ingredients may be used independently or in combination, and the ratio of these additives may also be appropriately selected by those skilled in the art.

[0072] In one aspect, the health functional food may further include a health functional food for preventing or improving ischemic heart disease in addition to aspartic acid or a food-based acceptable salt thereof and / or ornithine or a food-based acceptable salt thereof.

[0073] The above health functional food may be provided in combination with a health functional food for preventing or improving ischemic heart disease known in the past or a health functional food for preventing or improving ischemic heart disease that is newly developed.

[0074] When the above health functional food further includes a health functional food for preventing or improving ischemic heart disease, it is important to mix the amount that can achieve the maximum effect with the minimum amount without causing side effects, and this can be easily determined by a person skilled in the art.

[0075] In addition, in one aspect, the health functional food may be consumed alone or in combination with a health functional food for preventing or improving ischemic heart disease.

[0076] The above health functional food can be taken in conjunction with known compositions or newly developed health functional foods for the prevention or improvement of ischemic heart disease. These can be taken simultaneously, separately, or sequentially, and can be taken singly or in multiple doses. It is important to consider all of the above factors and take the amount that achieves maximum effect with the minimum amount possible without causing side effects, which can be readily determined by those skilled in the art.

[0077] According to one aspect, a health functional food significantly reduces the volume of an infarcted area in the myocardium, increases perfusion, increases the density of CD31-positive capillary lumen, and promotes angiogenesis by overexpressing VEGF by including aspartic acid or a food-based acceptable salt thereof, and when the health functional food further includes ornithine or a food-based acceptable salt thereof, it exhibits a more excellent effect of reducing the volume of an infarcted area in the myocardium and promoting angiogenesis, and therefore, the health functional food can be effectively utilized as an agent for preventing or improving ischemic heart disease.

[0078]

[0079] Another aspect provides a method for preventing or treating ischemic heart disease, comprising administering aspartic acid or a pharmaceutically acceptable salt thereof to a subject in need thereof.

[0080] The above “aspartic acid”, “pharmaceutically acceptable salt”, “substance”, “administration”, “ischemic heart disease”, “prevention”, “treatment”, etc. may be within the aforementioned scope.

[0081] In one aspect, in the step of administering the aspartic acid or a pharmaceutically acceptable salt thereof, the aspartic acid or a pharmaceutically acceptable salt thereof may be administered at 0.1 mg / kg to 1000 mg / kg. Specifically, the pharmaceutical composition comprises 0.1 mg / kg to 1000 mg / kg, 0.1 mg / kg to 800 mg / kg, 0.1 mg / kg to 500 mg / kg, 0.1 mg / kg to 300 mg / kg, 0.1 mg / kg to 100 mg / kg, 0.1 mg / kg to 10 mg / kg, 10 mg / kg to 1000 mg / kg, 10 mg / kg to 800 mg / kg, 10 mg / kg to 500 mg / kg, 10 mg / kg to 300 mg / kg, 10 mg / kg to 100 mg / kg, 100 mg / kg to 1000 mg / kg, 100 mg / kg to 800 mg / kg, 100 mg / kg to 500 mg / kg, 100 mg / kg to 300 It may be administered at mg / kg, 300 mg / kg to 1000 mg / kg, 300 mg / kg to 800 mg / kg, 300 mg / kg to 500 mg / kg, 500 mg / kg to 1000 mg / kg, 500 mg / kg to 800 mg / kg, or 800 mg / kg to 1000 mg / kg.

[0082] In one aspect, the method may further comprise the step of administering ornithine or a pharmaceutically acceptable salt thereof to the subject.

[0083] In one aspect, in the step of administering the ornithine or a pharmaceutically acceptable salt thereof, the ornithine or a pharmaceutically acceptable salt thereof may be administered at 0.1 mg / kg to 1000 mg / kg. Specifically, the pharmaceutical composition comprises 0.1 mg / kg to 1000 mg / kg, 0.1 mg / kg to 800 mg / kg, 0.1 mg / kg to 500 mg / kg, 0.1 mg / kg to 300 mg / kg, 0.1 mg / kg to 100 mg / kg, 0.1 mg / kg to 10 mg / kg, 10 mg / kg to 1000 mg / kg, 10 mg / kg to 800 mg / kg, 10 mg / kg to 500 mg / kg, 10 mg / kg to 300 mg / kg, 10 mg / kg to 100 mg / kg, 100 mg / kg to 1000 mg / kg, 100 mg / kg to 800 mg / kg, 100 mg / kg to 500 mg / kg, 100 mg / kg to 300 It may be administered at mg / kg, 300 mg / kg to 1000 mg / kg, 300 mg / kg to 800 mg / kg, 300 mg / kg to 500 mg / kg, 500 mg / kg to 1000 mg / kg, 500 mg / kg to 800 mg / kg, or 800 mg / kg to 1000 mg / kg.

[0084] Additionally, in one aspect, the aspartic acid or a pharmaceutically acceptable salt thereof may be administered simultaneously with ornithine or a pharmaceutically acceptable salt thereof.

[0085] In addition, in one aspect, when the aspartic acid or a pharmaceutically acceptable salt thereof is administered simultaneously with ornithine or a pharmaceutically acceptable salt thereof in the step of administering the aspartic acid or a pharmaceutically acceptable salt thereof, the aspartic acid or a pharmaceutically acceptable salt thereof; and ornithine or a pharmaceutically acceptable salt thereof may be administered at 0.1 mg / kg to 1000 mg / kg. Specifically, the pharmaceutical composition comprises 0.1 mg / kg to 1000 mg / kg, 0.1 mg / kg to 800 mg / kg, 0.1 mg / kg to 500 mg / kg, 0.1 mg / kg to 300 mg / kg, 0.1 mg / kg to 100 mg / kg, 0.1 mg / kg to 10 mg / kg, 10 mg / kg to 1000 mg / kg, 10 mg / kg to 800 mg / kg, 10 mg / kg to 500 mg / kg, 10 mg / kg to 300 mg / kg, 10 mg / kg to 100 mg / kg, 100 mg / kg to 1000 mg / kg, 100 mg / kg to 800 mg / kg, 100 mg / kg to 500 mg / kg, 100 mg / kg to 300 It may be administered at mg / kg, 300 mg / kg to 1000 mg / kg, 300 mg / kg to 800 mg / kg, 300 mg / kg to 500 mg / kg, 500 mg / kg to 1000 mg / kg, 500 mg / kg to 800 mg / kg, or 800 mg / kg to 1000 mg / kg.

[0086] A method for preventing or treating ischemic heart disease according to one aspect includes a step of administering aspartic acid or a pharmaceutically acceptable salt thereof, thereby significantly reducing the volume of an infarcted area in the myocardium, increasing perfusion, increasing the density of CD31-positive capillary lumen, and promoting angiogenesis by overexpressing VEGF. When the pharmaceutical composition further comprises ornithine or a pharmaceutically acceptable salt thereof, it exhibits a more excellent effect of reducing the volume of an infarcted area in the myocardium and promoting angiogenesis, and therefore, the pharmaceutical composition can be effectively utilized as a preventive, ameliorating, or a therapeutic agent for ischemic heart disease.

[0087]

[0088] Another aspect provides the use of aspartic acid or a pharmaceutically acceptable salt thereof for the manufacture of a medicament for the prevention or treatment of ischemic heart disease.

[0089] Specifically, it may provide the use of aspartic acid or a pharmaceutically acceptable salt thereof; and ornithine or a pharmaceutically acceptable salt thereof for the manufacture of a medicament for the prevention or treatment of ischemic heart disease.

[0090] The above “aspartic acid”, “pharmaceutically acceptable salt”, “substance”, “administration”, “ischemic heart disease”, “prevention”, “treatment”, etc. may be within the aforementioned scope.

[0091] In one aspect, aspartic acid or a pharmaceutically acceptable salt thereof for the manufacture of a drug for preventing or treating ischemic heart disease may be administered at 0.1 mg / kg to 1000 mg / kg. Specifically, aspartic acid or a pharmaceutically acceptable salt thereof for the manufacture of a drug for preventing or treating ischemic heart disease is used in an amount of 0.1 mg / kg to 1000 mg / kg, 0.1 mg / kg to 800 mg / kg, 0.1 mg / kg to 500 mg / kg, 0.1 mg / kg to 300 mg / kg, 0.1 mg / kg to 100 mg / kg, 0.1 mg / kg to 10 mg / kg, 10 mg / kg to 1000 mg / kg, 10 mg / kg to 800 mg / kg, 10 mg / kg to 500 mg / kg, 10 mg / kg to 300 mg / kg, 10 mg / kg to 100 mg / kg, 100 mg / kg to 1000 mg / kg, 100 mg / kg to 800 It may be administered at 100 mg / kg to 500 mg / kg, 100 mg / kg to 300 mg / kg, 300 mg / kg to 1000 mg / kg, 300 mg / kg to 800 mg / kg, 300 mg / kg to 500 mg / kg, 500 mg / kg to 1000 mg / kg, 500 mg / kg to 800 mg / kg, or 800 mg / kg to 1000 mg / kg.

[0092] In addition, in one aspect, aspartic acid or a pharmaceutically acceptable salt thereof for the manufacture of a drug for preventing or treating ischemic heart disease; and ornithine or a pharmaceutically acceptable salt thereof may be administered at 0.1 mg / kg to 1000 mg / kg. Specifically, aspartic acid or a pharmaceutically acceptable salt thereof for the manufacture of a drug for preventing or treating ischemic heart disease; And ornithine or a pharmaceutically acceptable salt thereof is 0.1 mg / kg to 1000 mg / kg, 0.1 mg / kg to 800 mg / kg, 0.1 mg / kg to 500 mg / kg, 0.1 mg / kg to 300 mg / kg, 0.1 mg / kg to 100 mg / kg, 0.1 mg / kg to 10 mg / kg, 10 mg / kg to 1000 mg / kg, 10 mg / kg to 800 mg / kg, 10 mg / kg to 500 mg / kg, 10 mg / kg to 300 mg / kg, 10 mg / kg to 100 mg / kg, 100 mg / kg to 1000 mg / kg, 100 mg / kg to 800 mg / kg, 100 mg / kg to 500 mg / kg, 100 mg / kg to It may be administered at 300 mg / kg, 300 mg / kg to 1000 mg / kg, 300 mg / kg to 800 mg / kg, 300 mg / kg to 500 mg / kg, 500 mg / kg to 1000 mg / kg, 500 mg / kg to 800 mg / kg, or 800 mg / kg to 1000 mg / kg.

[0093] Aspartic acid or a pharmaceutically acceptable salt thereof according to one aspect significantly reduces the volume of an infarcted area in the myocardium, increases perfusion, increases the density of CD31-positive capillary lumen, and promotes angiogenesis by overexpressing VEGF. When the aspartic acid or a pharmaceutically acceptable salt thereof is used together with ornithine or a pharmaceutically acceptable salt thereof, a more excellent effect of reducing the volume of an infarcted area in the myocardium and promoting angiogenesis are exhibited. Therefore, the aspartic acid or a pharmaceutically acceptable salt thereof; and / or the ornithine or a pharmaceutically acceptable salt thereof can be effectively utilized in the manufacture of a medicament for preventing, improving, or treating ischemic heart disease.

[0094]

[0095] A pharmaceutical composition according to one aspect significantly reduces the volume of an infarcted area in the myocardium, increases perfusion, increases the density of CD31-positive capillary lumen, and promotes angiogenesis by overexpressing VEGF by including aspartic acid or a pharmaceutically acceptable salt thereof, thereby exhibiting an excellent effect in preventing or treating ischemic heart disease. In addition, the composition further comprises ornithine or a pharmaceutically acceptable salt thereof, thereby inducing a better effect in reducing the volume of an infarcted area in the myocardium and angiogenesis than when administering aspartic acid or a pharmaceutically acceptable salt thereof alone, thereby exhibiting an excellent effect in preventing or treating ischemic heart disease.

[0096]

[0097] Figure 1 is a diagram confirming the change in myocardial infarction volume according to LOLA treatment in a rat model in which myocardial infarction was induced.

[0098] Figure 2 is a diagram showing the mechanism by which aspartic acid and ornithine act on angiogenesis.

[0099] Figure 3 is a diagram showing the results of performing a TUNEL assay and confirming damaged tissue using a confocal microscope.

[0100] Figure 4 shows a point on a linear plot of an experiment showing perfusion recovery (n = 5), with error bars representing the standard error of the mean.

[0101] Figure 5 shows the perfusion recovery of the three groups on POD 7 and 14. Specifically, the bar graph of the experiment represents the perfusion recovery rate, and the error bars represent the standard error of the mean. * indicates a significant difference at p < 0.05. The p value was calculated using a two-sample t-test. Group 1 (control group; n = 8); Group 2 (sorbitol; n = 8); Group 3 (LOLA; n = 8).

[0102] Figures 6 and 7 are diagrams showing angiogenesis in three groups after LOLA administration. Specifically, 1) Figure 6 shows the expression of CD31 protein detected by immunohistochemistry. The scale bar represents 100 μm. 2) Figure 7 shows the density of CD31-positive cells. The error bars represent the standard error of the mean. *** indicates a significant difference at p < 0.001.

[0103] Figures 8 to 10 are diagrams showing angiogenesis in three groups after LOLA administration. Specifically, 1) Figure 8 shows the expression of VEGF protein detected by immunohistochemistry. The scale bar represents 100 μm. 2) Figure 9 shows Western blotting of VEGF. The relative expression of the protein was normalized to the endogenous control GAPDH. 3) Figure 10 is a diagram showing the expression level of VEGF. * and *** indicate significant differences at p < 0.05 and p < 0.001, respectively.

[0104]

[0105] The present invention will be described in more detail below through examples. However, these examples are intended to exemplify the present invention and the scope of the present invention is not limited to these examples.

[0106]

[0107] <Reference Example 1> Experimental animal (rat model of myocardial ischemia / reperfusion injury)

[0108] Fifty healthy, age-matched adult male Wistar rats (Orientbio) weighing 400–430 g were used as experimental animals. All animals were acclimated for 1 week in sterile cages under a light / dark cycle (12 / 12 h), humidity of 50 ± 10%, and temperature of 22 ± 2 °C. A standard diet was provided during the acclimation period, and fresh water was provided ad libitum.

[0109] Afterwards, a tracheotomy was performed on the rat using an intravenous catheter (4712-020-116. IV Catheter 16G, Seun Medical). Specifically, the left anterior descending (LAD) coronary artery was ligated midway between the pulmonary artery and the apex of the rat using a 6-0 ethylene suture. The ligation was performed after placing a PE-10 tube (polyethylene tube, outer diameter 0.61 mm) between the LAD and the suture.

[0110] After LAD ligation, ischemia was confirmed by the development of myocardial cyanosis and movement disorders. Thirty minutes after LAD ligation, the PE-10 tube was removed to induce reperfusion. Control rats (Sham) underwent the same surgical procedure, except for ligation. Four hours after LAD ligation, the rats were anesthetized and euthanized.

[0111]

[0112] <Reference Example 2> Animal Experiment Protocol

[0113] Specifically, the control group (MI; n=4) and experimental group (MI+LOLA; n=4) of the animal experiment were set under the following conditions, and the experiment was conducted using four rats in each control and experimental group. During all experiments, the rectal temperature of the rats was maintained at 37.0 ± 0.5 °C using a feedback-controlled heating pad (HB 101, Harvard Apparatus):

[0114] - MI (control group): 15 minutes after LAD ligation, intraperitoneal injection of physiological saline (1.7 mL / kg)

[0115] - MI+LOLA (experimental group): LOLA (333 mg / kg) (Hepa-Merz infusion, Hanwha Pharma) was injected intraperitoneally 15 minutes after LAD ligation.

[0116]

[0117] <Reference Example 3> Infarct volume evaluation

[0118] To assess myocardial infarction, 2,3,5-triphenyltetrazolium chloride (TTC) staining (T8877, Sigma-Aldrich) was performed 4 or 24 hours after LAD I / R. The chests of anesthetized rats (sham or LAD I / R-operated rats) were reopened, and the hearts were excised and sectioned into 2-mm-thick sections on precooled coronary matrix devices (HSRA001-1, Zivic Instruments, Pittsburgh).

[0119] Coronal sections were immersed in a 1% TTC solution in sterile distilled water at 37°C for 30 min and then fixed in a 4% paraformaldehyde solution in phosphate-buffered saline for 48 h. Each stained section was scanned with a flatbed scanner (PERFECTION V800 PHOTO, EPSN).

[0120] To measure infarct volume, cardiac tissues between 0 and 8 mm from the apex were used. The anterior and posterior infarct areas in 2-mm-thick sections were measured using ImageJ 1.48v software. The infarct volume of each section was determined by multiplying the average of the anterior and posterior areas by the thickness (2 mm) ([thickness × (upper area + lower area) / 2]). The total infarct volume was calculated as the sum of the infarct volumes of each section, and the results were expressed as the mean ± SD (p = 0.001).

[0121]

[0122] <Reference Example 4> Apoptotic cell analysis

[0123] To identify the peri-infarct area of ​​the left ventricle, 2,3,5-TTC staining was performed. Two-mm-thick sections were selected from a region 4 to 6 mm from the apex, where normal and infarcted tissues were adequately mixed and the peri-infarct area was easily observed after TTC staining. These sections were fixed in 4% paraformaldehyde solution and embedded in paraffin.

[0124] Heart sections of 4 μm thickness were obtained by cutting the area including the infarct area using a microtome (LEICA RM 2335, Wetzlar).

[0125] Apoptotic cells were identified by TUNEL (terminal deoxynucleotidyl transferase (TdT)-mediated dUTP nick end labeling) assay using the DeadEnd™ Fluorometric TUNEL system (Promega, WI) according to the manufacturer's instructions. One slide from each animal was stained, and the peri-ischemic area was observed in the stained sections using a confocal microscope (LSM700, Carl Zeiss GmbG, Jena, Germany). TUNEL-positive cells in the peri-ischemic area were identified in the stained sections.

[0126]

[0127] <Reference Example 5> Experimental animals in which ischemic disease was induced to evaluate angiogenesis

[0128] Male Sprague-Dawley rats (Seongnam, South Korea), weighing 280–320 g (8 weeks old), were used. The rats were housed in a controlled environment (room temperature 20–24°C, humidity 40–60%) with ad libitum access to standard food and water throughout the experiment. Unilateral femoral artery ligation was then performed.

[0129] Perfusion imaging was performed before surgery. Immediately after surgery, perfusion imaging was performed (postoperative day 0). The rats were then randomly assigned to three groups, with eight rats per group:

[0130] - Group 1 (control group): Saline solution administered,

[0131] - Group 2 (Sorbitol experimental group): Sorbitol administration,

[0132] - Group 3 (LOLA experimental group): LOLA administration.

[0133] Specifically, Group 1 received saline (1.7 ml / kg), Group 2 received distilled water, sorbitol, and saline (20:2:30, 1.7 ml / kg), and Group 3 received LOLA (333 mg / kg) (Hepa-Merz infusion, Hanwha Pharma, Chuncheon, Republic of Korea) intraperitoneally five times on PODs 3, 5, 7, and 10 ALC. Since Hepa-Merz infusion consists of LOLA, sorbitol, and distilled water, Group 2 was administered the same amount of sorbitol and distilled water along with saline instead of LOLA. To create a normal control group, Group 2 was assigned the same amount of saline. Perfusion imaging was scanned at PODs 7 and 14. Immunohistochemical analysis and Western blotting of rats with induced ischemic disease were performed after the final perfusion imaging.

[0134]

[0135] <Reference Example 6> Perfusion Imaging

[0136] Rats were imaged for approximately 3 minutes using a laser Doppler imaging device (Moor LDI, Axminster, UK). Images were analyzed using Moor LDI Imaging Review. Perfusion rates were calculated by comparing preoperative and postoperative results.

[0137]

[0138] <Reference Example 7> Immunohistochemical analysis

[0139] The mice were euthanized and fixed by perfusion with 4% paraformaldehyde 14 days after surgery. They were then embedded in paraffin, sectioned at 5 μm using a microtome, and mounted on glass slides. The sections were blocked with Proteinase K (S3020, Agilent DAKO, Santa Clara, CA, USA) and then incubated with antibodies to VEGF (1:100, MA1-16629, Thermo Scientific, Waltham, MA, USA) and CD31 (1:100, MA1-80069, Thermo Scientific, Waltham, MA, USA) for 10 min. The samples were then incubated with secondary antibodies (Envision + system-HRP Labeled Polymer - Antimouse, K4001, Agilent DAKO, Santa Clara, CA, USA) for 20 min at room temperature. Capillary density was measured in mm 2 It was expressed as the number of CD31-positive cells per case and was written after consensus review by two doctors for each case.

[0140]

[0141] <Reference Example 8> Western blotting

[0142] Frozen muscle samples were homogenized in RIPA Lysis and Extraction Buffer (Thermo Fisher Scientific, Walthan, MA, USA) with 1% Halt™ Protease Inhibitor Cocktail (Thermo Fisher Scientific, Walthan, MA, USA) and centrifuged at 13,000 g for 15 min at 4°C. Protein concentrations were determined using the BCA assay kit (cat. no. 23250; Pierce; Thermo Fisher Scientific, Walthan, MA, USA) according to the manufacturer's instructions. Equal amounts of proteins were loaded onto 12% Tris-glycine SDS-polyacrylamide gels and separated by SDS-PAGE (Bio-Rad, Hercules, CA, USA). After electrophoresis, the gels were transferred to polyvinylidene difluoride (PVDF) membranes (GenDEPOT, Baker, TX, USA). Membranes were blocked in Tween-20 buffer (Biosoulution, Suwon, Korea) containing 5% bovine serum albumin (BSA) (bioWORLD, Dublin, OH, USA) for 1 h at room temperature in Tris-buffered saline (TBS). The membranes were incubated overnight at 4°C with primary antibodies against VEGF (MA1-16629; Thermo Fisher Scientific, Walthan, MA, USA) and GAPDH diluted 1:1,000 in 5% BSA-TBS-T. The membranes were washed five times for 5 min in TBS-T buffer, incubated with goat anti-mouse IgG antibody (HRP, GeneTex, Irvine, CA, USA) diluted 1:10,000 in 5% BSA-TBS-T at room temperature, and washed five times for 5 min in TBS-T buffer.Bands were detected using Clarity™ Western ECL Substrate (Bio-Rad, Hercules, CA, USA). Relative protein expression was normalized to the endogenous control GAPDH (Glyceraldehyde 3-phosphate dehydrogenase) using ImageJ software (National Institutes of Health, Bethesda, MD, USA).

[0143]

[0144] <Example 1> Evaluation of the effect of reducing myocardial infarction in rats induced with myocardial infarction

[0145] To determine the preventive or therapeutic effect of a pharmaceutical composition containing aspartic acid on ischemic heart disease, changes in myocardial infarction volume were evaluated in a rat model of myocardial infarction induced by L-ornithine-L-aspartate (LOLA).

[0146] Specifically, the rats used were prepared according to Reference Example 1, and the control and experimental groups were prepared according to Reference Example 2. Thereafter, the change in myocardial infarction volume according to drug treatment was measured according to Reference Example 3.

[0147] As shown in Table 1 and Figure 1 below, in the control group (MI; n=4) treated with only saline in rats with myocardial infarction induced through LAD I / R surgery, the infarct volume accounted for 21.8±4.04% of the whole heart, whereas in the experimental group treated with LOLA (MI+LOLA; n=4), the infarct volume accounted for only 7.1±3.44% of the whole heart (a decrease to about 1 / 3 of the control group), confirming that LOLA treatment exhibited a remarkably excellent preventive or therapeutic effect on rats with myocardial infarction induced.

[0148] That is, through the above results, it was confirmed that a pharmaceutical composition containing aspartic acid can be effectively utilized for the prevention or treatment of ischemic heart disease.

[0149] Total area infarction area infarction ratio (%)MI42067.3±3430.979243.5±2220.0221.8±4.04MI+LOLA43243.3±1124.403074.5±1493.707.1±3.44

[0150] * n=4; Results = mean±SD; p=0.001

[0151]

[0152] <Example 2> Apoptotic cell analysis

[0153] To determine the preventive or therapeutic effect of a pharmaceutical composition containing aspartic acid on ischemic heart disease, the peri-infarct area of ​​the left ventricle was identified using a TUNEL assay.

[0154] Specifically, the rats used were prepared according to Reference Example 1, and the control and experimental groups were prepared according to Reference Example 2. Thereafter, TUNEL-positive cells, i.e., apoptotic cells, were identified according to Reference Example 4 following drug treatment.

[0155] As shown in Fig. 3, in rats in which myocardial infarction was induced through LAD I / R surgery, in the control group (MI; n=3) treated with only saline, TUNEL-positive cells showing green fluorescence, i.e., apoptotic cells, were significantly increased, whereas in the experimental group treated with LOLA (MI+LOLA; n=3), TUNEL-positive cells were significantly decreased compared to the control group (MI) treated with only saline.

[0156] That is, through the above results, it was confirmed that treatment with a pharmaceutical composition containing aspartic acid significantly reduced apoptosis of cells around the infarct area of ​​the left ventricle in rats induced with myocardial infarction, thereby exhibiting a remarkably excellent preventive or therapeutic effect on cell damage caused by ischemic disease.

[0157]

[0158] <Example 3> Confirmation of perfusion changes

[0159] To determine the angiogenic effect of a pharmaceutical composition containing aspartic acid on ischemic disease, changes in perfusion were assessed. Specifically, the experiment was conducted according to Reference Examples 5 and 6.

[0160] As shown in Figure 4, immediate postoperative perfusion decreased by approximately 40% compared to the normal preoperative level. Perfusion then increased over 14 days, reaching a plateau with approximately 60% recovery between weeks 3 and 4. The period leading to this plateau was defined as the perfusion recovery period. To evaluate the effects of L-ornithine-L-aspartate (LOLA) on angiogenesis during the subacute phase of ischemic disease, changes in perfusion and the microenvironment were examined. LOLA was administered five times, starting on postoperative day (POD) three, over a two-week period.

[0161] On POD 7, the LOLA-administered group (group 3) showed a significantly higher perfusion recovery rate than the control group (group 1) and sorbitol-administered group (group 2) (group 1 vs group 2 vs group 3: 51.72 ± 13.81 vs 58.42 ± 15.95 vs 71.90 ± 18.66; p = 0.022) (Fig. 5). On POD 14, the perfusion recovery rate in group 3 showed a tendency to increase compared to the other groups (group 1 vs group 2 vs group 3: 62.97 ± 11.02 vs 60.69 ± 16.11 vs 72.76 ± 18.66; p = 0.204) (Fig. 5).

[0162] According to the above results, it was confirmed that a pharmaceutical composition containing aspartic acid exhibited a high perfusion recovery rate in rats induced with ischemic disease, and thus had a significantly excellent effect in preventing or treating ischemic disease.

[0163]

[0164] <Example 4> Histological evaluation of capillary density

[0165] For an in-depth analysis of perfusion differences between groups, immunohistochemistry targeting the endothelial cell marker CD31 was performed to quantify the vascular lumen density in muscle samples on POD 14. Specifically, the experiment was performed according to Reference Examples 5 and 7.

[0166] The density of CD31-positive luminal cells, indicating capillary angiogenesis, was significantly higher in LOLA-treated mice compared to the other groups (group 1 vs group 2 vs group 3: 157.25 ± 34.49 vs 162.50 ± 34.75 vs 352.25 ± 44.3; p < 0.001) (Figs. 6 and 7).

[0167] Through the above results, it was confirmed that when a pharmaceutical composition containing aspartic acid is administered, it promotes angiogenesis and exhibits a remarkably excellent preventive or therapeutic effect on ischemic disease, specifically, ischemic heart disease.

[0168]

[0169] <Example 5> Evaluation of VEGF expression

[0170] To further analyze the differences in perfusion between groups, the expression of VEGF, a factor associated with angiogenesis, was analyzed. Specifically, the experiment was performed according to Reference Examples 5, 7, and 8.

[0171] VEGF expression was significantly higher in LOLA-treated rats (group 3) than in the other groups (group 1 vs group 2 vs group 3: 0.76 ± 0.12 vs 0.63 ± 0.14 vs 1.00 ± 0.19; p < 0.001). In addition, immunohistochemical staining for VEGF was enhanced in group 3 compared to the other groups (Figs. 8 to 10).

[0172] Through the above results, it was confirmed that when a pharmaceutical composition containing aspartic acid is administered, it promotes angiogenesis and exhibits a remarkably excellent preventive or therapeutic effect on ischemic disease, specifically, ischemic heart disease.

Claims

1. A pharmaceutical composition for preventing or treating ischemic heart disease, comprising aspartic acid or a pharmaceutically acceptable salt thereof.

2. A pharmaceutical composition according to claim 1, wherein the aspartic acid is a compound represented by the following chemical formula 1: [Chemical Formula 1] .

3. A pharmaceutical composition according to claim 1, further comprising ornithine or a pharmaceutically acceptable salt thereof.

4. In claim 3, the pharmaceutical composition wherein the ornithine is a compound represented by the following chemical formula 2: [Chemical formula 2] .

5. A pharmaceutical composition according to claim 1, wherein the composition reduces the volume of an infarcted area in the myocardium.

6. A pharmaceutical composition according to claim 1, wherein the ischemic heart disease is at least one disease selected from the group consisting of coronary artery disease, stable angina, unstable angina, variant angina, myocardial infarction, heart attack, heart failure, arteriosclerosis, ischemic heart failure, and cardiomyopathy.

7. A pharmaceutical composition according to claim 1, wherein the composition is administered at a dose of 0.1 mg / kg to 1000 mg / kg.

8. Health functional food containing aspartic acid or a food-wise acceptable salt thereof for the prevention or improvement of ischemic heart disease.

9. A health functional food according to claim 8, further comprising ornithine or a food-wise acceptable salt thereof.

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