Methods and compositions for preventing, treating, or alleviating myocardial injury using neuregulins

Abstract

To provide a method and a composition for preventing, treating or delaying myocardial injury in mammals.SOLUTION: Provided are an administration method for, an administration frequency of and an administration dosage of a pharmaceutical formulation or composition for reducing myocardial damage, using neuregulin. It can be proved in a rat myocardial damage model that neuregulin can improve the cardiac function after myocardial infarction, suggesting that neuregulin can be used for preventing, treating or delaying myocardial infarction damage.SELECTED DRAWING: None

Description

[Technical Field]

[0001] FIELD OF THE INVENTION The present invention relates to the manufacture of a medicament for preventing, treating, or ameliorating myocardial damage in a mammal. The present invention also relates to the use of neuregulin (NRG) for cardiac function in mammals. Methods for administering pharmaceutical preparations or compositions containing NRG to prevent, treat, or alleviate muscle damage; In particular, the present invention relates to a method for preventing, treating, or alleviating myocardial damage. and a pharmaceutical composition comprising NRG for the purpose, as well as a method for preventing myocardial damage in mammals. The method, frequency, and administration of pharmaceutical preparations or compositions containing NRG for treating or alleviating and dosage. [Background technology]

[0002] BACKGROUND OF THE INVENTION Cardiovascular problems pose a serious threat to people's lives and health worldwide. There are many types of heart failure, myocardial infarction, coronary heart disease, arrhythmia, cardiomyopathy, and heart valve disease. Cardiovascular diseases include pericardial disease, infective endocarditis, pericardial disease, ischemic heart disease, and congenital heart disease. It tends to cause myocardial damage and affect cardiac function, which can lead to poor health and Myocardial infarction is a cardiovascular disease that seriously endangers human health. As the quality of life continues to improve, the incidence of ischemic myocardial infarction is steadily increasing. Coronary artery occlusion is a condition in which coronary blood supply is suddenly reduced or interrupted due to persistent occlusion of the coronary artery. Ischemic myocardial necrosis induced by severe and chronic myocardial ischemia caused by Ischemic myocardial infarction causes myocardial cell necrosis and scarring, which leads to cardiac Affects function.

[0003] When a myocardial infarction occurs, the coronary arteries are blocked for 20 to 30 minutes, resulting in a lack of blood, which causes myocardial infarction. Part of the cyst becomes necrotic, and the pathological process of myocardial infarction begins. After 2 hours, most of the affected myocardial cells gradually underwent coagulative necrosis, and at the same time, a large amount of inflammation occurred. The myocardial interstitium becomes congested and edematous with infiltration of necrotic cells. The effect is complete after 2 hours. Myocardial fibers dissolve after 1-2 weeks. The necrotic zone is finally dissolved into fibrils after 6 weeks. This is known as old or recovered myocardial infarction.

[0004] The normal function of the left ventricle is significantly affected after a myocardial infarction. When this occurs, the pumping function of the left ventricle is impaired, resulting in a decrease in cardiac output, stroke volume, and blood pressure. , and end-systolic volume may increase and end-diastolic volume may increase within weeks after infarction. .

[0005] Ventricular remodeling is a significant pathological event that occurs after myocardial infarction. The ring is a change that occurs in the structure and morphology of the infarcted and non-infarcted areas of the ventricle after myocardial infarction. The changes in the infarcted area mainly involve the expansion of the infarct, while the changes in the non-infarcted area mainly involve the expansion of the infarct. The classic sign of ventricular remodeling is decompensated ventricular hypertrophy. Changes in ventricular mass, volumetric expansion, and morphological changes lead to dysfunction of the ventricular pump. This can lead to heart failure and progression. The underlying mechanism is ventricular remodeling, a condition that occurs following a myocardial infarction and is persistent and progressive. The severity of these changes determines the patient's cardiac function and prognosis. Ventricular remodeling is a major risk factor affecting cardiac function and endangering human life. It is a type of cardiovascular disease.

[0006] Currently, the main treatment for myocardial infarction is reperfusion (thrombolytic therapy and interventional therapy) in the early stages. method), angiotensin (ANG) II receptor blockers, angiotensin converting enzyme (ACE ) inhibitors and beta-receptor blockers. These reduce infarct size and prevent recurrent myocardial ischemia. It is effective in reducing the risk of pulmonary embolism, improving revascularization, and suppressing excessive ventricular dilation. This will reduce the incidence of chronic heart failure.

[0007] Symptoms of myocardial infarction are closely related to the size and location of the infarct and the status of coronary collateral vessels. The main symptoms include pain, fever, tachycardia, nausea, vomiting, hypotension, shock, and arrhythmia. Major complications of myocardial infarction include papillary muscle dysfunction or rupture, cardiac rupture, and ventricular aneurysm. , embolism, and post-infarction syndrome (PMIS).

[0008] Most of the existing drugs or interventional therapies can only alleviate the symptoms of myocardial infarction, but Damage to the organ tissue cannot be repaired. For patients with advanced myocardial infarction, heart transplantation is recommended. is a last-ditch treatment option that can improve cardiac function and save terminally ill patients, but the donor's scarcity, surgical complexity, immune rejection, and high treatment costs have led to limited success in clinical practice. This makes it difficult to apply widely.

[0009] In summary, myocardial damage caused by cardiovascular diseases poses serious harm to human health. In particular, myocardial infarction, a fatal disease that seriously endangers human health, is a clinical There is a need for safer and more effective drug treatments in practice.

[0010] Neuregulin (NG) or heregulin (HRG), a member of the EGF-like family, binds to NR Structurally similar to each other, including G1, NRG2, NRG3, and NRG4, and their isomers It refers to a group of growth differentiation factors (GDFs), which stimulate breast cancer cell differentiation and milk protein secretion. (Lessor T et al., J Cell Biochem. 1998; 70 (4):587-595); Schwann cells of neural crest cells differentiation into bone marrow cells (Topilko et al., Mol Cell Neurosci, 1996; 8 (2-3): 71-75); Stimulation of acetylcholine receptor synthesis in skeletal muscle cells (Altiok N et al., EMBO.J. 1995; 14 (17): 4258-4266); promotion of cardiomyocyte survival and DM synthesis (Zhao YY et al., I Biol Che r. 1998; 273 (17): 10261-10269). In vivo studies performed on mouse embryos carrying the gene deficiency have demonstrated that NRG regulates cardiac and neural development. proved to be necessary.

[0011] NRG receptors are members of the EF receptor family, which also includes FR, ErbB2, ErbB, and ErbB4. NRG receptors play an important role in cell proliferation, differentiation, and survival. Tyrosine kinase consisting of a main domain, a transmembrane domain, and an intracellular tyrosine kinase domain When NRG binds to the extracellular domain of ErbB3 or ErbB4, it There, a transcriptional change occurs, resulting in ErbB3 / ErbB4 or ErbB2 / ErbB3 heterodimers. The ErbB4 / ErbB4 homodimer or ErbB4 / ErbB4 dimer is formed, and its C-terminus is phosphorylated. The modified C-terminus further binds to downstream signaling proteins within the cell, leading to AKT and / or activates the EK signaling pathway, ultimately leading to cell proliferation, cell differentiation, cell apoptosis, Can trigger a range of cellular responses, such as stimulating or inhibiting cell migration or cell adhesion Among these receptors, ErbB2 and ErbB4 are mainly expressed in cardiac tissue (Zhao YY et al. (References, Circ Res. 1999; 84 (12): 1380-1387).

[0012] Existing evidence suggests that the EGF-like domain of NRG-1, comprising amino acids 50-64, binds to the receptor and It has been shown that it has a high ability to activate ATP (Culousoou JM et al., J Biol Chem. 199 5; 270 (21): 12857-12863). NRG-1β can bind with high affinity to ErbB3 and ErbB4. B2 can form heterodimers with ErbB3 or ErbB4, and its affinity for its ligands The affinity of ErbB3 or ErbB4 homodimers for their ligands is higher than that of ErbB3 or ErbB4 homodimers. Studies have shown that signaling via NRG-1β, ErbB2, and ErbB3 is essential for the development of the sympathetic nervous system. It has been confirmed that (Britsch S et al., Dienes Dev. 1998; 12 (12): Loss of expression of NRG-1β or ErbB2 or ErbB4 results in cardiac developmental disorders. Embryonic lethality occurs (Gassmann M et al., Nature, 1995; 378(6555): 390-394). Recent studies have shown that NRG-1β, ErbB2, and ErbB4 are not only essential for cardiac development but also It has also been shown to play a crucial role in maintaining adult cardiac function (Kura Mochi Y et al., J Mol Cell Cardia 1. 2006; 41 (2): 228-235). NRG-1β is expressed in adults. It has been proven that it can enhance the formation of myocardial sarcomeres in various cardiac disorders. In all animal models, ingestion of the NRG-1β EGF-like domain improved cardiac function, It has been found that cardiac dysfunction can be prevented (Liu et al., J Am Coll Cardiol. 2006; 48: 1438-1447). Clinical trials have also shown that NRG is effective in treating chronic rheumatoid arthritis caused by various etiologies. It has been shown to have a therapeutic effect on chronic heart failure and significantly enhances cardiac function (CN200910057390.5). In an animal model of cerebral ischemia-reperfusion, NRG-1 showed a significant protective effect on brain cells. , inhibited apoptosis of brain cells, enhanced neuronal function, and reduced infarct size (Li (Q et al., Neurosci Lett. 2008; 443 (3): 155-159). Cardiac ischemia-reperfusion induces the upregulation of NRG-1. This induces release of ATP and activates the NRG / ErbB signaling pathway in cardiomyocytes (Kuramochi Y et al., J Biol Chem. 2004; 279 (49): 51141-51147), and NRG-1 inhibits cardiac ischemia-reperfusion There is evidence that it plays a role in the prevention, treatment or mitigation of injuries (WO2011091723) .

[0013] Myocardial injury is a fatal disease that seriously endangers human health. The method, frequency, and dosage of NRG-1 for this purpose have not yet been clearly determined. The present invention provides a method for addressing the above needs and a pharmaceutical composition containing NRG therefor. It is worth mentioning that the present invention provides a particularly optimized frequency of administration, and the present invention provides a particularly optimized method of administration. The present invention further provides a method for the preparation of a drug for preventing, treating, or alleviating myocardial damage in a mammal. The present invention relates to the use of NRG in the manufacture of a therapeutic agent for the treatment of myocardial infarction. The present invention provides a particularly optimized dosage and the present invention provides a particularly optimized It provides an optimized administration method. Summary of the Invention

[0014] (Detailed explanation) (A. Overview) The present invention relates to the manufacture of a medicament for preventing, treating, or ameliorating myocardial damage in a mammal. The present invention further provides a method for preventing and treating myocardial injury in a mammal. For the use of NRG for the manufacture of a medicament for the treatment or alleviation of NRG improves cardiac function affected by myocardial injury and reduces cardiac remodeling in humans. It can be done.

[0015] Many cardiovascular diseases, such as heart failure, myocardial infarction, coronary arteriosclerotic heart disease, arrhythmia, myocarditis, Heart valve disease, infective endocarditis, pericardial disease, ischemic heart disease, congenital heart disease, etc. can cause myocardial damage. Myocardial damage can affect cardiac function and put human health at risk. Infarction is accompanied by persistent coronary artery occlusion, usually accompanied by apoptosis and necrosis of myocardial cells, massive inflammation, and Myocardial infarction is accompanied by the infiltration of inflammatory cells and myocardial fibrosis, which induce myocardial injury. It tends to cause cardiac dysfunction and therefore affects human health.

[0016] The present invention demonstrates that NRG is essential for cardiac development and is crucial for maintaining cardiac function in adults. This invention is based on the scientific discovery that NRG plays a key role in the regulation of sarcomere in cardiomyocytes. Based on the scientific discovery that cellular cytoskeleton and cell-cell junction formation can be strengthened; The invention demonstrates that NRG inhibits the heart function of animals or patients with heart failure in various animal models and clinical trials. This invention is based on the scientific discovery that NRG can enhance cerebral ischemia-reperfusion Based on scientific findings that it exerts a protective effect on brain cells in animal models of aspiration; Akira has shown that NRG exerts a protective effect on brain cells in an animal model of cardiac ischemia-reperfusion. Based on scientific discoveries; NRG, NRG polypeptides, and mutants of NRG or with NRG-like functions All other conjugates that do so are within the scope of the present invention.

[0017] In a first aspect, the present invention provides a method for preventing, treating, or alleviating myocardial damage in a mammal. The present invention further provides a pharmaceutical preparation for preventing damage caused by myocardial infarction in a mammal. The present invention provides pharmaceutical preparations for the prevention, treatment, or palliative of a disease in a mammal, the mammal being preferably a human. The pharmaceutical preparation contains an effective amount of NRG or a functional fragment thereof, or a nucleic acid encoding NRG. or functional fragments thereof, or substances that increase the production and / or function of NRG, and pharmaceuticals The pharmaceutical preparations include carriers, excipients, etc. that are acceptable for the purpose of preventing, treating, or preventing myocardial injury. It can be used in combination with other drugs or therapies to alleviate or mitigate the symptoms of rheumatoid arthritis. In such cases, pharmaceutical preparations containing NRG are effective in increasing the EF value of the mammalian left ventricle. In another embodiment, the pharmaceutical preparation comprising NRG is administered to increase left ventricular end-diastolic volume (LVEDV) or left ventricle end-diastolic volume (LVEF). In another embodiment, the pharmaceutical preparation containing NRG is effective in reducing ventricular end-systolic volume (LVESV). The product is injected subcutaneously via a syringe or other device. In another embodiment, the product contains NRG. The pharmaceutical preparation is injected subcutaneously via a pump, such as an injection pump. In an embodiment, the syringe pump is a micropump. , the micropump is an insulin pump. It is worth mentioning that the present invention is The pharmaceutical preparation is suitable for any preparation available as NRG, and the pharmaceutical preparation comprises NRG as described above or and a pharmaceutically acceptable excipient, diluent, or carrier. Pharmaceutical preparations that may be used include, but are not limited to, those described herein.

[0018] In a second aspect, the present invention provides a method for preventing, treating, or alleviating myocardial damage in a mammal. The present invention further provides a method for preventing damage from myocardial infarction in a mammal, The present invention provides a method for treating or alleviating the symptoms of a rheumatoid arthritis, wherein the mammal is preferably a human. an effective amount of NRG or a functional fragment thereof, a nucleic acid encoding NRG or a functional fragment thereof or the production of NRG required for the prevention, treatment, or mitigation of myocardial injury in mammals. The method involves the use of substances that increase the function of other drugs, particularly effective amounts of NRG or its derivatives. a nucleic acid encoding NRG or a functional fragment thereof, or a nucleic acid encoding NRG or a functional fragment thereof, Combination of substances that increase the production and / or function of NRG to prevent, treat, or alleviate muscle damage. They can be used in combination.

[0019] In a third aspect, the present invention provides a method for preventing, treating, or alleviating myocardial damage in a mammal. The present invention further provides a composition for preventing damage from myocardial infarction in a mammal. The present invention provides a pharmaceutical composition for treating or ameliorating a disease characterized by a pulmonary embolism, in which the mammal is preferably a human. The pharmaceutical composition may comprise a compound according to the present invention for preventing, treating, or ameliorating myocardial damage in a mammal. and other drugs for preventing, treating, or ameliorating myocardial damage. The pharmaceutical composition contains an EGF-like domain, which binds to and activates the receptor. It has been proven that it is possible to However, the NRG provided by the present invention is, for example, a fragment of the NRG-1β2 isomer, 177-2 It contains 37 amino acids. The amino acid sequence of this fragment is as follows: [ka]

[0020] In a fourth aspect, the present invention provides a method for preventing, treating, or alleviating myocardial damage in a mammal. The present invention further provides a pharmaceutical preparation comprising NRG for use in administering to mammals. medicines containing NRG for use in preventing, treating, or mitigating damage caused by myocardial infarction in A dosage of the pharmaceutical preparation is provided, and the mammal is preferably a human. An effective dose is defined as the amount of NRG means that one or more beneficial effects can be achieved when the dose is applied to a mammal. The beneficial effects are thought to improve cardiac function in patients with myocardial damage and prevent further deterioration of that cardiac function. to prevent progression of cardiac dysfunction or to inhibit the progression of cardiac dysfunction that may be caused by myocardial damage; The dosage provided to mammals by the present invention is 0.1 μg / kg / day (protein / body). In one embodiment, the dose is 0.3 μg / kg to 360 μg / kg / day (protein / body weight). / day (protein / body weight) to 50 μg / kg / day (protein / body weight); in one embodiment In one embodiment, the effective dose is 7.5 μg / kg / day; in one embodiment, the effective dose is 15 μg / kg / day In another embodiment, the effective dose is 30 μg / kg / day.

[0021] In a fifth aspect, the present invention provides a method for preventing, treating, or alleviating myocardial damage in a mammal. The present invention further provides a method for administering a pharmaceutical preparation containing NRG to a mammal. Methods of administering pharmaceutical preparations containing NRG to prevent, treat, or alleviate damage caused by myocardial infarction The mammal is preferably a human. The pharmaceutical preparation may be administered orally, rectally, topically, or intravenously. Oral administration, inhalation administration, buccal administration (e.g., sublingual administration), parenteral administration (e.g., subcutaneous injection, intramuscular administration) Intake by intramuscular injection, intradermal injection, or intravenous injection), transdermal administration, or other suitable method In one embodiment, NRG is administered only once a day. In one embodiment, NRG is administered multiple times daily. In another embodiment, NRG is administered daily. In some embodiments, this tolerated dose of NRG is administered within a few days. In another embodiment, NRG is administered two days a week, In another embodiment, NRG is administered two days a week, one day a week, and one day a week. In another embodiment, NRG is administered subcutaneously three times daily for many consecutive weeks. In another embodiment, NRG is administered subcutaneously three times daily for a number of consecutive days. In another embodiment, NRG is administered subcutaneously three times daily for 38 consecutive days. In another embodiment, NRG is administered subcutaneously three times daily for 49 consecutive days. In another embodiment, NRG is administered subcutaneously three times daily for 60 consecutive days. In another embodiment, NRG is administered three times daily for more than 35 consecutive days. In another embodiment, NRG is administered subcutaneously multiple times per day for many consecutive days. In another embodiment, NRG is administered multiple times per day in a number of consecutive doses. Administered over consecutive days, then slowly discontinued over 3 weeks: Administered every other day during the first week and every 3 days for the second week; and every 4 days for the third week. NRG was administered subcutaneously three times daily for more than 38 consecutive days, then slowly discontinued. In another embodiment, NRG is administered subcutaneously three times daily for 49 consecutive days, followed by administration over a three-week period. Slowly discontinue: Week 1, every other day; Week 2, every 3 days; Week 3, every 4 days In another embodiment, NRG is administered subcutaneously multiple times per day for many consecutive days. In another embodiment, NRG is administered 3 times a day, followed by a slow daily taper. Once a day, over many consecutive days, the daily dose is then slowly reduced. In one embodiment, NRG is administered subcutaneously three times daily for more than 60 consecutive days, and then In another embodiment, NRG is administered three times a day for 60 consecutive days. Subcutaneous injection over a period of 1 week, then slowly discontinued over 3 weeks. The daily dose in week 1 is The daily dose in week 2 should be one-quarter of the continuous dose; the daily dose in week 3 should be one-quarter of the continuous dose. The daily dose should be one-eighth of the continuously administered dose.

[0022] The present invention also provides a kit for preventing, treating, or mitigating myocardial damage in a mammal. The present invention also provides a method for preventing, treating, or alleviating damage caused by myocardial infarction in a mammal. The kit includes a kit for preventing myocardial injury, the kit being for preventing myocardial injury, and the mammal is preferably a human. a single or multiple dose of said pharmaceutical preparation or composition for the prevention, treatment, or palliative of and instructions for use of the pharmaceutical preparation or composition.

[0023] The pharmaceutical preparations or compositions provided by the present invention may be administered before, during, or after the onset of heart disease. When used for prophylaxis, the pharmaceutical preparation or composition generally When used in therapy, the pharmaceutical preparation or composition generally is administered to treat a heart disease. In one embodiment, the pharmaceutical preparations provided herein are administered during or after the onset of the condition. In another embodiment, the product or composition is administered prior to the onset of heart disease. The provided pharmaceutical preparations or compositions are administered when a myocardial infarction occurs. In the present invention, the pharmaceutical preparation or composition is administered after the onset of heart disease. do.

[0024] The pharmaceutical preparations or compositions provided by the present invention can be administered orally, rectally, topically, Inhalation administration, buccal administration (e.g., sublingual administration), parenteral administration (e.g., subcutaneous injection, intramuscular injection) The drug may be administered by intradermal, intradermal, or intravenous injection), transdermal administration, or other suitable method. Subcutaneous injection can be performed using a syringe, a pump (microinjector pump), or another administration device. The dosage forms of the pharmaceutical preparations or compositions provided by the present invention include Examples include, but are not limited to, tablets, troches, cachets, dispersions, suspensions, solutions, and capsules. , ointments, and similar forms.

[0025] (B. Definition) Unless otherwise defined, all scientific and technical terms used herein are understood to be of ordinary skill in the art. All patent documents, patent application documents, Published patent documents and other publications are cited by reference. If a definition of a term has a different meaning from that explained in the document, the definition given in this section shall prevail. The explanation shall take precedence.

[0026] Unless otherwise specified, as used herein, "a" / "an" means "at least one" means "one" or "one or more."

[0027] As used herein, "mammal" refers to non-human primates (bovine, porcine, equine, feline, canine, It refers to mammals such as rats, mice, etc. or primates (monkeys, humans), preferably humans.

[0028] As used herein, "myocardial damage" refers to heart failure, myocardial infarction, coronary arteriosclerotic heart disease, arrhythmic heart disease, myocardial infarction ... Cardiac vein disease, cardiomyopathy, valvular heart disease, infective endocarditis, pericardial disease, ischemic heart disease, or congenital heart disease It refers to certain types of myocardial damage caused by pathological heart diseases such as Myocardial damage tends to cause dysfunction, thereby affecting human health. The causes are oxyradical production, calcium overload, and inflammatory reactions due to neutrophil infiltration into the injured area. response, apoptosis or necrosis of cardiomyocytes, tissue damage caused by energy supply imbalance metabolic disorders, abnormal cardiac signaling, cholesterol accumulation, and atherosclerotic plaque formation It is associated with multiple pathophysiological changes, including vascularization.

[0029] As used herein, "neuregulin" or "NRG" refers to any of ErbB2, ErbB3, ErbB4, or HerbB1. A protein or polypeptide that can bind to and activate a heterodimer or homodimer. NRG refers to peptides. NRG includes NRG isoforms, NRG EGF-like domains, and NRG EGF-like domains. and polypeptides containing NRG, mutants or derivatives of NRG, and those capable of activating the receptor. NRG also includes other gene products of NRG that can NRGs include polypeptides, fragments, and complexes with NRG-like functions. Preferably, NRGs are ErbB-like proteins. Proteins that can bind to and activate ErbB2 / ErbB4 or ErbB2 / ErbB3 heterodimers As an example, but not by way of limitation, The NRG (rhNRG) provided by the invention is a fragment of the NRG-1β2 isomer containing the EGF-like domain. The amino acid sequence of this fragment is as follows: R: [ka] The NRG used in the present invention activates the above receptor and regulates its biological function. For example, it can stimulate skeletal muscle cells to synthesize acetylcholine receptors; NRGs can promote differentiation, survival, and DNA synthesis. They have substantial effects on biological functions. As will be apparent to those skilled in the art, conservative NRG mutations that do not confer essential functions are also included. Mutation of a single amino acid in a sequence does not alter the biological function of a protein or polypeptide. No changes occur in the gene (Watson et al., "Molecular Biology of the Gene"). Gene), 4th edition, 1987, The Bejacmin / Cummings Pub. Co., p. 224). NRGs can be extracted from natural sources or obtained by recombinant technology, artificial synthesis, or other means. You can also obtain it.

[0030] As used herein, the term "EGF-like domain" refers to any of ErbB2, ErbB3, ErbB4, or heterodimers thereof. It is capable of binding to and activating a dimer or homodimer, and has been described in the following literature: WO 00 / 64400; Holmes et al., Science, 256: 1205-1210 (1992); U.S. Pat. No. 5,530,109 and 5,716,930; Hijazi et al., Int. J. Oncol., 13: 1061-1067 (1998); Chang et al., Nature, 387: 509-512 (1997); Carraway et al., Nature, 387: 512-516 (19 97); Higashiyama et al., J. Biochem., 122: 675-680 (1997); and WO 97 / 09425. The polypeptide encoded by the NRG gene has a structure similar to the EGF receptor binding zone described above. In some embodiments, the EGF-like domain refers to a peptide fragment of ErbB2 / ErbB4 or ErbB In some embodiments, E The GF-like domain comprises amino acids in the receptor binding zone of NRG-1. In the context of the invention, the EGF-like domain refers to amino acids 177-226, 177-237, or 177-240 of NRG-1. In some embodiments, the EGF-like domain is an amino acid sequence in the receptor binding zone of NRG-2. In some embodiments, the EGF-like domain comprises the receptor binding zone of NRG-3. In some embodiments, the EGF-like domain comprises the amino acid sequence of In some embodiments, the EGF-like domain comprises amino acids in the endothelial-binding zone. , including the amino acid sequence described in U.S. Pat. No. 5,834,229: Ala Glu Lys Glu Lys Thr Phe C ys Val Asn Gly Gly Glu Cys Phe Met Val Lys Asp Leu Ser Asn Pro.

[0031] NRG can be administered orally, rectally, topically, by inhalation, bucally (e.g., sublingually), Parenteral administration (e.g., subcutaneous, intramuscular, intradermal, or intravenous injection), transdermal administration, or other suitable means for incorporating the drug. The preferred route of administration of the moiety will depend on the condition and severity to be treated, and the characteristics of the particular NRG used. NRG can be administered alone, or more preferably, It may be administered with any pharmaceutically acceptable carrier or excipient. Suitable pharmaceutically acceptable carriers or excipients are available for use in this method (Remington: Pharmaceutical Sciences). The Science and Practice of Pharmacy (Remington: Alfonso R. Gennar) o (ed.) Mack Publishing Company, April 1997).

[0032] As used herein, a "pump" refers to a pump that delivers pharmaceutical fluids, drugs, proteins, and / or other compositions. A pump is a device for administering substances subcutaneously. It is used for continuous, accurate, and metered administration. The pump is equipped with a subcutaneous catheter for continuous subcutaneous infusion. The catheter can be placed externally or the catheter port can be connected to a pump mechanism. Microinfusion pumps are portable, easy-to-use devices that can be used for precise injections. For example, an insulin pump is a device that delivers insulin during the treatment of diabetes or other diseases. Insulin pumps are medical devices used to administer insulin or other medications. It is thought that insulin pumps are used to deliver insulin subcutaneously continuously. , a disposable thin-walled plastic pipe or catheter can be attached, so that A catheter is inserted under the skin to allow insulin or other drugs to enter the tissue. The pump can be an external device that can be connected to the patient, or it can be placed inside the patient's body. External pumps can be attached to devices that can be implanted in hospitals, clinics, etc. This refers to equipment designed for use in a fixed location, such as a lock, or similar location. This refers to ambulatory or portable devices such as pumps or similar devices that can be carried by the patient. The pump is a liquid medium capable of storing a liquid medium, including, but not limited to, NRG. Equipped with a reservoir.

[0033] The external pump may be connected to the patient, for example via fluid passage through suitable hollow tubing. The hollow tube can be connected to a hollow needle, which penetrates the patient's skin for injection. Alternatively, the hollow tube may be used to penetrate the patient through a cannula or similar object. An external pump can be worn by the patient or attached to the patient's clothing. The pump can be attached to the patient's body or underneath their clothing. Microinfusion pumps that can be used for high frequency infusions, such as, but not limited to, the MiniMed Paradigm 522 Insulin pump, MiniMedParadigm722 insulin pump, MiniMedParadigm515 insulin pump Insulin Pump, MiniMed Paradigm 715 Insulin Pump, MiniMed Paradigm 512R Insulin Pump MiniMed Paradigm 712R insulin pump, MiniMed Paradigm 508 insulin pump, and and MiniMedParadigm 508R insulin pump (Medtronic, Northridge, Canada), and This refers to other similar devices known in the art.

[0034] U.S. Patent 11 / 211,095 (filing date: 8 / 23 / 2005, publication number: US2006 / 0264894, registration number US768 6787) and published PCT applications WO01 / 70307 (PCT / US01 / 09139), WO04 / 030716 (PCT / US2003 / 028769 ), WO04 / 030717 (PCT / US2003 / 029019) and WO2013075622 (PCT / CN2012 / 0849 36), U.S. Pat. No. US2005 / 0065760 (Method for Advising Patients on Insulin Doses) (Visiting Patients Concerning Doses of Insulin) and US6,589,229 (Wearable Implants) Wearable Self-Containing Drug Infusion Device Examples of administration devices such as internal pumps are cited in this section.

[0035] As used herein, "a compound that can be used to prevent, treat, or mitigate myocardial damage" means a compound that can be used to prevent, treat, or mitigate myocardial damage. "Other drugs or therapies applicable to the treatment of myocardial injury" refers to drugs and interventional therapies generally applicable to the treatment of myocardial injury, as well as refers to drugs and interventional therapies generally applicable to the treatment of myocardial infarction and damage caused by myocardial infarction. Medications to treat myocardial infarction include antiplatelet drugs (aspirin, clopidogrel, etc.) , anticoagulants (heparin, bivalirudin, etc.), thrombolytic agents (alteplase, tenectep enzyme, urokinase, recombinant human pro-urokinase, etc.), lipid-lowering drugs (statins , cholesterol absorption inhibitors), angiotensin-converting enzyme inhibitors / ANG II receptor blockers , beta-receptor blockers, calcium channel blockers, nitrates, phosphatase inhibitors, Diuretics, renin-angiotensin-aldosterone system (RAS) antagonists, myocardial energy Energy optimizer, ischemic tissue metabolism improver, free radical scavenger Interventional therapies include coronary interventional therapies. [Brief explanation of the drawings]

[0036] BRIEF DESCRIPTION OF THE DRAWINGS [Figure 1] 1 shows echocardiographic results of the effect of chronic subcutaneous administration of NRG at various doses in the treatment of rats with acute myocardial infarction. [Figure 2] 1 shows echocardiographic results of the effect of NRG administered chronically subcutaneously at various frequencies in the treatment of rats with acute myocardial infarction. [Figure 3] 1 shows the therapeutic effect of NRG on acute myocardial infarction in rats through long-term subcutaneous administration followed by discontinuation due to reduced frequency. [Figure 4]1 shows the therapeutic effect of NRG on acute myocardial infarction in rats through chronic subcutaneous administration followed by discontinuation with dose reduction. [Example]

[0037] (Example) Example 1: Therapeutic effect of long-term subcutaneous administration of various doses of rhNRG on acute myocardial infarction in rats (Study on the effect-dose relationship of NRG) (1. Purpose) Effect of left coronary artery ligation on acute myocardial infarction in rats Treatment of acute myocardial infarction in rats by observing the therapeutic effects of various doses of rhNRG To investigate the dose-effect relationship of NRG on (2. Experimental Drugs) (2.1 Excipients: Developed by Zensun (Shanghai) Sci & Tech Co., Ltd.) (2.2 rhNRG: Developed by Zensun (Shanghai) Sci & Tech Co., Ltd.)

[0038] (3. Experimental Animals) 3.1 Strain and Source: Wistar rats, Shanghai Sippe-Bk Lab Animal Co., Ltd. (More supplied) (3.2 Sex, weight and certificate: Male, 200-270g)

[0039] (4. Experimental Materials and Equipment) Anesthesia equipment and isoflurane evaporator manufactured by MSS INTERNATIONAL LTD. Isoflurane, 100ml / bottle, manufactured by RWD Life Technologies Co., Ltd. Vivid E95 cardiac ultrasound detector. Ningbo Lingqiao suture needle (with thread) manufactured by Ningbo Medical Needle Co., Ltd.

[0040] (5. Experimental Methods) 5.1 Establishment of a rat model of heart failure induced by coronary artery ligation The rats were anesthetized with isoflurane through a gas anesthesia device. Then, the rats were placed in a supine position. After shaving the chest, the skin was disinfected with 75% alcohol. After incising the left anterior chest skin, The muscles were bluntly separated to expose the 4th and 5th ribs. Using a hemostat, the 4th and 5th ribs were The muscles between the ribs were bluntly cut. Using both hands, the heart was pushed out of the chest cavity, and lung expansion and The heart was fully exposed for observation of the heartbeat. The left atrial appendage and pulmonary artery cone were fully exposed. The left anterior descending coronary artery (LADCA) between them was ligated with a surgical suture. The organs were quickly returned to their original positions. Then, the pectoral muscles and skin were sutured. After the surgery, the rats were placed in a cage. The animals were returned to their cages, fed, and observed closely.

[0041] 5.2 Grouping and Administration Table 1. Grouping of experimental animals and administration schedule [Table 1] Administration was initiated on the day after the animal model of myocardial infarction was established.

[0042] (5.3 Observation Indicators) After anesthesia with 4% isoflurane, the rat was fixed on the operating table in the left lateral position. The animals were attached to a breathing mask attached to a gas anesthesia machine, and isoflurane was used to maintain anesthesia. The skin was disinfected with 75% alcohol and coated with a coupling agent. All echo signals from the rat left ventricle were detected using the left ventricle end-diastolic diameter and left ventricle end-systolic diameter. The left ventricular end-diastolic volume (EDV) and end-systolic volume (ESV) were calculated. Ejection fraction (EF) values ​​were also obtained: EF = (EDV-ESV) / EDV × 100%.

[0043] (5.3.2 Data Processing) All experimental data are expressed as ±SD.

[0044] (6. Experimental Results) (6.1 Echocardiography Results) Echocardiography was performed 60 days after continuous administration of NRG. The results showed that the LVEDd in the vehicle group, LVEDs and EF values ​​were 0.971±0.07cm, 0.832±0.08cm, and 34.6±7.00%, respectively; 15 The LVEDd, LVEDs, and EF values ​​of the μg / kg-NRG group were 0.975 ± 0.07 cm, 0.794 ± 0.10 cm, and 0.975 ± 0.07 cm, respectively. 42.9±11.32%; LVEDd, LVEDs, and EF values ​​in the 7.5 μg / kg-NRG group were 0.965±0.0, respectively. 7 cm, 0.808 ± 0.11 cm, and 38.4 ± 12.17% in the 3.75 μg / kg-NRG group; The EF values ​​were shown to be 0.994±0.08cm, 0.839±0.12cm, and 37.0±12.23%, respectively. According to the data on LVEDd and LVEDs, LVEDd and LVEDs may be decreased in the high-dose NRG group. EF According to the data, cardiac function of rats in the high-, medium-, and low-dose groups was significantly improved by continuous administration. The results were improved 60 days after the initial administration, and there was a dose-effect relationship between the three groups. For details, see Table 2 and Figure 1. Please refer to. Table 2. Efficacy of various doses of NRG administered subcutaneously for 60 days in the treatment of acute myocardial infarction in rats. Echocardiographic results of the patients (x±SD) [Table 2]

[0045] (7. Conclusion) After 60 days of treatment with rhNRG, 5 μg / kg, 2.5 μg / kg, or 1.25 μg / kg of NRG were administered subcutaneously three times daily. The EF values ​​of the irradiated treatment group were higher than those of the control group, and a specific dose-effect relationship was observed among the three doses. there were.

[0046] Example 2: Therapeutic effects of long-term subcutaneous administration of rhNRG at various frequencies on acute myocardial infarction in rats fruit) (1. Purpose) In a rat model of myocardial infarction induced by left coronary artery ligation, Long-term subcutaneous administration of rhNRG was used to evaluate the therapeutic effects of specific doses of rhNRG on acute myocardial infarction in rats. To research.

[0047] (2. Experimental Drugs) (2.1 Excipients: Developed by Zensun (Shanghai) Sci & Tech Co., Ltd.)

[0048] (2.2 rhNRG: Developed by Zensun (Shanghai) Sci & Tech Co., Ltd.)

[0049] (3. Experimental Animals) 3.1 Strain and Source: Wistar rats, Shanghai Sippe-Bk Lab Animal Co., Ltd. (More supplied)

[0050] (3.2 Sex, weight and certificate): Male, 200-270g.

[0051] (4. Experimental Materials and Equipment) This is the same as "4. Experimental materials and equipment" in Example 1.

[0052] (5. Experimental Methods) 5.1 Establishment of a rat model of heart failure induced by coronary artery ligation Same as "5.1 Establishment of a rat model of heart failure induced by coronary artery ligation" in Example 1 .

[0053] 5.2 Grouping and Administration Table 3. Grouping of experimental animals and administration schedule [Table 3]

[0054] All experimental animals were randomly divided into groups after coronary artery ligation. The rats were divided into four groups based on body weight: vehicle group (control group), NRG 30 μg / kg / day group, and NRG 30 μg / kg / BIW group. The animals were randomly divided into two groups: one receiving 30 μg / kg / day of NRG and the other receiving 30 μg / kg / day of NRG for 7 days. For the first three and four groups, rats were given the test substance three times a day for the first seven days. The animals were dosed according to their body weight at a daily dose of 30 μg / kg / day. For the fourth group, rats were injected with NRG once a week for the last four weeks at a daily dose of 30 μg / kg. It was decided.

[0055] (5.3 Observation Indicators) (5.3.1 Cardiac Function Tests) After anesthesia with 4% isoflurane, the rat was fixed on the operating table in the left lateral position. The animals were attached to a breathing mask attached to a gas anesthesia machine, and isoflurane was used to maintain anesthesia. The skin was disinfected with 75% alcohol and coated with a coupling agent. All echo signals from the rat left ventricle were detected using the left ventricle end-diastolic diameter and left ventricle end-systolic diameter. The left ventricular end-diastolic volume (EDV) and end-systolic volume (ESV) were calculated. The ejection fraction (EF) value was also obtained. EF-(EDV-ESV) / EDV×100%. The cardiac function of the rats was evaluated by the cardiac Echocardiography was performed at 1, 2, 3, and 5 weeks after the onset of myocardial infarction.

[0056] (5.3.2 Data Processing) All experimental data are expressed as ±SD.

[0057] (6. Experimental Results) (6.1 Echocardiography Results) Echocardiography was performed after 35 days of continuous NRG administration. , and EF values ​​were 0.925±0.084cm, 0.756±0.107cm, and 42.5±10.174%, respectively; NRG The LVEDd, LVEDs, and EF values ​​in the 30 μg / kg / day group were 0.879 ± 0.058 cm, 0.694 ± 0.077 cm, respectively. and 47.9±8.342%; LVEDd, LVEDs, and EF values ​​in the NRG / 30μg / kg / BIW group were 0.9 and 0.9%, respectively. 28±0.084cm, 0.746±0.110cm, and 45.2±10.248% in the NRG / 30μg / kg / day × 7 + QW group. VEDd, LVEDs, and EF values ​​were 0.931±0.070cm, 0.760±0.097cm, and 42.7±9.892%, respectively. It was.

[0058] As shown in the LVEDd and LVEDs data, after 35 days of continuous administration, NRG / 30μ g / kg / day significantly reduced LVEDd and LVEDs; The F value was significantly higher in the NRG / 30 μg / kg / day group than in the control group; the EF value was significantly higher in the NRG / 30 μg / kg / BIW group than in the control group. The NRG / 30μg / kg group showed a tendency to increase compared to the control group, and the NRG / 30μg / kg group showed a tendency to increase after the first 7 days of continuous administration. The cardiac function of rats in the g / day × 7 + QW group was improved to some extent compared with that of the control group, i.e. The cardiac function showed a tendency to improve. After that, injections were given every seven days to maintain the effect. See Table 4 and Figure 2 for the results.

[0059] Table 4. The therapeutic effect of NRG on myocardial infarction in rats through long-term subcutaneous administration at various frequencies. Echocardiogram results (x±SD) [Table 4]

[0060] Example 3: Long-term subcutaneous administration of α-amyloid to acute myocardial infarction in rats and subsequent reduction in the frequency of α-amyloid Therapeutic effect of rhNRG through discontinuation (1. Purpose) In a rat model of myocardial infarction induced by left coronary artery ligation, long-term administration and The therapeutic effect of rhNRG on acute myocardial infarction in rats through subsequent interruption by reduced frequency Observe the results.

[0061] (2. Experimental Drugs) (2.1 Excipients: Developed by Zensun (Shanghai) Sci & Tech Co., Ltd.) (2.2 rhNRG: Developed by Zensun (Shanghai) Sci & Tech Co., Ltd.)

[0062] (3. Experimental Animals) 3.1 Strain and Source: Wistar rats, Shanghai Sippe-Bk Lab Animal Co., Ltd. (More supplied) (3.2 Sex, weight and certificate): Male, 200-270g.

[0063] (4. Experimental Materials and Equipment) This is the same as "4. Experimental materials and equipment" in Example 1.

[0064] (5. Experimental Methods) 5.1 Establishment of a rat model of heart failure induced by coronary artery ligation This is the same as "5.1 Establishment of a rat model of heart failure induced by coronary artery ligation" in Example 1.

[0065] 5.2 Grouping and Administration After coronary artery ligation, rats were randomly divided into groups and administered medication. They were randomly divided into two groups, including a vehicle group and a NRG 30 μg / kg group. The vehicle group contained 19 rats. There were 18 rats in the NRG group, and 10 μg / kg of NRG was administered three times a day from the day after the model was created. Continuous administration of the drug was initiated by subcutaneous administration at a dose of 0.01 mg / kg. All animals were continuously dosed until day 38, and animals in the NRG group were examined by echocardiography. The animals in the NRG group were divided into two subgroups on average, with one subgroup receiving the induced The animals in the vehicle group continued to receive the drug, while the animals in the other subgroup were discontinued early. For the subgroup receiving continuous NRG, a 3-week withdrawal schedule was implemented on Day 49: NRG was administered subcutaneously every other day during the first week, every third day during the second week, and every fourth day during the third week. In terms of administration method, rats were injected subcutaneously with NRG three times a day in exactly the same manner as above. For the NRG-discontinued subgroup, rats were observed for clinical signs. All animals showed cardiac dysfunction. Echocardiography was performed weekly to monitor changes in cardiac function.

[0066] (5.3 Observation Indicators) (5.3.1 Cardiac Function Tests) After anesthesia with 4% isoflurane, the rat was fixed on the operating table in the left lateral position. The animals were attached to a breathing mask attached to a gas anesthesia machine, and isoflurane was used to maintain anesthesia. The skin was disinfected with 75% alcohol and coated with a coupling agent. All echo signals from the rat left ventricle were detected using the left ventricle end-diastolic diameter and left ventricle end-systolic diameter. The left ventricular end-diastolic volume (EDV) and end-systolic volume (ESV) were calculated. Ejection fraction (EF) values ​​were also obtained: EF = (EDV - ESV) / EDV × 100%.

[0067] (5.3.2 Data Processing) All experimental data were expressed as ±SD. One-way ANOVA analysis was performed using GraphPad Prism 6. P<0.05 indicates a significant difference between groups. P<0.01 indicates a significant difference between groups.

[0068] (6. Experimental Results) (6.1 Echocardiography Results) Echocardiography was performed after 35 days of continuous administration of NRG. LVEDd, LVED s, and EF values ​​were 0.988±0.08cm, 0.850±0.10cm, and 33.6±11.36%, respectively; in the NRG group The LVEDd, LVEDs, and EF values ​​were 0.953±0.05cm, 0.767±0.06cm, and 44.9±6.09%, respectively. The results showed that NRG significantly reduced LVEDd and LVEDs, enhancing cardiac contractile function. This indicates that left ventricular remodeling can be reversed. The LVEDd, LVEDs, and EF values ​​were 1.020±0.10cm, 0.881±0.15cm, and 33.1±14.55%, respectively. The LVEDd, LVEDs, and EF values ​​in the NRG-discontinued subgroup were 0.987 ± 0.05 cm, 0.807 ± 0.06 cm, and 4 2.2±5.48%, while the LVEDd, LVEDs, and EF values ​​in the NRG continuous administration subgroup were 0.973± 0.07 cm, 0.783 ± 0.08 cm, and 45.0 ± 5.51%; these results suggest that abrupt interruption of NRG may This indicates that NRG has some effect on cardiac function. Echocardiography was performed at week 2 after drug discontinuation. and EF values ​​were 1.043±0.06cm and 0.887±0.06, respectively. cm , 35.4±6.78%; during graded NRG The LVEDd, LVEDs, and EF values ​​of the dialysis subgroup were 0.989±0.07cm and 0.814±0.08cm, respectively. cm and 41.3±4. The rate was 92%. A significant difference was observed between the group receiving the vehicle and the control group. Echocardiography was performed three weeks after discontinuation. The LVEDd, LVEDs, and EF values ​​in the early NRG discontinuation subgroup were 1.010±0.06cm, 0.842±0.06cm, and 0.842±0.06cm, respectively. 38.9±5.04%; LVEDd, LVEDs, and EF values ​​in the gradual NRG discontinuation subgroup were 0.976±0. 0.06 cm, 0.805 ± 0.07 cm, and 40.8 ± 4.67%. Compared with the vehicle group, the cardiac function of rats was significantly improved. The effect of gradual discontinuation of NRG on the variability of variability was moderated. See Tables 5 and 6 and Figure 3 for the results. Please refer to. Table 5. Echocardiographic study of the efficacy of NRG administered subcutaneously for 35 days in the treatment of myocardial infarction in rats. Results (x±SD) [Table 5] ***: p<0.001 post-treatment subgroup compared with vehicle group; **: p<0.01 post-treatment subgroup compared with vehicle group; *: p<0.05 post-treatment subgroup compared with vehicle group Table 6. Efficacy of NRG in treating rats with myocardial infarction in two subgroups after 38 days of subcutaneous administration Echocardiography results (x±SD) [Table 6] ***: p<0.001 post-treatment subgroup compared with vehicle group; **: p<0.01 post-treatment subgroup compared with vehicle group; *: p<0.05 post-treatment subgroup compared with vehicle group

[0069] (7. Conclusion) Through long-term subcutaneous administration and subsequent discontinuation due to reduced frequency, rhNRG has been shown to be effective in preventing myocardial infarction. It can improve cardiac function and reduce cardiac remodeling in patients.

[0070] Example 4: Acute administration of steroids to rats through prolonged subcutaneous administration followed by dose reduction Therapeutic effect of rhNRG on chronic myocardial infarction (1. Purpose) In a rat model of myocardial infarction induced by left coronary artery ligation, long-term administration and The therapeutic effect of rhNRG on acute myocardial infarction in rats through subsequent interruption by reduced frequency Observe the results.

[0071] (2. Experimental Drugs) (2.1 Excipients: Developed by Zensun (Shanghai) Sci & Tech Co., Ltd.) (2.2 rhNRG: Developed by Zensun (Shanghai) Sci & Tech Co., Ltd.)

[0072] (3. Experimental Animals) 3.1 Strain and Source: Wistar rats, Shanghai Sippe-Bk Lab Animal Co., Ltd. (More supplied) (3.2 Sex, weight and certificate): Male, 200-270g.

[0073] (4. Experimental Materials and Equipment) This is the same as "4. Experimental materials and equipment" in Example 1.

[0074] (5. Experimental Methods) 5.1 Establishment of a rat model of heart failure induced by coronary artery ligation This is the same as "5.1 Establishment of a rat model of heart failure induced by coronary artery ligation" in Example 1. 5.2 Grouping and Administration

[0075] (5.3 Observation Indicators) After coronary artery ligation, rats were randomly divided into groups and administered medication. They were randomly divided into two groups based on body weight. Subcutaneous injections were administered three times daily, and the animals were weighed once daily. Echocardiography was performed on the 10th day after model creation. Echocardiograms were performed every 10 days for the 10-day treatment period, and weekly after the dose reduction and completion of the discontinuation. Echocardiography was performed every 2 weeks thereafter. All animals were continuously dosed until 60 days. A dose reduction discontinuation plan was implemented after 3 weeks. The doses were reduced to 100mg / kg in weeks 1, 2, and 3, respectively. The dose was then reduced to 15 μg / kg, 7.5 μg / kg, and 3.75 μg / kg. Clinical symptoms were observed for 3 weeks. After dose reduction, the drug was discontinued completely.

[0076] (5.3.1 Cardiac Function Tests) After anesthesia with 4% isoflurane, the rat was fixed on the operating table in the left lateral position. The animals were attached to a breathing mask attached to a gas anesthesia machine, and isoflurane was used to maintain anesthesia. The skin was disinfected with 75% alcohol and coated with a coupling agent. All echo signals from the rat left ventricle were detected using the left ventricle end-diastolic diameter and left ventricle end-systolic diameter. The left ventricular end-diastolic volume (EDV) and end-systolic volume (ESV) were calculated. Ejection fraction (EF) values ​​were obtained: EF = (EDV - ESV) / EDV × 100%.

[0077] (5.3.2 Data Processing) All experimental data were expressed as ±SD. One-way ANOVA analysis was performed using GraphPad Prism 6. P<0.05 indicates significant difference between groups; P<0.01 indicates significant difference between groups.

[0078] (6. Experimental Results) (6.1 Echocardiography Results) Echocardiography was performed after 60 days of continuous administration of NRG. LVEDd, LVED s, and EF were 1.048±0.07cm, 0.910±0.09cm, and 32.1±6.6%, respectively; NRG 30μg / kg / day group, LVEDd, LVEDs, and EF were 0.981±0.08cm, 0.794±0.08cm, and 43.8±8, respectively. 0%. Data on LVEDd and LVEDs showed that LVEDd and LVEDs were significantly higher in the daily NRG group. The EF value data showed a significant difference from the control group (p<0.001). The EF value of the NRG group increased significantly after 60 days of continuous administration, showing a significant difference from the control group. (p<0.001). After 60 days, treatment was performed at a lower dose without changing the frequency. An echocardiogram was performed. The LVEDd, LVEDs, and EF values ​​in the control group were 1.038±0.07cm, 0.8 99±0.10 cm and 32.4±9.5%; LVEDd, LVEDs, and EF values ​​in the NRG / 30 μg / kg / day group were 0.9 81±0.08 cm , 0.799±0.08cm and 42.3±11.2%. After 3 weeks of dose reduction for observation, The LVEDd, LVEDs, and EF values ​​in the control group were 1.065±0.07cm, 0.065±0.07cm, respectively. The LVEDd, LVEDs, and EF values ​​in the NRG / 30 μg / kg / day group were 0.942 ± 0.10 cm and 28.3 ± 9.4%. The results were 0.994±0.08cm, 0.826±0.10cm, and 39.3±12.7%, respectively. The LVEDd, LVEDs, and EF values ​​of the control group were 1.137±0.08cm, 1.006±0.08cm, respectively. 0.08 cm and 28.0 ± 5.7%; LVEDd, LVEDs, and EF values ​​in the NRG / 30 μg / kg / day group were 0.08 cm and 28.0 ± 5.7%, respectively. The results were 1.104±0.08cm, 0.950±0.09cm, and 33.4±7.6%. Nine weeks after discontinuation, NRG was administered daily. There was still a significant difference in LVEDd and LVEDs between the treatment and control groups; EF values ​​were significantly higher than those of the control group. The results still showed an upward trend compared to the control group. See Tables 7, 8, 9 and Figure 4 for the results.

[0079] (7. Conclusion) Considering a fixed dose and various administration frequencies, rhNRG suppressed myocardial infarction in rats during continuous administration. It exerted some therapeutic effects on the cardiac function of rats with acute myocardial infarction, rhNRG improved ventricular remodeling and delayed senescence due to myocardial infarction. It has a significant improving effect on cardiac function in rats with myocardial infarction for a long period after surgery. Table 7. Echocardiographic study of the efficacy of NRG administered subcutaneously for 60 days in the treatment of myocardial infarction in rats. Results (x±SD) [Table 7] ***: p<0.001 post-treatment subgroup compared with vehicle group; **: p<0.01 post-treatment subgroup compared with vehicle group; *: p<0.05 post-treatment subgroup compared with vehicle group Table 8. Treatment of rats with myocardial infarction after 60 days of subcutaneous administration of NRG followed by dose reduction 3 weeks later Echocardiographic results of the effect in (x ± SD) [Table 8] ***: p<0.001 post-treatment subgroup compared with vehicle group; **: p<0.01 post-treatment subgroup compared with vehicle group; *: p<0.05 post-treatment subgroup compared with vehicle group Table 9. Echocardiogram of the effect of NRG on the treatment of rats with myocardial infarction after discontinuation of subcutaneous administration on day 81 -Test results (x±SD) [Table 9] ***: p<0.001 post-treatment subgroup compared with vehicle group; **: p<0.01 post-treatment subgroup compared with vehicle group; *: p<0.05 post-treatment subgroup compared with vehicle group The present application provides the following aspects of the invention. (Aspect 1) In the manufacture of a medicament for preventing, treating, or ameliorating myocardial damage in a mammal, Use of neuregulin (NRG) or a functional fragment thereof. (Aspect 2) The use of embodiment 1, wherein the NRG is NRG-1, NRG-2, NRG-3, or NRG-4. (Aspect 3) The use of embodiment 1, wherein the NRG is NRG-1. (Aspect 4) Other compounds that can be used to prevent, treat, or reduce damage caused by myocardial infarction in mammals The use according to embodiment 1, comprising the use of a drug or treatment. (Aspect 5) 2. The method of claim 1, wherein the NRG enhances cardiac function and reduces cardiac remodeling in a mammal. Use of. (Aspect 6) The use according to embodiment 1, wherein the mammal is a human. (Aspect 7) 1. A composition for preventing, treating, or alleviating myocardial injury in a mammal, comprising an effective amount of The composition comprising NRG or a functional fragment thereof. (Aspect 8) A method for preventing, treating, or mitigating myocardial injury in a mammal, comprising: the method comprising subcutaneously administering NRG at a dose of 2.5 μg / kg / day to 50 μg / kg / day. 。 (Aspect 9) A method for preventing, treating, or mitigating myocardial injury in a mammal, comprising: The method comprises administering NRG multiple times per day for consecutive days. (Aspect 10) The NRG is administered multiple times daily for consecutive days, followed by a slow discontinuation of the NRG. 10. The method of embodiment 9, comprising:

Claims

1. 1. Use of neuregulin (NRG) in the manufacture of a medicament for use in a method for preventing, treating, or ameliorating myocardial injury in a mammal, comprising: the method comprising subcutaneously administering the NRG to the mammal multiple times daily at a dose of 15 μg / kg / day to 50 μg / kg / day for consecutive days; and The use, comprising slowly discontinuing the NRG after the administration.

2. 2. The use of claim 1, wherein the NRG is administered continuously over multiple days.

3. 2. The use of claim 1, wherein the NRG is administered multiple times daily for 30 to 60 consecutive days.

4. 4. The use of claim 3, wherein the NRG is administered three times daily for 35, 38, 49, or 60 days.

5. 4. The use of claim 3, wherein the NRG is administered three times a day for 60 days.

6. 1. A composition for use in a method for preventing, treating, or ameliorating myocardial injury in a mammal, the composition comprising neuregulin (NRG); the method comprising subcutaneously administering the NRG to the mammal multiple times daily at a dose of 15 μg / kg / day to 50 μg / kg / day for consecutive days; and the composition comprising slowly discontinuing the NRG after said administration.

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