Methods and compositions for preventing, treating, or mitigating myocardial injury using neuregulin.

Abstract

The present invention provides a pharmaceutical product for use in preventing, treating, or mitigating myocardial injury in mammals. [Solution] The use of neuregulin (NRG) in the manufacture of a pharmaceutical product for use in preventing, treating, or mitigating myocardial injury in mammals, wherein the method includes subcutaneous administration of the NRG to the mammal multiple times a day at a dose of approximately 30 μg / kg / day for several consecutive days, and then slowly discontinuing the NRG after the administration by reducing the daily dose over three weeks, wherein the daily dose in the first week is half of the administered dose, the daily dose in the second week is one-quarter of the administered dose, and the daily dose in the third week is one-eighth of the administered dose.

Description

Technical Field

[0001] (Field of the Invention) The present invention relates to the use of neuregulin (NRG) for the manufacture of a drug for preventing, treating, or alleviating myocardial injury in mammals. The present invention also relates to a method of administration, dosage frequency, and dosage of a pharmaceutical preparation or composition containing NRG for preventing, treating, or alleviating myocardial injury in mammals. In particular, the present invention relates to a method for preventing, treating, or alleviating myocardial injury, a pharmaceutical composition containing NRG for that purpose, and a method of administration, dosage frequency, and dosage of a pharmaceutical preparation or composition containing NRG for preventing, treating, or alleviating myocardial injury in mammals. dosage of a pharmaceutical preparation or composition containing NRG for preventing, treating, or alleviating myocardial injury in mammals.

Background Art

[0002] (Background of the Invention) Cardiovascular problems pose a serious threat to people's lives and health on a global scale. There are many types of cardiovascular diseases, including heart failure, myocardial infarction, atherosclerotic heart disease, arrhythmia, cardiomyopathy, heart valve disease, infective endocarditis, pericardial disease, ischemic heart disease, congenital heart disease, etc. Cardiovascular diseases tend to cause myocardial injury and affect heart function, thereby making the human body malfunction. Myocardial infarction is a cardiovascular disease that seriously endangers human health. As people's living conditions continue to improve, the incidence of ischemic myocardial infarction is steadily increasing. Myocardial infarction is mainly a type of ischemic myocardial necrosis induced by severe and chronic myocardial ischemia caused by the persistent occlusion of the coronary artery, resulting in a sharp decrease or interruption of coronary blood supply. Ischemic myocardial infarction causes myocardial cell necrosis and scarring, thereby affecting heart function. ​​​​​​​​

[0003] When a myocardial infarction occurs, the coronary arteries are blocked for 20 to 30 minutes, resulting in insufficient blood flow to the myocardium. A portion of the cell necrotizes, and the pathological process of myocardial infarction begins. As the myocardial necrosis becomes increasingly severe... Then, most of the affected cardiomyocytes gradually undergo coagulation necrosis after 2 hours, and simultaneously, massive flames erupt. The myocardial interstitium becomes congestive and edematous with symptomatic cell infiltration. The myocardial necrosis process is approximately 6-1 The process will end in 2 hours. The myocardial fibers will dissolve in 1-2 weeks. In other words, by macrophages It gets swallowed up, gradually breaks down into protofibrils, and after 6 weeks the necrotic zone finally becomes fibrous. It is completely replaced by a dense scar. This is known as chronic or regenerative myocardial infarction.

[0004] Normal left ventricular function is significantly affected after a myocardial infarction. The heart undergoes massive ischemia. When this occurs, the pumping function of the left ventricle is impaired, leading to a decrease in cardiac output, stroke volume, and blood pressure. Simultaneously, end-systolic volume may increase and end-diastolic volume may rise within a few weeks after infarction. .

[0005] Ventricular remodeling is a significant pathological behavior that occurs after myocardial infarction. A ring is a change in the structure and morphology of the ventricular infarcted and non-infarcted areas after a myocardial infarction. Changes refer to: Changes in the infarcted area mainly include the enlargement of the infarct, while changes in the non-infarcted area mainly This manifests as ventricular hypertrophy. A typical sign of ventricular remodeling is ventricular decompensated hypertrophy. This is because changes in ventricular mass, volume, and morphology lead to dysfunction of the ventricular pump. This can be triggered and may progress to heart failure. The underlying mechanism is ventricular remodeling. This condition is persistent and progressive, often occurring after a myocardial infarction. This is a significant change. The severity of this change determines the patient's cardiac function and prognosis. Ventricular remodeling is a major factor that affects a patient's cardiac function and endangers human life. It is a type of cardiovascular disease.

[0006] Currently, the main treatments for myocardial infarction are early-stage reperfusion (thrombolytic therapy and interventional therapy). (including the law), angiotensin (ANG) II receptor blockers, angiotensin-converting enzyme (ACE) These include inhibitors and beta-receptor blockers. These reduce infarct size and prevent recurrent myocardial ischemia. It is effective in reducing, improving revascularization, and suppressing excessive ventricular dilation, and The incidence of chronic heart failure decreases further.

[0007] The symptoms of myocardial infarction are closely related to the size and location of the infarction, as well as the condition of the coronary collateral vessels. The main symptoms include pain, fever, rapid heartbeat, nausea, vomiting, hypotension, shock, and arrhythmia. These include the following. The main complications of myocardial infarction include papillary muscle dysfunction or rupture, cardiac rupture, and ventricular artery aneurysm. These include embolism and post-infarction syndrome (PMIS).

[0008] Most existing drug or interventional therapies can only alleviate the symptoms of myocardial infarction, and the heart Damage to organ tissue cannot be repaired. In patients with advanced myocardial infarction, a heart transplant is necessary. It can improve cardiac function and save terminally ill patients as a last resort treatment option, but the donor Due to shortages, surgical complexity, immune rejection, and high treatment costs, clinical practice is limited. It's difficult to apply widely.

[0009] In summary, myocardial damage caused by cardiovascular disease poses a serious threat to human health. To cause. In particular, myocardial infarction, a fatal disease that seriously endangers human health, clinically There is a need to treat with a drug that is safer and more effective in practice.

[0010] Neuregulin (NG) or heregulin (HRG), which are members of the EGF-like family, are structurally similar to each other and include NR G1, NRG2, NRG3, and NRG4, as well as their isomers, referring to a group of growth differentiation factors (GDF). These exert a series of biological effects in (Lessor T et al., JCell Biochem. 1998; 70 (4):587-595); inducing the differentiation of neural crest cells into Schwann cells (Topilko et al., Mol Cell Neurosci, 1996; 8 (2-3): 71-75); stimulating the synthesis of acetylcholine receptors in skeletal muscle cells (Altiok N et al., EMBO.J. 1995; 14 (17): 4258-4266); promoting cardiomyocyte survival and DM synthesis (Zhao YY et al., I Biol Che r. 1998; 273 (17): 10261-10269). In vivo studies conducted on mouse embryos with severe NRG gene deficiency have demonstrated that NRG is necessary for heart and nerve development.

[0011] NRG receptors are members of the EF receptor family, including FR, ErbB2, ErbB, and ErbB4, and play important roles in cell proliferation, differentiation, survival, etc. NRG receptors are tyrosine kinase receptors composed of an extracellular ligand-binding domain, a transmembrane domain, and an intracellular tyrosine kinase domain. When NRG binds to the extracellular domain of ErbB3 or ErbB4, a conformational change occurs there, thereby forming an ErbB3 / ErbB4 or ErbB2 / ErbB3 heterodimer ​​​​​​​​​​ An imer or ErbB4 / ErbB4 homodimer is formed, and phosphorylation occurs at its C-terminus. The modified C-terminus further binds to downstream signaling proteins within the cell, such as AKT and / or This activates the EK signaling pathway, ultimately leading to cell proliferation, cell differentiation, and cell apoptosis. This can trigger a series of cellular responses, such as stimulating or inhibiting cell migration or cell adhesion. Of these receptors, ErbB2 and ErbB4 are mainly expressed in cardiac tissue (Zhao YY et al.). (Reference: Circ Res. 1999; 84 (12): 1380-1387).

[0012] Existing evidence suggests that the EGF-like domain of NRG-1, containing 50-64 amino acids, binds to the receptor and It has been shown to have a high ability to activate (Culousoou JM et al., J Biol Chem. 199) 5; 270 (21): 12857-12863). NRG-1β can bind to ErbB3 and ErbB4 with high affinity. B2 can form heterodimers with ErbB3 or ErbB4, and its affinity for its ligand The affinity is higher than that of homodimers of ErbB3 or ErbB4 for their ligands. Neurodevelopmental biology Previous studies have shown that the formation of the sympathetic nervous system involves signal transduction mediated by NRG-1β, ErbB2, and ErbB3. It has been confirmed that this is required (Britsch S et al., Dienes Dev. 1998; 12 (12): (1825-1836). Deficiency in the expression of NRG-1β, ErbB2, or ErbB4 leads to cardiac developmental disorders. Embryonic lethality is induced (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. It has also been shown to play a very important role in maintaining adult cardiac function (Kura (Reference by mochi Y et al., J Mol Cell Cardia1. 2006; 41 (2): 228-235). NRG-1β is present in adults. It has been proven that it can enhance the formation of myocardial sarcomeres in various cardiac conditions. In all animal models, ingestion of NRG-1β EGF-like domain improved cardiac function. It has been found that cardiac failure can be prevented (Liu et al., J Am Coll Cardiol.). (2006; 48: 1438-1447). In clinical trials, NRG is a chronic disease caused by various etiologies. It shows therapeutic effects against heart failure and significantly enhances cardiac function (CN200910057390.5). In an animal model of cerebral ischemia-reperfusion, NRG-1 showed a remarkable protective effect on brain cells. It inhibited apoptosis in brain cells, enhanced neuronal function, and reduced infarct size (Li Q et al. (Neurosci Lett. 2008; 443 (3): 155-159). NRG-1 due to cardiac ischemia-reperfusion. Release is induced, and the NRG / ErbB signaling pathway in cardiomyocytes is activated (Kuramochi Y et al. (J Biol Chem. 2004; 279 (49): 51141-51147), and NRG-1 is associated with cardiac ischemia-reperfusion. There is evidence that it plays a role in the prevention, treatment, or mitigation of injury (WO2011091723). .

[0013] Myocardial injury is a fatal disease that poses a serious risk to human health, but the treatment of myocardial injury The method, frequency, and dosage of NRG-1 administration for this purpose have not yet been clearly determined. This provides a method for meeting the above needs and a pharmaceutical composition containing NRG for that purpose. It is worth mentioning that the present invention provides a particularly optimized dosage frequency, and the present invention is particularly optimal The present invention provides a standardized dosage and a particularly optimized method of administration. The present invention further relates to a drug for preventing, treating, or mitigating myocardial injury in mammals. Regarding the use of NRG in manufacturing. For the treatment of myocardial infarction, the present invention is particularly optimized. The present invention provides a dosage frequency that is particularly optimized, and the present invention provides a dosage that is particularly optimized To provide a standardized administration method. [Overview of the project]

[0014] (Detailed explanation) (A. Overview) The present invention relates to the manufacture of drugs for preventing, treating, or alleviating myocardial injury in mammals. The present invention provides the use of NRG in relation to the prevention and treatment of myocardial injury in mammals. With regard to the use of NRG in the manufacture of drugs for the treatment or relief of illness, mammals are preferred It is a human. NRG improves cardiac function affected by myocardial damage and reduces cardiac remodeling. It can be done.

[0015] Many cardiovascular diseases, such as heart failure, myocardial infarction, coronary artery disease, arrhythmia, myocarditis, Heart valve disease, infective endocarditis, pericardial disease, ischemic heart disease, and congenital heart disease can all cause myocardial damage. It can cause myocardial damage, which affects heart function and endangers human health. Infarction is often accompanied by persistent coronary artery occlusion, typically involving apoptosis and necrosis of cardiomyocytes, and massive inflammation. It is accompanied by infiltration of symptomatic cells and myocardial fibrosis, which induce myocardial damage. Myocardial infarction damage is It tends to cause cardiac failure and therefore affects human health.

[0016] This invention demonstrates that NRG is essential for cardiac development and is extremely important in maintaining adult cardiac function. Based on the scientific discovery that NRG plays a crucial role; this invention relates to the sarcominescence of cardiomyocytes. A. Based on scientific discoveries that it can strengthen the formation of the cytoskeleton and intercellular junctions; The invention relates to the ability of NRG to the heart of animals or patients with heart failure in various animal models and clinical trials. Based on scientific discoveries that NRG can enhance organ function, this invention relates to the use of NRG in cerebral ischemia-reinfusion. Based on the scientific discovery that the flow exerts a protective effect on brain cells in animal models; Akira stated that NRG exerts a protective effect on brain cells in an animal model of cardiac ischemia-reperfusion. Based on scientific discovery; NRG, NRG polypeptide, and NRG mutants or NRG-like functions All other composites are within the scope of the present invention.

[0017] In a first embodiment, the present invention aims to prevent, treat, or mitigate myocardial injury in mammals. The present invention provides pharmaceutical preparations for treating myocardial infarction damage in mammals. The invention provides pharmaceutical preparations for prevention, treatment, or relief, and the mammal is preferably human. The pharmaceutical preparation contains an effective amount of NRG or its functional fragment, or nucleic acid encoding NRG. or its functional fragment, or substances that increase the production and / or function of NRG, as well as pharmaceuticals It includes carriers, excipients, etc. that may be acceptable. Pharmaceutical preparations are used to prevent and treat myocardial damage. Alternatively, it can be used in combination with other drugs or therapies to alleviate the condition. In this regard, pharmaceutical preparations containing NRG are effective in increasing the EF value of the left ventricle in mammals. In another embodiment, a pharmaceutical preparation containing NRG is used to measure left ventricular end-diastolic volume (LVEDV) or left It is effective in reducing end-systolic volume (LVESV). In another embodiment, a pharmaceutical compound containing NRG is used. The product is administered by subcutaneous injection via syringe or other device. In another embodiment, the product includes NRG. The pharmaceutical preparation is administered by subcutaneous injection via a pump such as an injection pump. In this embodiment, the syringe pump is a micropump. In a further embodiment... The micropump is an insulin pump. It is worth mentioning that this invention is a pharmaceutical Suitable for any preparation that can be used as such, the pharmaceutical preparation contains the above NRG or NRG and to include excipients, diluents, or carriers that are pharmaceutically acceptable. The pharmaceutical preparations include, but are not limited to, the contents of this application.

[0018] In a second embodiment, the present invention provides a method for preventing, treating, or mitigating myocardial injury in mammals. The present invention provides a method for preventing myocardial infarction-related injury in mammals. The present invention provides a method for treating or alleviating a condition, wherein the mammal is preferably a human. This refers to an effective amount of NRG or its functional fragment, or a nucleic acid or its functional fragment that encodes NRG. , or the production of NRG required for the prevention, treatment, or mitigation of myocardial injury in mammals and / or This includes the use of substances that enhance function. Other drugs, in particular effective amounts of NRG or other drugs, by means of Functional fragments of, nucleic acids or functional fragments thereof that encode NRG, or in mammals, cardiac Combined with substances that increase the production and / or function of NRG to prevent, treat, or alleviate muscle damage. They can be used together.

[0019] In a third embodiment, the present invention provides a method for preventing, treating, or mitigating myocardial injury in mammals. The present invention provides a composition for preventing damage caused by myocardial infarction in mammals. The invention provides a pharmaceutical composition for the treatment or relief of a disease, and the mammal is preferably a human. The pharmaceutical composition is for preventing, treating, or alleviating myocardial injury in mammals according to the present invention. This includes the type of NRG provided, and other drugs for preventing, treating, or mitigating myocardial damage. The pharmaceutical composition contains an EGF-like domain, which binds to and activates a receptor. It has been proven that it is possible. In particular, it is not intended to be limiting. 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 embodiment, the present invention provides a method for preventing, treating, or mitigating myocardial injury in mammals. The present invention provides dosages for pharmaceutical preparations containing NRG for use in mammals. Medical treatments including NRG used to prevent, treat, or alleviate damage caused by myocardial infarction in the following cases The dosage of the drug preparation is provided, and the mammal is preferably a human. The effective dose is the NRG This means that when the dose is applied to mammals, one or more beneficial effects can be achieved. The beneficial effect is to improve cardiac function in patients with myocardial damage and prevent further deterioration of that cardiac function. To prevent deterioration or to suppress the worsening of cardiac dysfunction that may be caused by myocardial damage. This can be achieved. The dose provided to mammals by this invention is 0.1 μg / kg / day (protein / body). The recommended daily intake is approximately 360 μg / kg / day (protein / body weight). In one embodiment, the dose is 0.3 μg / kg / day (protein / body weight) ~ 50 μg / kg / day (protein / body weight); 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 embodiment, the present invention provides a method for preventing, treating, or mitigating myocardial injury in mammals. The present invention provides a method for administering a pharmaceutical preparation containing NRG for use in mammals. Administration methods of pharmaceutical preparations containing NRG for the prevention, treatment, or mitigation of damage caused by myocardial infarction. The mammal is preferably human. The pharmaceutical preparation is administered orally, rectally, or ophthalmologically. Oral administration, inhalation administration, buccal administration (e.g., sublingual administration), parenteral administration (e.g., subcutaneous injection, intramuscular injection) Taken by intramuscular injection, intradermal injection, or intravenous injection, transdermal administration, or other appropriate method. This is possible. In one embodiment, NRG is administered only once a day. In another embodiment, NRG is administered multiple times a day. In one embodiment, NRG is administered daily. In another embodiment... In this embodiment, this tolerable dose of NRG is administered within a few days. In another embodiment, NRG is 1 It is administered multiple times a day for many consecutive days. In another embodiment, NRG is administered twice a week. It is administered multiple times a day for many consecutive weeks. In another embodiment, NRG is administered 2 days a week, It is administered by subcutaneous injection three times a day for many consecutive weeks. In another embodiment, NRG is administered three times a day. It is administered by subcutaneous injection over many consecutive days. In another embodiment, NRG is administered three times a day consecutively. In another embodiment, NRG is administered subcutaneously three times a day for 38 consecutive days. It is administered by subcutaneous injection over time. In another embodiment, NRG is administered three times a day for 49 consecutive days. It is administered by subcutaneous injection. In another embodiment, NRG is administered subcutaneously three times a day for 60 consecutive days. Inject as shown below. In another embodiment, NRG is administered three times a day for more than 35 consecutive days. It is administered by subcutaneous injection. In another embodiment, NRG is administered multiple times a day for many consecutive days. Agree, and then slowly interrupt. In another embodiment, NRG is multiple times a day, many consecutive Administer for several consecutive days, then gradually discontinue over the next three weeks: Administer every other day during the first week. In the second week, administer every three days; in the third week, administer subcutaneously every four days. In another embodiment NRG is administered subcutaneously three times a day for more than 38 consecutive days, and then gradually discontinued. In another embodiment, NRG is administered by subcutaneous injection three times a day for 49 consecutive days, followed by a 3-week course of treatment. Gradually discontinue: Administer every other day during the first week; every three days during the second week; every four days during the third week. It is administered by subcutaneous injection. In another embodiment, NRG is administered multiple times a day for many consecutive days. Administer the drug, and then slowly reduce the daily dose. In another embodiment, NRG is administered 3 times a day. The drug is administered multiple times over many consecutive days, and then the daily dose is slowly reduced. In terms of administration, NRG is administered by subcutaneous injection three times a day for more than 60 consecutive days, and thereafter... The daily dose is gradually reduced. In another embodiment, NRG is administered three times a day for 60 consecutive days. Administer subcutaneously for several weeks, then gradually discontinue over three weeks: The daily dose in the first week is continuous The dose administered shall be half of the dose given; the daily dose in the second week shall be one-quarter of the dose administered continuously; and the dose in the third week shall be one-quarter of the dose given continuously. The daily dose should be one-eighth of the dose administered continuously.

[0022] The present invention also provides a kit for preventing, treating, or mitigating myocardial injury in mammals. The present invention also provides a means to prevent, treat, or alleviate damage caused by myocardial infarction in mammals. The kit provides a procedure for which the mammal is preferably human. The kit is designed to prevent myocardial injury. Single doses or multiple doses of the pharmaceutical preparation or composition for prevention, treatment, or relief, and This includes instructions on how to use the pharmaceutical preparation or composition.

[0023] The pharmaceutical preparations or compositions provided by the present invention are administered before, during, or after the onset of heart disease. It can be administered. When used for prevention, pharmaceutical preparations or compositions generally have heart disease Administer before the onset of symptoms. When used for treatment, pharmaceutical preparations or compositions generally affect heart disease. It is administered during or after the occurrence of the disease. In one embodiment, the pharmaceutical preparation provided by the present invention The product or composition is administered before the onset of heart disease. In another embodiment, the present invention The provided pharmaceutical preparation or composition is administered when a myocardial infarction occurs. Another embodiment In this case, the pharmaceutical preparation or composition provided by the present invention is administered after the onset of heart disease. do.

[0024] Pharmaceutical preparations or compositions provided by the present invention, for oral administration, rectal administration, topical administration, Inhalation administration, buccal administration (e.g., sublingual administration), parenteral administration (e.g., subcutaneous injection, intramuscular injection) It can be taken in by intradermal injection, intravenous injection, transdermal administration, or other appropriate methods. Subcutaneous injection can be administered using a syringe, pump (micro-injection pump), or other administration device. It can be administered. The dosage forms of pharmaceutical preparations or compositions provided by the present invention include this While not limited to these, tablets, lozenges, cachets, dispersants, suspensions, solutions, and capsules are also used. It comes in the form of ointments and similar substances.

[0025] (B. Definition) Unless otherwise defined, all scientific and technical terms used herein are those of the United States. It has the same meaning as understood by the person concerned. All patent documents, patent application documents, Publicly available patent documents and other publications are cited as references. If the definition of "meaning" has a different meaning from the meaning explained in the above document, the meaning given in this section The explanation shall take precedence.

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

[0027] As used herein, "mammals" refers to non-human primates (cattle, pigs, horses, cats, dogs, This refers to rats, mice, etc., or primates (monkeys, humans), preferably humans.

[0028] As used herein, "myocardial injury" refers to heart failure, myocardial infarction, coronary artery atherosclerotic heart disease, and irregular heart disease. Pulse, cardiomyopathy, valvular heart disease, infective endocarditis, pericardial disease, ischemic heart disease, or congenital heart disease This refers to certain types of myocardial damage caused by pathological heart diseases such as [examples of pathological heart conditions]. Myocardial damage is a heart condition. It tends to cause dysfunction, thereby affecting human health. (Disease of myocardial injury) The causes include oxyradical production, calcium overload, and inflammatory reactions due to neutrophil infiltration into the injured area. In response, apoptosis or necrosis of cardiomyocytes, tissue caused by an imbalance in energy supply. Metabolic disorders, abnormalities in cardiac signaling, cholesterol accumulation, and atherosclerotic plaques This relates to multiple pathophysiological changes, including formation.

[0029] As used herein, "Neureglin" or "NRG" refers to ErbB2, ErbB3, ErbB4, or He Proteins or polyps that can bind to and activate telodimers or homodimers. This refers to the petidopropyl alcohol. NRG includes isoforms of NRG, EGF-like domains of NRG, and EGF-like domains of NRG. A polypeptide containing , a mutant or derivative of NRG, and the activation of the above receptor. It also contains other gene products of NRG that can be produced. In addition, NRG includes NRG-1, NRG-2, NRG-3, and NRG-4 These include polypeptides, fragments, and complexes having NRG-like function. Preferably, the NRG is ErbB Proteins that can bind to and activate 2 / ErbB4 or ErbB2 / ErbB3 heterodimers. It is a type of protein or polypeptide. For example, and this is not intended to be limiting, The NRG (rhNRG) provided by the invention is an isomer of NRG-1β2 containing an EGF-like domain. It is a fragment, or rather, a 177-237 amino acid fragment. The amino acid sequence of this fragment is as follows: ru: [ka] The NRG used in this invention activates the above receptor and modulates its biological function. This can be done. For example, stimulating skeletal muscle cells to synthesize acetylcholine receptors; and cardiomyocytes NRGs can promote differentiation, survival, and DNA synthesis. NRGs substantially affect biological functions. This also includes conservative NRG mutants that do not give [the specified trait]. As will be obvious to those skilled in the art, [the specified trait] is not important. Mutation of a single amino acid in the matrix does not affect the biological function of the protein or polypeptide. No change occurs (Watson et al., "Molecular Biology of the Genesis), 4th edition, 1987, The Bejacmin / Cummings Pub. co., p.224). Used in this invention NRGs can be extracted from natural sources, or synthesized through recombinant technology, artificial synthesis, or other means. It can also be obtained.

[0030] As used herein, "EGF-like domain" refers to ErbB2, ErbB3, ErbB4, or their heterozygotes. It can bind to and activate imers or homodimers, and the following references: WO 00 / 64400; Holmes et al., Science, 256: 1205-1210 (1992); US 5,530,109 Issues 5,716,930; Literature by Hijazi et al., Int. J. Oncol., 13: 1061-1067 (1998); Chang References by Carraway et al., Nature, 387: 509-512 (1997); References by Carraway et al., Nature, 387: 512-516 (19 97); See Higashiyama et al., J. Biochem., 122: 675-680 (1997); and WO 97 / 09425. Polypeptide encoded by the NRG gene has a structure similar to the EGF receptor binding zone shown. Refers to a ptide fragment. In some embodiments, the EGF-like domain is ErbB2 / ErbB4 or ErbB It binds to and activates the 2 / ErbB3 heterodimer. In some embodiments, E The GF-like domain contains amino acids in the receptor-binding zone of NRG-1. Several implementations In this context, 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 located in the receptor-binding zone of NRG-2. Contains an acid. In some embodiments, the EGF-like domain is the receptor binding zone of NRG-3. It contains amino acids in the following embodiment. In some embodiments, the EGF-like domain is the receptor for NRG-4. It contains amino acids in the body-binding zone. In some embodiments, the EGF-like domain is , containing the amino acid sequence described in U.S. Patent 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] NRGs can be administered orally, rectally, topically, by inhalation, or buccally (e.g., sublingually). Parenteral administration (e.g., subcutaneous injection, intramuscular injection, intradermal injection, or intravenous injection), transdermal administration, Alternatively, the drug can be taken up by other appropriate means. In all modes of administration, The preferred route of administration depends on the treatment conditions and severity, as well as the characteristics of the specific NRG used. Therefore, a choice needs to be made. NRG can be administered alone. Or, more preferably, NRG It can be administered with several pharmaceutically acceptable carriers or excipients. Applicable carriers or excipients suitable as pharmaceuticals are applied to this method (Remington: Pharmaceuticals) Science and Practice (Remington: The Science and Practice of Pharmacy), Alfonso R. Gennar o (ed.) Mack Publishing Company, April 1997).

[0032] As used herein, "pump" refers to a pump for pharmaceutical liquids, drugs, proteins, and / or other compositions. This is an administration device for subcutaneous injection of substances. The pump is used for continuous, accurate, and quantitative administration. It can be used. The pump is equipped with a subcutaneous catheter for continuous subcutaneous injection. The catheter can be placed externally, or the catheter port can be connected to a pump mechanism. It can also be implanted. The micro-injection pump is portable and easy to use for precise injection. It is a useful device. For example, an insulin pump is used during the treatment of diabetes or other diseases to deliver insulin. Or it is a medical device used to administer other drugs. Insulin pumps are also... It is thought to be used for continuous subcutaneous delivery of insulin. A disposable thin-walled plastic pipe or catheter can be attached, and as a result Insulin or other drugs can enter the tissue. The catheter is inserted subcutaneously as needed. It can be moved accordingly. The pump is an external device that can be connected to the patient, or inside the patient's body. It can be attached to a device that can be implanted. External pumps are used in hospitals and clinics. This refers to a device designed for use in a fixed location, such as a lock or similar place. This refers to a mobile or portable device such as a pump or similar device that can be carried by the patient. The lamp is capable of storing a fluid medium, for example, a fluid medium containing NRG, but not limited to this. It is equipped with a reservoir.

[0033] The external pump can be connected to the patient via a fluid through a suitable hollow tube, for example. Yes, it is possible. The hollow tube can be connected to a hollow needle, which can be used to inject into the patient's skin. It is used to pierce through. Alternatively, a hollow tube is used to insert a cannula or similar object into the patient. It can also be connected directly to the patient. The external pump can be worn by the patient or attached to the patient's clothing. It can be attached to the body or worn under the patient's clothing. A suitable pump will be used for these. While not limited to this, micro-infusion pumps suitable for high-frequency infusions, such as the MiniMedParadigm522, are also available. Insulin pumps, MiniMed Paradigm 722 insulin pump, MiniMed Paradigm 515 insulin pump Insulin pumps, MiniMed Paradigm 715 insulin pump, MiniMed Paradigm 512R insulin pump MiniMed Paradigm 712R insulin pump, MiniMed Paradigm 508 insulin pump, and The MiniMedParadigm508R insulin pump (Medtronic, Northridge, Canada), and the art of the art. This refers to other similar devices that are well known.

[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), US Special US2005 / 0065760 (Method for Advising Patients on Insulin Dosage) (Vising Patients Concerning Doses of Insulin) and US6,589,229 (Wearable Implantation) Except as described in the Wearable Self-Containing Drug Infusion Device Examples of administration devices, such as pumps, are cited in this section.

[0035] The term "can be used to prevent, treat, or alleviate myocardial damage" as used herein. "Other drugs or treatments" refers to drugs and interventions that are generally applicable to the treatment of myocardial injury. This refers to drugs and interventional therapies that are generally applicable to the treatment of damage caused by myocardial infarction. Drugs used to treat myocardial infarction include antiplatelet drugs (such as aspirin and clopidogrel). Anticoagulants (heparin, bivalirudin, etc.), thrombolytic agents (alteplase, tenectep) (Lase, urokinase, recombinant human prourokinase, etc.), lipid-lowering drugs (statins) (Cholesterol absorption inhibitors), angiotensin-converting enzyme inhibitors / ANG II receptor blockers β-receptor blockers, calcium channel blockers, nitrate esters, phosphatase inhibitors, Diuretics, renin-angiotensin-aldosterone system (RAS) antagonists, myocardial energy Energy optimizers, ischemic tissue metabolism improvers, free radical scavengers These include interventional therapies such as coronary artery intervention therapy. [Brief explanation of the drawing]

[0036] (Brief explanation of the drawing) [Figure 1] This paper presents echocardiographic results of the effects of long-term subcutaneous administration of NRG at various doses in the treatment of rats with acute myocardial infarction. [Figure 2] This paper presents echocardiographic results of the effects of long-term subcutaneous administration of NRG at various frequencies in the treatment of rats with acute myocardial infarction. [Figure 3] This study demonstrates the therapeutic effect of NRG on acute myocardial infarction in rats through long-term subcutaneous administration and subsequent discontinuation due to decreased frequency. [Figure 4]This study demonstrates the therapeutic effect of NRG on acute myocardial infarction in rats through long-term subcutaneous administration and subsequent dose reduction and discontinuation. [Examples]

[0037] (Examples) (Example 1: Therapeutic efficacy of long-term subcutaneous administration of various doses of rhNRG for acute myocardial infarction in rats) (Research on the dose-response relationship of NRGs) (1. Purpose) The effects of left coronary artery ligation on acute myocardial infarction in rats in a rat model of myocardial infarction. By observing the therapeutic effects of various doses of rhNRG, we can study the treatment of acute myocardial infarction in rats. To investigate the dose-response relationship of NRG for [specific condition]. (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. Laboratory animals) (3.1 Strain and source: Wistar rat, Shanghai Sippe-Bk Lab Animal Co., Ltd.) (More supply) (3.2 Sex, weight, and certificate: Male, 200-270g)

[0039] (4. Experimental materials and equipment) An anesthesia device, an 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 needles (with thread) manufactured by Ningbo Medical Needle Co., Ltd.

[0040] (5. Experimental Method) (5.1 Establishment of a rat model of heart failure induced by coronary artery ligation) The rats were anesthetized with isoflurane via a gas anesthesia device. Then, the rats were placed in a supine position. It was fixed in place. After shaving the chest, the skin was disinfected with 75% alcohol. After incision of the skin of the left anterior chest, the chest The muscles were bluntly separated, exposing the 4th and 5th ribs. Using hemostatic forceps, the 4th and 5th ribs were... The muscles between the ribs were bluntly cut. Using both hands, the heart was pushed out of the thoracic cavity, and the lungs expanded and The heart was fully exposed for observation of the heartbeat. The left atrial appendage and pulmonary cone were also fully exposed. The left anterior descending coronary artery (LADCA), located between them, was ligated by surgical suture. After ligation, the heart The organs were quickly returned to their original positions. Next, the pectoral muscles and skin were sutured. After the surgery, the rat was cadaver. They were returned to their enclosure, 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 an animal model of myocardial infarction was established.

[0042] (5.3 Observational Indicators) After anesthesia with 4% isoflurane, the rat was fixed to the operating table in a left lateral position. The rat's head was then... The patient was secured to the breathing mask of a gas anesthesia machine, and isoflurane was used to maintain anesthesia. After shaving the chest. The skin was disinfected with 75% alcohol and covered with a coupling agent. An echocardiogram probe was used. Then, all echo signals from the rat left ventricle were detected. Left ventricular end-diastolic diameter and left ventricular end-systolic diameter The diurnal diameter (D) was measured. The left ventricular end-diastolic volume (EDV) and end-systolic volume (ESV) were calculated. Furthermore, the ejection fraction (EF) value was obtained. EF = (EDV - ESV) / ​​EDV × 100%.

[0043] (5.3.2 Data Processing) All experimental data were expressed using ±SD.

[0044] (6. Experimental Results) (6.1 Results of echocardiography) Echocardiography was performed 60 days after the start of continuous administration of NRG. From the results, the LVEDd group of the excipient group The LVEDs and EF values ​​were 0.971±0.07 cm, 0.832±0.08 cm, and 34.6±7.00%, respectively;15 In the μg / kg-NRG group, the LVEDd, LVEDs, and EF values ​​were 0.975±0.07 cm, 0.794±0.10 cm, and , respectively. The values ​​were 42.9±11.32%; and the LVEDd, LVEDs, and EF values ​​for the 7.5μg / kg-NRG group were 0.965±0.0, respectively. The measurements were 7 cm, 0.808 ± 0.11 cm, and 38.4 ± 12.17%; LVEDd, LVEDs, and 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 data from LVEDd and LVEDs, LVEDd and LVEDs may decrease in the high-dose NRG group. EF According to the data, cardiac function in rats in the high-dose, medium-dose, and low-dose groups was all achieved through continuous administration. Improvement was observed after 60 days, and a dose-response relationship was observed among the three groups. For details, see Table 2 and Figure 1. Please refer to this. Table 2 Efficacy of subcutaneous administration of various doses of NRG for 60 days in rats treating acute myocardial infarction. Echocardiogram results for the patient. (x±SD) [Table 2]

[0045] (7. Conclusion) After 60 days of treatment with rhNRG, NRG at doses of 5 μg / kg, 2.5 μg / kg, and 1.25 μg / kg is administered subcutaneously three times a day. The EF values ​​in the injected treatment group were higher than those in the control group, and there was no specific dose-response relationship among the three doses. there were.

[0046] (Example 2: Therapeutic efficacy of long-term subcutaneous administration of rhNRG at various frequencies for acute myocardial infarction in rats) fruit) (1. Purpose) In a rat model of myocardial infarction caused by left coronary artery ligation, various frequencies Long-term subcutaneous administration of rhNRG has shown therapeutic effects of specific doses of rhNRG on acute myocardial infarction in rats. To investigate.

[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. Laboratory animals) (3.1 Strain and source: Wistar rat, Shanghai Sippe-Bk Lab Animal Co., Ltd.) (More supply)

[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 apparatus" in Example 1.

[0052] (5. Experimental Method) (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 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 assigned to groups after coronary artery ligation. Based on survival after ligation, rats were selected. The subjects were divided into four groups based on body weight: excipient group (control group), NRG 30 μg / kg / day group, and NRG 30 μg / kg / BIW group. They were randomly divided into two groups: one receiving NRG 30 μg / kg / day × 7 + QW group. On the day an animal model of myocardial infarction was established. Administration was initiated later. For the first three and fourth groups, rats were given the drug three times a day for the first seven days. The drug was administered by injection and weighed once daily. The medication was given to the animals according to their body weight, with a dose of 30 μg / kg / day. For the fourth group, rats were injected with NRG once a week for the last four weeks, with a daily dose of 30 μg / kg. That's what I decided.

[0055] (5.3 Observational Indicators) (5.3.1 Cardiac function tests) After anesthesia with 4% isoflurane, the rat was fixed to the operating table in a left lateral position. The rat's head was then... The patient was secured to the breathing mask of a gas anesthesia machine, and isoflurane was used to maintain anesthesia. After shaving the chest. The skin was disinfected with 75% alcohol and covered with a coupling agent. An echocardiogram probe was used. Then, all echo signals from the rat left ventricle were detected. Left ventricular end-diastolic diameter and left ventricular end-systolic diameter The diurnal diameter (D) was measured. The left ventricular end-diastolic volume (EDV) and end-systolic volume (ESV) were calculated. Furthermore, the ejection fraction (EF) value was obtained: EF - (EDV - ESV) / ​​EDV × 100%. This value represents the cardiac function of rats. Echocardiography was performed in the first, second, third, and fifth weeks following the onset of myocardial infarction.

[0056] (5.3.2 Data Processing) All experimental data were expressed using ±SD.

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

[0058] As shown in the LVEDd and LVEDs data, after 35 days of continuous administration, NRG / 30μ g / kg / day can significantly reduce LVEDd and LVEDs; according to EF value data, E The F value was significantly higher in the NRG / 30μg / kg / day group than in the control group; the EF value was higher in the NRG / 30μg / kg / BIW group. In comparison with the control group, an upward trend was observed, and during continuous administration for the first 7 days, NRG / 30μg / k The cardiac function of rats in the g / day x 7 + QW group was improved to some extent compared to the control group, that is, Cardiac function showed an improving trend. Subsequently, injections were administered every 7 days to maintain the effect. For the results, please refer to Table 4 and Figure 2.

[0059] Table 4. Therapeutic effects of NRG on myocardial infarction in rats through long-term subcutaneous administration at various frequencies. Ultrasound examination results (x±SD) [Table 4]

[0060] (Example 3: Long-term subcutaneous administration to rats with acute myocardial infarction and subsequent decrease in frequency) (Therapeutic effect of rhNRG through interruption) (1. Purpose) In a rat model of myocardial infarction caused by left coronary artery ligation, long-term administration and The therapeutic effect of rhNRG on acute myocardial infarction in rats, through subsequent interruptions due to decreased frequency. Observe the fruit.

[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. Laboratory animals) (3.1 Strain and source: Wistar rat, Shanghai Sippe-Bk Lab Animal Co., Ltd.) (More supply) (3.2 Sex, weight, and certificate): Male, 200-270g.

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

[0064] (5. Experimental Method) (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 by coronary artery ligation" in Example 1.

[0065] (5.2 Grouping and Administration) Rats were randomly divided into groups after coronary artery ligation and administered medication. Based on survival after ligation, the rats were selected. The rats were randomly divided into two groups: an excipient group and an NRG 30 μg / kg group. The excipient group consisted of 19 rats. There were 18 rats in the NRG group. From the day after the model was created, 10 μg / kg was administered three times a day. Continuous administration of the drug was initiated by subcutaneous administration at the prescribed dose. Echocardiography was performed after model creation. The procedure was carried out on the 14th. All animals were given medication continuously until the 38th day, and the animals in the NRG group underwent echocardiography. The study was conducted by dividing the animals in the NRG group into two subgroups on average, and the animals in one subgroup were subjected to The medication was continued in the other subgroup of animals, while the medication was discontinued early. The excipient group continued to receive the medication. Medication was administered. For the subgroup receiving continuous NRG administration, a 3-week interruption plan was implemented on day 49. In the first week, the medication is administered every other day; in the second week, it is administered every three days; and in the third week, NRG is administered subcutaneously every four days. Injection was administered. Regarding the method of administration, NRG was subcutaneously injected into rats three times a day, exactly as described above. In the NRG interruption subgroup, the clinical symptoms of rats were observed. All animals underwent cardiac function testing. Echocardiograms were performed weekly to monitor changes in brain function.

[0066] (5.3 Observational Indicators) (5.3.1 Cardiac function tests) After anesthesia with 4% isoflurane, the rat was fixed to the operating table in a left lateral position. The rat's head was then... The patient was secured to the breathing mask of a gas anesthesia machine, and isoflurane was used to maintain anesthesia. After shaving the chest. The skin was disinfected with 75% alcohol and covered with a coupling agent. An echocardiogram probe was used. Then, all echo signals from the rat left ventricle were detected. Left ventricular end-diastolic diameter and left ventricular end-systolic diameter The diurnal diameter (D) was measured. The left ventricular end-diastolic volume (EDV) and end-systolic volume (ESV) were calculated. Furthermore, the ejection fraction (EF) value was 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 Prism6. The study was conducted. A P<0.05 indicates a statistically significant difference between groups. A P<0.01 indicates a significant statistically significant difference between groups.

[0068] (6. Experimental Results) (6.1 Results of echocardiography) Echocardiography was performed after 35 days of continuous administration of NRG. LVEDd and LVED in the excipient group. The 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. Yes; this result indicates that NRG significantly reduces LVEDd and LVEDs and enhances cardiac contractility. This demonstrates that left ventricular remodeling can be reversed. After 49 days of continuous administration, the excipient group 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 ​​for the NRG interruption subgroup were 0.987±0.05cm, 0.807±0.06cm, and 4, respectively. The values ​​were 2.2±5.48%, while the LVEDd, LVEDs, and EF values ​​in the NRG continuous administration group were 0.973± The measurements were 0.07 cm, 0.783 ± 0.08 cm, and 45.0 ± 5.51%; these results suggest that abrupt discontinuation of NRG in rats This indicates that it has some effect on cardiac function. A gradual discontinuation plan was implemented for the NRG continuous administration group. The procedure was performed, and echocardiography was conducted two weeks after drug discontinuation. The early NRG discontinuation subgroup consisted of LVEDd and LVEDs. The EF values ​​were 1.043±0.06 cm, 0.887±0.06 cm, and 35.4±6.78%, respectively; gradual NRG discontinuation. The LVEDd, LVEDs, and EF values ​​for the subgroups were 0.989±0.07 cm, 0.814±0.08 cm, and 41.3±4.92%, respectively. A significant difference was observed compared to the group receiving excipients. Echocardiography was performed in the third week after discontinuation. In the early NRG discontinuation subgroup, the LVEDd, LVEDs, and EF values ​​were 1.010±0.06 cm, 0.842+0.06 cm, and 38 cm, respectively. The values ​​were 0.9 ± 5.04%; the LVEDd, LVEDs, and EF values ​​in the stepwise NRG discontinuation subgroup were 0.976 ± 0.0, respectively. The measurements were 6 cm, 0.805 ± 0.07 cm, and 40.8 ± 4.67%. Compared to the excipient group, the cardiac function of rats was improved. Conversely, the effects of gradual discontinuation of NRG were mitigated. See Tables 5 and 6 and Figure 3 for the results. I want to be illuminated. Table 5 Echocardiographic results of the efficacy of 35-day subcutaneous administration of NRG in treating myocardial infarction in rats. Results (x±SD) [Table 5] ***: p<0.001 Subgroup after treatment compared to the excipient group; **: p<0.01 Subgroup after treatment compared to the excipient group; *: p<0.05 Subgroup after treatment compared to the excipient group Table 6. Efficacy of NRG in the treatment of myocardial infarction in two subgroups of rats after 38 days of subcutaneous administration. Echocardiography results (x±SD) [Table 6] ***: p<0.001 Subgroup after treatment compared to the excipient group; **: p<0.01 Subgroup after treatment compared to the excipient group; *: p<0.05 Subgroup after treatment compared to the excipient group

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

[0070] (Example 4: Rapid changes in rats through long-term subcutaneous administration and subsequent dose reduction and interruption) Therapeutic effects of rhNRG on dysentery myocardial infarction) (1. Purpose) In a rat model of myocardial infarction caused by left coronary artery ligation, long-term administration and The therapeutic effect of rhNRG on acute myocardial infarction in rats, through subsequent interruptions due to decreased frequency. Observe the fruit.

[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. Laboratory animals) (3.1 Strain and source: Wistar rat, Shanghai Sippe-Bk Lab Animal Co., Ltd.) (More supply) (3.2 Sex, weight, and certificate): Male, 200-270g.

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

[0074] (5. Experimental Method) (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 by coronary artery ligation" in Example 1. (5.2 Grouping and Administration)

[0075] (5.3 Observational Indicators) Rats were randomly divided into groups after coronary artery ligation and administered medication. Based on survival after ligation, the rats were selected. The animals were randomly divided into two groups based on body weight. Subcutaneous injections were administered three times a day, and the animals were weighed once a day. Medication was administered according to body weight. Echocardiography was performed on the 10th day after model creation. All animals Echocardiography will be performed every 10 days, and after dose reduction, echocardiography will be performed weekly to complete discontinuation. Echocardiography was performed every two weeks thereafter. All animals were given medication continuously until day 60, and A plan to discontinue the medication by reducing the dosage over the next three weeks was implemented. The dosage was adjusted for each of the following weeks: Week 1, Week 2, and Week 3. The dosage was reduced to 15 μg / kg, 7.5 μg / kg, and 3.75 μg / kg. Clinical symptoms were observed for 3 weeks. After reducing the dosage, the medication was completely discontinued.

[0076] (5.3.1 Cardiac function tests) After anesthesia with 4% isoflurane, the rat was fixed to the operating table in a left lateral position. The rat's head was then... The patient was secured to the breathing mask of a gas anesthesia machine, and isoflurane was used to maintain anesthesia. After shaving the chest. The skin was disinfected with 75% alcohol and covered with a coupling agent. An echocardiogram probe was used. Then, all echo signals from the rat left ventricle were detected. Left ventricular end-diastolic diameter and left ventricular end-systolic diameter The diurnal diameter (D) was measured. The left ventricular end-diastolic volume (EDV) and end-systolic volume (ESV) were calculated. The ejection fraction (EF) value was 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 Prism6. The study was conducted. P<0.05 indicates a statistically significant difference between groups; P<0.01 indicates a significant difference between groups.

[0078] (6. Experimental Results) (6.1 Results of echocardiography) Echocardiography was performed after 60 days of continuous administration of NRG. LVEDd and LVED in the excipient group. s and EF were 1.048±0.07 cm, 0.910±0.09 cm, and 32.1±6.6%, respectively; NRG 30 μg In the / kg / day group, LVEDd, LVEDs, and EF were 0.981±0.08 cm, 0.794±0.08 cm, and 43.8±8 cm, respectively. It was 0%. According to the data for LVEDd and LVEDs, LVEDd and LVEDs in the daily NRG administration group had The EF value was significantly reduced, showing a remarkable statistical difference compared to the control group (p<0.001). The EF value in the NRG group increased significantly after 60 days of continuous administration, showing a remarkable statistically significant difference compared to the control group. (p<0.001). After 60 days, the treatment was performed with reduced dose but no change in frequency. In week 3, cardiac An ultrasound examination was performed. The LVEDd, LVEDs, and EF values ​​of the control group were 1.038 ± 0.07 cm and 0.8 cm, respectively. The values ​​were 99±0.10 cm and 32.4+9.5%; the LVEDd, LVEDs, and EF values ​​in the NRG / 30μg / kg / day group were 0.9 The values ​​were 81±0.08, 0.799±0.08 cm, and 42.3±11.2%. For observation, after a 3-week dose reduction... The medication was completely discontinued. The LVEDd, LVEDs, and EF values ​​in the control group were 1.065 ± 0.07 cm and 0.9 cm, respectively. The values ​​were 42±0.10 cm and 28.3±9.4%; the LVEDd, LVEDs, and EF values ​​in the NRG / 30μg / kg / day group were The measurements were 0.994±0.08cm, 0.826±0.10cm, and 39.3±12.7%, respectively. Echocardiography was performed 9 weeks after the interruption. The tests were performed. The LVEDd, LVEDs, and EF values ​​in the control group were 1.137±0.08 cm and 1.006±0, respectively. The values ​​were 0.8 cm and 28.0 + 5.7%; the LVEDd, LVEDs, and EF values ​​for the NRG / 30 μg / kg / day group were respectively The measurements were 1.104±0.08cm, 0.950±0.09cm, and 33.4±7.6%. Nine weeks after the interruption, NRG was administered daily. Significant differences in LVEDd and LVEDs between the treatment group and the control group still exist; EF values ​​are compared to the control group. The trend continued to rise. For the results, please refer to Tables 7, 8, and 9 and Figure 4.

[0079] (7. Conclusion) Considering a fixed dose and various administration frequencies, rhNRG can cause myocardial infarction in rats during continuous administration. It exerts some therapeutic effect on, and as a result enhances cardiac function in rats with acute myocardial infarction. It improved ventricular remodeling and delayed aging due to myocardial infarction. rhNRG is still moderate. Even after discontinuation, it exerts a significant improvement in cardiac function in rats with myocardial infarction over a long period of time. Table 7 Echocardiographic results of the efficacy of 60-day subcutaneous administration of NRG in treating myocardial infarction in rats. Results (x±SD) [Table 7] ***: p<0.001 Subgroup after treatment compared to the excipient group; **: p<0.01 Subgroup after treatment compared to the excipient group; *: p<0.05 Subgroup after treatment compared to the excipient group Table 8 Treatment of rats with myocardial infarction following 60 days of subcutaneous administration of NRG and subsequent dose reduction for 3 weeks. Echocardiographic results of the effect in (x±SD) [Table 8] ***: p<0.001 Subgroup after treatment compared to the excipient group; **: p<0.01 Subgroup after treatment compared to the excipient group; *: p<0.05 Subgroup after treatment compared to the excipient group Table 9 Efficacy of NRG in the treatment of myocardial infarction in rats after discontinuation of subcutaneous administration on day 81. - Test results (x ± SD) [Table 9] ***: p<0.001 Subgroup after treatment compared to the excipient group; **: p<0.01 Subgroup after treatment compared to the excipient group; *: p<0.05 Subgroup after treatment compared to the excipient group

Claims

1. In the manufacture of pharmaceuticals for the prevention, treatment, or mitigation of myocardial injury in mammals, Use of uregrin (NRG) or its functional fragments.

2. The use according to claim 1, wherein the NRG is NRG-1, NRG-2, NRG-3, or NRG-4.

3. The use according to claim 1, wherein the NRG is NRG-1.

4. Other uses available to prevent, treat, or alleviate damage caused by myocardial infarction in mammals The use according to claim 1, including the use of a drug or treatment.

5. The NRG enhances mammalian cardiac function and reduces cardiac remodeling, as described in claim 1. Use on a vehicle.

6. The use according to claim 1, wherein the mammal is a human.

7. A composition for preventing, treating, or mitigating myocardial injury in mammals, in an effective amount The composition comprising NRG or a functional fragment thereof.

8. A method for preventing, treating, or mitigating myocardial injury in mammals, wherein neurate The method comprising subcutaneously administering grin (NRG) at a dose of 2.5 μg / kg / day to 50 μg / kg / day. 。

9. A method for preventing, treating, or mitigating myocardial injury in mammals, wherein neurate The method comprising administering gurin (NRG) multiple times a day for several consecutive days.

10. The aforementioned NRG is administered multiple times a day for several consecutive days, and then the NRG is slowly discontinued. The method according to claim 9, including the following.

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