Methods for preventing, treating, or delaying heart failure with preserved ejection fraction, and neureglin compositions.

Abstract

Medicaments and methods for preventing, treating, or delaying heart failure with preserved ejection fraction in mammals are provided. The present invention provides the use of neuregulin in the manufacture of a medicament for preventing, treating, or delaying heart failure with preserved ejection fraction in a mammal, and a pharmaceutical preparation for preventing, treating, or delaying heart failure with preserved ejection fraction in a mammal, the pharmaceutical preparation comprising an effective amount of neuregulin.

Description

Technical Field

[0001] (Field of the Invention) The present invention relates to the use of neuregulin protein in the manufacture of a medicament for preventing, treating or delaying heart failure with preserved ejection fraction in mammals, and to the use of said medicament for preventing, treating or delaying heart failure with preserved ejection fraction in mammals. More particularly, the present invention provides a method for preventing, treating or delaying heart failure with preserved ejection fraction in mammals, said method comprising administering to a mammal having heart failure with preserved ejection fraction or a particular population at risk of having heart failure with preserved ejection fraction a medicament comprising neuregulin protein. In particular, the present invention relates to a new use of neuregulin in the treatment of cardiovascular disease, namely heart failure with preserved ejection fraction.

Background Art

[0002] (Background of the Invention) Neuregulin (NRG; heregulin, HRG) is a type of glycoprotein with a molecular weight of 44 KD, also known as glial growth factor (GGF) and novel differentiation factor (NDF). As a ligand of the ErbB family of tyrosine kinase receptors, neuregulin contributes to cell signaling. The NRG family has four members: NRG1, NRG2, NRG3, and NRG4 (Falls et al., Exp Cell Res. 284:14-30, 2003). NRG1 plays an important role in the nervous system, heart and chest. Also, it has been demonstrated that NRG1 signaling plays a role in the development and function of other organ systems, as well as in the pathology of human diseases (including schizophrenia and breast cancer). There are many (including schizophrenia and breast cancer). There are many There are isomers. This is based on studies using genetically modified mice (gene knockout mice). isomers having different N-terminal regions or different EGF-like domains have different in vivo functions. It has been shown that it possesses [the characteristic]. This invention is based on NRG-1β.

[0003] NRG-1β is a transmembrane protein (Holmes et al., Science 256, 1205-1210, 1992). The extracellular domain is the N-terminal region, which contains an immunoglobulin-like domain (Ig-like domain) and an EGF-like domain. It contains a domain. The intracellular region is the C-terminal region. Extracellular matrix metallopro Under the action of teinase, the extracellular domain of NRG is cleaved by the enzyme and becomes free, Therefore, it promotes binding to ErbB3 receptors on the cell surface and activation of related cell signaling pathways. ru.

[0004] The EGF receptor family is classified into four classes, including ErbB1, ErbB2, ErbB3, and ErbB4. These can be transmembrane proteins with a molecular weight of approximately 180-185 kD. All except rbB2 contain an extracellular ligand-binding domain in the N-terminal region. These include ErbB3. All of them possess protein tyrosine kinase activity in their intracellular C-terminal region. ErbB1 While epidermal growth factor receptors are the primary receptors, ErbB3 and ErbB4 are neuroregulin receptors. Of these neuroregulin receptors, only ErbB2 and ErbB4 are highly expressed in the heart (Yarden et al.). (Reference: Nat Rev Mol Cell Biol, 2: 127-137, 2001).

[0005] When NRG binds to the extracellular domain of ErbB3 or ErbB4, it interacts with ErbB3, ErbB4, and other ErbB receptors. The formation of a heterodimer with (usually containing ErbB2) or a homodimer of ErbB4 is induced, As a result, phosphorylation occurs in the intracellular regions of these receptors (Yarden et al., Nat Rev Mol Ce (BioL, 2: 127-137, 2001). The phosphorylated intracellular domain then acts as an intracellular signaling pathway. It binds to signaling proteins, thereby activating the downstream AKT or ERK signaling pathway. and a series of cellular responses, such as cell proliferation, cell apoptosis, cell migration, cell differentiation, or fine-grained cells. It stimulates or reduces cell adhesion.

[0006] NRG plays a particularly important role in cardiac development (WO0037095, CN1276381, WO0309930) 0, WO9426298, US6444642, WO9918976, WO0064400, Zhao et al., J. Biol. Chem. 273, 1 0261-10269, 1998). In the early stages of embryonic development, NRG expression is limited to the endocardium, and Afterward, NRG is released to peripheral cardiomyocytes via paracrine linkage, and the protein tyrosine on the cell membrane is released. It binds to the extracellular domain of the kinase receptor ErbB4. ErbB4 then heterogeneously binds to ErbB2. It forms a mer. The formation and activation of the ErbB4 / ErbB2 complex is in the early stages of the sponge-like heart. Essential for trabeculae formation. Genes for three proteins: NRG protein, ErbB4, and ErbB2. If either of these is absent, the embryo will lack a column and die in the uterus during the early stages of development. It is likely that in WO0037095, a certain concentration of neuregulin can sustain the ERK signaling pathway. It activates sarco, promoting the differentiation and growth of cardiomyocytes, and at the site where cardiomyocytes adhere to other cells, sarco It induces the remodeling of the mare and cytoskeleton, improving the structure of cardiomyocytes and enhancing cardiomyocyte contraction. It has been shown that it is possible. Also, in WO0037095 and WO003099300, NRG is various It has been shown to be usable for the detection, diagnosis, and treatment of cardiovascular disease.

[0007] The following is a list of some prior art documents related to the present invention: 1. "Function of the myocardium and "Manipulation (Cardiac muscle function and manipulation)": WO0037095; 2. "Neuregri "New application of neuregulin and its analogs": C N1276381;3. "Neureglin-based methods and compositions for the treatment of cardiovascular disease (Neu regulin based methods and composition for treating cardiovascular diseases): WO03099300;4. Zhao YY, Sawyer DR, Baliga RR, Opel DJ, Han X, Marchionni MA, and The literature by Kelly RA states that "Neuregulin promotes the survival and growth of cardiomyocytes." s Promote Survival and Growth of Cardiac Myocytes)” J. Biol.Chem. 273, 10261-1 0269 (1998); 5. "Methods for treating muscle diseases and disorders" Ases and disorders: WO9426298:6. "Formation or development of myotubes using Neuroglin" Methods of increasing myotube formation or survival or muscle cell mitogenesis, differentiation or survival using a neuregulin 」: US6444642. 7. 「Therapeutic methods comprising use of a neuregulin 」: WO9918976; 8. 「Methods for treating congestive heart failure 」: WO0064400; 9. Holmes WE, Sliwkowski MX, Akita RW, Henzel WJ, Lee J, Park JW, Yansur a D, Abadi N, Raab H, Lewis GD et al., 「Identification of heregulin, a specific activator p185erbB2 」 Science 256, 1205 - 1210 (1992); 10. Falls DL, 「Neuregulins: functions, forms and signaling strategies 」 Experimental Cell Research, 284, 14 - 30 (2003). 11. Yarden Y, Sliwkowski X, 「Untangling the ErbB signaling Network 」 Science 287, 1948 - 1952 (2000). 12. 「Methods of increasing myotube formation or survival or muscle cell mitogenesis, differentiation or survival using a neuregulin 」: US6444642. 13. 「Therapeutic methods comprising use of a neuregulin 」: WO9918976; 14. 「Methods for treating congestive heart failure 」: WO0064400; 15. Holmes WE, Sliwkowski MX, Akita RW, Henzel WJ, Lee J, Park JW, Yansur a D, Abadi N, Raab H, Lewis GD et al., 「Identification of heregulin, a specific activator p185erbB2 It should be noted that there seems to be some inaccuracies or incompleteness in the original text, especially in the repeated and fragmented content. The above translation is based on the existing text as accurately as possible.」Nature Reviews: Molecular Cell Biology, 2127-137 (2001).

[0008] Heart failure (HF) is a cardiac dysfunction syndrome caused by various heart diseases, including systolic heart failure (SHF) and diastolic heart failure (DHF). In 2008, the European Society of Cardiology (ESC) issued the " Diagnosis and treatment guidelines of acute / chronic heart failure", and defined DHF as heart failure with preserved ejection fraction (HF-PEF). Systolic heart failure is a pathological condition in which the heart with reduced myocardial contractility fails to meet the metabolic requirements, resulting in insufficient cardiac output, low perfusion of tissues or organs, and congestion of the pulmonary circulation and / or systemic circulation. Heart failure with preserved ejection fraction (HF-PEF) often refers to diastolic heart failure caused by abnormal diastolic relaxation of the left ventricle, reduced myocardial compliance, myocardial cell hypertrophy, and interstitial fibrosis with increased left ventricular stiffness. This leads to abnormal diastolic filling, reduced cardiac output, increased end-diastolic pressure of the left ventricle, and the occurrence of heart failure. Epidemiological data from the US National Heart, Lung, and Blood Institute in 2006 showed that heart failure with preserved ejection fraction or diastolic heart failure accounted for more than 50% of the total number of patients with heart failure. Heart failure with preserved ejection fraction can exist alone or appear together with systolic dysfunction. It is more likely to occur in elderly women with hypertension, diabetes, and left ventricular hypertrophy. Diastolic heart failure and systolic heart failure have similar symptoms and signs. Patients usually have hypertension and other underlying diseases. In the early stage of heart failure, fatigue of unknown cause and reduced exercise tolerance are common. Furthermore, as the disease progresses, symptoms such as shortness of breath, orthopnea, paroxysmal nocturnal dyspnea, and peripheral edema may occur. These symptoms significantly affect the patient's quality of life and prognosis. At present, the treatment of heart failure mainly focuses on relieving symptoms, improving cardiac function, and preventing the progression of the disease. However, there is still a lack of effective treatment methods for heart failure with preserved ejection fraction.

[0009] Diastolic heart failure and systolic heart failure have similar symptoms and signs. Patients usually have hypertension and other underlying diseases. In the early stage of heart failure, fatigue of unknown cause and reduced exercise tolerance A persistent increase in heart rate of 15-20 beats per minute can be an early sign of left ventricular dysfunction. Symptoms include dyspnea at the time of breathing, paroxysmal nocturnal dyspnea, and high pillow sleep. It may be present later. Ascites or lower extremity edema may occur in the patient as a major or sole symptom. This can occur, and at the same time, impaired exercise endurance gradually develops in the patient.

[0010] Diastomas is a more complex physiological process than systomas, involving multiple factors. Therefore, Diagnosing heart failure with preserved ejection fraction or diastolic heart failure is more difficult than diagnosing systolic heart failure. A diagnosis can be made if the following conditions are met: 1. Typical symptoms and signs of heart failure; 2. Normal LVEF (or slightly decreased ≥45%), normal left ventricular morphology; 3. There is evidence of an underlying heart condition. For example, in patients with hypertension, ultrasound cardiac examination The diagnosis shows evidence of left ventricular hypertrophy, left atrial enlargement, and left ventricular diastolic dysfunction; 4. Increased BNP / NT-proBNP; 5. Cardiac ultrasound does not show valvular heart disease, and there is no pericardial disease, hypertrophic cardiomyopathy, or restrictive (invasive) cardiomyopathy. Cardiomyopathy (including lump-forming cardiomyopathy) and other conditions were ruled out.

[0011] Heart failure with preserved ejection fraction or diastolic heart failure is associated with various causes, and in these causes, the left heart Ventricular blood pressure / volume mechanisms are relatively well-recognized etiologies. Hypertension, hypertrophic cardiomyopathy, aortic valve Patients with stenosis showed a significantly increased ventricular end-diastolic blood pressure and a significantly decreased left ventricular volume. These have the effect of affecting ventricular filling, causing a leftward shift and concentricity of the blood pressure-volume curve. This leads to the formation of reconstructive processes. Prolonged stress overload can result in diastolic heart failure.

[0012] Ventricular diastolic function has two stages: relaxation (the dominant energy-consuming process) and This includes ventricular muscle compliance. Ventricular muscle relaxation is the dominant energy consumption process. This is the change in cardiac chamber blood pressure per unit time during a certain diastolic period. Ventricular muscle compliance This is caused by the change in unit volume during diastole, which is a passive filling process of the heart chamber. This refers to changes in blood pressure. Relaxation is the dominant ventricular muscle diastole in the early stages of diastole, and the cardiac retraction of myocardial fibers. The length of the pre-systolic phase and the ability to return to blood pressure, and energy-dependent Ca 2+ Leading energy transport It is an energy consumption process and includes isovolumetric relaxation and the early diastolic rapid filling phase. Left ventricular relaxation is , duration of isovoluted intravenous respiration (IVRT), maximum rate of blood pressure decrease (-dp / dt), mitral valve E-peak deceleration time ( This is reflected by parameters including DT. Used in two-dimensional ultrasound cardiac diagnosis and hemodynamic testing. These parameters obtained can be used to some extent to assess cardiac diastolic function. It is possible.

[0013] Furthermore, there is no specific treatment for heart failure with preserved ejection fraction. Current treatment guidelines Dryn is a standard medication that controls blood pressure, reduces heart rate, and decreases fluid retention. For example, angiotensin-converting enzyme inhibitors / angiotensin II receptor inhibitors, beta-blockers This includes the use of (and diuretics), but these drugs can improve the symptoms of systolic heart failure. It is not possible to improve the clinical symptoms and prognosis of heart failure with preserved ejection fraction. Patients with preserved heart failure or diastolic heart failure have a poor prognosis and a relatively high rate of re-injury. The patient has hospitalizations and repeated hospitalizations, which increases the burden on the overall healthcare system. Systolic Heart failure is the outcome of developing diastolic heart failure. How the heart in the early stages of diastolic heart failure Whether it improves diastolic outcomes and prevents further deterioration of diastolic heart failure remains to be seen. This is a major challenge in the treatment of heart failure.

[0014] Prior art literature includes information on neureglins in heart failure or diastolic heart failure with preserved ejection fraction. There are no reports on the role of proteins. In this invention, the neuregry of mammals The administration of natriol significantly improves the symptoms of heart failure with preserved ejection fraction, and natriol Grin is used to prevent, treat, or delay preserved ejection fraction heart failure in mammals. It was discovered that it could be used in the manufacture of eye medicine. [Overview of the project]

[0015] (Content of the invention) (A. Summary of the Invention) This invention states that NRG is extremely important for cardiac development and maintaining the function of a mature heart. Based on scientific discoveries, this invention relates to NRG in the sarcomeres, cytoskeleton, and cellular indirections of cardiomyocytes. This invention is based on the scientific discovery that the formation of the compound can be strengthened. Furthermore, the present invention is based on the fact that NRG is active Animals with heart failure in physical models, and the hearts of patients with heart failure in clinical trials. Based on scientific discoveries that it can improve function. Neureglin, Neureg Phosphate polypeptides, neuregulin derivatives, or compounds that mimic the activity of neuregulin. All of these fall within the scope of the present invention.

[0016] NRG proteins bind to ErbB receptors on the surface of cardiomyocytes and activate the ERK signaling pathway within the cell. By continuously activating them, the structure of cardiomyocytes can be altered, thereby changing the function of cardiomyocytes To improve one's abilities.

[0017] In a first embodiment of the present invention, in mammals, particularly humans, heart failure with preserved ejection fraction is achieved. Methods for preventing, treating, or delaying heart failure with preserved ejection fraction Effective in mammals, especially humans, that require or desire treatment or delay of treatment. A quantity of NRG or its functional fragment, or nucleic acids encoding NRG or its functional fragment, The drug administered a substance that improves the production and / or functionality of NRG, and as a result the ejection fraction was maintained. A method is provided that includes achieving an effect of preventing, treating, or delaying heart failure. It can be done.

[0018] In a second embodiment, the present invention provides a method for maintaining ejection fraction in heart failure in mammals, particularly humans. A pharmaceutical preparation for the purpose of preventing, treating, or delaying, which contains an effective amount of NRG or Functional fragments, or nucleic acids encoding NRG or its functional fragments, or production of NRG and / or This includes substances that improve functionality, as well as carriers, excipients, etc., that are acceptable as pharmaceuticals. The following pharmaceutical preparation is provided. This pharmaceutical preparation prevents and treats heart failure with preserved ejection fraction. Alternatively, it can be used in combination with other drugs(s) to delay the onset of action.

[0019] In another embodiment, the present invention relates to heart failure with preserved ejection fraction in mammals, particularly humans. A composition for preventing, treating, or delaying infant feeding, provided by the present invention. Pharmaceutical preparations for preventing, treating, or delaying heart failure with preserved ejection fraction in animals. Substances, as well as other drugs for preventing, treating, or delaying heart failure with preserved ejection fraction. The present invention provides the composition containing (multiple) of the above-mentioned components.

[0020] The present invention further aims to prevent and treat heart failure with preserved ejection fraction in mammals, particularly humans. A kit for treating or delaying heart failure with one or more doses that preserve ejection fraction. Pharmaceutical preparations or compositions used to prevent, treat, or delay, and said pharmaceutical preparation The kit includes instructions on how to use the substance or composition. [Modes for carrying out the invention]

[0021] (B. Definition) Unless otherwise defined, all technical and scientific terms used herein are defined in accordance with the present invention. It has the same meaning as is generally understood by those skilled in the art of the field to which it belongs. All patents, applications, published applications, and other publications cited herein are incorporated in their entirety by reference. The definitions set forth in this section are incorporated herein by reference in patents, applications, and publications. If the definition is contrary to, or otherwise contradicts, the definition stated in the application and other publications. The definitions set forth in this section shall supersede any definitions incorporated herein by reference.

[0022] As used herein, the singular forms "a," "an," and "the" are Unless the context clearly indicates otherwise, "at least one" or "1 It means "more than or equal to".

[0023] As used herein, "neureglin" or "NRG" refers to ErbB2, ErbB3, ErbB4, or Proteins that can bind to and activate those heterodimers or homodimers. Refers to a substance or peptide, such as a neureglin isoform or neureglin EGF-like domain. polypeptides containing a neuregulin EGF-like domain, neuregulin variants, or derivatives , and all kinds of neuregulin-like gene production that can activate the above receptors It contains substances. Also, neuregrin contains NRG-1, NRG-2, NRG-3, and NRG-4 proteins, neuregrin The present embodiment includes peptides, fragments, and compounds having the function of regulin. Neuregrin binds to ErbB2 / ErbB4 or ErbB2 / ErbB3 heterodimers and activates them. A protein or peptide that can be used to limit, for example, Although not the same, the peptide of the present invention is a fragment of the NRG-1 β2 isoform, namely 177-237 It contains an amino acid fragment, which includes an EGF-like domain having the following amino acid sequence: [ka] The NRG protein of the present invention activates the above receptors and modulates their biological functions. This can, for example, stimulate the synthesis of acetylcholine receptors in skeletal muscle cells, and in cardiac muscle cells. It promotes cell differentiation and survival, as well as DNA synthesis. Furthermore, NRG proteins play a vital role in biological functions. Includes NRG variants with conservative mutations that do not affect quality. Those skilled in the art will generally know that A single amino acid mutation in an unintended region can result in a protein or polypeptide. It is well known that it will not alter the biological activity of (for example, Watson et al.'s literature). "Molecular Biology of the Gene," 4th edition, 1987, The Bejacm (See in / Cummings Pub.co., p.224). The NRG protein of this invention is derived from natural sources. It can be isolated, or obtained through recombinant technology, artificial synthesis or other means. Cut.

[0024] The terms "epidermal growth factor-like domain" or "EGF-like domain" used herein refer to ErbB2, It binds to ErbB3, ErbB4, or their heterodimers or homodimers, and activates them. Encoded by the neuregulin gene, all of its contents are referenced herein. This is incorporated into WO 00 / 64400, Holmes et al.'s literature, Science, 256:1205-1210 (1992); US National Patent Nos. 5,530,109 and 5,716,930; Literature by Hijazi et al., Int. J. Oncol., 13:1061-10 67 (1998); Chang et al., Nature, 387:509-512 (1997); Carraway et al., Nature, 387:512-516 (1997); Higashiyama et al., J. Biochem., 122:675-680 (1997); and WO This refers to polypeptide fragments structurally similar to the EGF receptor binding domain, as described in 97 / 09425. In certain embodiments, the EGF-like domain is an ErbB2 / ErbB4 or ErbB2 / ErbB3 heterodimer. It binds to and activates them. In a particular embodiment, the EGF-like domain is a receptor for NRG-1. Includes the amino acid sequence of the encapsulation domain. In some embodiments, the EGF-like domain This refers to amino acid residues 177-226, 177-237, or 177-240 of NRG-1. Specific embodiments In this case, the EGF-like domain contains the amino acid sequence of the receptor-binding domain of NRG-2. In this embodiment, the EGF-like domain contains the amino acid sequence of the receptor-binding domain of NRG-3. In one embodiment, the EGF-like domain contains the amino acid sequence of the receptor-binding domain of NRG-4. In a particular embodiment, the EGF-like domain is described in U.S. Patent No. 5,834,229. like, [ka] It contains the amino acid sequence.

[0025] As used herein, "heart failure with preserved ejection fraction (HF-PEF)" refers to heart failure with a normal left ventricular ejection fraction. Heart failure accompanied by heart failure (HFNEF), heart failure with preserved left ventricular ejection fraction (HF-PLVEF), heart failure with preserved systolic function Also known as heart failure with elevated heart rate (HF-PSF) or diastolic heart failure (DHF), it involves myocardial cell hypertrophy and interstitial Fibrosis causes abnormal left ventricular diastolic relaxation and decreased myocardial compression Lyan, primarily due to increased stiffness, causes abnormal left ventricular diastolic filling. This leads to decreased cardiac output, increased left ventricular end-diastolic blood pressure, and the development of normal heart failure. This refers to a slightly reduced left ventricular ejection fraction (LVEF). Heart failure with preserved ejection fraction can exist on its own. It may occur, or may appear simultaneously with systolic dysfunction.

[0026] As used herein, "isovolumetric relaxation time (IVRT)" refers to the period during which the ventricles are in an isovolumetric closure state with decreasing blood pressure. In some cases, the ventricles begin to relax while the aortic valve and atrioventricular valves are closed. This means that if left ventricular relaxation does not function properly, IVRT will be prolonged. When this happens, IVRT decreases.

[0027] As used herein, "blood pressure decrease rate (-dp / dt)" refers to the left ventricular blood pressure during isovolumetric relaxation time. This refers to the rate of decrease. The larger the value, the faster the rate of decrease in left ventricular blood pressure, and the more the cardiac diastolic function decreases. It improves. The rate of blood pressure decrease is one of the reliable indicators for assessing myocardial relaxation.

[0028] As used herein, "mitral valve E peak" refers to the early stage of cardiac mitral valve orifice. This refers to the diastolic peak (E), which is the maximum blood flow velocity through the valve orifice during the rapid filling period of the left ventricle. Reflected. The E peak in the mitral valve orifice flow curve represents early diastolic active relaxation of the left ventricle. This reflects relaxation of the left ventricle.

[0029] As used herein, "E-peak descent time (DT)" refers to the deceleration time of the descent of the mitral valve E-peak. In other words, it refers to the slowdown of blood flow to the left atrium caused by premature diastolic mitral valve movement. This reflects rapid changes in left atrial blood pressure during the filling period. A smaller value indicates a more rapid change in blood pressure. The decrease in active relaxation usually occurs in the early stages of the disease, and the early diastolic filling volume of the left ventricle. This manifests as a decrease in E, a decrease in the E peak, and an extension of DT by >240 ms.

[0030] As used herein, “other drugs for the treatment of heart failure with preserved ejection fraction” Angiotensin-converting enzyme inhibitors / angiotensin II receptor inhibitors, β-receptor inhibitors Tagonist, calcium antagonist, cyclic adenosine monophosphate, catecholamine, Nitrate esters, phosphatase inhibitors, diuretics, renin, angiotensin, aldosterone This includes RAS antagonists and myocardial energy optimizers, which are used in cardiac patients with preserved ejection fraction. This refers to known drugs for the treatment of all conditions. [Examples]

[0031] (Examples) (Example 1: Effects of recombinant human neureglins on cardiac function in hypertensive heart failure rats) test)

[0032] This study involved recombinant human neuregulin (rhNRG) in SHR hypertensive rats with heart failure. This study concerns the therapeutic effect of ). Methods: SHR hypertensive rat strain, standard diet, diet Monitoring of changes in cardiac function during the period. After 16 months, ejection fraction (EF) decreased to 70%, indicating a high level of heart failure. The successful establishment of a rat blood pressure model was suggested. Rats with hypertensive heart failure were randomly selected. The subjects were divided into a negative control group, an NRG-treated group, and a captopril-treated group. rh NRG was administered for 5 consecutive days, followed by a 2-day rest period, and the NRG group underwent 3 treatment cycles. At the end of the third treatment cycle, each group of rats was examined using cardiac ultrasound to determine changes in cardiac function. After the third treatment cycle, the hemodynamics of the rats were measured.

[0033] (1. Laboratory animals) 1.1 Strain, origin: SHR hypertensive rat strain developed at the Animal Center of the Chinese Academy of Sciences It was purchased from the Chinese Academy of Sciences. Additionally, the WKY strain was used as a control for SHR in China. I purchased it from the National Institute of Animal Sciences' Animal Center.

[0034] 1.2 Sex, age: Male, 6 weeks old.

[0035] 1.3 Feeding: Standard rodent feed, free access to water, 12-hour light-dark cycle.

[0036] (2. Test drug)

[0037] Details: Neucardin®, 61 amino acids, produced by Shanghai Zensun Sci & Tech Co., Ltd. .

[0038] (3. Test materials)

[0039] 3.1 Cardiac ultrasound diagnostic equipment: Philips Sonos 5500

[0040] 3.2 Captopril: Sino American Shanghai Squibb Pharmaceuticals

[0041] (4. Experimental Method)

[0042] 4.1 Establishment of a rat model of hypertensive heart failure

[0043] SHR hypertensive rat strain, normal feeding, monitoring of cardiac function changes during feeding. 16 months later. In SHR rats, the ejection fraction (EF) decreased to 70%, and LVDd and LVDs significantly increased, leading to hypertensive heart disease. This suggests that a defective rat model has been successfully established.

[0044] 4.2 Group Classification and Administration

[0045] After the model was successfully established, rats were randomly assigned to a negative control group, an NRG treatment group, and a captive group. Patients were divided into the topril treatment group. rhNRG was administered for 5 consecutive days, followed by a 2-day rest period. This constituted one treatment cycle, and the drug was administered intravenously once daily at a dose of 6.5 μg / kg. A total of three treatment cycles were performed. The drug was administered in a tethered manner. Simultaneously, the NRG group was given drinking water twice daily via intragastric administration. Pril was administered intragastricly at a dose of 10 mg / kg twice daily. The NRG excipient was used for three treatment cycles. It was administered by tail vein infusion over time. The negative control group was given drinking water by intragastric administration, and NR The G excipient was administered by tail vein injection.

[0046] 4.3 Cardiac Ultrasound Diagnostic Test

[0047] Before the procedure and at the end of the second and third treatment cycles, cardiac ultrasound was performed after ketamine anesthesia. Then, the changes in cardiac function were determined.

[0048] 4.4 Hemodynamic Measurement

[0049] At the end of the third treatment cycle, the rats were anesthetized by intraperitoneal infusion of 3% pentobarbital. A midline incision is made in the neck, the left common carotid artery is isolated and intubated, and the arterial and left ventricular hemodynamic parameters are measured. The temperature was measured.

[0050] (5. Experimental Results)

[0051] Compared to the negative control group, NRG significantly improved hemodynamics in hypertensive rats. This was achieved, and -dp / dt showed a statistically significant difference between these groups (-7467.6±715.8 and -, respectively). 5488.1±1340.3, P=0.016); and captopril significantly reduced blood pressure in hypertensive rats. This was possible (174.5±33.0 vs. 216.5±23.2, and 228.0±26.0; p=0.029, p=0.017). ).

[0052] (6. Conclusion)

[0053] rhNRG was administered to hypertensive heart failure rats at a dose of 6.5 μg / kg for 5 consecutive days in one treatment cycle. Then, a two-day drug-free period was established. After two or three cycles of treatment, rhNRG was administered at left ventricular end-diastolic and Furthermore, it can prevent terminal volume enlargement and improve hemodynamics, resulting in Cardiac function is improved in hypertensive heart failure rats. As shown in Table 1, captopril Lowering blood pressure can improve cardiac function in hypertensive heart failure rats. rhNRG does not lower blood pressure, but rather the rate of left ventricular descent during isovolumetric diastole -d By increasing the p / dt ratio, cardiac function in hypertensive heart failure rats can be improved.

[0054] Table 1. Hemodynamic parameters of each group after 3 treatment cycles [Table 1]

[0055] (Example 2: Recombinant human neureglins for cardiac function in heart failure patients with preserved ejection fraction) (Test of action)

[0056] The effect of recombinant human neureglins on cardiac function in patients with heart failure who have preserved ejection fraction. To evaluate its use, a preliminary clinical trial was conducted at the Sixth People's Hospital affiliated with Shanghai Jiao Tong University. The test was conducted at (Pital Affiliated to Shanghai Jiao Tong University). The study included two patients in the placebo group and two patients in the experimental group.

[0057] (1. Main participation criteria)

[0058] 1.1 Left ventricular ejection fraction (LVEF) ≥ 50% (two-dimensional echocardiography);

[0059] 1.2 New York City Cardiac Function (NYHA) level II or III;

[0060] 1.3 Clear diagnosis of chronic heart failure (including history, symptoms, and signs), clinical symptoms in the past month It was stable;

[0061] 1.4 Patients receiving standard treatment for heart failure receiving the target dose for at least one month. Alternatively, the maximum tolerable dose has been reached, or the dose has not been changed within the past month;

[0062] 1.5 Understand and sign the informed consent form.

[0063] (2. Test drug)

[0064] Name: Recombinant human neuregulin for injection

[0065] Details: 250 g / vial

[0066] Dosage form: Lyophilized powder for injection

[0067] Route of administration: Intravenous infusion

[0068] Placebo (zero dose):

[0069] Name: Excipient of lyophilized recombinant human neureglins

[0070] Dosage form: Lyophilized powder for injection

[0071] Route of administration: Intravenous infusion

[0072] (3. The route of administration, dosage, and course of treatment are shown in Table 2.) Table 2 Dosage, route of administration, and course of treatment [Table 2]

[0073] (4. Data collection:) The mitral valve flow spectrum in two-dimensional ultrasound cardiac diagnosis is used during the screening period, days 11-13. , and detected on the 30th.

[0074] (5. Results and Discussion:) Table 3. Numerical changes in IVRT and DT in the mitral valve flow spectrum. [Table 3]

[0075] Table 3 shows that IVRT and DT levels gradually increased in patients who received a placebo, whereas NRG was administered to patients who received a placebo. The IVRT and DT values ​​of the treated patients were shown to be significantly reduced, indicating a degree of improvement in diastolic function. This was shown to be the case.

[0076] The above embodiments do not limit the scope of protection of the invention. This does not deviate from the object and scope of the present invention. Those skilled in the art can adjust and modify the present invention without doing so. Therefore, the present invention The scope of protection shall be defined by the claims, not by specific examples. ru. This application provides an invention having the following configuration: (Composition 1) Medical treatments to prevent, treat, or delay heart failure with preserved ejection fraction in mammals. The use of neuregulin in the manufacture of pharmaceuticals. (Configuration 2) The use according to configuration 1, wherein the aforementioned neuregulin is NRG-1. (Composition 3) The use of the neuregulin according to configuration 1, wherein the neuregulin contains the amino acid sequence of sequence number 1. (Composition 4) The use according to configuration 1, wherein the mammal is a human. (Composition 5) Medical treatments to prevent, treat, or delay heart failure with preserved ejection fraction in mammals. A pharmaceutical preparation comprising an effective amount of NRG. (Composition 6) The pharmaceutical preparation according to configuration 5, wherein the aforementioned neuregulin is NRG-1. (Composition 7) The pharmaceutical preparation according to Constituent 5, wherein the neuregulin contains the amino acid sequence of Sequence ID No. 1. (Composition 8) The pharmaceutical preparation according to composition 5, wherein the mammal is a human. (Composition 9) A set of devices for preventing, treating, or delaying heart failure with preserved ejection fraction in mammals. A product, which is a pharmaceutical preparation described in component 5 and for treating heart failure with preserved ejection fraction. The composition comprising the drug(s) (or more). (Composition 10) Used in mammals to prevent, treat, or delay heart failure with preserved ejection fraction. A kit to be used, comprising the pharmaceutical preparation described in component 5 and instructions for the use of the pharmaceutical preparation. The kit includes the above.

Claims

1. A pharmaceutical preparation for use in a method for preventing or treating heart failure with preserved ejection fraction in humans, The pharmaceutical preparation contains an effective amount of neuregulin, The pharmaceutical preparation wherein the neuregulin contains the amino acid sequence of Sequence ID No.

1.

2. The pharmaceutical preparation for use according to claim 1, wherein the neuregulin is NRG-1.

3. The pharmaceutical preparation for use according to claim 1, wherein the human has a left ventricular ejection fraction (LVEF) ≥ 50% and / or New York function (NYHA) level II or III.

4. A pharmaceutical preparation for use according to any one of claims 1 to 3, wherein the method comprises the administration of one or more further agents for treating heart failure with preserved ejection fraction.

5. The pharmaceutical preparation for use according to claim 4, wherein the further agent is an angiotensin-converting enzyme inhibitor / angiotensin II receptor inhibitor, a β-receptor antagonist, a calcium antagonist, a cyclic adenosine monophosphate, a catecholamine, a nitrate ester, a phosphatase inhibitor, a diuretic, a renin-angiotensin-aldosterone system (RAS) antagonist, or a myocardial energy optimizer.

6. A kit for use in a method for preventing or treating heart failure with preserved ejection fraction in humans, The kit comprises the pharmaceutical preparation described in claim 1.

7. The kit for use according to claim 6, wherein the human has a left ventricular ejection fraction (LVEF) ≥ 50% and / or New York function (NYHA) level II or III.

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