RU2795320C2 - Therapeutic agent for the treatment of cardiomyopathy, previous myocardial infarction and chronic heart failure - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: group of inventions relates to the prevention or treatment of cardiomyopathy and past myocardial infarction, as well as the resulting chronic heart failure. Methods are provided for treating cardiomyopathy and past myocardial infarction, preventing chronic heart failure caused by cardiomyopathy or past myocardial infarction, inhibiting a structural abnormality of the heart selected from cardiomegaly, cardiomyocyte hypertrophy, and myocardial fibrosis in a subject with cardiomyopathy or past myocardial infarction. The methods include administering to a subject with cardiomyopathy or prior myocardial infarction an effective amount of the following: (a) a peptide consisting of SEQ ID NO: 1; (b) a peptide consisting of an amino acid sequence in which one amino acid has been replaced, deleted, inserted or added in SEQ ID NO: 1, and having the activity of promoting migration of PDGFR α-positive and CD29-positive mesenchymal stem cells; or (c) a peptide consisting of an amino acid sequence having an identity of 97% or more with SEQ ID NO: 1 and having promoting activity of migration of PDGFR α-positive and CD29-positive mesenchymal stem cells.

EFFECT: inventions make it possible to improve heart functions by inhibiting cardiomegaly, cardiomyocyte hypertrophy and myocardial fibrosis and stimulating angiogenesis in dilated, ischemic and/or hypertensive cardiomyopathy.

27 cl, 23 dwg, 4 ex

Description

The field of technology to which the invention belongs

The present application relates to pharmaceutical compositions comprising a peptide fragment of the HMGB1 protein for the prevention and/or treatment of cardiomyopathy, past myocardial infarction and chronic heart failure.

Prerequisites for the creation of the invention

Cardiomyopathy is defined as "myocardial disease associated with cardiac dysfunction", often accompanied by structural abnormalities of the heart such as cardiomegaly, cardiomyocyte hypertrophy, and myocardial fibrosis; and shows symptoms of chronic heart failure as it progresses. Secondary cardiomyopathy can be alleviated by treating the underlying disease, but there is still no effective treatment for idiopathic cardiomyopathy.

Myocardial infarction, which causes myocardial necrosis due to coronary artery occlusion, is a major serious heart disease that is the leading cause of death in developed countries (the first in the United States and the second in Japan). While acute outcomes have been improved by advances in diagnostic technology, catheter therapy, and coronary artery bypass grafting, progression of cardiomegaly and cardiac hypertrophy and chronic heart failure often occur in cases of extensive infarction, severe reperfusion failure, and in cases where the opportunity for cardiac arrest is missed. treatment.

When chronic heart failure caused by cardiomyopathy or previous myocardial infarction becomes severe, existing drugs for chronic heart failure (ACE inhibitors, β blockers, etc.) in some cases cannot achieve sufficient relieving effects, and heart transplantation may be required . However, the waiting period for transplantation in Japan is up to about 3 years (Non-Patent Document 1), and complications such as cerebral infarction and device-related infection often occur during the waiting period. In addition, even if a heart transplant is obtained, there are problems such as an increased risk of infectious diseases due to the side effects of immunosuppressive drugs, which then must be constantly administered, and possible complications, such as coronary artery lesions; and 10-year survival after heart transplantation is about 50% (non-patent document 2). Under such circumstances, the development of new therapeutic agents for the treatment of cardiomyopathy and past myocardial infarction, as well as the resulting chronic heart failure, is desirable.

Recently, regenerative medicine using mesenchymal stem cells has attracted attention, and, regarding chronic heart failure after myocardial infarction, it has been reported that the effect of improving cardiac function can be achieved by direct transplantation of a cell sheet consisting of mesenchymal stem cells isolated from ovarian membrane, bone marrow, or adipose tissue, in the heart in an animal model of chronic myocardial infarction (Non-Patent Document 3 and Patent Document 1). However, cell sheet transplantation is very burdensome for the patient because it requires a thoracotomy and cannot be used for patients in whom surgery is difficult due to factors such as advanced age.

The mechanism by which damaged tissue releases bone marrow pluripotent stem cell recruitment factors into the blood to stimulate the regeneration of damaged tissue has also been confirmed. In previous studies, the present inventors have found that peptide fragments of the high mobility group 1 protein (HMGB1) mobilize mesenchymal bone marrow stem cells from the bone marrow to the peripheral blood and that administration of the peptide fragments in the acute phase of myocardial infarction leads to the accumulation of mesenchymal stem cells occurring from the bone marrow, at and near the infarction site, resulting in improved cardiac function (Patent Documents 2 and 3).

However, in the acute phase of myocardial infarction, a large number of cardiomyocytes are quickly necrotized by ischemia and a strong inflammatory reaction occurs, whereas cardiomyopathy usually does not cause such phenomena and is characterized by the fact that structural abnormalities such as cardiomegaly, cardiomyocyte hypertrophy and myocardial fibrosis progress slowly. The pathologies of these two diseases are very different. Necrosis and inflammation of cardiomyocytes are also localized in myocardial infarction, and this disease is characterized by the progression of cardiomegaly and cardiac hypertrophy. Therefore, it is unclear whether peptide fragments of the HMGB1 protein have therapeutic effects on cardiomyopathy and prior myocardial infarction.

List of reference documents

[Non-Patent Documents]

Non-Patent Document 1: Fukushima et al., "Registry Report on Heart Transplantation in Japan (June 2016)," Circulation Journal (Advance Publication, January 6, 2017, Article ID: CJ-16-0976)

Non-Patent Document 2: Lund et al., ʺThe Registry of the International Society for Heart and Lung Transplantation: Thirty-third Adult Heart Transplantation Report-2016; Focus Theme: Primary Diagnostic Indications for Transplant, J Heart Lung Transplant. 2016 Oct;35(10):1158-1169

Non-Patent Document 3: Ishikane Shin, ʺDevelopment of multi-growth factor secreted fetal membrane-derived mesenchimal stem cell sheetsʺ June 11, 2014, Grants-in-Aid for Scientific Research, Research report

[Patent Documents]

Patent Document 1: WO2006/080434

Patent Document 2: WO2012/147470

Patent Document 3: WO2014/065347

The essence of the invention

[Problems to be solved by the present invention]

The purpose of the present application is to provide new pharmaceuticals effective for the prevention and/or treatment of cardiomyopathy and past myocardial infarction, as well as the resulting chronic heart failure. Another object of the present application is to provide new pharmaceuticals effective for the prevention and/or treatment of heart diseases associated with structural abnormalities and/or dysfunction of the heart.

[Ways to solve problems]

As a result of studying the effects of HMGB1 peptide fragments in cardiomyopathy, the present inventors have found that HMGB1 peptide fragments having certain amino acid sequences exhibit the effects of improving cardiac function, inhibiting cardiac structural abnormalities (cardiomyocyte hypertrophy and myocardial fibrosis), and promoting angiogenesis in an animal model of dilated cardiomyopathy. The present inventors also found that in an animal model of ischemic cardiomyopathy resulting from myocardial infarction, specific peptide fragments of HMGB1 exhibit effects of improving cardiac function, inhibiting cardiac structural abnormalities (cardiomegaly, cardiomyocyte hypertrophy and myocardial fibrosis), and promoting angiogenesis. In addition, the present inventors have found that specific HMGB1 peptide fragments exhibit an inhibitory effect on cardiac structural abnormalities (cardiomyocyte hypertrophy and myocardial fibrosis) in an animal model of hypertensive cardiomyopathy. Thus, the present application provides pharmaceutical compositions comprising specific HMGB1 peptide fragments for the prevention and/or treatment of cardiomyopathy and past myocardial infarction and resulting chronic heart failure. Specific peptide fragments of HMGB1 also inhibit structural abnormalities and/or dysfunction of the heart in cardiomyopathy and inhibit structural abnormalities and dysfunction of the heart in myocardial infarction. Thus, the present application provides pharmaceutical compositions comprising specific HMGB1 peptide fragments for the prevention and/or treatment of heart diseases associated with structural abnormalities and/or dysfunction of the heart.

Thus, the present application represents the following inventions:

[1] A pharmaceutical composition for the prevention and/or treatment of a heart disease associated with a structural abnormality and/or dysfunction of the heart, which comprises a substance described in any of (a) to (c) below (referred to below as substance A):

(a) a peptide fragment of HMGB1 comprising the amino acid sequence described in SEQ ID NO: 1;

(b) a peptide comprising an amino acid sequence wherein one or more amino acids are replaced, deleted, inserted or added in the amino acid sequence described in SEQ ID NO: 1 and has cell migration promoting activity; And

(c) a peptide comprising an amino acid sequence having a sequence identity of about 80% or more with the amino acid sequence described in SEQ ID NO: 1 and having cell migration promoting activity.

[2] The pharmaceutical composition according to [1], wherein the heart disease associated with a structural abnormality and/or dysfunction of the heart is cardiomyopathy.

[3] The pharmaceutical composition according to [2], wherein the cardiomyopathy is idiopathic cardiomyopathy.

[4] The pharmaceutical composition of [3], wherein the idiopathic cardiomyopathy is selected from the group consisting of dilated cardiomyopathy, hypertrophic cardiomyopathy, restrictive cardiomyopathy, and arrhythmogenic right ventricular cardiomyopathy.

[5] The pharmaceutical composition according to [2], wherein the cardiomyopathy is a secondary cardiomyopathy.

[6] The pharmaceutical composition according to [5], wherein the secondary cardiomyopathy is selected from the group consisting of ischemic cardiomyopathy, hypertensive cardiomyopathy, valvular cardiomyopathy, drug-induced cardiomyopathy, alcoholic cardiomyopathy, mitochondrial cardiomyopathy, cardiac sarcoidosis-induced cardiomyopathy, cardiac amyloidosis, myocarditis-induced cardiomyopathy cardiomyopathy caused by muscular dystrophy cardiomyopathy caused by Fabry's disease cardiomyopathy and cardiomyopathy of pregnancy.

[7] The pharmaceutical composition according to [1], wherein the heart disease associated with a structural abnormality and/or dysfunction of the heart is a past myocardial infarction.

[8] Pharmaceutical composition comprising substance A, which is intended for the prevention and/or treatment of chronic heart failure caused by cardiomyopathy or past myocardial infarction.

[9] A pharmaceutical composition comprising Substance A for inhibiting a structural abnormality of the heart selected from the group consisting of cardiomegaly, cardiomyocyte hypertrophy, and myocardial fibrosis in a patient with cardiomyopathy or myocardial infarction.

[10] A pharmaceutical composition comprising substance A, which is intended for the prevention and/or treatment of cardiomyopathy selected from the group consisting of dilated cardiomyopathy, ischemic cardiomyopathy and hypertensive cardiomyopathy.

[11] The pharmaceutical composition according to [10], wherein the ischemic cardiomyopathy is caused by a previous myocardial infarction.

[12] A pharmaceutical composition comprising substance A, which is intended for the prevention and/or treatment of chronic heart failure caused by cardiomyopathy selected from the group consisting of dilated cardiomyopathy, ischemic cardiomyopathy and hypertensive cardiomyopathy.

[13] The pharmaceutical composition according to [12], wherein the ischemic cardiomyopathy is caused by a previous myocardial infarction.

[14] The pharmaceutical composition according to [12], wherein the chronic heart failure is caused by dilated cardiomyopathy or ischemic cardiomyopathy.

[15] The pharmaceutical composition of [14], wherein the chronic heart failure caused by dilated cardiomyopathy or ischemic cardiomyopathy is HFrEF.

[16] The pharmaceutical composition according to [12], wherein the chronic heart failure is caused by hypertensive cardiomyopathy.

[17] The pharmaceutical composition of [16], wherein the chronic heart failure caused by hypertensive cardiomyopathy is HFpEF.

[A1] A method for preventing and/or treating a heart disease associated with a structural abnormality and/or dysfunction of the heart, which comprises administering to a subject an effective amount of Substance A.

[A2] The method of [A1], wherein the heart disease associated with a structural abnormality and/or dysfunction of the heart is cardiomyopathy.

[A3] The method of [A2], wherein the cardiomyopathy is idiopathic cardiomyopathy.

[A4] The method of [A3], wherein the idiopathic cardiomyopathy is selected from the group consisting of dilated cardiomyopathy, hypertrophic cardiomyopathy, restrictive cardiomyopathy, and arrhythmogenic right ventricular cardiomyopathy.

[A5] The method of [A2], wherein the cardiomyopathy is a secondary cardiomyopathy.

[A6] The method of [A5], wherein the secondary cardiomyopathy is selected from the group consisting of ischemic cardiomyopathy, hypertensive cardiomyopathy, valvular cardiomyopathy, drug-induced cardiomyopathy, alcoholic cardiomyopathy, mitochondrial cardiomyopathy, cardiac sarcoidosis cardiomyopathy, cardiac amyloidosis cardiomyopathy, cardiomyocardial myocarditis party muscular dystrophy-induced cardiomyopathy, Fabry disease-induced cardiomyopathy, and cardiomyopathy of pregnancy.

[A7] The method of [A1], wherein the heart disease associated with a structural abnormality and/or dysfunction of the heart is a past myocardial infarction.

[A8] A method for preventing and/or treating chronic heart failure caused by cardiomyopathy or past myocardial infarction, which comprises administering to a subject an effective amount of Substance A.

[A9] A method for inhibiting a structural heart abnormality selected from the group consisting of cardiomegaly, cardiomyocyte hypertrophy, and myocardial fibrosis in a patient with cardiomyopathy or prior myocardial infarction, which comprises administering to the subject an effective amount of Substance A.

[A10] A method for preventing and/or treating a cardiomyopathy selected from the group consisting of dilated cardiomyopathy, ischemic cardiomyopathy, and hypertensive cardiomyopathy, which comprises administering to a subject an effective amount of Substance A.

[A11] The method of [A10], wherein the ischemic cardiomyopathy is caused by a previous myocardial infarction.

[A12] A method of preventing and/or treating chronic heart failure caused by cardiomyopathy selected from the group consisting of dilated cardiomyopathy, ischemic cardiomyopathy, and hypertensive cardiomyopathy, which comprises administering to a subject an effective amount of Substance A.

[A13] The method of [A12], wherein the ischemic cardiomyopathy is caused by a previous myocardial infarction.

[A14] The method of [A12], wherein the chronic heart failure is caused by dilated cardiomyopathy or ischemic cardiomyopathy.

[A15] The method of [A14], wherein the chronic heart failure caused by dilated cardiomyopathy or ischemic cardiomyopathy is HFrEF.

[A16] The method of [A12], wherein the chronic heart failure is caused by hypertensive cardiomyopathy.

[A17] The method of [A16], wherein the chronic heart failure caused by hypertensive cardiomyopathy is HFpEF.

[B1] Substance A for use in the prevention and/or treatment of a heart disease associated with a structural abnormality and/or dysfunction of the heart.

[B2] The substance A of [B1], wherein the heart disease associated with a structural abnormality and/or dysfunction of the heart is cardiomyopathy.

[B3] Substance A according to [B2], wherein the cardiomyopathy is idiopathic cardiomyopathy.

[B4] Substance A according to [B3], wherein the idiopathic cardiomyopathy is selected from the group consisting of dilated cardiomyopathy, hypertrophic cardiomyopathy, restrictive cardiomyopathy, and arrhythmogenic right ventricular cardiomyopathy.

[B5] Substance A according to [B2], wherein the cardiomyopathy is a secondary cardiomyopathy.

[B6] The substance A of [B5], wherein the secondary cardiomyopathy is selected from the group consisting of ischemic cardiomyopathy, hypertensive cardiomyopathy, valvular cardiomyopathy, drug-induced cardiomyopathy, alcoholic cardiomyopathy, mitochondrial sarcoidosis cardiomyopathy, cardiac amyloidosis cardiomyopathy, myocardial cardiomyopathy volume cardiomyopathy caused by muscular dystrophy cardiomyopathy caused by Fabry's disease cardiomyopathy and cardiomyopathy of pregnancy.

[B7] Substance A according to [B1], wherein the heart disease associated with a structural abnormality and/or dysfunction of the heart is a past myocardial infarction.

[B8] Substance A for use in the prevention and/or treatment of chronic heart failure caused by cardiomyopathy or past myocardial infarction.

[B9] Substance A for use in inhibiting a structural abnormality of the heart selected from the group consisting of cardiomegaly, cardiomyocyte hypertrophy, and myocardial fibrosis in a patient with cardiomyopathy or prior myocardial infarction.

[B10] Substance A for use in the prevention and/or treatment of cardiomyopathy selected from the group consisting of dilated cardiomyopathy, ischemic cardiomyopathy and hypertensive cardiomyopathy.

[B11] Substance A according to [B10], wherein the ischemic cardiomyopathy is caused by a previous myocardial infarction.

[B12] Substance A for use in the prevention and/or treatment of chronic heart failure caused by cardiomyopathy selected from the group consisting of dilated cardiomyopathy, ischemic cardiomyopathy and hypertensive cardiomyopathy.

[B13] Substance A according to [B12], wherein the ischemic cardiomyopathy is caused by a previous myocardial infarction.

[B14] Substance A according to [B12], wherein the chronic heart failure is caused by dilated cardiomyopathy or ischemic cardiomyopathy.

[B15] Substance A according to [B14], wherein the chronic heart failure caused by dilated cardiomyopathy or ischemic cardiomyopathy is HFrEF.

[B16] Substance A according to [B12], wherein the chronic heart failure is caused by hypertensive cardiomyopathy.

[B17] Substance A according to [B16], wherein chronic heart failure caused by hypertensive cardiomyopathy is HFpEF.

[C1] The use of Substance A for the manufacture of a medicament for the prevention and/or treatment of a heart disease associated with a structural abnormality and/or dysfunction of the heart.

[C2] The use of [C1], wherein the heart disease associated with a structural abnormality and/or dysfunction of the heart is cardiomyopathy.

[C3] The use of [C2], wherein the cardiomyopathy is idiopathic cardiomyopathy.

[C4] The use of [C3], wherein the idiopathic cardiomyopathy is selected from the group consisting of dilated cardiomyopathy, hypertrophic cardiomyopathy, restrictive cardiomyopathy, and arrhythmogenic right ventricular cardiomyopathy.

[C5] The use of [C2], wherein the cardiomyopathy is a secondary cardiomyopathy.

[C6] The use of [C5] wherein the secondary cardiomyopathy is selected from the group consisting of ischemic cardiomyopathy, hypertensive cardiomyopathy, valvular cardiomyopathy, drug-induced cardiomyopathy, alcoholic cardiomyopathy, mitochondrial cardiomyopathy, cardiac sarcoidosis cardiomyopathy, cardiac amyloidosis cardiomyopathy, cardiomyocardial myocarditis party muscular dystrophy-induced cardiomyopathy, Fabry disease-induced cardiomyopathy, and cardiomyopathy of pregnancy.

[C7] The use of [C1], wherein the heart disease associated with a structural abnormality and/or dysfunction of the heart is a past myocardial infarction.

[C8] The use of Substance A for the preparation of a medicament for the prevention and/or treatment of chronic heart failure caused by cardiomyopathy or past myocardial infarction.

[C9] The use of Substance A for the preparation of a medicament for inhibiting a structural heart abnormality selected from the group consisting of cardiomegaly, cardiomyocyte hypertrophy and myocardial fibrosis in a patient with cardiomyopathy or prior myocardial infarction.

[C10] The use of Substance A for the manufacture of a medicament for the prevention and/or treatment of cardiomyopathy selected from the group consisting of dilated cardiomyopathy, ischemic cardiomyopathy and hypertensive cardiomyopathy.

[C11] The use of [C10], wherein the ischemic cardiomyopathy is caused by prior myocardial infarction.

[C12] The use of Substance A for the manufacture of a medicament for the prevention and/or treatment of chronic heart failure caused by cardiomyopathy selected from the group consisting of dilated cardiomyopathy, ischemic cardiomyopathy and hypertensive cardiomyopathy.

[C13] The use of [C12], wherein the ischemic cardiomyopathy is caused by prior myocardial infarction.

[C14] Use according to [C12], wherein the chronic heart failure is caused by dilated cardiomyopathy or ischemic cardiomyopathy.

[C15] The use of [C14], wherein the chronic heart failure caused by dilated cardiomyopathy or ischemic cardiomyopathy is HFrEF.

[C16] The use of [C12], wherein the chronic heart failure is caused by hypertensive cardiomyopathy.

[C17] The use of [C16], wherein the chronic heart failure caused by hypertensive cardiomyopathy is HFpEF.

Brief description of the drawings

Fig. 1 is a graph showing the measurement results of left ventricular ejection fraction (LVEF), left ventricular end diastolic diameter (LVDd) and left ventricular end systolic diameter (LVDs) in the HMGB1 peptide (1-44) and PBS administration groups before, after 4 weeks and 6 weeks after administration.

Fig. 2 is a photograph showing the result of Sirius red staining of myocardial tissue sections and a graph showing the percentage of positively stained area.

Fig. 3 is a photograph showing the result of immunostaining (anti-CD31 antibody) of myocardial tissue sections and a graph showing capillary density.

Fig. 4 is a photograph showing the result of PAS staining of myocardial tissue sections and a graph showing the short diameter of cardiomyocytes.

Fig. 5 is a photograph showing the result of immunostaining using antibodies against PDGFRα and CD29. PDGFRα - green; CD29 - red; DAPI - blue.

Fig. 6 is a graphical representation of the number of PDGFRα-positive and CD29-positive cells in the HMGB1 (1-44) peptide and PBS administration groups.

Fig. 7 is a graphical representation of VEGF and TSG-6 expression levels in the RT-PCR assay. RQ=relative quantity.

Fig. 8 is a photograph showing the results of electron microscope observation of cardiomyocytes.

Fig. 9 is a graph showing the results of measuring the left ventricular ejection fraction (LVEF) in the time period before administration - 14 weeks after administration in the HMGB1 peptide administration group (1-44) and the PBS group.

Fig. 10 is a graphical representation of viability in the HMGB1 peptide group (1-44) and the PBS group.

Fig. 11 is a graph showing the measurement results of left ventricular ejection fraction (LVEF) in the HMGB1 (1-44) peptide administration group and the PBS group before and 4 weeks after administration.

Fig. 12 is a graph showing the measurement results of left ventricular end diastolic diameter (LVDd) and left ventricular end systolic diameter (LVDs) in the HMGB1 peptide (1-44) administration group and the PBS group before and 4 weeks after administration.

Fig. 13 is a graph showing the results of measuring the left ventricular ejection fraction (LVEF) in the time period before administration - 4 weeks after administration in the HMGB1 peptide (1-44) and PBS administration groups.

Fig. 14 is a photograph showing the result of Sirius red staining of myocardial tissue sections and a graph showing the percentage of positively stained area.

Fig. 15 is a graph showing capillary density at the infarction margin and a photograph showing the result of von Willebrand factor staining at the infarct margin.

Fig. 16 is a graph showing the short diameter of cardiomyocytes at the border of an infarct.

Fig. 17 is an image of fluorescent immunostaining of the infarct margin in the HMGB1 peptide administration group (1-44). PDGFRα - green; CD90 - red; DAPI - blue. The arrows in (a) indicate PDGFRα- and CD90-double positive cells, and the dotted line marks the border of the infarction.

Fig. 18 is an image of fluorescent immunostaining of the infarct margin in the HMGB1 peptide administration group (1-44). PDGFRα - green; CD105 - red; DAPI - blue. Arrows indicate PDGFRα- and CD105-double-positive cells, and the dotted line indicates the border of the infarction.

Fig. 19 is an image of fluorescent immunostaining of the infarct margin in the HMGB1 (1-44) peptide administration group and in the PBS group. PDGFRα - green; CD90 - red; DAPI - blue. Arrows indicate PDGFRα- and CD90-double positive cells, and the dotted line marks the border of the infarct.

Fig. 20 is a graph showing heart weight at 15 weeks of age.

Fig. 21 is a graph showing the change in LVAWd from 6 weeks to 15 weeks of age (however, no measurements were taken at 9 and 14 weeks of age).

Fig. 22 is a photograph showing the results of Sirius red staining of myocardial tissue sections.

Fig. 23 is a graph showing the percentage of area positively stained with Sirius red in sections of myocardial tissue. All p-values between groups are 0.1 or greater, except for those shown in the figure.

Method for carrying out the invention

The present application provides pharmaceutical compositions for the prevention and/or treatment of heart diseases accompanied by a structural anomaly and/or dysfunction of the heart, which include an HMGB1 peptide fragment comprising the amino acid sequence described in SEQ ID NO: 1.

In the present application, structural abnormalities of the heart include, but are not limited to, cardiomegaly, cardiomyocyte hypertrophy, myocardial fibrosis, and the like. In one aspect, the structural heart abnormality is selected from the group consisting of cardiomegaly, cardiomyocyte hypertrophy, and myocardial fibrosis.

In the context of the present application, dysfunction of the heart (cardiac dysfunction) refers to a violation of the pumping function by which the heart receives and sends blood, including impaired contractility and impaired diastolic function. In one aspect, cardiac dysfunction is selected from the group consisting of impaired contractility and impaired diastolic function. In another aspect, the cardiac dysfunction is selected from the group consisting of impaired ventricular contractility and impaired ventricular diastolic function. In another aspect, the dysfunction of the heart is selected from the group consisting of impaired contractility and impaired diastolic function of the left ventricle.

The present application provides pharmaceutical compositions for the prevention and/or treatment of cardiomyopathy which comprise an HMGB1 peptide fragment comprising the amino acid sequence described in SEQ ID NO: 1.

As used herein, "cardiomyopathy" refers to myocardial disease accompanied by cardiac dysfunction. Also, cardiomyopathy is often accompanied by structural abnormalities of the heart, such as cardiomegaly, cardiomyocyte hypertrophy, and myocardial fibrosis.

Cardiomyopathies that have no apparent cause, such as hypertension and coronary heart disease, and have lesions predominantly in the myocardium are called idiopathic cardiomyopathy, and those that have a clear association with a causative disease or systemic disease are called secondary cardiomyopathy.

Idiopathic cardiomyopathies include, but are not limited to, dilated cardiomyopathy, hypertrophic cardiomyopathy, restrictive cardiomyopathy, arrhythmogenic right ventricular cardiomyopathy, and unclassified cardiomyopathy. "Dilated cardiomyopathy" refers to a disease characterized by left ventricular enlargement and impaired left ventricular contractility, without overlapping with coronary heart disease or an overworking condition (hypertension or valvular disease) that can lead to diffuse impaired contractility.

Secondary cardiomyopathies include, but are not limited to, ischemic cardiomyopathy, hypertensive cardiomyopathy, valvular cardiomyopathy, drug-induced cardiomyopathy, alcoholic cardiomyopathy, mitochondrial cardiomyopathy, cardiac sarcoidosis cardiomyopathy, cardiac amyloidosis cardiomyopathy, myocarditis cardiomyopathy, cardiomyopathy muscular dystrophy Fabry disease-induced cardiomyopathy and cardiomyopathy in pregnancy. In one aspect, muscular dystrophy-induced cardiomyopathy is a Duchenne, Becker, or Emery-Dreyfus-type muscular dystrophy cardiomyopathy.

As used herein, "ischemic cardiomyopathy" refers to a disease in which coronary heart disease (previous myocardial infarction or angina pectoris) causes cardiomegaly and impaired myocardial contractility. Peptide fragments of HMGB1 may prevent and/or treat ischemic cardiomyopathy because the peptides have an inhibitory effect on cardiomegaly and improve the effect on myocardial contractility. In one aspect, ischemic cardiomyopathy is caused by a previous myocardial infarction.

As used herein, "hypertensive cardiomyopathy" refers to a disease in which hypertension causes cardiomyocyte hypertrophy and cardiac dysfunction (impaired diastolic function or impaired contractility). Peptide fragments of HMGB1 can prevent and/or treat hypertensive cardiomyopathy since the peptides can suppress hypertrophy of cardiomyocytes caused by hypertension.

In this application, the term "pharmaceutical composition" is used interchangeably with the terms "pharmaceutical agent", "drug", and "pharmacological composition".

The present application provides pharmaceutical compositions that include an HMGB1 peptide fragment comprising the amino acid sequence described in SEQ ID NO:1 for the treatment of myocardial infarction.

As used herein, "post-myocardial infarction" refers to a condition in which 30 or more days have passed since the onset of myocardial infarction in humans (14 or more days after the onset of myocardial infarction in rats), and which is accompanied by a structural abnormality or functional heart disease. Past myocardial infarction with cardiomegaly and impaired cardiac contractility is classified as ischemic cardiomyopathy. Also, in the context of the present application, the term "post-myocardial infarction" is used interchangeably with "chronic myocardial infarction".

The peptide fragments of HMGB1 in the present application inhibit structural abnormalities (eg, cardiomegaly, cardiomyocyte hypertrophy, and myocardial fibrosis) and dysfunction (eg, impaired contractility) of the heart in myocardial infarction. Also, the HMGB1 peptide fragments of the present application have an improving effect on cardiac function (eg, contractility) in cardiomyopathy. Thus, the HMGB1 peptide fragments of the present application are believed to have broad therapeutic effects on cardiac diseases associated with structural abnormality and/or cardiac dysfunction, such as past myocardial infarction and cardiomyopathy.

The present application also provides pharmaceutical compositions for the prevention and/or treatment of chronic heart failure caused by cardiomyopathy, which include an HMGB1 peptide fragment comprising the amino acid sequence described in SEQ ID NO: 1.

Cardiomyopathy leads to a state of chronic heart failure where heart function deteriorates due to the progression of the disease state. Peptide fragments of HMGB1 can prevent and/or treat chronic heart failure in patients with cardiomyopathy because they can suppress structural cardiac abnormalities such as cardiomegaly, cardiomyocyte hypertrophy and myocardial fibrosis and improve cardiac function in cardiomyopathy.

The present application also provides pharmaceutical compositions for the prevention and/or treatment of chronic heart failure caused by myocardial infarction, which include an HMGB1 peptide fragment comprising the amino acid sequence described in SEQ ID NO: 1.

Postponed myocardial infarction leads to a state of chronic heart failure, since the heart function deteriorates due to the progression of a structural heart abnormality or the like. Peptide fragments of HMGB1 may prevent and/or treat chronic heart failure in patients with prior myocardial infarction, as they may suppress structural cardiac abnormalities such as cardiomegaly, cardiomyocyte hypertrophy, and myocardial fibrosis, and improve cardiac function in prior myocardial infarction.

In the present application, "chronic heart failure" refers to a disease state in which the pumping function of the heart is impaired due to chronic myocardial disease, and the absolute and relative volume of blood, proportional to the oxygen demand of the major peripheral organs, cannot be pumped, causing congestion in the lungs, systemic venous system or both systems, thereby interfering with everyday routine activities. Types of chronic heart failure include heart failure with reduced left ventricular ejection fraction (heart failure with reduced ejection fraction: HFrEF) and heart failure with preserved left ventricular ejection fraction (heart failure with preserved ejection fraction: HFpEF). HFrEF is characterized by <50% left ventricular ejection fraction and has impaired contractility. HFpEF is characterized by a left ventricular ejection fraction of 50% or greater and has impaired diastolic function. In one aspect, HFrEF is due to dilated cardiomyopathy or ischemic cardiomyopathy. In one aspect, HFpEF is due to hypertensive cardiomyopathy.

The present application also provides pharmaceutical compositions for inhibiting a structural heart abnormality selected from the group consisting of cardiomegaly, cardiomyocyte hypertrophy, and myocardial fibrosis in patients with cardiomyopathy and past myocardial infarction, which comprise an HMGB1 peptide fragment comprising the amino acid sequence described in SEQ ID NO : 1. In one aspect, the pharmaceutical compositions of the present application are compositions used for patients with cardiomyopathy and past myocardial infarction to (i) inhibit a structural abnormality of the heart selected from the group consisting of cardiomegaly, cardiomyocyte hypertrophy and myocardial fibrosis, (ii) promote angiogenesis in the heart; or (iii) improving contractility or diastolic function of the heart. Cardiac contractility includes, but is not limited to, for example, ventricular contractility such as left ventricular contractility. Cardiac diastolic function includes, but is not limited to, for example, ventricular diastolic function, such as left ventricular diastolic function.

In the present application, an HMGB1 peptide fragment comprising the amino acid sequence described in SEQ ID NO:1 refers to a peptide consisting of a portion of the HMGB1 protein and comprising the amino acid sequence described in SEQ ID NO:1. Such peptides can be obtained as genetic recombinants (recombinants) by incorporating the DNA encoding the peptide into an appropriate expression system, or they can be synthesized artificially.

As used herein, the HMGB1 protein includes, but is not limited to, proteins comprising the amino acid sequence described in SEQ ID NO:2 and proteins encoded by DNA comprising the base sequence described in SEQ ID NO:3.

The peptide fragment of HMGB1, comprising the amino acid sequence described in SEQ ID NO: 1, in this application includes, without limitation, the following:

1) an HMGB1 peptide fragment comprising the amino acid sequence described in SEQ ID NO: 1 and having cell migration promoting activity;

2) an HMGB1 peptide fragment comprising the amino acid sequence described in SEQ ID NO: 1 and having an activity that stimulates the migration of mesenchymal stem cells; And

3) HMGB1 peptide fragment consisting of the amino acid sequence described in SEQ ID NO: 1.

As used herein, cells whose migration is stimulated by the HMGB1 peptide fragment include, but are not limited to, bone marrow or bone marrow-derived cells (eg, bone marrow stem cells or bone marrow-derived stem cells).

In the present application, "bone marrow cells" refers to cells present in the bone marrow, while "cells derived from the bone marrow" refer to "bone marrow cells" mobilized from the bone marrow to the outside of the bone marrow. "Bone marrow cells" may also include undifferentiated cells such as stem cells and progenitor cells present in the bone marrow.

In the present application, cells whose migration is stimulated by the HMGB1 peptide fragment also include, but are not limited to, mesenchymal stem cells. "Mesenchymal stem cells" are cells isolated from bone marrow or other tissues (e.g., blood such as cord blood, but also skin, adipose tissue, dental pulp, etc.) that can be cultured and propagated as adherent cells on culture dishes (made of plastic or glass) and have the potential to differentiate into mesenchymal tissues such as bone, cartilage, adipose tissue and muscle. In one aspect, mesenchymal stem cells also have the ability to differentiate into epithelial and neural tissue. Mesenchymal stem cells in the present application may exist as a heterogeneous population of cells containing not only stem cells in the narrow sense, but also progenitor cells, and they may include, under culture conditions, differentiated cells in addition to stem cells in the narrow sense and/ or progenitor cells. In one aspect, mesenchymal stem cells may consist of only stem cells in the narrow sense, or they may be a cell population consisting of several types of progenitor cells.

In the present invention, progenitor cells are defined as cells having the potential to unidirectionally differentiate into cells of a specific tissue other than the blood system, and include cells having the potential to differentiate into mesenchymal tissue, epithelial tissue, nervous tissue, parenchymal organ, or vascular endothelium.

As used herein, cells whose migration is stimulated by the HMGB1 peptide fragment also include, but are not limited to, bone marrow-derived mesenchymal stem cells and bone marrow-derived mesenchymal stem cells. "Bone marrow mesenchymal stem cells" are cells present in the bone marrow and are cells characterized in that they are harvested from bone marrow and can be cultured and rapidly propagated as adherent cells on culture dishes (made of plastic or glass), and they have the potential to differentiate into mesenchymal tissues such as bone, cartilage, adipose tissue and muscle, as well as nervous and epithelial tissue. In this application, the term "mesenchymal bone marrow stem cells" is used interchangeably with the terms "mesenchymal bone marrow stromal cells", "pluripotent bone marrow stem cells", and "pluripotent bone marrow stromal cells".

"Bone marrow-derived mesenchymal stem cells" refers to mesenchymal bone marrow stem cells mobilized from the bone marrow to the outside of the bone marrow, and they are cells that can be obtained by sampling peripheral blood and harvesting from mesenchymal tissues such as adipose tissue , epithelial tissues such as skin, and nerve tissues such as the brain. In this application, the term "bone marrow-derived mesenchymal stem cell" is used interchangeably with the terms "bone marrow-derived mesenchymal stromal cell", "bone marrow-derived pluripotent stem cell", and "bone marrow-derived pluripotent stromal cell".

Bone marrow mesenchymal stem cells and bone marrow-derived mesenchymal stem cells are also characterized in that they are able to differentiate into epithelial tissues such as keratinocytes, which form the skin, and nervous system tissues, which make up the brain, when injected into an injured part. living organism immediately after collection or immediately after adhesion to the culture dish.

Bone marrow-derived mesenchymal stem cells and bone marrow-derived mesenchymal stem cells preferably have the ability to differentiate into osteoblasts (identified by observing calcification or the like when differentiation is induced), chondrocytes (identified by alcian blue positive staining, safranin-positive staining). O or the like), adipocytes (identified by positive staining with Sudan III, etc.), as well as mesenchymal cells such as fibroblasts, smooth muscle cells, skeletal muscle cells, stromal and tendon cells, nerve cells, pigment cells cells, epidermal cells, hair follicle cells (expressing the cytokeratin family, hair keratin family or the like), epithelial cells (for example, epidermal keratinocytes and intestinal epithelial cells expressing the cytokeratin family or the like), endothelial cells, and furthermore, parenchymal organ cells such as the liver, kidney, and pancreas. However, cells after differentiation are not limited to the above cells.

Examples of human mesenchymal stem cell markers include, but are not limited to, all or some of PDGFRα positive, PDGFRβ positive, Lin negative, CD45 negative, CD44 positive, CD90 positive, CD29 positive, Flk-1 negative, CD105 positive, CD73 positive, CD90 positive, CD71 positive, Stro-1 positive, CD106 positive, CD166 positive, CD31 negative, CD271 positive and CD11b negative.

Examples of mouse mesenchymal stem cell markers include, but are not limited to, all or some of CD44 positive, PDGFRα positive, PDGFRβ positive, CD45 negative, Lin negative, Sca-1 positive, c-kit negative, CD90 positive, CD105 positive, CD29 positive , Flk-1 negative, CD271 positive and CD11b negative.

Examples of rat mesenchymal stem cell markers include, but are not limited to, all or some of PDGFRα positive, CD44 positive, CD54 positive, CD73 positive, CD90 positive, CD105 positive, CD29 positive, CD271 positive, CD31 negative, and CD45 negative.

In the present application, cells whose migration is stimulated by the HMGB1 peptide fragment also include, but are not limited to, PDGFRα-positive cells. Examples of PDGFRα-positive cells whose migration is stimulated by the HMGB1 peptide fragment include, but are not limited to, PDGFRα-positive mesenchymal stem cells, PDGFRα-positive bone marrow-derived mesenchymal stem cells, and PDGFRα-positive cells derived from bone marrow, which are obtained as adherent cells by culturing the cells of the blood monocyte fraction obtained by harvesting bone marrow (collecting bone marrow cells) or collecting peripheral blood. Examples of PDGFRα-positive mesenchymal stem cells include cells that are positive for PDGFRα and CD44, cells that are positive for PDGFRα and CD90, cells that are positive for PDGFRα and CD105, cells that are positive for PDGFRα and CD29, and the like. In one aspect, the PDGFRα-positive mesenchymal stem cells may be cells that are negative for CD44.

In the pharmaceutical compositions of the present application, peptides that comprise an amino acid sequence with one or more amino acid residues modified (substitutions, deletions, insertions or additions) in the amino acid sequence described in SEQ ID NO: 1 and have cell migration promoting activity can be used in place of or together with an HMGB1 peptide fragment comprising the amino acid sequence described in SEQ ID NO: 1. Examples of such peptides include, but are not limited to, the following:

i) a peptide comprising an amino acid sequence in which one or more amino acids (e.g., one-ten, one-nine, one-eight, one-seven, one-six, one-five, one-four, one-three, or one or two) have been replaced, deleted, inserted or added in the amino acid sequence described in SEQ ID NO: 1, and having the activity of promoting cell migration;

ii) a peptide consisting of an amino acid sequence in which one or more amino acids (for example, one-ten, one-nine, one-eight, one-seven, one-six, one-five, one-four, one-three or one or two) have been replaced, removed, inserted or added in the amino acid sequence described in SEQ ID NO: 1, and having the activity of promoting cell migration;

iii) a peptide comprising an amino acid sequence having a sequence identity of about 80% or more, for example, about 85% or more, about 90% or more, about 91% or more, about 92% or more, about 93% or more, about 94% or more, about 95% or more, about 96% or more, about 97% or more, about 98% or more, or about 99% or more cell migration; And

iv) a peptide consisting of an amino acid sequence having a sequence identity of about 80% or more, for example, about 85% or more, about 90% or more, about 91% or more, about 92% or more, about 93% or more, about 94% or more, about 95% or more, about 96% or more, about 97% or more, about 98% or more, or about 99% or more with the amino acid sequence described in SEQ ID NO:1, and having activity stimulation of cell migration.

Examples of cells whose migration is stimulated by these peptides include, but are not limited to, mesenchymal stem cells; bone marrow-derived mesenchymal stem cells, PDGFRα-positive cells, PDGFRα-positive mesenchymal stem cells, PDGFRα-positive bone marrow-derived mesenchymal stem cells, and PDGFRα-positive bone marrow-derived cells obtained as adherent cells by culturing the cells of the blood monocyte fraction obtained by harvesting bone marrow (collecting bone marrow cells) or collecting peripheral blood.

An effective amount of a peptide of the present application or a pharmaceutical composition comprising a peptide (hereinafter referred to as a peptide or the like) is administered to a subject for the treatment or prevention of the disease or symptom described in this application.

An effective amount, in the context of this application, refers to an amount sufficient to treat or prevent the disease or symptom described in this application. Treatment, in the context of the present application, includes, but is not limited to, relief, delay, blockade, improvement, remission, cure, complete cure, and the like. Prevention, in the context of the present application, also includes, but is not limited to, relief, delay, blockade, and the like.

Subjects, in the context of this application, include, without limitation, mammals, birds, fish, and the like. Mammals include, but are not limited to, humans and non-human animals such as humans, mice, rats, monkeys, pigs, dogs, rabbits, hamsters, guinea pigs, horses, sheep, whales, and the like. In this application, the term "subject" is used interchangeably with the terms "patient", "individual", and "animal".

There are no restrictions on the site of administration of the peptide or the like of the present application, and the peptides and the like of the present application may exert their effects when administered to any site, such as a site with a structural or functional tissue abnormality or a site near it, other than a site (except of these sites), a site remote from a site with a structural or functional tissue abnormality, a site remote from a site with a structural or functional tissue abnormality, or a site distant and displaced relative to a site with a structural or functional tissue abnormality.

For example, when a peptide or the like of the present application is administered at or near a site with a structural or functional abnormality of the heart, cells (e.g., mesenchymal stem cells) are recruited to the site of administration to induce or stimulate regeneration of cardiac tissue or improve structural or functional impairment of the heart. Also, for example, when the peptide or the like of the present application is administered at a site other than a site with a structural or functional abnormality of the heart or its surroundings, bone marrow cells (for example, bone marrow mesenchymal stem cells) are mobilized from the bone marrow to a site with a structural or functional abnormality of the heart or its surroundings through the peripheral circulation to induce or promote the regeneration of cardiac tissue or improve the structural or functional abnormality of the heart. In the present application, "peripheral blood flow" is also referred to as "blood circulation" and "peripheral circulating blood flow".

The peptide or the like of the present application may also exhibit its effects when administered to any tissue, such as tissue other than cardiac tissue, tissue remote from cardiac tissue, tissue distal to cardiac tissue, or tissue distal to and displaced from cardiac tissue. fabrics. Thus, the peptide or the like of the present application is effectively used to regenerate cardiac tissue into which it is difficult to directly administer a drug from outside the body, or to improve structural or functional abnormalities of the heart.

Methods for administering the peptide or the like of the present application include, but are not limited to, oral administration and parenteral administration, including intravascular (intra-arterial, intravenous, and the like), intramuscular, subcutaneous, intradermal, intraperitoneal, nasal, pulmonary, and transdermal administration. Also, the peptide or the like according to the present application can be administered systemically or locally (for example, subcutaneously, intradermally or into the surface of the skin, into the eyeball or conjunctiva of the eyelid, nasal mucosa, oral and gastrointestinal mucosa, vaginal and endometrial mucosa or damaged area) by injection, for example, intravenous injection, intramuscular injection, intraperitoneal injection and subcutaneous injection.

In addition, instead of the peptide or the like of the present application, cells secreting the peptides of the present application, gene therapy vectors into which DNA encoding the peptides have been inserted, and including pharmaceutical compositions thereof, can be used.

In addition, the route of administration can be appropriately selected according to the age and symptoms of the patient. For example, when administering the peptides of the present application, the dosage may be selected in the range of 0.0000001 to 1000 mg per kilogram of body weight per administration. Alternatively, the dosage may be selected, for example, in the range of 0.00001 to 100,000 mg/patient body. Also, when a cell secreting the peptide of the present application or a gene therapy vector into which the DNA encoding the peptide has been inserted is administered, the administration can be carried out so that the amount of the peptide is in the above range. However, the pharmaceutical compositions in the present application are not limited to these doses.

Pharmaceutical compositions of the present application may be formulated according to conventional methods (eg, Remington's Pharmaceutical Science, latest edition, Mark Publishing Company, Easton, U.S.A.) and may contain pharmaceutically acceptable carriers along with additives. Examples include, but are not limited to, surfactants, excipients, colorants, flavoring agents, preservatives, stabilizers, buffers, suspending agents, isotonizing agents, binders, disintegrants, lubricants, flow agents, and flavoring agents, and can also be used other commonly used media as needed. Specific examples include, light anhydrous silicic acid, lactose, crystalline cellulose, mannitol, starch, carmellose calcium, carmellose sodium, hydroxypropyl cellulose, hydroxypropyl methylcellulose, polyvinyl acetal diethylaminoacetate, polyvinyl pyrrolidone, gelatin, medium chain fatty acid triglycerides, polyoxyethylene hydrogenated castor oil 60, white sugar, carboxymethyl cellulose, corn starch, inorganic salts.

All prior art documents cited in this application are incorporated into this application by reference.

The present invention is further illustrated by, but not limited to, the following examples.

Examples

[Example 1] Evaluation of the effectiveness of the HMGB1 peptide fragment for the treatment of dilated cardiomyopathy

(1) Materials and methods

J2N-k hamsters (18 weeks old, male, 20 animals in total), which are animal models of dilated cardiomyopathy, were obtained from Japan SLC, allowed to acclimate for 2 weeks, and then used for experiments. J2N-k hamsters spontaneously developed dilated cardiomyopathy due to deletion mutations in the δ-sarcoglycan gene (in particular, cardiomyocyte shedding and fibrosis began at about 5 weeks of age, cardiomegaly and cardiac dysfunction appeared at about 20 weeks of age, and eventually animals eventually died of congestive heart failure after about 1 year J Biochem 2003 Aug;134(2):269-76). Also, a peptide consisting of amino acid residues 1-44 (SEQ ID NO: 1) of the human HMGB1 protein was chemically synthesized by the solid phase method. Hereinafter, this peptide is referred to as the HMGB1(1-44) peptide and is abbreviated as "1-44" in the drawings corresponding to the Examples.

J2N-k hamsters were divided into HMGB1 peptide treatment (1-44) (n=10) and PBS treatment (control, n=10) and treatment started at 20 weeks of age (body weight about 120 g). The test substance was administered by injecting a solution of the HMGB1 (1-44) peptide, which was adjusted to a concentration of 1 mg/ml using PBS as a carrier, into the external jugular vein at a dose of 3 ml/kg (3 mg/kg based on the dose of the peptide ) once a day for four consecutive days. In the control group, PBS was injected into the external jugular vein at a dose of 3 ml/kg once a day for four consecutive days. 6 weeks after administration, a thoracotomy was performed under deep anesthesia and the heart was removed. The middle part of the heart was separated for cryopreservation and for fixation in paraffin and carried out histopathological examination. Also carried out molecular biological studies using the apical myocardium to evaluate the effects of the introduction of the peptide HMGB1 (1-44).

(2) Measured indicators

i) Cardiac function

Echocardiography was done before administration and 4 and 6 weeks after administration; and left ventricular end-diastolic diameter (LVDd), left ventricular end-systolic diameter (LVDs), and left ventricular ejection fraction (LVEF) were measured and calculated to evaluate cardiac function.

ii) Myocardial fibrosis

Sections of myocardial tissue were stained with Sirius red and the proportion of positively stained area in the entire myocardial area was calculated as the percentage of fibrosis (%).

iii) Angiogenesis

Immunostaining of myocardial tissue sections was performed using an anti-CD31 antibody to determine the number of blood vessels (number of vascular endothelial cells). This measurement was carried out in five different areas and the average value was calculated.

iv) Hypertrophy of cardiomyocytes

Staining with Schiff's reagent (Periodic Acid (PAS)) of myocardial tissue sections was performed to measure the short diameter of cardiomyocytes retaining nuclear architecture, and the mean value of the measurements was obtained. This measurement was carried out in five different areas and the average value was calculated.

v) Recruitment of mesenchymal stem cells

Immunostaining of myocardial tissue sections was performed using antibodies against PDGFRα and CD29, which are surface markers of mesenchymal stem cells, to assess whether or not there is an accumulation of mesenchymal stem cells. Nuclear staining was performed using DAPI.

vi) RT-PCR

Intramyocardial expression of VEGF, an angiogenic factor secreted by mesenchymal myeloid stem cells, and TSG-6, an anti-inflammatory cytokine, was assessed by RT-PCR using apical myocardium.

vii) Mitochondrial structure

The structure of cardiomyocytes was observed using electron microscopy. In particular, full-thickness mid-section myocardium was sampled and pre-fixed with 1/2 Karnovsky's fixative, followed by fixation with 2% osmium tetroxide, block-staining with 0.5% aqueous uranium acetate, and dehydration with ethanol, and then immersed in infiltrating epoxy. (Quetol812) and polymerized. Ultrathin sections (70-110 nm) were made and electron stained with uranium acetate and lead stain solution, and then observed by electron microscopy (H-7500, Hitachi High Technologies).

(3) Results

i) Cardiac function

Cardiac function assessments 4 and 6 weeks after administration showed significantly higher levels of LVEF in the HMGB1 (1-44) peptide administration group compared to the PBS administration group, and inhibition of left ventricular contractility reduction (FIG. 1). These results indicate an improving effect of the HMGB1(1-44) peptide on cardiac function. There were no significant differences in LVDd and LVDs at any time point across 4 and 6 weeks of dosing.

ii) Myocardial fibrosis

The analysis showed that the percentage of area positively stained with Sirius red was significantly less in the HMGB1 peptide (1-44) group than in the PBS group, indicating inhibition of myocardial fibrosis (Fig. 2).

iii) Angiogenesis

The immunostaining results showed that in the middle layer, the HMGB1 (1-44) peptide administration group had significantly more blood vessels and improved angiogenesis than the PBS group (FIG. 3). In the subendocardial layer, the number of blood vessels in the HMGB1 (1-44) peptide administration group was higher than in the PBS group.

iv) Hypertrophy of cardiomyocytes

The results of PAS staining showed that the short diameter of cardiomyocytes in both the subendocardial layer and the middle layer was significantly smaller in the HMGB1 peptide (1-44) group than in the PBS group, indicating inhibition of cardiomyocyte hypertrophy (Fig. 4).

v) Recruitment of mesenchymal stem cells

Immunostaining showed that cells positive for both PDGFRα and CD29 were recruited to myocardial tissue in the HMGB1(1-44) peptide administration group (FIG. 5). In addition, significantly more PDGFRα-positive and CD29-positive cells were present in myocardial tissue in the HMGB1 (1-44) peptide administration group than in the PBS group (FIG. 6).

vi) RT-PCR

The analysis showed that both VEGF and TSG-6 had significantly higher expression levels in the HMGB1 (1-44) peptide administration group than in the PBS group (FIG. 7).

vii) Mitochondrial structure

Electron microscopic examination of cardiomyocytes revealed a loss of mitochondrial crist structure in the PBS group, while the crist structure was preserved in the HMGB1 (1-44) peptide administration group (Fig. 8). These results confirm the effect of administration of the HMGB1(1-44) peptide on the maintenance of cardiac function.

[Example 2] Evaluation of the effectiveness of the HMGB1 peptide fragment for the treatment of dilated cardiomyopathy (long-term follow-up)

(1) Materials and methods

As in Example 1, twenty J2N-k hamsters were prepared and divided into HMGB1(1-44) peptide treatment group (n=11) and PBS treatment group (control, n=9) and treatment started at 20 weeks of age. The test substance was administered by injecting a solution of the HMGB1 (1-44) peptide, which was adjusted to a concentration of 1 mg/ml using PBS as a carrier, into the external jugular vein at a dose of 3 ml/kg (3 mg/kg based on the dose of the peptide ) once a day for four consecutive days. In the control group, PBS was injected into the external jugular vein at a dose of 3 ml/kg once a day for four consecutive days. Thereafter, cardiac function and survival were assessed by rearing the hamsters under normal conditions.

(2) Measured indicators

i) Cardiac function

Echocardiography was done before administration, 4 weeks after administration, and every 2 weeks thereafter to measure LVEF.

ii) Survival

After the last dose, the animals were kept under normal conditions and survival was assessed.

(3) Results

i) Cardiac function

The LVEF in the HMGB1 (1-44) peptide administration group remained significantly higher than in the PBS group up to 6 weeks post-administration (FIG. 9).

ii) Survival

Long-term follow-up showed a trend towards increased viability in the HMGB1(1-44) peptide administration group (FIG. 10).

[Example 3] Evaluation of the effectiveness of the HMGB1 peptide fragment for improving cardiac function after myocardial infarction

(1) Materials and methods

SD rats (7 weeks old, male, body weight approximately 250 g) were anesthetized with the inhalational anesthetic sevofran (or isoflurane) and intubated endotracheally after adequate suppression was achieved, and deep anesthesia was maintained with the inhaled anesthetic. A thoracotomy was performed in the supine position at the left fourth intercostal space, and the proximal left anterior descending coronary artery was ligated with 6-0 prolene sutures to create a wide area myocardial infarction model. Cardiac function was assessed by echocardiography 2 weeks after infarction, and cases in which extensive infarction was obtained (LVEF<50%) were used as a rat model of myocardial infarction (a total of 17 animals). As in the above examples, the HMGB1(1-44) peptide consisting of SEQ ID NO: 1 was used.

Rat models of past myocardial infarction were divided into HMGB1(1-44) peptide treatment group (n=9) and PBS treatment group (control, n=8) and treatment started 2 weeks after infarction. The test substance was administered by injecting a solution of the HMGB1 (1-44) peptide, which was adjusted to a concentration of 1 mg/ml using PBS as a carrier, into the femoral veins at a dose of 3 ml/kg (3 mg/kg per peptide dose) once a day for four consecutive days. In the control group, PBS was injected at a dose of 3 ml/kg once a day through the femoral vein for four consecutive days. Four weeks after injection, the heart was removed by repeated thoracotomy under deep anesthesia. At the time of removal, blood of 5 ml or more was collected by puncture from the right atrium towards the inferior vena cava, and the infarcted area of the heart was cut into four short-axis slices of the same thickness and separated for cryopreservation and fixation in paraffin. Carried out a comparative study of both groups using physiological, histopathological and molecular biological methods to assess the effects of the introduction of the peptide HMGB1 (1-44). The state after 2 weeks (14 days) or more after the onset of myocardial infarction in the rat model is equivalent to the state after 30 days or more after the onset of myocardial infarction in humans. In addition, the rat model of past myocardial infarction obtained in this study can be considered as a model of ischemic cardiomyopathy, since LVEF decreased to 42% at the time before the start of administration, and progression of cardiomegaly was observed in the control group.

(2) Measured indicators

i) Cardiac function

Echocardiography was done at 1, 2 and 4 weeks after administration and left ventricular end diastolic diameter (LVDd), left ventricular end systolic diameter (LVDs) and left ventricular ejection fraction (LVEF) were measured and calculated to assess cardiac function.

ii) Myocardial fibrosis

Sections of myocardial tissue were stained with Sirius red, and the proportion of the positively stained area in the total area of the left ventricular myocardium was calculated as the percentage of fibrosis (%).

iii) Angiogenesis

Von Willebrand factors were stained at the border of the infarct to determine the number of blood vessels (number of vascular endothelial cells). This measurement was carried out in 10 different areas and the average value was calculated.

iv) Hypertrophy of cardiomyocytes

Staining with Schiff's reagent (periodic acid (PAS)) was performed at the border of the infarction and the short diameter of cardiomyocytes retaining nuclear architecture at the border of the infarct was measured and the average value of the measurements was obtained. This measurement was carried out in 10 different areas and the average value was calculated.

v) Recruitment of mesenchymal stem cells at the border of infarction

Immunostaining of myocardial tissue was performed using antibodies against PDGFRα, CD90 and CD105, which are surface markers of mesenchymal stem cells, to assess whether there was an accumulation of mesenchymal stem cells at the edge of the infarction. Nuclear staining was performed using DAPI.

(3) Results

i) Cardiac function

Assessment of cardiac function 4 weeks after administration showed significantly higher levels of LVEF in the HMGB1 (1-44) peptide administration group compared to the PBS administration group (FIG. 11). In addition, LVDd and LVD 4 weeks post-dose showed lower values in the HMGB1 (1-44) peptide administration group compared to the PBS group, and inhibition of cardiomegaly was observed (FIG. 12).

In addition, regarding LVEF change over time, the PBS group showed a decrease in LVEF with time post-dose, while the HMGB1 peptide (1-44) group showed an increase in LVEF up to 4 weeks post-dose, with an average improvement in LVEF of about 7% compared to pre-dose value (FIG. 13).

ii) Myocardial fibrosis

The analysis showed that the percentage of Sirius red positive area was significantly less in the HMGB1 peptide (1-44) group than in the PBS group, indicating inhibition of myocardial fibrosis (FIG. 14).

iii) Angiogenesis

The results of von Willebrand factor staining at the border of the infarction showed that in the HMGB1 (1-44) peptide administration group there were significantly more vessels than in the PBS group (P=0.05), indicating an increase in angiogenesis (Fig. 15).

iv) Hypertrophy of cardiomyocytes

The results of PAS staining at the border of the infarction showed that the short diameter of cardiomyocytes was significantly smaller in the HMGB1 peptide administration group (1-44) than in the PBS group (P=0.05), indicating inhibition of cardiomyocyte hypertrophy (Fig. 16).

v) Recruitment of mesenchymal stem cells at the border of infarction

Myocardial tissue immunostaining showed that both PDGFRα and CD90 positive cells and both PDGFRα and CD105 positive cells were recruited near the border of the infarction in the HMGB1(1-44) peptide administration group (Fig. 17 and 18). Also, there were more PDGFRα-positive and CD90-positive cells near the border of the infarction in the HMGB1 (1-44) peptide administration group than in the PBS group (Fig. 19).

[Example 4] Evaluation of the effectiveness of the HMGB1 peptide fragment for hypertensive cardiomyopathy

(1) Materials and methods

Salt-sensitive Dahl rats (hereinafter also referred to as DIS/Eis rats) were used in the experiments. DIS/Eis rats are hypertensive when fed a high-salt diet and represent a model in which heart failure develops at a high rate (e.g., consuming a diet enriched with 8% NaCl from 6 weeks of age has a blood pressure of 250 mm Hg at 9 weeks of age, deaths begin at 12 weeks of age, and death is very likely by 16 weeks).

Ten DIS/Eis rats (6 weeks old, male, body weight about 200g, Japan SLC) were prepared and divided into two groups, a high-salt diet group (n=7) and a low-salt diet group (n=3), and feeding was continued. until 15 weeks of age, when the first group received food enriched with 8% NaCl and the last group received food enriched with 0.3% NaCl. Other than food, nothing interfered with free movement, and the animals were kept clean and rested without restrictions on drinking water, food, etc. in their cells.

At the age of 11 weeks, the high salt diet group (n=7) was divided into the HMGB1 peptide treatment group (1-44) (n=3) and the control group (n=4) and treatment was started. The test substance was administered by injecting a solution of the HMGB1 (1-44) peptide, which was adjusted to a concentration of 1 mg/ml using PBS as a carrier, into the tail vein at a dose of 3 ml/kg (3 mg/kg per peptide dose) once a day for four consecutive days. The control group was injected with PBS at a dose of 3 ml/kg once a day via the tail vein for four consecutive days. Similar to the control group, the low salt diet group (n=3) also received PBS via the tail vein for four consecutive days. 4 weeks after administration (at 15 weeks of age), the rat heart was removed by thoracotomy under general anesthesia and body weight was measured and then histological sections of the left ventricular myocardium at the papillary muscle level were prepared and subjected to histological analysis. In the following description, the HMGB1 (1-44) peptide administration group, which was maintained on a high salt diet, is briefly referred to as the "High-1-44 group"; the control group, which was kept on a high-salt diet, is abbreviated as "High-PBS group", and the group, which was kept on a low-salt diet, is abbreviated as "Low-PBS group".

(2) Measured indicators

i) Heart weight

At 15 weeks of age, hearts were removed from rats in each group and weighed.

ii) Cardiac function and ventricular wall thickness

Echocardiography was done before administration and every week after the start of administration to determine left ventricular end diastolic diameter (LVDd), left ventricular end systolic diameter (LVDs), left ventricular ejection fraction (LVEF), left ventricular anterior wall end diastolic size (LVAWd ) and end-diastolic size of the posterior wall of the left ventricle (LVPWd).

iii) Myocardial fibrosis

Sections of myocardial tissue were stained with Sirius red and the proportion of positively stained area in the area of the whole heart or left ventricle was calculated as the percentage of fibrosis (%).

(3) Results

i) Heart weight

Compared to the Low-PBS group, the High-1-44 and High-PBS groups had larger heart masses. In addition, the High-1-44 group had a lower heart mass than the High-PBS group, indicating that administration of the HMGB1(1-44) peptide inhibited cardiac hypertrophy (FIG. 20).

ii) Cardiac function

During the follow-up period, LVEF remained at 70% or more in all groups (no significant differences between groups).

iii) Ventricular wall thickness

Changes in LVAWd during the observation period are shown in FIG. 21. At 15 weeks of age, LVAWd in the High-PBS group and High-1-44 group were greater than in the Low-PBS group, but LVAWd in the High-1-44 group was less than in the High-PBS group, showing a trend inhibition of an increase in the thickness of the anterior wall of the left ventricle with the introduction of the HMGB1 peptide (1-44).

iv) Myocardial fibrosis

The results of Sirius red staining of myocardial tissue sections are shown in Fig. 22. Compared to the Low-PBS group, the High-PBS group had increased fibrosis around the coronary arteries. The percentage of Sirius red positive area (percent fibrosis) in the whole heart was less in the High-1-44 group than in the High-PBS group, showing a trend of inhibition of myocardial fibrosis with the administration of the HMGB1(1-44) peptide (Fig. 23A). The percentage of fibrosis in the left ventricle was less in the High-1-44 group than in the High-PBS group, indicating that administration of the HMGB1(1-44) peptide inhibited myocardial fibrosis (FIG. 23B).

(4) Discussion

DIS/Eis rats fed a high salt diet developed hypertension. It was also confirmed that rats did indeed develop cardiac hypertrophy and cardiomyocyte hypertrophy. Therefore, DIS/Eis rats kept on the high salt diet used in the examples can be evaluated as a model for hypertensive cardiomyopathy.

Cardiac hypertrophy/cardiomyocyte hypertrophy is a structural abnormality that results in decreased diastolic function of the heart. The DIS/Eis rats fed the high salt diet used in the present examples can also be evaluated as a model for HFpEF as they develop cardiac hypertrophy/cardiomyocyte hypertrophy while maintaining normal left ventricular contractility.

The HMGB1(1-44) peptide of the present invention inhibits cardiac hypertrophy and cardiomyocyte hypertrophy in DIS/Eis rats fed a high salt diet, and thus can be used to prevent and/or treat hypertensive cardiomyopathy and HFpEF.

Industrial Applicability

Pharmaceutical compositions comprising the peptide of the present application are useful as pharmaceutical compositions for the prevention and/or treatment of heart diseases accompanied by structural abnormalities and/or dysfunction of the heart. Pharmaceutical compositions comprising the peptide of the present application are expected to provide greater benefit to patients with cardiomyopathy and previous myocardial infarction and resulting chronic heart failure, for whom surgery is difficult due to factors such as advanced age, and who cannot receive sufficient effect of existing drugs for the treatment of chronic heart failure. In addition, pharmaceutical compositions comprising the peptide of the present application exhibit effects such as improving cardiac function and inhibiting cardiomyocyte hypertrophy in many models of cardiomyopathy, which are dilated cardiomyopathy, ischemic cardiomyopathy, and hypertensive cardiomyopathy, and thus can be expected that they will have broad therapeutic effects on various types of cardiomyopathy, including idiopathic and secondary cardiomyopathies.

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SEQUENCE LIST

<110> Genomix Co., Ltd.

Osaka University

<120> Medicine for the treatment of cardiomyopathy, myocardial infarction

myocardial and chronic heart failure

<130> G6-A1601P

<150> JP 2017-013293

<151> 2017-01-27

<150> JP 2017-151788

<151> 2017-08-04

<160> 3

<170>PatentIn version 3.5

<210> 1

<211> 44

<212> Protein

<213> Artificial sequence

<220>

<223> Artificially synthesized peptide sequence

<400> 1

Met Gly Lys Gly Asp Pro Lys Lys Pro Arg Gly Lys Met Ser Ser Tyr

1 5 10 15

Ala Phe Phe Val Gln Thr Cys Arg Glu Glu His Lys Lys Lys His Pro

20 25 30

Asp Ala Ser Val Asn Phe Ser Glu Phe Ser Lys Lys

35 40

<210> 2

<211> 215

<212> Protein

<213> Homo sapiens

<400> 2

Met Gly Lys Gly Asp Pro Lys Lys Pro Arg Gly Lys Met Ser Ser Tyr

1 5 10 15

Ala Phe Phe Val Gln Thr Cys Arg Glu Glu His Lys Lys Lys His Pro

20 25 30

Asp Ala Ser Val Asn Phe Ser Glu Phe Ser Lys Lys Cys Ser Glu Arg

35 40 45

Trp Lys Thr Met Ser Ala Lys Glu Lys Gly Lys Phe Glu Asp Met Ala

50 55 60

Lys Ala Asp Lys Ala Arg Tyr Glu Arg Glu Met Lys Thr Tyr Ile Pro

65 70 75 80

Pro Lys Gly Glu Thr Lys Lys Lys Phe Lys Asp Pro Asn Ala Pro Lys

85 90 95

Arg Pro Pro Ser Ala Phe Phe Leu Phe Cys Ser Glu Tyr Arg Pro Lys

100 105 110

Ile Lys Gly Glu His Pro Gly Leu Ser Ile Gly Asp Val Ala Lys Lys

115 120 125

Leu Gly Glu Met Trp Asn Asn Thr Ala Ala Asp Asp Lys Gln Pro Tyr

130 135 140

Glu Lys Lys Ala Ala Lys Leu Lys Glu Lys Tyr Glu Lys Asp Ile Ala

145 150 155 160

Ala Tyr Arg Ala Lys Gly Lys Pro Asp Ala Ala Lys Lys Gly Val Val

165 170 175

Lys Ala Glu Lys Ser Lys Lys Lys Lys Glu Glu Glu Glu Asp Glu Glu

180 185 190

Asp Glu Glu Asp Glu Glu Glu Glu Glu Asp Glu Glu Asp Glu Asp Glu

195 200 205

Glu Glu Asp Asp Asp Asp Glu

210 215

<210> 3

<211> 648

<212> DNA

<213> Homo sapiens

<400> 3

atgggcaaag gagatcctaa gaagccgaga ggcaaaatgt catcatatgc attttttgtg 60

caaacttgtc gggaggagca taagaagaag cacccagatg cttcagtcaa cttctcagag 120

ttttctaaga agtgctcaga gaggtggaag accatgtctg ctaaagagaa aggaaaattt 180

gaagatatgg caaaagcggga caaggcccgt tatgaaagag aaatgaaaac ctatatccct 240

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Claims (57)

1. A method of treating a heart disease associated with a structural abnormality and/or dysfunction of the heart in a subject with heart disease, wherein the method comprises administering to the subject with heart disease an effective amount of a substance described in any of (a) to (c) below: (a) a peptide consisting of the amino acid sequence described in SEQ ID NO: 1; (b) a peptide consisting of an amino acid sequence in which one amino acid has been replaced, deleted, inserted or added in the amino acid sequence described in SEQ ID NO: 1, and has the activity of promoting the migration of PDGFRα-positive and CD29-positive mesenchymal stem cells; And (c) a peptide consisting of an amino acid sequence having a sequence identity of 97% or more with the amino acid sequence described in SEQ ID NO: 1, and having the activity of promoting the migration of PDGFRα-positive and CD29-positive mesenchymal stem cells, wherein the heart disease associated with a structural abnormality and/or dysfunction of the heart is selected from cardiomyopathy and prior myocardial infarction. 2. A method of treating a heart disease associated with a structural abnormality and/or dysfunction of the heart in a subject with heart disease, wherein the method comprises administering to the subject with heart disease an effective amount of a peptide consisting of an amino acid sequence in which one or two amino acids have been added to the amino acid the sequence described in SEQ ID NO: 1 and having the activity of promoting the migration of PDGFRα-positive and CD29-positive mesenchymal stem cells, and where the heart disease associated with a structural abnormality and/or dysfunction of the heart is selected from cardiomyopathy and past myocardial infarction. 3. A method of treating a heart disease associated with a structural abnormality and/or dysfunction of the heart in a subject with heart disease, wherein the method comprises administering to the subject with heart disease an effective amount of a peptide consisting of an amino acid sequence in which one amino acid is added to the amino acid sequence described in SEQ ID NO: 1, and having the activity of stimulating the migration of PDGFRα-positive and CD29-positive mesenchymal stem cells, and the heart disease associated with a structural abnormality and/or dysfunction of the heart is selected from cardiomyopathy and past myocardial infarction. 4. A method of treating a heart disease associated with a structural abnormality and/or dysfunction of the heart in a subject with heart disease, wherein the method comprises administering to the subject with heart disease an effective amount of a peptide consisting of the amino acid sequence described in SEQ ID NO: 1, and where heart disease associated with a structural abnormality and/or dysfunction of the heart, selected from cardiomyopathy and past myocardial infarction. 5. The method according to any one of paragraphs. 1-4, wherein the heart disease associated with a structural abnormality and/or dysfunction of the heart is a cardiomyopathy. 6. The method of claim 5 wherein the cardiomyopathy is idiopathic cardiomyopathy. 7. The method of claim 6, wherein the idiopathic cardiomyopathy is selected from the group consisting of dilated cardiomyopathy, hypertrophic cardiomyopathy, restrictive cardiomyopathy, and arrhythmogenic right ventricular cardiomyopathy. 8. The method of claim 5, wherein the cardiomyopathy is a secondary cardiomyopathy. 9. The method of claim 8, wherein the secondary cardiomyopathy is selected from the group consisting of ischemic cardiomyopathy, hypertensive cardiomyopathy, valvular cardiomyopathy, drug-induced cardiomyopathy, alcoholic cardiomyopathy, mitochondrial cardiomyopathy, cardiac sarcoidosis-induced cardiomyopathy, cardiac amyloidosis-induced cardiomyopathy, myocarditis, muscular dystrophy cardiomyopathy, Fabry disease cardiomyopathy, and peripartum cardiomyopathy. 10. The method according to any one of paragraphs. 1-4, where the heart disease associated with a structural abnormality and/or dysfunction of the heart is a past myocardial infarction. 11. The method according to any one of paragraphs. 1-4, where the administration is the systemic administration of a substance to a site other than cardiac tissue. 12. A method for preventing chronic heart failure caused by cardiomyopathy or past myocardial infarction in a subject with cardiomyopathy or past myocardial infarction, where the method includes administering to the subject with cardiomyopathy or past myocardial infarction an effective amount of a substance described in any of points (a) - (c ) below: (a) a peptide consisting of the amino acid sequence described in SEQ ID NO: 1; (b) a peptide consisting of an amino acid sequence in which one amino acid has been replaced, deleted, inserted or added in the amino acid sequence described in SEQ ID NO: 1, and has the activity of promoting the migration of PDGFRα-positive and CD29-positive mesenchymal stem cells; And (c) a peptide consisting of an amino acid sequence having a sequence identity of 97% or more with the amino acid sequence described in SEQ ID NO: 1, and having the activity of promoting the migration of PDGFRα-positive and CD29-positive mesenchymal stem cells. 13. A method for preventing chronic heart failure caused by cardiomyopathy or past myocardial infarction in a subject with cardiomyopathy or past myocardial infarction, where the method includes administering to a subject with cardiomyopathy or past myocardial infarction an effective amount of a peptide consisting of an amino acid sequence in which one or two amino acids added to the amino acid sequence described in SEQ ID NO: 1, and having the activity of promoting the migration of PDGFRα-positive and CD29-positive mesenchymal stem cells. 14. A method for preventing chronic heart failure caused by cardiomyopathy or past myocardial infarction in a subject with cardiomyopathy or past myocardial infarction, where the method includes administering to a subject with cardiomyopathy or past myocardial infarction an effective amount of a peptide consisting of an amino acid sequence in which one amino acid is added to the amino acid sequence described in SEQ ID NO: 1 and having the activity of promoting the migration of PDGFRα-positive and CD29-positive mesenchymal stem cells. 15. A method for preventing chronic heart failure caused by cardiomyopathy or past myocardial infarction in a subject with cardiomyopathy or past myocardial infarction, wherein the method comprises administering to the subject with cardiomyopathy or past myocardial infarction an effective amount of a peptide consisting of the amino acid sequence described in SEQ ID NO: 1. 16. The method according to any one of paragraphs. 12-15, where administration is systemic administration of a substance to a site other than cardiac tissue. 17. A method for inhibiting a structural heart abnormality selected from the group consisting of cardiomegaly, cardiomyocyte hypertrophy, and myocardial fibrosis in a subject with cardiomyopathy or prior myocardial infarction, wherein the method comprises administering to the subject with cardiomyopathy or prior myocardial infarction an effective amount of a substance described in any one of paragraphs (a) - (c) below: (a) a peptide consisting of the amino acid sequence described in SEQ ID NO: 1; (b) a peptide consisting of an amino acid sequence in which one amino acid has been replaced, deleted, inserted or added to the amino acid sequence described in SEQ ID NO: 1, and has the activity of promoting the migration of PDGFRα-positive and CD29-positive mesenchymal stem cells; And (c) a peptide consisting of an amino acid sequence having a sequence identity of 97% or more with the amino acid sequence described in SEQ ID NO: 1, and having the activity of promoting the migration of PDGFRα-positive and CD29-positive mesenchymal stem cells. 18. A method for inhibiting a structural heart abnormality selected from the group consisting of cardiomegaly, cardiomyocyte hypertrophy, and myocardial fibrosis in a subject with cardiomyopathy or prior myocardial infarction, wherein the method comprises administering to the subject with cardiomyopathy or prior myocardial infarction an effective amount of a peptide consisting of an amino acid sequence , in which one or two amino acids are added to the amino acid sequence described in SEQ ID NO: 1, and having the activity of promoting the migration of PDGFRα-positive and CD29-positive mesenchymal stem cells. 19. A method for inhibiting a structural heart abnormality selected from the group consisting of cardiomegaly, cardiomyocyte hypertrophy, and myocardial fibrosis in a subject with cardiomyopathy or a history of myocardial infarction, wherein the method comprises administering to a subject with a cardiomyopathy or a history of myocardial infarction an effective amount of a peptide consisting of an amino acid sequence , in which one amino acid is added to the amino acid sequence described in SEQ ID NO: 1, and having the activity of promoting the migration of PDGFRα-positive and CD29-positive mesenchymal stem cells. 20. A method for inhibiting a structural heart abnormality selected from the group consisting of cardiomegaly, cardiomyocyte hypertrophy, and myocardial fibrosis in a subject with cardiomyopathy or prior myocardial infarction, wherein the method comprises administering to the subject with cardiomyopathy or prior myocardial infarction an effective amount of a peptide consisting of an amino acid sequence described in SEQ ID NO: 1. 21. The method according to any one of paragraphs. 17-20, where the administration is systemic administration of a substance to a site other than cardiac tissue. 22. The use of a substance for the treatment of a heart disease associated with a structural abnormality and/or dysfunction of the heart in a subject with heart disease, wherein the substance is intended to be administered to a subject with heart disease and is described in any of (a) to (c) below: (a) a peptide consisting of the amino acid sequence described in SEQ ID NO: 1; (b) a peptide consisting of an amino acid sequence in which one amino acid has been replaced, deleted, inserted or added in the amino acid sequence described in SEQ ID NO: 1, and has the activity of promoting the migration of PDGFRα-positive and CD29-positive mesenchymal stem cells; And (c) a peptide consisting of an amino acid sequence having a sequence identity of 97% or more with the amino acid sequence described in SEQ ID NO: 1, and having the activity of promoting the migration of PDGFRα-positive and CD29-positive mesenchymal stem cells, wherein the heart disease associated with a structural abnormality and/or dysfunction of the heart is selected from cardiomyopathy and prior myocardial infarction. 23. The use of a substance for the prevention of chronic heart failure caused by cardiomyopathy or past myocardial infarction in a subject with cardiomyopathy or past myocardial infarction, where the substance is intended for administration to a subject with cardiomyopathy or past myocardial infarction and is described in any of points (a) - (c ) below: (a) a peptide consisting of the amino acid sequence described in SEQ ID NO: 1; (b) a peptide consisting of an amino acid sequence in which one amino acid has been replaced, deleted, inserted or added in the amino acid sequence described in SEQ ID NO: 1, and has the activity of promoting the migration of PDGFRα-positive and CD29-positive mesenchymal stem cells; And (c) a peptide consisting of an amino acid sequence having a sequence identity of 97% or more with the amino acid sequence described in SEQ ID NO: 1, and having the activity of promoting the migration of PDGFRα-positive and CD29-positive mesenchymal stem cells. 24. Use of a substance for inhibiting a structural abnormality of the heart selected from the group consisting of cardiomegaly, cardiomyocyte hypertrophy, and myocardial fibrosis in a subject with cardiomyopathy or prior myocardial infarction, wherein the substance is intended to be administered to a subject with cardiomyopathy or prior myocardial infarction and is described in any of paragraphs (a) - (c) below: (a) a peptide consisting of the amino acid sequence described in SEQ ID NO: 1; (b) a peptide consisting of an amino acid sequence in which one amino acid has been replaced, deleted, inserted or added in the amino acid sequence described in SEQ ID NO: 1, and has the activity of promoting the migration of PDGFRα-positive and CD29-positive mesenchymal stem cells; And (c) a peptide consisting of an amino acid sequence having a sequence identity of 97% or more with the amino acid sequence described in SEQ ID NO: 1, and having the activity of promoting the migration of PDGFRα-positive and CD29-positive mesenchymal stem cells. 25. Use of a substance in the manufacture of a medicament for the treatment of a heart disease associated with a structural abnormality and/or dysfunction of the heart in a subject with heart disease, where the substance is intended to be administered to a subject with heart disease and is described in any of (a) to (c ) below: (a) a peptide consisting of the amino acid sequence described in SEQ ID NO: 1; (b) a peptide consisting of an amino acid sequence in which one amino acid has been replaced, deleted, inserted or added in the amino acid sequence described in SEQ ID NO: 1, and has the activity of promoting the migration of PDGFRα-positive and CD29-positive mesenchymal stem cells; And (c) a peptide consisting of an amino acid sequence having a sequence identity of 97% or more with the amino acid sequence described in SEQ ID NO: 1, and having the activity of promoting the migration of PDGFRα-positive and CD29-positive mesenchymal stem cells, wherein the heart disease associated with a structural abnormality and/or dysfunction of the heart is selected from cardiomyopathy and prior myocardial infarction. 26. The use of a substance in the manufacture of a medicament for the prevention of chronic heart failure caused by cardiomyopathy or past myocardial infarction in a subject with cardiomyopathy or past myocardial infarction, where the substance is intended for administration to a subject with cardiomyopathy or past myocardial infarction and is described in any of paragraphs (a ) - (c) below: (a) a peptide consisting of the amino acid sequence described in SEQ ID NO: 1; (b) a peptide consisting of an amino acid sequence in which one amino acid has been replaced, deleted, inserted or added in the amino acid sequence described in SEQ ID NO: 1, and has the activity of promoting the migration of PDGFRα-positive and CD29-positive mesenchymal stem cells; And (c) a peptide consisting of an amino acid sequence having a sequence identity of 97% or more with the amino acid sequence described in SEQ ID NO: 1, and having the activity of promoting the migration of PDGFRα-positive and CD29-positive mesenchymal stem cells. 27. The use of a substance in the manufacture of a medicament for the inhibition of a structural abnormality of the heart selected from the group consisting of cardiomegaly, cardiomyocyte hypertrophy and myocardial fibrosis in a subject with cardiomyopathy or a history of myocardial infarction, where the substance is intended to be administered to a subject with cardiomyopathy or a history of myocardial infarction and described in any of paragraphs (a) - (c) below: (a) a peptide consisting of the amino acid sequence described in SEQ ID NO: 1; (b) a peptide consisting of an amino acid sequence in which one amino acid has been replaced, deleted, inserted or added in the amino acid sequence described in SEQ ID NO: 1, and has the activity of promoting the migration of PDGFRα-positive and CD29-positive mesenchymal stem cells; And (c) a peptide consisting of an amino acid sequence having a sequence identity of 97% or more with the amino acid sequence described in SEQ ID NO: 1, and having the activity of promoting the migration of PDGFRα-positive and CD29-positive mesenchymal stem cells.

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