EP3920957A1

EP3920957A1 - Cardio-protective effect of vasoconstriction-inhibiting factor (vif)

Info

Publication number: EP3920957A1
Application number: EP20703987.6A
Authority: EP
Inventors: Elisa-Anamaria LIEHN; Joachim Jankowski; Vera Jankowski
Original assignee: Rheinisch Westlische Technische Hochschuke RWTH
Current assignee: Rheinisch Westlische Technische Hochschuke RWTH
Priority date: 2019-02-05
Filing date: 2020-02-05
Publication date: 2021-12-15
Also published as: WO2020161158A1; US20220096597A1; DE102019102786A1

Abstract

The invention relates to a vasoconstriction-inhibiting factor (VIF) or a nuclei acid encoding same, for the prophylaxis and/or treatment of the effects of a heart disease. The invention further relates to pharmaceutical compositions that contain the VIF and to viable (combination) therapies, in particular using the pharmaceutical compositions according to the invention. The invention also relates to a kit for non-therapeutic in-vitro use, containing the VIF or a nucleic acid encoding same.

Description

Cardioprotective effect of vasoconstriction-inhibiting factor (VIF)

Technical area

The invention lies in the field of cardiovascular medicaments, in particular with regard to those therapeutic agents which can be used specifically in the prevention and / or therapy of the consequences of a heart disease, in particular a heart attack.

Background of the invention

High blood pressure and its consequences are one of the most common causes of death worldwide. High blood pressure often remains undetected as a hidden danger until it manifests itself in critical secondary diseases and / or damage. High blood pressure puts a particular strain on the cardiovascular system. On the one hand, high blood pressure puts a particular strain on the heart itself, especially the left ventricle, which applies / has to apply the high pressure and therefore has to do a constant extra work. To further ensure this, the thick muscle layer of the heart (myocardium) is enlarged further. With increasing thickness, however, the oxygen supply to the inner muscle layers becomes increasingly difficult. Over a longer period of time this can ultimately lead to cardiac insufficiency, in which the heart is no longer able to supply the body with a sufficient amount of oxygen-rich blood. On the other hand, due to the high pressure in the blood vessels, the blood vessels wear out or harden and thus enable arteriosclerosis to develop, in which in particular cholesterol esters and other fats are deposited in the vessel wall. Through these deposits, the blood vessels constrict, which in turn can lead to a further increase in blood pressure due to the associated increased vascular resistance. In the longer term, in particular, this dramatically increases the risk of coronary heart disease, angina pectoris, myocardial infarction and stroke.

The "American College of Cardiology" (ACC) and the "American Heart Association" (AHA) have been describing the following classification of blood pressure values ("New ACC / AHA High Blood Pressure Guidelines Lower Definition of Hypertension") since November 13, 2017.

Systolic [mmHg] Diastolic [mmHg]

Normal <120 and <80

Increased <130 and <80

Stage i 130-139 or 80-89

Stage 2 ^ 140 or ^ 90

Hypertensive crisis ^ 180 or § 20

The “European Society of Cardiology” (ESC) and the European Society of Hypertension (ESH) continue to refer to the classification published in 2003 (Tran et Giang, “Changes in blood pressure Classification, blood pressure goals and pharmacological treatment of essential hypertension in medical guidelines from 2003 to 2013 ", IJC Metabolie & Endocrine 2 (2014), 1-10) \

Systolic diastolic

Blood pressure value blood pressure value

[mmHg] [mmHg]

Optimal <120 <80

Normal 120-129 80-84

Increases 130-139 85-89

Stage i 140-159 90-99

Stage 2 160-170 100-109

Stage 3 ^ 180 ^ 1 10

Isolated systolic ^ 140 <90

hypertension The renin-angiotensin system (or the entire renin-angiotensin-aldosterone system) and in particular the underlying angiotensin peptides are essential for the regulation of blood pressure.

The enzyme renin activates a cascade in which renin converts the previously inactive angiotensinogen into angiotensin I through its protease function. Angiotensin I is finally converted into angiotensin I I by the angiotensin converting enzyme (ACE), which has a strong vasoconstrictive effect and the release of other substances, e.g. of the hormone vasopressin, which in turn have an antihypertensive effect.

Salem et al., Identification of the Vasoconstriction-Inhibiting Factor (VIF), “A Potent Endogenous Cofactor of Angiotensin II Acting on the Angiotensin II Type 2 Receptor”, Circulation, 2015 describes a new peptide, vasoconstriction-inhibiting factor (VIF), which can intervene in the vasoconstrictive properties of angiotensin II through an effect on the angiotensin II type 2 receptor. The VIF peptide as such and its formation in the human body is described in principle. However, possible effects on human pathophysiology, in particular with regard to cardiovascular diseases, and thus the therapeutic applicability and utilization of the peptide are not studied.

In addition, it was found that in patients with heart failure the plasma concentration of i.a. VIF was increased (Figure 1). It was unclear, however, whether and how the occurring phenomenon could be made technically usable. Furthermore, Salem et al. only the effect of VIF on vasoconstriction. Specific effects or the targeted use of VIF for specific illnesses are not disclosed.

The object of the present invention is the targeted uncovering of the mechanisms of action and the molecular mechanisms of the VIF, in particular in the context of cardiac events such as angina pectoris and myocardial infarction, in order to develop its therapeutic potential. A special focus here is on targeted studies to utilize the VIF in a possible treatment of patients with cardiac diseases. In particular, specific technical modifications of the VIF and its smaller peptides should be used to investigate further properties and VIF mutants. The aim of this discovery and investigation is to provide a preparation or combination preparation that can be used in the treatment and prevention of cardiac diseases. The previous treatment of heart diseases, especially a heart attack, often includes chronic lowering of blood pressure and thus lifelong drug therapy. Examples of this are therapy with beta blockers, statins, ACE inhibitors or peptides such as serelaxin. However, for serelaxin, for example, the phase III study RELAX-AHF-2 was unable to provide evidence of a clinical benefit of the corresponding active ingredient RLX030 (serelaxin). Possible side effects or long-term consequential damage from all established therapies cannot be ruled out. Based on this, the primary object of the present invention was to discover new, improved forms of therapy for patients with cardiac diseases to facilitate therapy, for which VIF was not previously described.

Definitions

The terms “amino acid molecule / sequence”, “protein”, “peptide” or polypeptide are used interchangeably herein, without going into the length of a specific amino acid sequence. The term “amino acid” or “amino acid sequence” or “amino acid molecule” includes all natural or chemically synthesized proteins, peptides or polypeptides or a modified protein, peptide, polypeptide and enzyme (polypeptide with a catalytic activity), where the term “ modified "includes any recombinant, chemical or enzymatic modification of the protein, peptide, polypeptide and enzyme or the nucleic acid sequence that encodes them.

The terms “sequence (s)” and “molecule (s)” are used interchangeably herein when referring to nucleic acid or amino acid sequences / molecules.

As used herein, the term "pharmaceutically acceptable" refers to those ingredients, materials, compositions and / or dosage forms which, within the scope of a medical consideration or within the definition of any medical regulatory and / or regulatory agency, are suitable for contact with cells, tissues, or the like - that of the constituent parts of a subject, ie humans and animals, including contact with malignant cells or tissues of a subject, without excessive toxicity, irritation, allergic reaction or other complications or side effects with an appropriate risk-benefit ratio for a subject / patient is. According to a preferred embodiment, one or more auxiliaries, as described below, are used. The term "subject" as used herein refers to a human or a non-human animal. The term includes mammals (e.g. humans, other primates, pigs, rodents (e.g. mice, rats or hamsters), rabbits, guinea pigs, cows, horses, cats, dogs, sheep and goats), but is not limited to this. In one embodiment the subject is a human.

In addition to classic diseases such as coronary heart disease, the term "heart disease" preferably also includes pathological conditions and events of the heart and thus among others also in particular a heart attack, angina pectoris and ischemia in the heart tissue.

The term "consequences of heart disease" as used herein does not include the occurrence of the disease itself, e.g. the occurrence of a heart attack, but rather the associated functional and / or pathological phenomena, e.g. a restriction in cardiac output or the area affected by the ischemia of the infarct.

The term “heart tissue” includes, but is not limited to, the pericardium, the epicardium, the pericardium, the fat layer located under the heart (Te / a subepicardiaca), the myocardium with the heart muscle cells and the endocardium, as well as the arterial and venous vascular accesses to the heart tissue, especially the coronary arteries.

The term “infarction” describes the destruction of tissue - especially through necrosis - as a result of an insufficient supply of oxygen (hypoxia), preferably through insufficient blood flow (ischemia).

The terms "treating", "treating", "treatment" and "therapy" as used herein describe treatment in a mammal, e.g. in a human including (a) preventing the consequences of a disease, i. to stop their development; (b) alleviating the consequences of illness, i.e. to bring about a declining development of the functions impaired by the disease or of the tissue affected by it; and / or (c) curing the consequences of the disease. The terms “treatment” and “therapy” are used interchangeably and include any form of preventive and / or curative treatment or therapy.

The terms "preventive,""preventive," or "prevention" mean that prophylactic treatment was given before the onset of the disease or before the symptoms associated with a disease to be prevented appeared. However, prevention does not always lead to a complete one Absence of the disease and its symptoms; ameliorating or delaying the disease or its symptoms is thus likewise encompassed by prevention, as described herein.

The term “partial sequence”, as used herein in the context of nucleic acid, amino acid and / or peptide sequences, relates to a coherent / contiguous fragment that can be derived from a matrix sequence according to the present application. A partial sequence therefore usually comprises 3, 4, 5, 6, 7, 8, 9, 10 or more contiguous positions according to the matrix sequence, possibly including additional modifications.

If a percentage of the homology or identity of nucleic acid or amino acid sequences is referred to in the present application, these values define those values that can be obtained by using the EMBOSS Water Pairwise Sequence Alignments (nucleotide) program (http: //www.ebi. ac.uk/Tools/psa/emboss_water/nucleotide.html) for nucleic acids or the EMBOSS Water Pairwise Sequence Alignments (protein) program (http://www.ebi.ac.uk/Tools/psa/emboss_water/) for amino acids were. These programs, which are provided by the European Molecular Biology Laboratory (EMBL) European Bioinformatics Institute (EBI) for local sequence alignments, use a modified Smith-Waterman algorithm (see http://www.ebi.ac.uk/Tools/ psa / and Smith, TF & Waterman, MS “Identification of common molecular subsequences” Journal of Molecular Biology, 1981 147 (1): 195-197). When performing an alignment, the standard parameters defined by EMBL-EBI are used. These parameters are (i) for amino acid sequences at: Matrix = BLOSUM62, gap open penalty = 10 and gap extend penalty = 0.5 or (ii) for nucleic acid sequences at: Matrix = DNAfull, gap open penalty = 10 and gap extend penalty = 0.5.

The pharmaceutical composition, as described herein, can be applied systemically or, if relevant, also locally. In the case of systemic use, the pharmaceutical composition or its active ingredients are transferred via direct (eg intravenous injection) or indirect (eg orally via the gastrointestinal tract) route into the blood and / or lymphatic system, which means that it is distributed throughout the whole Body or areas not separated by a special barrier (e.g. blood-brain barrier). When used locally, the pharmaceutical composition is applied to the tissue in which it is intended to act. For example, topical application or injection can take place. In some embodiments, local application can also take place in an adjacent tissue. In one embodiment, the pharmaceutical composition is in an orally administrable form. The known pharmaceutical forms are particularly preferred for such an application, e.g. tablets (non-coated as well as coated tablets, e.g. with enteric coatings), capsules, coated tablets, sprays, gels, bars, dosing bags, granules, pellets, syrups, solid mixtures, Dispersions in liquid phases, emulsions, solutions, pastes or other swallowable or chewable pharmaceutical preparations and aqueous or oily suspensions. In particular, but not exclusively, in a preventive therapy, an orally administrable form is particularly advantageous, since there is a high level of patient compliance.

In another embodiment, the pharmaceutical composition can be present in a form which can be administered intravenously, for example as a solution. If applicable, forms which can be administered can be obtained from a mixture of the active ingredient and auxiliaries. Such auxiliaries can, for example, fillers (such as sugar, sugar alcohols and cyclodextrins, thus for example sucrose, lactose, fructose, maltose, raffinose, sorbitol, lactitol, mannitol, maltitol, erythritol, inositol, trehalose, isomalt, inulin, maltodextrin, ß- Cyclodextrin, hydroxypropyl-ß-cyclodextrin, sulfobutyl ether-cyclodextrin or combinations thereof; calcium phosphate); Carriers (such as polyethylene glycol (PEG), polyethylene oxide (PEO), polyvinylpyrrolidone (PVP), polyvinyl alcohol (PVA), hydroxypropyl methyl cellulose (HMPC), hydroxypropyl cellulose (HPC), carboxymethyl ethyl cellulose (CMEC), hydroxypropylmethyl cellulose phthalate (HPMethyl cellulose), polymethacrylate Urea and sugar (e.g. mannitol)); Polymers (such as polyvinylpyrrolidone, vinylpyrrolidone / vinyl acetate copolymer, polyalkylene glycol (e.g. polyethylene glycol), hydroxyalkyl cellulose (e.g. hydroxypropyl cellulose), hydroxyalkylmethyl cellulose (e.g. hydroxypropylmethyl cellulose), carboxymethyl cellulose, sodium carboxymethyl cellulose, sodium carboxymethyl cellulose, polyvinyl acrylate, e.g. polyvinyl acrylate, e.g. , Vinyl alcohol / vinyl acetate copolymer, polyglycosylated glycerides, xanthan gum, carrageenan, chitosan, chitin, polydextrin, dextrins, starch and starch derivatives, proteins and combinations thereof); Surfactants (such as sodium dodecyl sulfate, Brij 96, Tween 80); Disintegrants (such as starch such as sodium starch glycolate, corn starch or their derivatives); Binders (such as, for example, povidone, crosspovidone, polyvinyl alcohols, hydroxypropylmethyl cellulose, microcrystalline cellulose, polyvinylpyrrolidone); Lubricants (such as stearic acid or its salts such as magnesium stearate, silicon dioxide, talc); Sweeteners (such as aspartame); Flavorings (such as ß-carotene); Plasticizers (such as triethyl citrate, dibutyl phthalate); Coating material (such as polyvinyl acetate phthalate, hydroxypropylmethylcellu- losephtalat); Coolants (such as menthol derivatives (e.g. L-mentyl lactate, L-menthyl alkyl carbonate, menthone ketals); re- / recrystallization inhibitors; flux; defoamers; antioxidants; adsorbents; dyes; substances that change the pH value.

A pharmaceutical composition according to the invention can also contain preservatives, solvents, stabilizers, wetting agents, emulsifiers, salts for adjusting the osmotic pressure, buffers or other components and substances customary for pharmaceutical compositions.

Brief description of the figures

FIG. 1 shows the increased plasma concentration of VIF in patients with heart failure (NYHA levels III and IV) compared to control persons (NYHA level <II). The NYHA classification is a scheme originally published by the New York Heart Association for classifying heart diseases according to their severity. Most often it is used to classify heart failure into different stages according to the patient's ability to perform. NYHA I: heart disease without physical limitations. Daily physical exertion does not cause inadequate exhaustion, arrhythmia, shortness of breath or angina pectoris. NYHA II: Heart disease with slight physical impairment. No complaints in peace. Daily physical exertion causes exhaustion, arrhythmias, shortness of breath or angina pectoris. NYHA III: heart disease with severe impairment of physical performance during normal activity. No complaints in peace. Low physical exertion causes exhaustion, arrhythmias, shortness of breath or angina pectoris. NYHA IV: Heart disease with discomfort in all physical activity and at rest. Bedridden.

FIG. 2 shows the antihypertensive effect of VIF on male Wistar rats after subcutaneous application of angiotensin II (0.4 mg per kg and day) with (A) or without (■) intraperitoneal application of VIF (1 mg per ml).

FIG. 3 shows the size of the area affected by an induced myocardial infarction (as a percentage of the ventricle) and the ejection fraction after the induced myocardial infarction (as a percentage of the ejection fraction before myocardial infarction) in mice with prior and subsequent 2-day treatment with VIF and mice without treatment with VIF.

FIG. 4 shows the size of the area affected by an induced myocardial infarction in the form of histological sections. FIG. 5 shows (A) the influence of the VIF treatment (A) on the ejection fraction of the heart after a myocardial infarction and (B) the influence of VIF on the blood pressure. CTR = control; preOP = control measurement before the surgical intervention in the animal model, as explained in more detail in the examples, in particular example 6.

FIG. 6 (A) to (E) shows the results, explained in more detail in Example 7, of immunohistochemical analyzes using VIF, which in (D) and (E) clearly demonstrate the positive influence of VIF on the formation of new vessels.

FIG. 7 (A) and (B) show the results of promoting the mitochondrial oxygen consumption rate as induced by VIF, which were explained in more detail in Example 8.

Detailed description

The primary object is achieved according to the invention by the provision of a vasoconstriction-inhibiting factor (VIF) or a nucleic acid encoding it, for use in the prevention and / or treatment of the consequences of a heart disease, preferably selected from the group consisting of coronary heart diseases, Myocarditis, myocardial infarction, myocardial ischemia, and myocardial hypoxia.

Previous therapies for heart disease specialize in lowering blood pressure over the long term in order to avoid recurrence of the same or similar heart diseases. Acute treatment strategies as well as compatible and safe prevention strategies for high-risk patients are urgently needed. There is also a great need for maintenance therapies after heart disease, which for example prevent the deterioration of the condition of the heart tissue.

In the case of a heart attack, for example, the blood flow to parts of the heart muscle is disturbed or interrupted. As a result, hypoxia occurs as a result of the lack of oxygen supply, which can lead to the death of cardiac muscle cells and thus to inflammatory reactions in the heart tissue. Such damage that has already occurred cannot, however, be treated satisfactorily and sustainably by the previous forms of therapy. In addition, treatment with previous forms of therapy is still unsatisfactory today. Patients often show no improvement in cardiac function, which is why further, invasive therapies are often necessary. Such therapies include cardiac resynchronization therapy (CRT), a biventricular pacemaker or an implantable cardioverter / defibrillator (ICD), which in turn carry a high risk of bleeding and infection.

It was surprisingly found that VIF, in addition to its influence on the renin-angiotensin system - and thus also on the regulation of blood pressure - has a protective effect in, for example, a heart attack. Surprisingly, a protective effect of VIF on heart muscles during persistent circulatory disorders was discovered and further characterized. The protective effect was shown by the fact that in pilot studies (pre-) treatment with VIF significantly reduced the area affected by an infarction. Such an effect is not described in the prior art for VIF and represents an enormous potential in the prevention of the consequences of a heart disease, in particular through a new approach which is aimed specifically at prevention or attenuation of the consequences of heart disease. By reducing the affected area, the resulting consequences are mitigated and ultimately the therapy for the affected patients is also facilitated (e.g. by a lower drug dose or by weaker drugs with fewer side effects and thus higher compliance for the patient).

It was also found that VIF leads to an improvement in the ejection fraction (ejection fraction) of the heart after a persistent circulatory disorder. The ejection fraction serves as a measure of the heart function. Such an effect is also not described in the prior art for VIF and represents a great advantage for a possible treatment, since previous therapies have so far failed to improve cardiac function. E.g. a clinical benefit of the active ingredient RLX030 (Serelaxin) cannot be provided. In the RELAX-AHF-2 study by Novartis, Serelaxin could neither reduce cardiovascular mortality in the first 180 days nor any further deterioration in initially stabilized patients in hospital in the first 5 days after the first episode of heart failure.

Studies in animal models have also shown that VIF surprisingly leads to the formation of new vessels as well as to an increased mitochondrial oxygen consumption rate after an induced infarction. It is known that after an acute infarction, significant metabolic changes occur not only in the infarcted but also in the surviving, non-infarcted segment (Mathes et al., 1974 Decreased contractility of the non-infarcted myocardium after an experimental infarction. In: Thauer R. , Pleschka K. (Ed.) The Arterial System, Edition 40), among others to a lower contractility due to a decreased oxygen supply. As part of the data collection for the present invention, it has now surprisingly been shown that VIF does not only play a role in vasoconstriction. Rather, VIF also have specific properties that can play an important role in therapy both in the prevention and treatment of heart disease. It could be shown that VIF increases the oxygen consumption rate of the mitochondrial respiratory chain in relevant cell types of the heart muscle, and thus can contribute to an increased contractility of the heart muscle.

At the same time, the administration of VIF does not lead to an increased inflammatory reaction, which makes the peptide interesting for both prophylactic and curative use. These specific effects of VIF have not been described to date and were not to be expected in view of the global mechanisms of action of VIF already described. A vasoconstriction-inhibiting factor (VIF) is also preferred for use according to the invention, the VIF having an amino acid sequence according to SEQ ID NO: 1 (from homo sapiens) or an amino acid sequence with at least 90%, 91%, 92%, 93%, 94% , 95%, 96%, 97%, 98%, 99% sequence identity to the sequence according to SEQ ID NO: 1. An amino acid sequence according to SEQ ID NO: 1 or an amino acid sequence with at least 95% sequence identity to the sequence according to SEQ ID NO: 1 is preferred. Also preferred is an amino acid sequence in which an amino acid has been specifically substituted for the sequence according to SEQ ID NO: 1, for example in order to examine the effect / the mechanism of action of VIF. SEQ ID NO: 1 describes the amino acid sequence of the vasoconstriction-inhibiting factor (VIF) in its entire length.

In another embodiment, the VIF preferably contains an amino acid sequence according to SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7 or SEQ ID NO : 8 or an amino acid sequence with at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% sequence identity to the sequence according to SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7 or SEQ ID NO: 8. An amino acid sequence according to SEQ ID NO: 2, SEQ ID NO : 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7 or SEQ ID NO: 8 or an amino acid sequence with at least 95% sequence identity to the sequence according to SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7 or SEQ ID NO: 8. Also preferred is an amino acid sequence in which one amino acid is specifically opposite to the sequence according to SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7 or SEQ ID NO: 8 was substituted, for example to investigate the effect / the mechanism of action of VIF. The SEQ ID NOs: 2 to 8 describe the amino acid sequences of individual peptides within the VIF (SEQ ID NO: 1).

The present invention also relates to a nucleic acid for use according to the invention, the nucleic acid having an amino acid sequence according to SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO : 6, SEQ ID NO: 7 or SEQ ID NO: 8 or an amino acid sequence with at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% sequence Identity to the sequence according to SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7 or SEQ ID NO : 8 coded. An amino acid sequence according to is preferred SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7 or SEQ ID NO: 8, or one Amino acid sequence with at least 95% sequence identity to the sequence according to SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7 or SEQ ID NO: 8. Further preferred is an amino acid sequence in which one amino acid is specifically opposite to the sequence according to SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7 or SEQ ID NO: 8 was substituted, for example in order to investigate the effect / the mechanism of action of VIF.

The present invention preferably relates to a vasoconstriction-inhibiting factor (VIF) or a nucleic acid encoding it, for use according to the invention, the prevention and / or treatment reducing the area of the heart tissue affected by the heart disease and / or reducing the restriction of the Cardiac output involves heart disease.

In one embodiment, the present invention relates to a vasoconstriction-inhibiting factor (VIF) or a nucleic acid encoding it for use according to the invention, the prevention and / or treatment of the consequences of a heart disease being achieved by increased neovascularization as a result of the administration of VIF.

In a further embodiment, the present invention relates to a vasoconstriction-inhibiting factor (VIF) or a nucleic acid encoding it for use according to the invention, the prevention and / or treatment of the consequences of a heart disease being achieved by an increased mitochondrial oxygen consumption rate as a result of the administration of VIF becomes.

In yet another embodiment, the present invention relates to a vasoconstriction-inhibiting factor (VIF) or a nucleic acid encoding it, for use according to the invention, the prevention and / or treatment of the consequences of a heart disease through an increased contractility of the heart muscle cells as a result of the VIF Gift is achieved. In particular, the property of VIF, through its influence on the respiratory chain of the mitochondria and thus the oxygen turnover of a cell, of directly influencing the metabolism of cardiac muscle cells, suggests dual therapeutic use of VIF: on the one hand for prevention in patients with a risk and / or predisposition the development of coronary heart disease, as well as treatment after a heart attack to strengthen the cells affected by the infarct. In a further embodiment, the present invention relates to a vasoconstriction-inhibiting factor (VIF) or a nucleic acid encoding this for use according to the invention, the treatment of the consequences of a heart disease being achieved by increased monocyte infiltration into the infarcted tissue as a result of administration of VIF becomes.

It is particularly advantageous for the prevention and / or treatment of the consequences of a heart disease to keep the affected area of the heart tissue as small as possible, especially after ischemia, so that only little tissue has to be treated and the expected loss of function is as small as possible hold. This makes it possible to keep the required amount of medication and thus the potential side effects or long-term consequences low. The restriction of the cardiac output is one of the most important and therefore most dangerous impairments, since a restricted cardiac output can endanger the oxygen supply to the entire body. The less the cardiac output is restricted, the less the loss of function of the heart is.

A vasoconstriction-inhibiting factor (VIF) or a nucleic acid encoding it is further preferred for use according to the invention, the amino acid sequence or the nucleic acid sequence encoding it being produced by a fully synthetic process, by a biotechnological process or a combination of such processes, or wherein the production of the amino acid sequence, or the nucleic acid sequence encoding it, comprises a fully synthetic process, a biotechnological process or a combination of such processes.

In addition, a vasoconstriction-inhibiting factor (VIF) or a nucleic acid that codes for this is preferred for use according to the invention, the VIF as a peptide, protein, optionally as a partial sequence of the VIF amino acid sequence and / or corresponding mimetics or as nucleic acid and / or a mixture thereof is used or applied, optionally together with at least one further pharmaceutically acceptable agent.

The dosage form of the VIF or its form when used, whether as a peptide, protein or nucleic acid, optionally as a partial sequence of the VIF amino acid sequence and / or corresponding mimetics and / or a mixture thereof, can have an influence on the spatial distribution of the VIF, its Have concentration in blood or its half-life. Preferably, auxiliaries, as described herein, are used as pharmaceutically acceptable agents, in particular to achieve the properties of the VIF described in to influence the corresponding interest. The properties described have an influence in particular on the amount of VIF available in the blood or in the heart tissue. The too fast or too slow degradation of VIF, or its half-life, plays a major role. It is preferred that larger fluctuations in the VIF concentration are avoided in order to avoid possible over- or under-dosing.

The nucleic acid sequence or the amino acid sequence optionally contains at least one additional sequence as a partial sequence of the VIF amino acid sequence and / or corresponding mimetics, preferably where the at least one additional sequence has a stabilizing function, a marker function, an interaction function, a modulation function, or a Has a localizing function. The additional sequence in the direction from the 5 'to the 3' end of the nucleic acid, or in the direction from the C-terminal to the N-terminal end, is preferred before or after the sequence of the VIF or the sequence encoding the VIF , and not within this sequence. According to a preferred embodiment of the present invention, the at least one additional sequence does not adversely affect the activity of VIF. According to another embodiment of the present invention, the one or more of the at least one additional sequence (s) can positively or negatively influence the activity of VIF. According to a further embodiment of the present invention, the at least one additional sequence does not influence the activity of VIF.

Additional functions such as a stabilizing function are of great advantage in order to counteract possible degradation or degradation processes in the desired area of application (e.g. the human body, especially in the blood vessels and heart tissue), which results in longer-lasting and greater availability in terms of area. Marker functions allow tracking and thus marking of the already treated tissue, or the distribution in the application area can also be studied and analyzed by tracking. An interaction function can enable interaction with previously selected other substances or fabrics. Modulation functions allow an influence on the activity, for example, which can for example be tied to a certain length of stay in the application area, whereby an activity before and / or after a previously selected time frame is no longer possible or is only then possible. A localizing function is, for example, a signal peptide, or a nucleic acid or amino acid sequence that encodes it, which triggers or initiates transport into a previously selected tissue. Through such functions the applicability, the availability and the activity of the VIF or the nucleic acid or amino acid sequence can be specifically influenced in order to obtain an optimal result.

The present invention further relates to a pharmaceutical composition containing or consisting of a) vasoconstriction-inhibiting factor (VIF) as defined above, b) optionally at least one auxiliary and / or additive, preferably wherein the at least one auxiliary and / or additive is selected from the group consisting of fillers, carriers, polymers, surfactants, disintegrants, binders, lubricants, sweeteners, flavorings, plasticizers, coating materials, coolants, re / recrystallization inhibitors, fluxes, defoamers, antioxidants, adsorbents, colorants, pH value changes Substances, preservatives, solvents, stabilizers, wetting agents, emulsifiers, salts for setting the osmotic pressure or buffers, and c) at least one further pharmaceutically active substance, the at least one further pharmaceutically active substance being selected from statins, anticoagulants, beta blockers n, ACE inhibitors, platelet aggregation inhibitors, sartans, calcium antagonists, diuretics, the pharmaceutical composition for use for the prevention and / or treatment of the consequences of a heart disease, preferably selected from the group consisting of coronary heart disease, myocarditis, myocardial infarction, and myocardial ischemia myocardial hypoxia, is suitable, preferably wherein the prevention and / or treatment includes a reduction in the area of the heart tissue affected by the heart disease and / or a reduction in the restriction of the cardiac output by the heart disease.

The protective effects of VIF on cardiac muscle cells described above can advantageously be used in therapy for the prevention and / or treatment of the consequences of a heart disease. It is particularly advantageous to combine the therapy with other pharmaceutically active substances. For example, the antihypertensive effects of the substances used to date can be combined with the protective effects of VIF. On the one hand, the occurrence of heart diseases, such as described above, prevented or delayed. In addition, the consequences of heart disease, as described above, can be reduced or even prevented. Often, high blood pressure - an important cause of heart disease - is not discovered in time before heart disease occurs. The risk of heart disease can then be reduced by lowering blood pressure, but it is by no means eliminated. An additional, preferably joint therapy with VIF, preferably as part of a pharmaceutical composition according to the invention, can in this situation at least reduce or even prevent the consequences of a heart disease, as described above, for example as part of long-term therapy, but also as part of short-term treatment .

In one embodiment, the pharmaceutical composition for use in treating the consequences of a heart disease is one that is used in the context of post-infarction secondary prevention, VIF in combination with a statin and / or a beta blocker and / or an anticoagulant and / or optional is given an ACE inhibitor. In maintenance therapy after a heart attack, the administration of an ACE inhibitor and / or an angiotensin II receptor blocker can preferably be reduced with the simultaneous administration of VIF.

In a further embodiment, a pharmaceutical composition for use in the prevention of a heart disease, in particular in high-risk and high-risk patients for the development of a coronary heart disease, or a heart failure, is provided, wherein VIF in combination with a statin and / or a beta blocker and / or a Anticoagulant and / or optionally an ACE inhibitor is administered. In prophylaxis, VIF can in particular be administered together with a statin, or another lipid-lowering agent, in order to counteract synergistic atherosclerosis and plaque formation caused by the statin, as well as directly the reduced activity of the cells of the heart tissue due to VIF. This can prevent the risk of a heart attack, but also the risk of a perioperative heart attack as a complication.

The person skilled in the art is aware that the selection of the at least one further medicinally active ingredient administered in combination with VIF, as well as the concentration of VIF itself and the at least one further medicinally active ingredient and the treatment and dosage regimen, of any underlying diseases, such as hyper- cholesterolemia, can depend.

The current therapies for the treatment of a heart attack are not all able to alleviate the acute necrosis in the heart tissue and the regeneration of the infarcted Promote tissue. Hypoxia leads to an induced death of cardiomyocytes as a result of an infarction. This direct damage is currently only insufficiently treated by secondary therapies after a heart attack.

Patients who have suffered a heart attack are usually treated simultaneously with different drug groups as a result of a heart attack, usually in a combination of four or more preparations, often in a four-fold combination of an anticoagulant to inhibit blood clotting, such as ASA or clopidogrel , a statin to lower cholesterol, an ACE inhibitor or angiotensin II receptor blocker to lower blood pressure and a beta blocker to lower your heart rate. The additional administration of VIF can, on the one hand, reduce the use of antihypertensive agents because of its vasodilatory properties. At the same time, through its influence on the formation of new blood vessels, as well as on the mitochondrial respiratory chain, and the recruitment of monocytes, VIF can significantly contribute to the faster regeneration of infarcted and neighboring heart tissue and thus have a direct positive influence on the treatment of acute coronary heart disease.

Likewise, cardiac risk patients can be identified and adequately treated long before a heart attack. In one embodiment, VIF is therefore used preventively for cardio- or vasculoprotective strategies, alone or in combination.

Therapy with VIF, preferably in a pharmaceutical composition according to the invention, can be carried out in the context of acute treatment, prevention and / or maintenance therapy.

In a pharmaceutical composition according to the invention, components a) and c) are preferably used in a pharmaceutically effective amount. This amount is typically about a concentration of 1 to 1,000 pg / kg body weight. In one embodiment, the dosage / dose of a VIF peptide according to the present invention is 1, 10, 30, 50, 100 or 250 pg / kg / day. In the case of acute administration, the use of a higher concentration per day (in the range from about 250 pg / kg to 15,000 pg / kg, depending on the extent of the acute symptoms to be treated and furthermore depending on individual patient factors, the concentration can also be about 500 pg / kg to 10,000 pg / kg, about 750 pg / kg to 7,500 pg / kg, or about 500 pg / kg to 5,000 pg / kg) are preferred, for long-term or maintenance therapy a lower dose per day (<5,000 pg / kg , or also (<1000 pg / kg) The dose can vary from application form to application form, as is known to the person skilled in the art. The pharmaceutical composition according to the invention can be applied systemically or locally. Preferred systemic applications are oral or parenteral such as intravenous, subcutaneous or endobronchial applications, applications per os, or an injection directly into the target tissue to be treated, preferably to induce a topical effect.

Preferably the pharmaceutical composition of the invention is in solid form, e.g. as powder, in liquid form, e.g. as a solution for injection or as an aerosol.

In addition, the present invention relates to a kit for non-therapeutic in-vitro use, containing the vasoconstriction-inhibiting factor (VIF) or a nucleic acid that encodes this, as defined above.

Such a kit is used in particular in uncovering the mechanisms of action and the molecular mechanisms of VIF. In addition to the VIF or the nucleic acid that encodes it, other content can also be present. These include, for example, further compounds, substances or reagents that can be used for the detection work.

A kit can be provided in such a way that the contents are present in already measured amounts and / or concentrations so that they can be used directly or can simply be diluted to a usable concentration. If further contents are present in addition to the VIF or the nucleic acid encoding it, these are preferably provided in a quantity or weight ratio to the VIF or the nucleic acid encoding it, in which they are actually or approximately used.

The present invention will now be further described by the accompanying non-limiting examples, the drawings and the sequence listing.

Examples

Example 1: Synthesis of the VIF

The VIF was automatically synthesized using the solid-phase method and standardized fluorene-nylmethyloxycarbonyl chloride chemistry via continuous-flow peptide synthesis. Example 2: Blood pressure lowering effect of the VIF

Male Wistar rats were used to investigate the in vivo effect of the VIF. All animals had free access to standard rat chow and tap water ad libitum. The mean arterial pressure was measured consciously using a tail-cuff sphygmomanometer. Five determinations were carried out, the mean value serving as the basal value. The animals then received VIF intraperitoneally (1 mg per ml) and angiotensin II subcutaneously (0.4 mg per kg and day). The control group, on the other hand, received only angiotensin II.

The blood pressure was then measured every 5 minutes for 30 to 45 minutes using a microcatheter and determined using the ADInstruments software (Miliar, Germany).

The results of the blood pressure measurements can be seen in FIG. 2, the groups with (▲) and without (■) application of VIF being plotted.

Example 3: Protective effect of the VIF on cardiac tissue

To investigate the protective effects of the VIF, mice were treated with VIF for 2 days before and 2 days after the investigation. Control animals received no treatment.

A heart attack was triggered on the day of the examination. For this purpose, the mice were anesthetized and ventilated by an intraperitoneal injection of 100 mg / kg body weight ketamine and 10 mg / kg body weight xylazine. The heart attack was triggered by an occlusion of the LAD (left anterior descending artery).

The area of the affected tissue was then determined via histological sections. For this, the heart was removed, perfused with 1% Evans Blue, frozen for 2 h at -20 ° C. and then cut into 5 sections. The sections were incubated for 10 min with preheated TTC solution and fixed in formalin. Then recordings were made and the infarct area was calculated using DISKUS (Hilgard, Germany).

It was found that in the animals treated with VIF the infarct size was only about half as large as in the control group. In animals treated with VIF, the tissue affected by the infarct was found to occupy approximately 25% of the ventricle. In control animals the affected tissue was about 50% of the ventricle. The results are shown in FIGS. 3 and 4.

The subsequent 2-day treatment with VIF also significantly improved the ejection fraction after the infarction. When measured, the ejection fraction of the control animals was almost 20% of the function in the healthy state, whereas the animals treated with VIF had an ejection fraction averaging 30% of the function in the healthy state. The results are shown in FIG. 3.

Example 4: Pharmaceutical Composition

The formulation of peptide-containing pharmaceutical compositions is conditioned by the solubility profile of the particular peptide of interest, its stability and the isoelectric point of the peptide as active ingredient. These characteristics also determine the optimal pH value, which is used in the context of development and formulation. In particular, the choice of the correct buffer system can be of great importance. Depending on the application route, peptide-containing pharmaceutical compositions are then dissolved in a suitable physiologically compatible buffer / solvent system immediately prior to their use, provided that they are provided in powder form or in lyophilized form. The addition of stabilizers and preservatives is also important, for example to prevent contamination of the peptide active ingredient. Stabilization can be of great importance in particular for non-parenteral administration if a certain half-life of the peptide in the patient must be achieved so that the peptide active ingredient can develop its activity over a given period of time. In addition, other aids may be available. The use of aids for delayed release can also be of importance in the context of the VIF peptides of the present invention, in particular when these are used in long-term therapy. Suitable pharmaceutical compositions based on peptides are familiar to pharmacologists (see Pharmaceutical Formulation Development of Peptides and Proteins, edited by Lars Hovgaard, Sven Frokjaer, Marco van de Weert, Taylor & Francis, 2012).

For a pharmaceutical composition according to the invention, the substances were provided in powder form, solution or emulsion and mixed one after the other and optionally brought into solution. Different buffer systems were used under physiological conditions, depending on whether a full-length VIF peptide or a of the truncated variants was used (cf. Swain et al., Recent Patents on Biotechnology, 2013, 7). The mixture was then sterile filtered. The stability and functionality of the peptides was checked over time by means of analytical in vitro tests.

Example 5: Heart Attack Animal Model Study

In order to study the newly identified effects of VIF (according to SEQ ID NO: 1) in more detail, and in particular also in vivo, male wild-type C57BL / 6N mice (Charles River, Germany) were 8 to 10 weeks old under anesthesia (100 mg / kg ketamine, 10 mg / kg xylazine, ip) and analgesia (0.1 mg / kg buprenorphine) intubated. The mice were ventilated using a rodent respirator (Harvard Apparatus, Germany) with positive pressure and oxygen. A left thoracotomy was then performed and the Ml ("myocardial infarction") was performed by occlusal ligature of the left anterior descending artery (LAD) with 0/7 silk, as previously described in Curaj et al. (Minimally invasive surgical procedure of inducing myocardial infarction in mice. J Vis Exp. 2015: e52197). The rib, muscle and skin incisions were closed with separate sutures. Analgesia was continued for five days after the indicated infarction using 0.1 mg / kg buprenorphine every eight hours. The hearts were then removed at defined times (after 0, 1, 4, 7, 14, 21, 28 days) and prepared for further analysis.

VIF was dissolved at 6.7 pg / ml (1 mmol / l) in NaCl and loaded into 100 μl osmotic pumps of the Alzet type 1002 (0.25 ml / hour, Charles River, Cologne, Germany), resulting in one dose of 0.8 pg / kg per 24 hours. The Alzet pumps were implanted 24 hours prior to MI induction. The pumps for the controls were accordingly only filled with NaCl. All mice were kept under standardized conditions in the specially designated animal rooms at the University Hospital Aachen (Germany). All animal experiments and test protocols were approved by the local authorities in compliance with the European and German animal welfare laws (84-02.04.2016.A315). All mice were included in the analysis, unless the animals had died in the course of the experiment.

Example 6: Echocardiography

Two-dimensional and M-mode echocardiography measurements were made with an ultrasound image converter especially for small animals (Vevo 770, FUJIFILM Visualsonics, Toronto, Canada). Both measurements were taken before and after a heart attack. For this purpose, the mice were anesthetized with 1.5-2% isoflurane and placed on their backs on a heat pad. The ejection fraction, cardiac output volume and heart rate were analyzed. The results are shown in FIG.

It can be seen from the results that VIF, on the one hand, significantly increases the ejection fraction of the heart after treatment post-infarction (FIG. 5A), which on this scale was not expected whether the properties described for VIF and a significant contribution to future treatments after a myocardial infarction can enable. There is also a slight decrease in blood pressure (FIG. 5B).

Example 7: Histology and Immunohistochemistry

To determine the size of the infarct, Gömöri trichrome staining was carried out. Subsequently, with the help of ImageJ. three sections per mouse from 3 to 5 different mice after one of anti-SMA antibodies (actin of smooth muscles, DAKO, FIG. 6D), anti-ADC3 antibodies (BD Pharmingen, FIG. 6B), anti-MPO antibodies (Neomarkers, FIG. 6A) and anti-CD31 antibodies (Santa Cruz Biotechnology, FIG. 6E) staining, followed by staining with fluorescein isothiocyanate (FITC) or a Cy3-conjugated secondary antibody (DAKO, Germany), analyzed. Positive-stained cells or double-positive-stained cells (FIG. 6C for anti-ADC3 / MPO) were counted in three different fields per section and indicated as cells per visual field (200-fold magnification).

The results are shown in FIG. On the one hand, these data clearly demonstrate (FIGS. 6A and 6C) that the VIF treatment does not lead to an increased infiltration of neutrophils visualized with anti-MPO compared to untreated controls (CTR). This is an important indication for the therapeutic applicability of VIF in that it does not cause acute cell-mediated inflammatory reactions. After the invasion of neutrophils, a special subpopulation of monocytes is invaded in a second phase after a heart attack. These Gr1 -high expressing (high) monocytes (Gr1 + CCR2 + CX3CR1 low) are characterized like human CD14 ^hi9h CD16 monocytes, dominate the early phase of myocardial infarction and show phagocytic, proteolytic and inflammatory functions. Gr1 -highly expressing monocytes digest the infarct heart tissue and remove cell debris from this area. The number of monocytes (anti-ADC3 visualized) in VIF-treated animals was predominantly the same as in the control group. However, on day 7, a slight, but statistically significant, increase was found in the individual coloration (FIG. 6B). This can be assessed positively in that the infiltrating monocytes contribute to the elimination of dead tissue and consequently to an improved regeneration, which can be directly influenced by the administration of VIF. This confirms that VIF can be used as a therapeutic agent after an acute heart attack to promote and accelerate cell regeneration.

In principle, the results for VIF-treated animals and for the control group in the observed period of 28 days after induced infarction were almost the same.

With regard to the function originally described for the VIF peptide as a vasoconstrictive factor, the newly discovered effect of the formation of new vessels, which could be achieved after an induced infarction through the use of VIF (FIG. 6D SMA and E CD31), was surprising. 6D and E show the number of SMA- and CD31-positive cells in the field of view, CD31 being used as a marker for endothelial cells and SMA as a marker for smooth muscle cells. For both markers, a statistically significant amount of increased myofibroblast count and angiogenesis, both as an indication of vascularization, could be observed in the groups treated with VIF on day 7.

In this way, surprisingly, in the critical phase after an infarction, which can damage large parts of the heart tissue, the VIF administration leads to accelerated neovascularization as the basis for healing the tissue damaged by the infarction. This makes VIF an interesting candidate in the treatment of an acute heart attack in order to specifically promote the formation of new blood vessels and thus minimize the damage that occurs.

Example 8: Energy catabolism measurements

In order to further the role of VIF post-infarct to investigate the above-described animal model, a measurement was the 0 ₂ consumption in the tissue (OCR for "oxygen consumption rate") of VIF-treated and non-treated animals according to the induced infarct performed (Agilent Seahorse XF Cell Mito Stress Test Kit; Seahorse Bioanalyzer, Seahorse Bioscience). The mitochondrial function of cells is determined. FCCP (carbonyl cyanide-4 (trifluoromethoxy) phenylhydrazone) is used as a modulator. The experiments were carried out using both 5 pM FCCP (FIG. 7A) and 2.5 pM FCCP (FIG. 7B) on HL-1 cells (heart muscle cell line) as performed. The administration of FCCP leads to an induced breakdown of the proton gradient and thereby interrupts the mitochondrial membrane potential. FCCP-stimulated OCR can therefore be used to determine the delta between maximal and basal activity. This delta, in turn, is a measure of how well a cell can react to increased energy requirements (e.g. after stress).

As FIGS. 7A and B illustrate, VIF-treated cells (VIF in each case titrated from 0.1 to 1 mM) were always able to achieve significantly higher OCR values than the untreated control cells (CTRL).

This means that VIFs increase the maximum myocardial oxygen turnover and thereby the contractility, among other things. of heart muscle cells can increase. This can make a decisive contribution both to the prophylaxis as well as the therapy of coronary heart diseases, since it can specifically influence the relevant OCR values by modulating the respiratory chain reaction. These results show that VIF, in addition to its role as a vasoconstrictive factor, can unexpectedly influence other metabolic physiological processes as a specific modulator. These effects are currently also being further checked for the other VIF variants (according to SEQ ID NOs: 2 to 8) by means of in vitro and in vivo analyzes.

Example 9: Statistical Analysis

The statistical data shown in the figures represent the mean ± SEM (standard error of the mean). Statistical analysis was performed using Prism 7 software (GraphPad). The means of two groups were compared using Student's unpaired t-test using Welch's correction by significant variance. More than two groups were assessed using a one-way ANOVA analysis of variance followed by a Newman-Keuls post-hoc test, or a two-way ANOVA analysis of variance followed by a Bonferroni multiple comparison test, in the case of more than two variable parameters as indicated analyzed. P values of <0.05 were considered significant.

Claims

Expectations

1. Vasoconstriction-inhibiting factor (VIF) or nucleic acid encoding it, for use in the prevention and / or treatment of the consequences of a heart disease, preferably selected from the group consisting of coronary heart disease, myocarditis, myocardial infarction, myocardial ischemia and myocardial hypoxia .

2. Vasoconstriction-inhibiting factor (VIF) for use according to claim 1, wherein the VIF has an amino acid sequence according to SEQ ID NO: 1 or an amino acid sequence with at least 90%, 91%, 92%, 93%, 94%, 95%, 96 %, 97%, 98%, 99% sequence identity to the sequence according to SEQ ID NO: 1.

3. Vasoconstriction-inhibiting factor (VIF) for use according to claim 1, wherein the VIF has an amino acid sequence according to SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6 , SEQ ID NO: 7 or SEQ ID NO: 8 or an amino acid sequence with at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% sequence identity the sequence according to SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7 or SEQ ID NO: 8.

4. Nucleic acid for use according to claim 1, wherein the nucleic acid has an amino acid sequence according to SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6 , SEQ ID NO: 7 or SEQ ID NO: 8 or an amino acid sequence with at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% sequence identity the sequence according to SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7 or SEQ ID NO: 8 coded.

5. Vasoconstriction-inhibiting factor (VIF) or nucleic acid encoding the same for use according to one of the preceding claims, wherein the prevention and / or treatment is a reduction in the area of the heart tissue affected by the heart disease and / or a reduction in the impairment of cardiac output by heart disease.

6. Vasoconstriction-inhibiting factor (VIF) or nucleic acid encoding this, for use according to one of the preceding claims, wherein the prevention and / or treatment of the consequences of heart disease through increased formation of new blood vessels as a result of the administration of VIF is achieved.

7. Vasoconstriction-inhibiting factor (VIF) or nucleic acid encoding it, for use according to one of claims 1 to 5, wherein the prevention and / or treatment of the consequences of a heart disease is achieved by an increased mitochondrial oxygen consumption rate as a result of the VIF administration.

8. Vasoconstriction-inhibiting factor (VIF) or nucleic acid encoding this for use according to claim 7, wherein the prevention and / or treatment of the consequences of a heart disease is achieved by an increased contractility of the heart muscle cells as a result of the administration of VIF.

9. Vasoconstriction-inhibiting factor (VIF) or nucleic acid encoding it, for use according to one of claims 1 to 5, wherein the treatment of the consequences of a heart disease is achieved by increased monocyte infiltration into the infarcted tissue as a result of the administration of VIF.

10. Vasoconstriction-inhibiting factor (VIF) or nucleic acid encoding this for use according to one of the preceding claims, wherein the amino acid sequence or the nucleic acid sequence encoding it is produced by a fully synthetic method, by a biotechnological method or a combination of such methods , or wherein the production of the amino acid sequence, or the nucleic acid sequence encoding it, comprises a fully synthetic process, a biotechnological process or a combination of such processes.

11. Vasoconstriction-inhibiting factor (VIF) or nucleic acid encoding this for use according to one of the preceding claims, wherein the VIF is used as a peptide, protein, or as nucleic acid and / or a mixture thereof, or is administered, optionally together with at least one further pharmaceutically acceptable agent.

12. Vasoconstriction-inhibiting factor (VIF) or nucleic acid encoding it for use according to one of the preceding claims, wherein the nucleic acid sequence or the amino acid sequence contains at least one additional sequence, preferably wherein the at least one additional sequence has a stabilizing function, a marker Function, an interaction function, a modulation function, or a localizing function.

13. A pharmaceutical composition containing or consisting of a) vasoconstriction-inhibiting factor (VIF) as defined in one of the preceding claims, b) optionally at least one auxiliary and / or additive, preferably wherein the at least one auxiliary and / or additive is selected from the group consisting of fillers, carriers, polymers, surfactants, disintegrants, binders, lubricants, sweeteners, flavorings, plasticizers, coating materials, coolants, re / recrystallization inhibitors, fluxes, defoamers, antioxidants, adsorbents, colorants, pH value changes Substances, preservatives, solvents, stabilizers, wetting agents, emulsifiers, salts for setting the osmotic pressure or buffers, and c) at least one further pharmaceutically active substance, the at least one further pharmaceutically active substance being selected from statins, anticoagulants, beta blockers, ACE Inhibitors , Platelet aggregation inhibitors, sartans, calcium antagonists, diuretics, the pharmaceutical composition being suitable for use for the prevention and / or treatment of the consequences of a heart disease, preferably selected from the group consisting of coronary heart disease, myocarditis, myocardial infarction, myocardial ischemia and myocardial hypoxia , preferably wherein the prevention and / or treatment includes a reduction in the area of the heart tissue affected by the heart disease and / or a reduction in the restriction of the cardiac output by the heart disease.

14. Pharmaceutical composition according to claim 13, wherein the pharmaceutical composition is used for the treatment of the consequences of a heart disease in the context of post-infarction secondary prevention, wherein VIF in combination with a statin and / or a beta blocker and / or an anticoagulant and / or optionally an ACE inhibitor is administered, or wherein the pharmaceutical composition is used for the prevention of a heart disease, wherein VIF is administered in combination with a statin and / or a beta blocker and / or an anticoagulant and / or optionally an ACE inhibitor, preferred, VIF being administered in combination with a statin.

15. Kit for non-therapeutic in-vitro use, containing the vasoconstriction-inhibiting factor (VIF) or a nucleic acid which encodes this, as defined in one of the preceding claims.