Compositions for preventing atrial and ventricular fibrillation

[0001] Arrhythmias are a significant health problem. Arrhythmias include, but are not limited to, ventricular tachycardia, supraventricular tachycardia and atrial fibrillation. Of these, atrial fibrillation is the most common arrhythmia. It has been estimated that over one million people in the United States alone suffer from atrial fibrillation. The prevalence of atrial fibrillation is predicted to increase over the next few decades as populations in the United States and Europe age, because atrial fibrillation tends to be more common with age.

[0002] Cardiac rhythm disturbances are an important cause of morbidity and mortality. The ability to tolerate arrhythmias is less in the post-operative period than for similar arrhythmias in the pre-operative period. Haemodynamic instability is more likely due to the possibility of myocardial dysfunction. The extracorporeal circulation, trauma of the conductive system during surgery, metabolic and electrolyte abnormalities, especially hypokalaemia and hypomagnesaemia, contribute to the increased incidence of post-operative arrhythmias. Stress associated with surgery with increased sympathetic tone and the use of inotropic support are additional factors. A delayed arrhythmia may occur as a result of a recurrent wave associated with scarring.

[0003] Atrial fibrillation can be treated with drugs intended to maintain normal sinus rhythm and / or to reduce ventricular response rates. In particular, many of the older trials were limited to pharmacotherapy, radio-frequency ablation, the use of implantable devices and related strategies. Although drug therapy remains a popular way of reducing certain arrhythmia events, it has been noted that systemic effects are often poorly tolerated. In addition, it is considered that the tendencies conducive to arrhythmia, as demonstrated by many drugs, can increase mortality in many situations. It would be desirable to provide more effective treatments for or prevention of arrhythmias.

SUMMARY OF THE INVENTION [0004] In accordance with the purpose of the present invention, as exemplified and broadly described herein, the present invention relates to a composition for use in preventing a cardiac arrhythmia disorder in a subject. Additional advantages of the disclosed compositions will be set forth in the description below and in part will result from the description or may be learned in the practice of the disclosed compositions. The advantages of the disclosed compositions will be realized and achieved with the aid of elements and connections particularly indicated in the accompanying claims. It should be noted that both the above general description and the detailed description below have only exemplary and explanatory character and do not limit the invention as claimed.

DETAILED DESCRIPTION [0005] The disclosed compositions may be more readily understood with reference to the following detailed description of specific embodiments and the Example and the following description incorporated herein.

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[0006] Materials, compositions and elements that can be used for, can be used in combination with, can be used in the production or which are the products of the disclosed compositions are disclosed. These and other materials are disclosed herein and it is assumed that when combinations, subsets, interactions, groups, etc. of these materials are disclosed herein, a specific reference to each of the various individual and collective combinations and permutations of these compounds may not be clearly disclosed, everyone is specifically included and described here. For example, if a peptide is disclosed and discussed and a number of modifications that can be made to a number of molecules, including a peptide, are contemplated, specifically and all combinations and permutations of the peptide and modifications that are possible, are included unless specifically stated otherwise. Thus, if the class of molecules A, B and C is disclosed, as well as the class of molecules D, E and F and discloses the example of a combined AD molecule, then, even if each is not individually listed, each is individually and collectively included. Thus, in this example, each combination of AE, AF, BD, BE, BF, CD, CE and CF is specifically considered and should be considered disclosed from disclosure A, B and C; D, E and F; and an example combination of AD. Similarly, any subset or combination thereof is also specifically considered and disclosed. Thus, for example, the subgroup AE, BF and CE is specifically considered and should be considered disclosed in disclosure A, B and C; D, E and F; and an example of the AD combination. This idea applies to all aspects of this application, including, but not limited to, steps in the methods of making and using the disclosed compositions. Thus, if there are a number of additional steps that can be performed, it is recognized that each of these additional steps may be performed with any particular embodiment or combination of embodiments of the disclosed methods, and that each such combination is specifically considered and should be considered disclosed.

[0007] Those skilled in the art will recognize or will be able to determine, using no more than routine experimentation, many equivalents for specific embodiments of the compositions described herein. Such equivalents are intended to be covered by the following claims.

[0008] It should be noted that the disclosed compositions are not limited to the specific methodology, protocols and reagents described, as they may vary. It should also be noted that the terminology used herein is only intended to describe specific embodiments and is not intended to limit the scope of the present invention, which will be limited only by the appended claims.

A. DEFINITIONS [0009] Unless otherwise defined, all technical and scientific terms used herein have the same meanings as commonly understood by one of ordinary skill in the art to which the disclosed compositions belong. Although any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present compositions, particularly useful methods, devices and materials are as described. Nothing herein should be construed as an admission that the present invention does not give rise to a date prior to such disclosure as a result of the prior invention. Nothing here is an admission that any reference is state of the art. Discussion of references defines what

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[0010] It should be noted that as used herein and in the appended claims, the singular forms "a," "an" and "the" include plural references unless the context clearly dictates otherwise. Thus, for example, reference to "a compound" includes the plural of such compounds, a reference to "a compound" is a reference to one or more compounds and their equivalents known to those skilled in the art, and so on.

[0011] "Optional" or "optionally" means that the event, situation or material described above may appear or be present or not, and that the description includes instances where an event, situation or material appears or is present and cases where they do not appear or are not present.

[0012] Ranges can be expressed herein as from "about" one particular value and / or to "about" another specific value. When such a range is expressed, the other embodiment includes from one particular value and / or another specific value. Similarly, when values ​​are expressed as approximations, by using the preceding "about", it is assumed that the value specified is another embodiment. In addition, it should be noted that the endpoints of each range are relevant to both the other endpoint and independently of the other endpoint. It should also be noted that there are a number of values ​​disclosed herein, and that each value is also disclosed here as "about" a particular value, except for the value itself. For example, if the value "10" is disclosed, "about 10" is also disclosed. It should also be noted that when the value is disclosed, "less than or equal to _" values ​​are also disclosed - values, "greater than or equal values" and possible ranges between values, according to the respective understanding of the person skilled in the art. For example, if the value "10" is disclosed, then "less than or equal to 10" as well as "greater than or equal to 10" are also disclosed. It should also be noted that throughout the application data is provided in a number of different formats, and the data reflects the endpoints and starting points and ranges for any combination of these data points. For example, if a particular data point & quot; 10 & quot; and a specific data point is disclosed, it is meant that greater than, greater than or equal to, less than, less than or equal to and equal to 10 and 15 are considered disclosed, as well as between 10 and 15. It should also be noted that each unit between two specific units is also disclosed. For example, if 10 and 15 are disclosed, 11, 12, 13 and 14 are also disclosed.

Throughout the description and claims of the present description, the word & quot; comprise & quot; and variants of the word, such as & quot; comprising & quot; and & quot; comprising & quot; means & quot; including but not limited to & quot; and is not intended to exclude, for example, other additional ingredients. , components, integers or steps.

B. COMPOSITIONS [0014] Disclosed herein are compositions for use in the prevention of a arrhythmia disorder in a subject. The use comprises administering to the myocardium a subject a therapeutically effective amount of a composition comprising a mammalian extracellular matrix (ECM).

[0015] In some aspects, the mammalian ECM is from a native source. In some aspects, the mammalian ECM is produced in vitro using mammalian cells. In some aspects, the mammalian ECM is extracted directly from mammalian tissue / organs. In some aspects, the mammalian ECM containing composition further comprises a synthetic ECM.

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[0016] In certain aspects, a composition comprising a mammalian ECM inhibits scarring. In some aspects, a composition comprising a mammalian ECM stimulates the regeneration of damaged tissue. In some aspects, a composition comprising a mammalian ECM inhibits inflammation.

[0017] By "preventing" or "preventing" is meant a reduction in the incidence or severity of a disease or condition. This term does not require complete exclusion of the disease or condition. The term includes rather a reduction in the likelihood of the disease occurring. Thus, compositions for use in reducing the occurrence and / or severity of a arrhythmia disorder in a subject comprising a mammalian ECM are disclosed.

[0018] The term "therapeutically effective" means that the amount of composition used is an amount sufficient to alleviate one or more of the causes, symptoms or sequelae of the disease or disorder. Such mitigation requires only a reduction or alteration, not necessarily elimination, of the cause, symptom or consequence of the disease or disorder.

[0019] As used herein, the term "cardiac tissue" includes the myocardium, the endocardium, the pericardium and the pericardium of the heart. The term as used herein also refers to large vessels leading to and from the heart. The term used here also refers to the part of the vagus nerve that innervates the heart.

[0020] Thus, the use comprises administering a mammalian ECM-containing composition to the subject's heart. In some aspects, the use comprises administering a mammalian ECM-containing composition to the myocardium of a subject. The cardiac muscle may be the cardiac muscle of the chamber. The cardiac muscle may be the atrial myocardium. In some aspects, the methods comprise administering a mammalian ECM-containing composition to a subject's endocardium. In some aspects, the use comprises administering a mammalian ECM-containing composition to the subject's endocardium. In some aspects, the use comprises administering a mammalian ECM-containing composition to the subject's pericardium.

[0021] As used herein, the term "subject" means any subject that is the target of administration. The subject may be a vertebrate, e.g. a mammal. Thus, an individual can be a human. The term does not indicate a specific age or gender. Thus, the term is intended to include adult and newborn individuals as well as fetuses, whether male or female. The patient refers to a subject afflicted with the disease or disorder. The term "patient" includes human and animal subjects. The terms "patient" and "subject" as used herein may be used interchangeably.

[0022] In some aspects, the subject with the use disclosed is identified as being at risk of arrhythmias. In some aspects, the subject undergoing the disclosed use has undergone heart surgery, including, but not limited to, open heart surgery. In certain aspects, a subject with the disclosed use has undergone a number of complex procedures performed at the heart, including, but not limited to, open-heart procedures. In some aspects, the subject under the disclosed use has undergone heart valve surgery. In some aspects, the subject with the use disclosed has at least 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, or 85 years. In some aspects, the composition is administered to a subject having a myocardial infarction. In some aspects, the subject has disclosed emphysema with the use disclosed

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EP 3 000 472 B1 or chronic obstructive pulmonary disease. In some aspects, the subject with the use disclosed has a history of arrhythmias.

[0023] In some aspects, the disclosed use does not include administering a patch comprising the small intestinal submucosal (SIS) to the orifice of the cardiac heart. In some aspects, the disclosed use does not consist of administering a patch containing the small intestinal submucosal (SIS) to the orifice of the cardiac atrial sac. In some aspects, the mammalian ECM is not SIS. Thus, in some aspects, a composition comprising a mammalian ECM is not composed of SIS. In some aspects, a composition comprising a mammalian ECM is not a patch. In some aspects, the disclosed use does not comprise administering a mammalian ECM-containing composition in the form of a patch to the orifice in the pericardial sac. In some aspects, the cardiac tissue in the disclosed application is not an oculi. In some aspects, the disclosed use does not include administering the composition to the pericardium.

[0024] In one aspect, the disclosed use comprises administering a patch comprising the small intestinal submucosal (SIS) to the orifice of the cardiac atrial sac. The compositions used may contain an additional agent, such as anti-arrhythmic drugs or non-native cells. In other aspects, the disclosed use comprises administering a patch comprising the small intestinal submucosal (SIS) to the orifice of the pericardial sac, but the use further comprises additional steps.

[0025] Also disclosed herein is a composition for use in preventing a cardiac arrhythmia in a subject comprising administering to the myocardium a therapeutically effective amount of a mammalian extracellular matrix composition and further comprising a drug for arrhythmia, a lipid lowering drug, a cell, a protein or a combination thereof. .

1. Heart Rhythm Disorder. [0026] A heart rhythm disorder (also referred to as dysrhythmia) is a term for any of a large and heterogeneous group of conditions in which there is abnormal electrical activity in the heart. A heart beat (pulse) may be too fast or too slow and may be regular or irregular.

[0027] Some arrhythmias are medical life-threatening cases that can result in cardiac arrest and sudden death. Others cause symptoms such as abnormal awareness of heartbeats (palpitations) and can only be annoying. Others may not be associated with any symptoms at all, but they are a predisposition to a potentially life-threatening stroke or an embolism.

[0028] The term sinus arrhythmia refers to the usual phenomenon of mild acceleration and deceleration of heart rate that occurs with inspiration and exhalation. It is usually quite clearly marked in children and gradually decreases with age. It can also appear during breathing exercises during meditation, which include patterns of deep breathing and breath holding.

[0029] Each heartbeat arises as an electrical impulse from a small tissue surface in the right atrium of the heart called the sinus node or the sinoatrial node (SA). The pulse initially causes contraction of both atria, then activates the atrioventricular node (or AV), which is usually only an electrical connection between the atria and the chambers or the main pumping chambers. The pulse is then spread through both chambers through the Purkin's His bundle resulting in a synchronized myocardial contraction.

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[0030] Heart rate less than 60 beats per minute is bradycardia. This may be caused by a slowed sinus node signal (referred to as sinus bradycardia), a break in normal sinus node activity (referred to as sinus arrest) or by blocking an electrical pulse on its way from the atria to the ventricles (referred to as an AV block or cardiac conduction block) ). The cardiac conduction block occurs in varying degrees and severity. It can be caused by reversible intoxication of the AV node (drugs that interfere with conductivity) or by irreversible damage to the node.

[0031] Heart rate faster than 100 beats per minute is tachycardia. Tachycardia may result in palpitations; however, tachycardia is not necessarily a heart rhythm disorder. Increased heart rate is the usual answer to physical exercise or emotional stress. It mediates the influence of the sympathetic nervous system on the sinus node, and this is referred to as sinus tachycardia. Other factors that increase the activity of the sympathetic nervous system in the heart include ingested or injected substances such as caffeine or amphetamines and a hyperactive thyroid gland (hyperthyroidism). Heart rate can be increased by sympathomimetic drugs.

[0032] Tachycardia, which is not sinus tachycardia, is usually the result of the addition of abnormal pulses that can be initiated in one of three mechanisms: automatism, reversal wave or triggered automatism.

[0033] Automism refers to a cardiac muscle cell that itself triggers an impulse. All of the cells in the heart have the ability to initiate the action potential; however, only some of these cells are intended for continuous triggering of heartbeats. These cells are in the cardiac conduction system and include the SA node, the AV node, the His bunch and the Purkinje fiber. The SA node is the only specialized place in the vestibule that has higher automatism (faster pacing) than the rest of the heart, and therefore usually is responsible for setting the heart rate and initiating each heartbeat. Any part of the heart that initiates an impulse without waiting for a SA node is called an ectopic focus and is, by definition, a pathological phenomenon. This may occasionally cause a single premature blow or, if the ectopic focus triggers more often than the SA node, it can produce a prolonged abnormal rhythm. Conditions that increase automatism include stimulation of the sympathetic nervous system and hypoxia. The resulting heart rate depends on where the first signal begins. If it is a SA node, the rhythm remains correct but fast; if it is an ectopic focus, many types of arrhythmias may arise.

[0034] The arrhythmias resulting from the reversal wave appear when the electrical pulse cyclically circulates in a tight periphery in the heart and does not move from one end of the heart to the other, and then stops. Each heart cell is capable of transmitting impulses in any direction, but it can do so only once within a short period of time. Usually, the action potential of the pulse will spread in the heart quickly enough, so that every cell will respond only once. However, if the conductivity is incorrectly slow in certain areas, part of the impulse will arrive late and potentially be treated as a new impulse. Depending on the time, it may create a persistent abnormal circulating rhythm. Circulating reversal waves are responsible for the flutter of the atria, most paroxysmal supraventricular tachycardia and severe ventricular tachycardia. When the whole chamber

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The heart is involved in many phenomena of the micro-circulating reversal wave, and thus is introduced into trembling through chaotic electrical impulses, it is said about flickering.

[0035] Flicker may be at the atrium (atrial fibrillation) or at the ventricle (ventricular fibrillation). If left untreated, ventricular fibrillation (VF or V-fib) can lead to death in a few minutes.

[0036] Triggered strikes occur when problems at the ion channel level in individual cardiac cells may result in aberrant propagation of electrical activity and may lead to persistent abnormal rhythms. Triggered strokes are relatively rare, but they can be the result of drugs against arrhythmias.

[0037] A rhythm disorder can be classified according to the speed (normal, tachycardia, bradycardia) or mechanism (automatism, reversal wave, flicker).

[0038] It is also appropriate to classify the arrhythmias according to the origin. For example, atrial arrhythmias include premature atrial contractions (PAC), atrial shunt pacemaker, multifocal atrial tachycardia, atrial flutter, and atrial fibrillation (Afib). Disorders of the nodal rhythm include supraventricular tachycardia (SVT), recurrent nodal tachycardia AV (the most common cause of paroxysmal supraventricular tachycardia (PSVT)), nodal rhythm, nodular tachycardia and premature nodal arousal. Atrio-ventricular arrhythmias include recurrent AV tachycardia (occurs when the circulating reversal wave passes between the atrium and the ventricles in a place other than the AV node).

[0039] Ventricular arrhythmias include pre-premature ventricular agitation (PVC) (sometimes referred to as additional ventricular contractions (VEB)), accelerated ventricular rhythm, monomorphic ventricular tachycardia, polymorphic ventricular tachycardia, and ventricular fibrillation.

[0040] Heart blocks (also known as AV blocks, the most common causes of bradycardia) include a first degree heart block (PR interval longer than 200 ms in ECG), a second degree heart block (Type 1 and 2) and a third degree heart block ( also known as total heart block).

[0041] Heart rhythm disorders are often first detected by auscultating heart beats with a stethoscope or by sensing peripheral pulses. With these methods, you can not usually diagnose specific arrhythmias, but they can give a general indication of heart rate and whether it is regular or irregular. Not all of the heart's electrical pulses produce audible or perceptible impacts; in the case of many arrhythmias, premature or abnormal impacts do not result in effective pumping action and are experienced as "missed" hits.

[0042] The simplest specific diagnostic test for assessing the rhythm of the heart is an electrocardiogram (ECG or ECG for short). The Holter Monitor is an ECG record recorded over a 24 hour period to detect arrhythmias that may occur briefly and unpredictably over a 24-hour period.

[0043] Arousal sudden death syndrome (SADS) is a term used to describe sudden death due to cardiac arrest due to a rhythm disorder. Often, the subject has no symptoms prior to sudden death. The most common cause of sudden death in the United States is coronary artery disease. Every year in the United States for this reason

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EP 3 000 472 B1 suddenly dies around 300,000 people. SADS can also be caused, for example, by many inherited heart conditions and diseases that can affect young people.

[0044] In children, SADS can cause, for example, viral myocarditis, QT prolongation syndrome, Brugada syndrome, catecholaminergic polymorphic ventricular tachycardia and hypertrophic cardiomyopathy, and arrhythmogenic right ventricular dysplasia.

[0045] In the present invention, the atrial fibrillation is atrial fibrillation or ventricular fibrillation.

2. Administration [0046] In some aspects, the ECM mammal is a patch in a form such as a sheet, plug, laminate, weave, polymer matrix, multiple strands or one or more strands. Thus, in some aspects, the ECM mammal is placed in direct contact with the subject's cardiac tissue during cardiac surgery. In the present invention, a composition comprising a mammalian ECM is administered to an orifice in the cardiac case. In some aspects, the composition overlaps the orifice in the pericardial sac. Thus, a mammalian ECM-containing composition can be administered to the surgical orifice during or after cardiac surgery.

[0047] When the ECM mammal is in a solid form, such as a sheet, plug, laminate weave, polymer matrix, multiple strand, or one or more strands, the composition may be attached to cardiac tissue using standard means available in the art. For example, a composition comprising a mammalian ECM can be attached to the cardiac tissue via sutures, bioadhesive agents such as fibrin glue, mechanical sutures and the like.

[0048] The disclosed compounds and compositions comprising a mammalian ECM may be administered in any suitable manner. For example, the compositions can be administered parenterally (e.g., intramuscular injection), locally or similar. Thus, in some aspects, a composition comprising a mammalian ECM is suitable for injection. The disclosed compositions can be injected into cardiac tissue using standard means. For example, a composition comprising a mammalian ECM may be delivered to the cardiac tissue via a syringe or a cardiac or coronary catheter. Heart catheterization is the insertion of a catheter into the heart or vessel. This can be done for both diagnostic and / or intervention purposes. Cannulation of coronary vessels is a subgroup of this technique that involves coronary catheterization.

[0049] Thus, in some aspects, a mammalian ECM containing composition can be injected into the myocardium. In some aspects, a mammalian ECM-containing composition can be injected into the cardiac region. In some aspects, a mammalian ECM-containing composition can be injected into the heart's endocardium. In some aspects, a composition comprising a mammalian ECM can be injected into the cardiac pericardium. In some aspects, a mammalian ECM-containing composition can be injected between the layers of the heart, e.g. between the pericardium and the papillary region, between the papillary region and the myocardium, and between the myocardium and the endocardium.

[0050] In some aspects, the mammalian ECM-containing composition may be administered to the intracorporal or intraventricular septal defect. For example, in some aspects, a composition

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EP 3 000 472 B1 containing a mammalian ECM can be administered to the defect of the interventricular septum. Ventricular septal defect (VSD) is a defect in the ventricular septum, the wall separating the left and right ventricles. The intraventricular septum consists of a lower muscular and a higher membrane part and is largely innervated with conductive cardiomyocytes. The membrane part, which is close to the atrioventricular junction, is most commonly affected in adults and older children. Congenital VSDs are the most common congenital heart defects.

[0051] In some aspects, the mammalian ECM-containing composition may be administered to the atrial septal defect (ASD). ASD is a form of congenital heart disease that allows blood to flow between the left and right atrium through the atrial septum. The atrial septum is the tissue that separates right and left atrium. Without this barrier, or if there is a cavity in this compartment, it is possible to pass blood from the left side of the heart to the right side of the heart or vice versa. [1] Regardless of the two directions of inter-atrial communication, this results in the mixing of arterial and venous blood. Mixing arterial and venous blood can be hemodynamically significant or not, even if clinically relevant. This mixing of blood may result or not what is known as "rolling". The amount of rolling present, if present, is haemodynamic (see Pathophysiology below). "Right-to-left transfusion" is usually a more dangerous scenario (see Pathophysiology below).

[0052] The ECM manger may be in the form of an aerosol. Thus, in some aspects, the ECM mammal may be sprayed onto the subject's cardiac tissue.

[0053] The ECM mammalian can be present in the form of particles. The particulate ECM can be administered by injecting the composition in an emulsion, spraying, applying as a layer, packing, sprinkling, spreading or other similar types of administration of dry particles, a liquid composition or semi-solid compositions.

[0054] In certain aspects, the composition is administered to the epicardial surface of the heart. Thus, in some aspects, the composition is injected, sprayed or attached to the epicardial surface of the heart.

The exact amount of composition required may vary between subjects, depending on the species, age, body weight and general condition of the subject, the severity of the disorder being treated. Thus, it is not possible to determine the exact amount for each composition. However, an appropriate amount can be determined by one of ordinary skill in the art using only routine experimentation, taking into account the present disclosure. Thus, effective dosages and schedules for administering the compositions can be determined empirically, and such determination falls within the skill of the art. Dose ranges for administering the compositions are those that are large enough to achieve the desired effect at which the symptom or disorder is affected. The dose should not be so large as to cause unwanted side effects, such as undesirable cross reactions, anaphylactic reactions and the like. In general, the dose will vary depending on the age, condition, sex and extent of the disease in the patient, the route of administration, or whether the schedule includes other drugs, and may be determined by a person skilled in the art. Doses may vary and may be administered in one or more daily doses for one or several days. Guidance can be found in the literature on the appropriate dosage for given classes of pharmaceutical products. Doses may vary and may be administered in one or more daily doses for one or several days. Guidance can be found in the literature on the appropriate dosage for given classes of pharmaceutical products. Doses may vary and may be administered in one or more daily doses for one or several days. Guidance can be found in the literature on the appropriate dosage for given classes of pharmaceutical products.

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[0056] Following administration of the disclosed composition to prevent arrhythmia, the efficacy of use can be assessed in various ways well known to the skilled person. For example, one of ordinary skill in the art will recognize that the composition disclosed herein is effective in preventing a cardiac arrhythmia in an individual using an electrocardiogram.

[0057] The compositions disclosed herein may be administered prophylactically to subjects who are at risk of arrhythmias. The disclosed compositions may also be used, for example, as tools for isolating and testing candidates for new medicaments for treating or preventing a arrhythmias. The disclosed compositions can also be used in various ways as research tools. Other uses are disclosed, result from the disclosure and / or will be understood by those skilled in the art.

3. Combination treatment [0058] The uses disclosed herein may further comprise treating the subject with the aid of conventional therapies for arrhythmias. For example, there are many classes of drugs against arrhythmias with different mechanisms of action and many different individual medications within these classes. Thus, the use can further comprise administering to the subject one or more drugs against rhythm disorders.

[0059] Some arrhythmias, e.g. atrial fibrillation, cause blood to clot in the heart and increase the risk of embolism and stroke. Anticoagulants such as warfarin and heparin, and antiplatelet agents such as aspirin may reduce the risk of coagulation. Thus, the use may further comprise administering an anticoagulant to the subject.

[0060] Arrhythmias can also be treated electrically, by means of a cardiac shock, externally to the chest or internally to the heart by implanted electrodes or during surgery. Cardioversion can be achieved pharmacologically or by applying a shock synchronized with the appropriate heart beat. This is used for the treatment of supraventricular tachycardia. In planned cardioversion, the recipient is usually sedated or lightly anaesthetized into the procedure. For example, atrial flutter can be treated with the help of cardioversion. Thus, the use may further comprise treating the subject with the aid of cardioversion.

[0061] Along with the synchronized cardioversion, a reverse shock is provided by means of washers or paddles for the selected electrical current value over a predetermined number of milliseconds, at the optimal time of the heart cycle, which corresponds to the R-wave of the QRS complex at the ECG. The shock synchronization with R-wave prevents shock during the wounded phase (or relative refractory period) of the heart cycle, which could induce ventricular fibrillation.

[0062] Defibrillation differs from cardioversion in that the shock is not synchronized with the heart cycle. It is necessary for the chaotic rhythm of ventricular fibrillation, and it is also used in ventricular tachycardia without pulse. It is often required in more electricity for defibrillation than for cardioversion. Since most individuals with ventricular fibrillation are unconscious, there is essentially no need for sedation. Thus, the use may further comprise treating the subject with the aid of defibrillation.

[0063] Defibrillation or cardioversion can be performed with the help of an implantable cardioverteradefibrilator (ICD). Thus, the use may further comprise administering to the subject an ICD.

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[0064] Electrical treatment of arrhythmias also includes cardiac stimulation. Immediate stimulation may be necessary for reversible causes of very slow heartbeats or bradycardia (for example due to drug overdose or myocardial infarction). A fixed pacemaker can be placed in situations where it is not expected to exit bradycardia. Thus, the use may further comprise administering to the subject a pacemaker.

[0065] Small sensors may in certain aspects be introduced through blood vessels to mimic electrical activity from the interior of the heart. This enables very accurate location of abnormal conduction surfaces, followed by destruction with heat, cold, electrical or laser probes.

C. COMPOSITIONS [0066] An ECM mammal patch has been shown to act as a mechanical scaffolding while the body recruits cells necessary for remodeling and repairing cardiac tissue. This discloses the surprising ability of the mammalian ECM to further prevent arrhythmias. Thus, compositions comprising a mammalian ECM for use in the prevention of a arrhythmia disorder in a subject are disclosed herein. The disclosed compositions may be natural or synthetic. The compositions may be cell-free or may contain cells, such as stem cells.

[0067] The mammalian ECM-containing compositions disclosed herein may be in the form of, for example, a patch, emulsion, injectable solution, gel, lotion, paste, powder, thread, strand, spray, vapor, aerosol, cream or coating. The composition may further comprise one or more additional ingredients, including for example a cell, peptide, polypeptide, protein or other biological moieties. When the composition is in the form of a patch, it may be in a form selected from a sheet, laminate, weave, polymer matrix, multiple strands, one or more strands or combinations thereof.

[0068] The mammalian ECM containing compositions disclosed herein can be made in particulate form and liquefied, as described in US Patent No. 5,275,826 to Badylak, US Patent No. 6,579,538 to Spievack, and US Patent No. 6,933,326 to Griffey. The liquefied or emulsified compositions (liquid or semi-solid forms) may be present at a certain concentration, e.g. in a concentration of extracellular matrix of greater than about 0.001 mg / ml. The concentration of these liquid or semi-solid components of the extracellular matrix composition may range from about 0.001 mg / ml to about 200 mg / ml. The concentrations may furthermore be in more specific ranges, such as, for example, the following set of ranges: about 5 mg / ml to about 150 mg / ml, about 10 mg / ml to about 125 mg / ml, about 25 mg / ml to about 100 mg / ml, about 20 mg / ml to about 75 mg / ml, about 25 mg / ml to about 60 mg / ml, about 30 mg / ml to about 50 mg / ml, and about 35 mg / ml to about 45 mg / ml, and about 40 mg / ml to about 42 mg / ml. This set of scopes is exemplary and not intended to be exhaustive. It is contemplated that any value within any of these specifically mentioned ranges is a reasonable and useful value for the concentration of a liquid, emulsion, gel, paste or other liquid or semi-solid component of the composition.

1. Mammalian extracellular matrix [0069] Extracellular matrix materials act as a natural scaffold for repairing soft tissues in the body. Animal studies have shown that the extracellular matrix material is reconstructed and replaced by the host tissue. The ECM is an ECM

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This is a resorptive biological material used successfully as xenograft tissue transplant that induces structural remodeling of various animal tissues including blood vessels, bladder, dura mater, abdominal wall, tendons and ligaments. Examples of mammalian ECM include small intestinal submucosa (SIS), bladder infiltrate (UBS), gastric submucosa (SS), and hepatic submucosal (LS) or liver basal membrane (LBM).

[0070] The remodeling process involves rapid neovascularization and a strong accumulation of mesenchymal and epithelial cells that support the extensive deposition of the new extracellular matrix. The non-collagenous part, for example, the extracellular matrix SIS is composed of various glycoproteins, such as hyaluronic acid, heparin, dermatan and chondroitin sulfate A, as well as growth factors FGF-2 and TGF-β.

[0071] After processing, the ECM mammal can stop many endogenous proteins that act as growth and differentiation factors. These factors stimulate the local environment to settle the mammalian ECM with cells that are then capable of differentiating into the primary tissue that replaces the ECM mammal.

[0072] The ECM sucker is a scaffolding matrix of polymerized "structural" proteins that belong to three groups: collagens, glycoproteins and proteoglycans (which contain the entire length of the glycosaminoglycan repeats). These molecules actually polymerize to form a scaffold or protein matrix that occurs in dynamic interaction with cells and nearby functional proteins (either on the cells or bound to the structural protein). Thus, the ECM mammal also includes in its scaffold matrix the "functional" proteins that interact with structural proteins and with migrating or recruited cells, such as stem cells. Functional proteins of the matrix also interact with cells expressing proteins during the duration and maintenance of the matrix scaffolding itself, as it rebuilds and sustains its elements. Some proteins can be both structural and functional protein depending on the protein configuration and distribution throughout the matrix.

[0073] The ECM of, for example, cardiac tissue is formed from type I collagen (predominantly), III, IV, V and VI, which together constitute 92% of the dry weight of the matrix. ECM of cardiac tissue is also formed by glycosaminoglycans (GAGs), which include chondroitin sulfate A and B, heparan, heparin and hyaluronic acid. Glycoproteins such as fibronectin and entactin, proteoglycans such as decorin and perlexide, and growth factors such as transforming growth factor beta (TGF-β), fibroblast growth factor 2 (FGF-2) and vascular endothelial growth factor (VEGF) play a key role in the activity of the myocardial regenerative matrix. In addition, the exact chemical composition of the matrix seems to play a role in its functioning, including, for example, which type of collagen prevails in the matrix.

[0074] Cell adhesion molecules (CAMs) facilitate cell adhesion in the ECM. CAMs may be available endogenously or added as an additional component of the composition. CAMs are glycoproteins located on the surface of the cell membrane or transmembrane connected to the cytoskeleton components of the cell. Specific CAMs include cadherins which are calcium dependent and more than 30 types are known. Also interacting as CAM are integrins, which are proteins that connect the cytoskeleton of the cell in which they are placed, to the extracellular matrix or to other cells by subunits

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Transcription alpha and beta on the integrin protein. Cell migration, embryogenesis, hemostasis and wound healing are facilitated by integrins in the matrix. Syndecans are proteoglycans that bind to ligands to initiate cell motility and differentiation. Immunoglobulins provide any necessary immune and inflammatory responses. Selektins stimulate cell-cell interactions.

i. Native Sources and Preparations [0075] In some aspects, the ECM mammal is from a native source. Native scaffolds of the extracellular matrix and proteins that form them can be found in their natural environment, i.e. matrices of extracellular mammals. These materials may be prepared for use in mammals in tissue transplantation procedures.

[0076] In some aspects, the mammalian ECM is extracted from mammalian tissue / organs. For example, in some aspects, the ECM mammal comprises a basal membrane (or transient epithelial layer), a membrane, a submucosal membrane, a muscle membrane, a serous membrane, or a combination thereof from a mammalian tissue source. Thus, in some aspects, the ECM mammal comprises a basal membrane (or transient epithelial layer) from a mammalian tissue source. In some aspects, the ECM mammal comprises an adhering specific membrane from a mammalian tissue source. In some aspects, the ECM mammal comprises a submucosal membrane from a mammalian tissue source. In some aspects, the ECM mammal comprises muscle membrane from a mammalian tissue source. In some aspects, the ECM mammal comprises a serous membrane from a mammalian tissue source.

[0077] For example, small intestinal submucosa (SIS) is described in US Patent No. 5,275,826; Urinary bladder infiltration (UBS) is described in US Patent No. 5,554, 389; Gastric submucosal (SS) is described in US Patent No. 6,099,567; and hepatic submucosal (LS) or basal liver membrane (LBM) is described in US Patent No. 6,379,710, each of which is incorporated herein by reference for guidance on how to make and use these native extracellular matrices.

[0078] Thus, in some aspects, the ECM mammal of the disclosed compositions is small intestinal submucosa (SIS). In some aspects, the ECM mammal of the disclosed compositions is urinary bladder (UBS). In some aspects, the ECM mammal of the disclosed compositions is gastric submucosa (SS). In some aspects, the ECM mammal of the disclosed compositions is a submucosal liver (LS). In some aspects, the ECM mammal of the disclosed compositions is a basal liver membrane (LBM).

[0079] In some aspects, the ECM mammal of the disclosed compositions is of dermis. For example, AlloDerm®, produced by LifeCell Corporation, is a cell-free tissue matrix that is made of normal human skin using processing techniques developed to remove the epidermis and cells in the dermis without significantly altering the normal biochemistry and molecular architecture of the connective tissue matrix. The product obtained is in lyophilized form allowing for extended storage and easy transport without degradation or loss of normal elements of the tissue matrix. AlloDerm® can retain decorin, hyaluronic acid, chondroitin sulfates, nidogen, growth factors and other biochemical proteins present in normal soft tissues. Additionally,

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[0080] In some aspects, the ECM mammal of the disclosed compositions is fascia. In some aspects, the ECM mammal of the disclosed compositions is from parenchymal tissue. In some aspects, the ECM mammal of the disclosed compositions is from the pericardium. In some aspects, the ECM mammal of the disclosed compositions is the extracellular matrix of the myocardium, in some aspects, the ECM mammal of the disclosed compositions is cell-free cardiac tissue, produced for example by coronary artery perfusion with detergents (Ott, HC, et al Nat Nat. February 2008, 14 (2): 213-21).

[0081] In certain aspects, the ECM mammal comprises a collagen scaffold derived from a tissue or mammalian organ source. The collagen scaffold from a mammalian source may in some aspects contain a basal membrane of the source mammalian tissue.

[0082] In certain aspects, the mammalian ECM is produced in vitro. For example, the ECM mammal may be produced from a mammalian cell culture. The ECM can be produced from proteins extracted from mammalian tissue / organs. For example, in some aspects, the ECM mammal contains an artificial collagen scaffold synthesized from collagen extracted from the source tissue or organ of the mammal. Collagen from mammalian sources can be derived from matrix-containing tissues and used to make matrix compositions. The extracellular matrices can be synthesized from cell cultures as in the product produced by Matrigel ™. In addition, the material of the extracellular matrix of the dermis, the subcutaneous matrix material extracellular, the material of the extracellular matrix of the large intestine, material matrix extracellular placenta, the matrix material of the extracellular network, the material of the extracellular matrix of the heart and the extracellular matrix material of the lungs, can be used, obtained and stored similar to those described herein for SIS, SS, LBM and UBS materials. Other organ and tissue basement membrane sources for use with the disclosed compositions include spleen, lymph nodes, salivary glands, prostate gland, pancreas and other secretory glands. In general, any mammalian tissue that includes the extracellular matrix can be used to develop the extracellular matrix element. Other organ and tissue basement membrane sources for use with the disclosed compositions include spleen, lymph nodes, salivary glands, prostate gland, pancreas and other secretory glands. In general, any mammalian tissue that includes the extracellular matrix can be used to develop the extracellular matrix element. Other organ and tissue basement membrane sources for use with the disclosed compositions include spleen, lymph nodes, salivary glands, prostate gland, pancreas and other secretory glands. In general, any mammalian tissue that includes the extracellular matrix can be used to develop the extracellular matrix element.

[0083] The collagen matrix can be selected from a range of commercially available collagen matrices or it can be prepared from a wide range of natural sources of collagen. The collagen matrix for use with the disclosed compositions can contain highly conserved collagens, glycoproteins, proteoglycans and glycosaminoglycans in their natural configuration and in their natural concentration. Collagens may be derived from animal sources, from plant sources or from synthetic sources, all of which are available and standard in the field.

[0084] The proportion of scaffold material in the composition when the native scaffold is used can be large, since the natural balance of extracellular matrix proteins in native scaffolds typically constitutes more than 90% of the extracellular matrix material on a dry basis. Thus, the scaffold element in the composition by weight can be substantially more than 50% of the total dry weight of the composition. The scaffold may comprise an amount in the composition by weight greater than 60%, greater than 70%, greater than 80%, greater than 82%, greater than 84%, greater than 86%, greater than 88%, greater than 90%, greater than 92% %, greater than 94%, greater than 96% and greater than 98% of the total composition.

[0085] Native extracellular matrices can be prepared with caution so that their biological activity to prevent arrhythmia is preserved in the largest possible way.

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EP 3 000 472 B1 degree. Key functions that can be retained include control or initiation of cell adhesion, cell migration, cell differentiation, cell proliferation, cell death (apoptosis), angiogenesis stimulation, proteolytic activity, enzymatic activity, cell motility, protein and cell modulation, activation of transcription events , providing translational events, inhibiting certain biological activities, e.g. inhibiting coagulation, attracting stem cells and chemotaxis. Tests for determining these activities are standard in the field. For example, material analysis can be used to identify molecules present in the material composition. In vitro cell adhesion tests can also be performed to ensure that the material or composition is capable of adhering to the cells.

[0086] The mammalian ECM containing compositions disclosed may be cell-free to render them non-immunogenic. In some aspects, the cell debridement process is completed while maintaining certain functions of key proteins, by replacing proteins accidentally extracted with cells or by adding exogenous cells to matrix compositions after cell extraction, which cells produce or transfer proteins involved in the prevention of arrhythmias.

[0087] When adding proteins to the extracellular matrix composition, proteins can simply be added to the composition, or each protein can be covalently attached to the molecule in the matrix. Standard procedures for linking proteins to molecules can be used to obtain covalent attachment.

[0088] For the removal of cells, at the exit from the source tissue / organ as an ECM mammalian source, the tissue / organ source perfusion process may be used. The source tissue / organ can be perfused with a cell-removing agent, e.g. 0.1% peracetic acid, which deprives the organs of the cells. The source tissue / organ can then be cut into parts and stored (e.g., in an aqueous environment, liquid nitrogen, cold, lyophilized or compressed under vacuum) for later use. Any suitable cell-removing agent may be used for the tissue / organ source perfusion process.

[0089] With respect to the submucosal tissue, the extractions can be carried out at a pH close to neutral (ranging from about pH 5.5 to about pH 7.5) in order to preserve the presence of growth factors in the matrices. Alternatively, acidic conditions (i.e., pH less than 5.5) may be used to preserve the presence of glycosaminoglycan elements in a temperature range between 0 and 50 degrees Celsius. To control the acidic or alkaline environment for these aqueous extracts, a buffer and a chaotropic agent (generally at a concentration of about 2M to about 8M), such as urea (at a concentration of about 2M to 4M), guanidine (at a concentration of about 2M) may be selected. up to about 6M, usually around 4M), sodium chloride, magnesium chloride and non-ionic or ionic surfactants. Urea at 2M at pH 7.4 ensures the extraction of FGF-2 and glycoprotein of fibronectin.

[0090] Due to the collagen structure of the basement membrane and the desire to minimize the degradation of the membrane structure during cell dissociation, the collagen-specific enzyme activity in the enzyme solutions used at the cell dissociation step can be minimized. For example, the source tissue / organ may be subjected to a calcium chelating agent or a chaotropic agent, such as a mild detergent such as Triton 100. The cell dissociation step may also be carried out

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EP 3 000 472 B1 using a calcium chelating agent or a chaotropic agent, in the absence of enzymatic treatment of the tissue / organ. The cell dissociation step may be performed by suspending the source tissue sections in a mixed solution containing about 0.05 to about 2%, more usually about 0.1 to about 1% by weight protease, optionally containing a chaotropic agent or a calcium chelating agent in an amount effective to optimize release and separation of cells from the basement membrane without significant degradation of the membrane matrix.

[0091] After contacting the source tissue / organ with the cell dissociation solution for a sufficient time to release all cells from the matrix, the resulting tissue / organ base membrane can be rinsed once or more times with saline and optionally stored frozen in a hydrated or partially dehydrated state. for use as described below. The cell dissociation step may require several treatments with the cell dissociation solution to release essentially all of the cells from the basal membrane. The source tissue / organ may be treated with a protease solution to remove constituent cells, and the obtained extracellular matrix material is further treated to remove or inhibit any residual enzymatic activity. E.g,

The basal membrane or other native scaffolds of the extracellular matrix can be sterilized using conventional sterilization techniques including tanning using glutaraldehyde, tanning using formaldehyde at acidic pH, treatment with ethylene oxide, propylene oxide treatment, gas plasma depleting, gamma irradiation, and juicing with peracetic acid. The sterile technique is preferably used, which does not significantly impair the mechanical strength and biotropic properties of the material. For example, it is believed that strong gamma radiation can cause loss of strength in the transplanted material. Examples of sterilization techniques include transplantation to peracetic acid, gamma-irradiated at low doses,

ii. Synthetic ECM [0093] Also disclosed are compositions containing a synthetic ECM for the disclosed use.

Synthetic ECM for use in the disclosed compositions can be made using synthetic molecules that polymerize largely similar to native collagen, and which form a scaffolding environment that mimics the native mammalian scaffold environment. Accordingly, materials such as polyethylene terephthalate (Dacron®), polytetrafluoroethylene (PTFE), crosslinked glutaraldehyde, polylactate (PLA), polyglycol (PGA), hyaluronic acid, polyethylene glycol (PEG), polyethylene, nitinol can be used. , and collagen from non-animal sources (such as plant or synthetic collagens) as components of the synthetic scaffold of the extracellular matrix. These synthetic materials are standard in the art, and the formation of hydrogels and matrix-like materials from them is also standard.

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[0094] Materials similar to the extracellular matrix are described generally in Rosso et al. (Journal of Cellular Physiology 199: 174-180, 2004), which is incorporated herein by reference for guidance on how to create and use these materials. In addition, some materials similar to the extracellular matrix are mentioned herein. Particularly useful biodegradable and / or bioabsorbable polymers include polylactides, polyglycolides, polycaprolactone, polydioxane and their random and block copolymers. Examples of specific polymers include poly-D, L-lactide, polylactide-co-glycolide (85:15) and polylactide-co-glycolide (75:25). Biodegradable and / or bioabsorbable polymers used in the fibrous matrix of the disclosed compositions may have a molecular weight in the range of from about 1,000 to about 8,000,000 g / mol, including about 4,000 to about 250,000 g / mol. The biodegradable and / or bioabsorbable fibrous material can be a biodegradable and bioabsorbable polymer. Examples of suitable polymers can be found in Bezzwada, Rao S et al. (1997) Poly (p-Dioxanone) and its copolymers, in Handbook of Biodegradable Polymers, AJ Domb, J. Kost and DM Wiseman, editors, Hardwood Academic Publishers, The Netherlands, pp. 29-61. The biodegradable and / or bioabsorbable polymer may contain a monomer selected from the group consisting of a glycoside, lactide, dioxanone, caprolactone, trimethylene carbonate, ethylene glycol and lysine. The material may be a random copolymer, a block copolymer or a mixture of monomers, homopolymers, copolymers and / or heteropolymers that contain these monomers. Biodegradable and / or bioabsorbable polymers can contain bioabsorbable and biodegradable linear aliphatic polyesters such as polyglycolide (PGA) and its random copolymer poly (glycolide-co-lactide-) (PGA-co-PLA). The FDA approved these polymers for use in surgical applications, including medical sutures. The advantage of these synthetic absorbable materials is their ability to degrade by simply hydrolyzing the ester backbone in aqueous media such as body fluids. Degradation products are ultimately metabolised to carbon dioxide and water or can be excreted through the kidneys. These polymers are very different from cellulose-based materials that can not be absorbed by the body. The FDA approved these polymers for use in surgical applications, including medical sutures. The advantage of these synthetic absorbable materials is their ability to degrade by simply hydrolyzing the ester backbone in aqueous media such as body fluids. Degradation products are ultimately metabolised to carbon dioxide and water or can be excreted through the kidneys. These polymers are very different from cellulose-based materials that can not be absorbed by the body. The FDA approved these polymers for use in surgical applications, including medical sutures. The advantage of these synthetic absorbable materials is their ability to degrade by simply hydrolyzing the ester backbone in aqueous media such as body fluids. Degradation products are ultimately metabolised to carbon dioxide and water or can be excreted through the kidneys. These polymers are very different from cellulose-based materials that can not be absorbed by the body. Degradation products are ultimately metabolised to carbon dioxide and water or can be excreted through the kidneys. These polymers are very different from cellulose-based materials that can not be absorbed by the body. Degradation products are ultimately metabolised to carbon dioxide and water or can be excreted through the kidneys. These polymers are very different from cellulose-based materials that can not be absorbed by the body.

[0095] Other examples of suitable biocompatible polymers are polyhydroxyalkyl methacrylates, including ethyl methacrylate and hydrogels such as polyvinylpyrrolidone, polyacrylamides, etc. Other suitable bioabsorbable materials are biopolymers that include collagen, gelatin, alginic acid, chitin, chitosan, fibrin, hyaluronic acid, dextran, polyamino acids, polylysine and copolymers of these materials. Any glycosaminoglycan (GAG) polymer can be used. GAGs may include, e.g., heparin, chondroitin A or B sulphate and hyaluronic acid or their synthetic analogs. Any combination, copolymer, polymer or mixture thereof of the above examples is included for use in the disclosed compositions. Such bioabsorbable materials can be prepared by known methods.

[0096] Nucleic acids from any source can be used as a polymer biomaterial. Sources include naturally occurring nucleic acids as well as synthesized nucleic acids. Nucleic acids suitable for use in the disclosed compositions include naturally occurring forms of nucleic acids, such as DNA (including structures A, B and Z), RNA (including mRNA, tRNA and rRNA together or separately) and cDNA, as well as any synthetic or artificial forms of polynucleotides. Nucleic acids used in the disclosed compositions can be modified in various ways, including cross-linking, internal chain modifications such as methylation and cap addition, and by copolymerization. In addition, other preferred molecules can be attached to the nucleic acid chains.

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EP 3 000 472 B1

The nucleic acid sequence may be irrelevant to many aspects of the disclosure. However, specific sequences may be used to prevent any significant effect due to the coding of information by nucleic acids, to eliciting specific cellular responses, or to affect the physical structure of the molecule. Nucleic acids can be used in various crystal structures, both in completed biomaterials and during their production processes. The nucleic acid crystal structure may be influenced by the salts used with the nucleic acid. For example, the Na, K, Bi and Ca salts of DNA all have different precipitation rates and different crystal structures. In addition, pH affects the crystal structure of nucleic acids.

[0097] The physical properties of nucleic acids may also be affected by the occurrence of other physical properties. For example, the inclusion of a hairpin loop may result in more flexible biomaterials or may provide specific cutting sites. The produced nucleic acid polymers and copolymers can be used for a variety of tissue engineering applications, including to increase tensile tissue strength, improve wound healing, promote wound healing, as tissue dies, to direct tissue formation, to stimulate nerve growth, improving vascularization in tissues, as a biodegradable adhesive agent, as a coating of a device or implant, or to improve the function of a tissue or body part. Polymers can also be more specifically used as seams, scaffoldings and wound dressings. The type of nucleic acid polymer or copolymer used may influence the resulting chemical and physical structure of the polymeric biomaterial.

iii. Combinations [0098] The composition disclosed herein may comprise mammalian ECM combinations from two or more sources or in two or more separate forms. Thus, the disclosed compositions may comprise any combination of native and / or synthetic mammalian ECMs disclosed herein.

[0099] Thus, for example, the composition may comprise mammalian ECM combinations from sources such as, but not limited to, small intestinal submucosa, basal membrane, gastric fluid, bladder infiltrate, basal membrane, basal pancreatic membrane, submucosa large intestine, interstitial lung membrane, respiratory tract fluid, extracellular matrix of the heart, dermis matrix and total extracellular matrix from any mammalian fetal tissue. Any of these tissue sources can provide an extracellular matrix that can be processed into the indicated form (liquid, semi-solid or solid form) for use in the composition.

[0100] The mammalian ECM junctions from two or more sources can be mixed in solids, mixed liquids, mixed emulsions, mixed gels, mixed pastes or mixed solid particles. All of these compositions are mixtures of extracellular matrices from two or more sources, e.g. mixtures of powders or particles from two or more extracellular matrices, mixtures of pastes from two or more extracellular matrices, mixtures of emulsions or gels from two or more extracellular matrices and mixtures of liquids from two or more extracellular matrices.

[0101] The compositions may be formed from three mammalian source tissues, four mammalian source tissue, five mammalian source tissue, six source mammalian tissues and possibly in

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EP 3 000 472 B1 of ten or more source tissues. These source tissues can come from the same mammal (for example the same cow, the same pig, the same rodent, the same man, etc.), the same mammal species (eg cows, pigs, rodents, human) or different species mammals (for example, a hepatic matrix from a pig, submucosa of the small intestine from a cow, and bladder infiltrate from a dog, all mixed together in a composition).

[0102] The compositions may contain two or more liquid matrices (from different source tissues) attached together. The compositions may be two or more emulsion matrices (from different source tissues) attached together. The compositions may be two or more matrices of particles (from different source tissues) attached together. The composition may be a liquid mixture of two or more extracellular matrices.

[0103] For example, the composition may comprise a combination of SIS in the form of a sheet, particles, emulsion, gel or liquid with SS or LBM or UBS in the form of a sheet, particles, emulsion, gel or liquid. For example, the composition may comprise a combination of SS in the form of a sheet, particles, emulsion, gel or liquid with SIS or LBM or UBS in the form of a sheet, particles, emulsion, gel or liquid. For example, the composition may comprise a combination of LBM in the form of a sheet, particles, emulsion, gel or liquid with SS or SIS or UBS in the form of a sheet, particles, emulsion, gel or liquid. For example, the composition may comprise a combination of UBS in the form of a sheet, particles, emulsion, gel or liquid with SS or SIS or LBM in the form of a sheet, particles, emulsion, gel or liquid.

[0104] The disclosed compositions may comprise mammalian ECM combinations from one or more sources, but in two or more separate embodiments. For example, the composition may comprise a gel matrix connected to a matrix of particles. In some aspects, the mammalian ECM in particulate form may be spotted on a mammalian ECM in the form of a sheet.

[0105] In some aspects, the composition may comprise a combination of SIS, SS or LBM or UBS in the form of a sheet, emulsion, gel or liquid with SIS, SS or LBM or UBS in particulate form. In some aspects, the composition may comprise a combination of SIS, SS or LBM or UBS in the form of particles, emulsions, gels or liquids with SIS, SS or LBM or UBS in the form of a sheet. In some aspects, the composition may comprise a combination of SIS, SS or LBM or UBS in the form of a sheet, particle, gel or liquid with SIS, SS or LBM or UBS in the form of an emulsion. In some aspects, the composition may comprise a combination of SIS, SS or LBM or UBS in the form of a sheet, particles, emulsion or liquid with SIS, SS, LBM or UBS in the form of a gel. In some aspects, the composition may comprise a combination of SIS, SS or LBM or UBS in the form of a sheet, particles, emulsion or gel with SIS, SS or LBM or UBS in liquid form.

[0106] As disclosed herein, a mammalian ECM-containing composition may be formulated to have a preferred consistency. For example, the ECM mammal can be prepared as a combination of gel and particles in an optimal ratio, to prevent disperse in the bloodstream. For example, a composition comprising a mammalian ECM may contain about 40% ECM in the form of a gel and about 60% ECM in the form of dry particles. Thus, there is disclosed a composition comprising a mammalian ECM in both a gel and a dry particle form, wherein the gel form is about 10, 15, 20, 25, 30, 35, 36, 37, 38, 39, 40, 41, 42 , 43, 44, 45, 50% ECM in the composition. Thus, the form of the dry particles can be about 90, 85, 80, 75, 70, 65, 64, 63, 62, 61, 60, 59, 58, 57, 56, 55, 50% ECM in the composition.

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[0107] The choice of concentrations of liquid or semi-solid compositions (liquids, gels, emulsions or pastes) is important. For example, liquid forms may be in the concentration range, from highly diluted at about 0.001 mg / ml, to higher concentrations up to about 200 mg / ml. The concentrations may furthermore be in more specific ranges such as, for example, the following set of ranges: from about 5 mg / ml to about 150 mg / ml, from about 10 mg / ml to about 125 mg / ml, from about 25 mg / ml up to about 100 mg / ml, from about 20 mg / ml to about 75 mg / ml, from about 25 mg / ml to about 60 mg / ml, from about 30 mg / ml to about 50 mg / ml, from about 35 mg / ml to about 45 mg / ml, and from about 40 mg / ml to about 42 mg / ml. This set of scopes is exemplary and not intended to be exhaustive. taken into account,

[0108] The emulsion may be more concentrated than the liquid form and may retain shape, which may be useful when applying the matrix composition to certain parts of the body, hence its characterization as "semi-solid". The emulsion can be concentrated enough to form shapes, such as a plug or other configuration adapted to the location where the matrix composition is used. The thick emulsion may be spread or otherwise applied to the spot as a paste and sprinkled with solid particles on top of the emulsion. The solid particles may be reconstituted in water to form an emulsion or may be used dry as a powder of particles to which the region of the treated subject may be scattered.

[0109] Dry particles or reconstituted particles that form an emulsion of two or more mammalian ECMs can be mixed together in certain proportions. For example, 50% SIS can be mixed with 50% SS in a vial. The mixture can then be liquefied by hydrating it in a suitable buffer, e.g. saline. The hydration may be performed to the desired concentration of the mammalian ECM mixture, for example in the range of from about 0.001 mg / ml to about 200 mg / ml. The concentrations may furthermore be in more specific ranges, such as for example the following set of ranges: from about 5 mg / ml to about 150 mg / ml, from about 10 mg / ml to about 125 mg / ml, from about 25 mg / ml up to about 100 mg / ml, from about 20 mg / ml to about 75 mg / ml, from about 25 mg / ml to about 60 mg / ml, from about 30 mg / ml to about 50 mg / ml, from about 35 mg / ml to about 45 mg / ml and from about 40 mg / ml to about 42 mg / ml. This set of scopes is exemplary and not intended to be exhaustive. It is contemplated that any value of any of these specifically mentioned ranges is a reasonable and useful value for the concentration of a liquid or semi-solid element of the composition.

[0110] The lower the concentration of extracellular matrix, the more fluid the composition will be. The higher the concentration of the extracellular matrix, the more the composition approaches a consistency similar to an emulsion or semi-solid gel.

[0111] The ratio of mixtures of two (or more) extracellular matrices in any given composition from different sources (or the same source) may be uneven. So, for example, LMB can occur in 75% and SIS can occur in 25%, i.e. a ratio of 3: 1). Any suitable ratio can be used: 1: 1, 1: 2, 1: 3, 1: 4, 1: 5, and so on. When 3 or more of the source tissues of the extracellular matrix are represented in the composition, there may be the same type of balance or imbalance in the amounts of the matrix. For example, for an extracellular matrix from 3 sources, each source can exist in equal proportions, i.e. 1: 1: 1 (33% / 33% / 33%). Alternatively, incommensurate

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The amount of particles may be from a single source, e.g. 2: 1: 1 (50% / 25% / 25%). Similarly, all three sources can be disproportionate, e.g. 50% / 30% / 20%.

[0112] Two or more mammalian ECMs may be liquefied (or emulsified) separately, and liquefied or emulsified compositions mixed together. In another alternative, two or more mammalian ECMs may be liquefied or emulsified separately and administered separately. In addition, two or more mammalian ECMs may remain in the form of solid particles and be mixed together in a vial for administration as a combination of solid particles. Re-hydration of the mammalian ECM mixture in the form of dry particles can be obtained immediately before use.

2. Proteins [0113] The disclosed mammalian ECM-containing compositions may further comprise exogenous proteins, such as those typically found in mammalian ECMs. The protein may be collagen, proteoglycan, glycosaminoglycan chain (GAG), glycoprotein, growth factor, cytokine, cell-surface-related protein, cell adhesion molecule (CAM), angiogenic growth factor, endothelium ligand, matricin, matrix metalloprotease, cadherin, immunoglobulin, collagen fibrous, non-fibrous collagen, basal membrane collagen, multiplexin, small leukin-rich proteoglycan, decorin, biglikan, fibromodulin, keratocan, lumican, epiphate, heparin sulfate proteoglycan, perlekan, agrin, testicane, syndecan, glypican, serglycin, selectin, lektican, aggrecan , vornican, neurocan, brewican, cytoplasmic domain-44 (CD-44), macrophage stimulant, amyloid precursor protein, heparin, chondroitin B sulphate (dermatan sulfate), chondroitin A sulfate, heparan sulphate, hyaluronic acid, fibronectin (Fn), tenascin, elastin, fibrillin, laminin, nidogen / entactin, fibulina I, fibulin II, integrin, molecule transmembrane, platelet derived growth factor (PDGF), epithelial growth factor (EGF), transforming growth factor alpha (TGF-alpha), transforming growth factor beta (TGF-β), fibroblast growth factor 2 (FGF-2) (also called basic factor fibroblast growth (bFGF)), thrombospondin, osteopontin, angiotensin converting enzyme (ACE) or vascular endothelial growth factor (VEGF). This list is not intended to be exhaustive. fibronectin (Fn), tenascin, elastin, fibrillin, laminin, nidogen / entactin, fibulin I, fibulin II, integrin, transmembrane molecule, platelet-derived growth factor (PDGF), epithelial growth factor (EGF), transforming growth factor alpha (TGF-alpha ), transforming growth factor beta (TGF-β), fibroblast growth factor 2 (FGF-2) (also called basic fibroblast growth factor (bFGF)), thrombospondin, osteopontin, angiotensin converting enzyme (ACE) or vascular endothelial growth factor (VEGF) ). This list is not intended to be exhaustive. fibronectin (Fn), tenascin, elastin, fibrillin, laminin, nidogen / entactin, fibulin I, fibulin II, integrin, transmembrane molecule, platelet-derived growth factor (PDGF), epithelial growth factor (EGF), transforming growth factor alpha (TGF-alpha ), transforming growth factor beta (TGF-β), fibroblast growth factor 2 (FGF-2) (also called basic fibroblast growth factor (bFGF)), thrombospondin, osteopontin, angiotensin converting enzyme (ACE) or vascular endothelial growth factor (VEGF) ). This list is not intended to be exhaustive. transforming growth factor beta (TGF-β), fibroblast growth factor 2 (FGF-2) (also called basic fibroblast growth factor (bFGF)), thrombospondin, osteopontin, angiotensin converting enzyme (ACE) or vascular endothelial growth factor (VEGF). This list is not intended to be exhaustive. transforming growth factor beta (TGF-β), fibroblast growth factor 2 (FGF-2) (also called basic fibroblast growth factor (bFGF)), thrombospondin, osteopontin, angiotensin converting enzyme (ACE) or vascular endothelial growth factor (VEGF). This list is not intended to be exhaustive.

[0114] Thus, the mammalian ECM containing compositions disclosed herein may contain collagen I and III, elastin, laminin, CD44, hyaluronan, syndecan, bFGF, HGF, PDGF, VEGF, Fn, tenascin, heparin, heparan sulfate, chondroitin B sulfate, integrin, decoration, TGF-β, or a combination thereof.

3. Cells [0115] In some aspects, the mammalian ECM-containing compositions disclosed herein further comprise one or more cells, wherein the cells do not include human embryonic stem cells. In some aspects, the cells are not native, i.e. they are heterologous to a mammalian ECM. In some aspects, the cells are stem cells. A non-exhaustive list of stem cells includes fetal cardiomyocytes, myofibroblasts, mesenchymal stem cells, autonsplanted cardiomyocytes, adipocytes, totipotent cells, pluripotent cells, blood stem cells, myoblasts, adult stem cells, bone marrow cells, mesenchymal cells, non-human embryonic stem cells. , parenchymal cell, epithelial cell, endothelial cell, mesothelial cell, fibroblast, osteoblast,

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A pluripotent stem cell, a bone marrow derived progenitor, a progenitor cell, a myocardium cell, a skeletal cell, a fetal cell, a non-human germ cell, an undifferentiated cell, a multipotent progenitor cell, a unipotent progenitor cell, a monocyte, a cardiomyocyte, cardiac myoblast, skeletal myoblast, macrophage, capillary cell endothelial cell, xenogenic cell, allogeneic cell, adult stem cell and postnatal stem cell.

[0116] In some aspects, the stem cells have the potential to differentiate into cardiac tissue cells. Thus, in some aspects, the stem cells are pluripotent. In some aspects, the stem cells are angioblasts or hemangioblasts. In some aspects, the stem cells are myoblasts. Stem cells can be obtained and maintained using standard methods for growing stem cells.

4. Pharmaceuticals [0117] The mammalian ECM-containing compositions disclosed herein may further comprise any known or newly discovered substance that can be administered to the subject's heart. For example, the mammalian ECM-containing compositions disclosed herein may further comprise any agent against arrhythmias. Anti-arrhythmic agents are a group of pharmaceutical agents that are used to inhibit fast and / or irregular heart rhythms (arrhythmias).

[0118] The Vaughan Williams classification, introduced in 1970, is one of the most widely used classifications of drugs against arrhythmias. This scheme classifies drugs based on their primary mechanism of action against arrhythmias. There are five major classes of anti-arrhythmic drugs in the Vaughan Williams classification: Class I measures block the Na channel (Na +); Class II agents are anti-sympathetic nervous system (most agents in this class are beta blockers); Class III measures affect the outflow through the potassium channel (K +); Class IV measures affect calcium channels and AV node; and Class V measures work through other or unknown mechanisms.

[0119] Class Ia agents include Quinidine, Procainamide and Disopyrin. Class Ib agents include Lidocaine, Phenytoin and Mexidin. Class Ic agents include Flekainide, Propafenone and Morycysin. Class II agents include Propranolol, Esmolol, Timolol, Metoprolol and Atenolol. Class III agents include Amiodarone, Sotalol, Ibutylid and Dofetilide. Class IV measures include Werapamil and Diltiazem. Class V agents include Adenosine and Digoxin.

[0120] Thus, the mammalian ECM-containing compositions disclosed herein may further comprise one or more of Quinidine, Procainamide, Dizopramamide, Lidocaine, Phenytoin, Mexiletin, Flekainide,

Propafenone, Moriczyn, Propranolol, Esmolol, Timolol, Metoprolol, Atenolol, Amiodarone, Sotalol, Ibutylid, Dofetilide, Verapamil, Diltiazem, Adenosine and Digoxin.

[0121] The provided compositions may further comprise one or more antibiotics (e.g., Aminoglycosides, Cephalosporins, Chloramphenicol, Clindamycin, Erythromycin, Fluoroquinolones, Macrolides, Azolides, Metronidazole, Penicillins, Tetracyclines, Trimethoprim-Sulfomethoxazol and Vancomycin).

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[0122] The provided compositions may further comprise one or more steroids (e.g., Andrany (e.g., Testosterone), Cholestans (e.g., Cholesterol), Cholic Acids (e.g., Cholic Acid), Corticosteroids (e.g.

Dexamethasone), Estrans (e.g. Estradiol) and Pregnane (e.g. Progesterone).

[0123] The provided compositions may further comprise one or more classes of narcotic and non-narcotic analgesics, including, but not limited to, Morphine, Codeine, Heroin, Hydromorphone, Leworfanol, Meperidine, Methadone, Oxidone, Propoxyphene, Fentanyl, Methadone, Naloxone, Buprenorphine Butorphanol, Nalbufin and Pentazocine.

[0124] Provided compositions may further comprise one or more anti-inflammatory agents, including but not limited to Aclofenac, Alclometasone Dipropionate, Algestone Acetonide, Alpha Amylase, Amcinephal, Amcinephide, Amphozide, Amiprilose Hydrochloride, Anakinra, Aniolac, Anitrazafen, Apazon, Disodium Balsalazide, Bendazac, Benoxaprofen, Benzydamine Hydrochloride, Bromelain, Broperamol, Budesonide, Carprofen, Cycloprofen, Cyntazone, Cliprofen, Clobetasol Propionate, Clobetasone Butterbone, Klopirac, Klotykazon Propionate, Kormetasone Acetate, Cortodoxone, Decanoate, Deflazakort, Delatestryl, Depo-Testosterone, Dezonide, Dexamethasone, Dexamethasone Dipropionate, Diclofenac Potassium, Diclofenac sodium, Diflorazone diacetate, Diflumidone sodium, Diflunisal, Difluprednat, Diftalon, Dimethylsulfoxide, Drocinonide, Endryson, Enlimomab, Enolikam sodium,Epiryzol, Etodolak, Etofenamat, Felbinak, Fenamol, Fenbufen, Fenklofenak, Fenklorak, Fendosal, Fenpipalon, Fentiazak, Flazalon, Fluzakort, Flufenamic acid, Flumizol, Flunizolide acetate, Fluniksin, Meglumine flunixin, Butyl fluoxyn, Fluoromethyl acetate, Flukvana, Flurbiprofen, Fluretofen , Fluticasone Propionate, Furaprofen, Furobufen, Halcinonide, Halobetazole Propionate, Haloprednone Acetate, Ibufenac, Ibuprofen, Ibuprofen Aluminum, Ibontrofen Pikonol, Ilonidap, Indometacin, Indomethacin sodium, Indoprofen, Indoxol, Intrasol, Isoflupredon Acetate, Izoksepak, Isoxicam, Ketoprofen, Lofemizol hydrochloride , Lomoxicam, Loteprednol Etabonian, Sodium Meklofenamate, Meclofenamic Acid, DimaślanFlumisole, Flunizolide Acetate, Flunixin, Meglumine flunixin, Butyl Fluoxide, Fluorometholon Acetate, Flucvazone, Flurbiprofen, Fluretofen, Fluticasone Propionate, Furaprofen, Furobufen, Halcinonide, Halobetazole Propionate, Haloprednone Acetate, Ibufenac, Ibuprofen, Ibuprofen Aluminum, Ibuprofen Pikonol, Ilonidap, Indomethacin , Indomethacin Sodium, Indoprofen, Indoxol, Intrasol, Isoplupredon Acetate, Izoksepak, Izoxicam, Ketoprofen, Lofemizol hydrochloride, Lomoxicam, Loteprednol Etabonian, Sodium Meklofenamate, Meclofenamic acid, DimaślanFlumisole, Flunizolide Acetate, Flunixin, Meglumine flunixin, Butyl Fluoxide, Fluorometholon Acetate, Flucvazone, Flurbiprofen, Fluretofen, Fluticasone Propionate, Furaprofen, Furobufen, Halcinonide, Halobetazole Propionate, Haloprednone Acetate, Ibufenac, Ibuprofen, Ibuprofen Aluminum, Ibuprofen Pikonol, Ilonidap, Indomethacin , Indomethacin Sodium, Indoprofen, Indoxol, Intrasol, Isoplupredon Acetate, Izoksepak, Izoxicam, Ketoprofen, Lofemizol hydrochloride, Lomoxicam, Loteprednol Etabonian, Sodium Meklofenamate, Meclofenamic acid, DimaślanIndomethacin, Indomethacin Sodium, Indoprofen, Indoxol, Intrasol, Isoplupredon Acetate, Izoksepak, Izoxicam, Ketoprofen, Lofemizol hydrochloride, Lomoxicam, Loteprednol Etabonian, Sodium Meklofenamate, Meclofenamic acid, DimaślanIndomethacin, Indomethacin Sodium, Indoprofen, Indoxol, Intrasol, Isoplupredon Acetate, Izoksepak, Izoxicam, Ketoprofen, Lofemizol hydrochloride, Lomoxicam, Loteprednol Etabonian, Sodium Meklofenamate, Meclofenamic acid, Dimaślan

Mekloryzone, Mefenamic acid, Mezalamin, Mezeklazon, Mesterolon, Metandrostenolone, Metenolone, Metenolone acetone, Methylprednisolone acetone, Momiflumat, Nabumetone, Nandrolone, Naproxen, Sodium Naproxen, Naproxol, Nimazone, Sodium Olsalazine, Orgoteine, Orpanoxin, Oxandrolone, Oxaprozin, Oxyfenbutazone, Oxymetolone , Paraniline Hydrochloride, Sodium Polysulfate, Pentosan,

Glycerylate Fenbutazone Sodium, Pirfenidone, Piroxicam, Piroxicam cinnamate, Piroxicam olamate, Pirprofen, Prednazat, Prifelon, Prodolowa acid, Prokwazon, Proksazol, Proksazol Citrate, Rimeksolon, Romazarit, Salkoleks, Salnacedin, Salsalan, Sangwinarium Chloride, Seklazon, Sermetacin, Stanozolol, Sudoxicam , Sulindak, Suprofen, Talmetacin, Talniflumat, Talsolan, Tebufelon, Tenidap, Tenodap Sodium, Tenoxicam, Tesykam, Tezimide, Testosterone, Testosterone Mixtures, Tetidamine, Thiopinac, Tiksokortol Povalate, Tolmetin, Sodium Tolmethine, Triclonid, Triflumidate, Zydomethacin and Sodium Zomepirac.

[0125] The provided compositions may further comprise one or more lipid-lowering drugs. The term "lipid-lowering drug" as used herein refers to a drug that can be administered to a subject to reduce serum levels of various lipids associated with cardiac diseases, including, but not limited to, cholesterol, low-density lipoprotein (LDL), lipoprotein very low density (VLDL) and triglycerides.

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[0126] For example, lipid-lowering drugs may be statins, including but not limited to Lovastatin, Simvastatin, Atorvastatin, Fluvastatin, Pravastatin, Rosuvastatin, Cerivastatin and Pitavastatin. It has been considered that any statin drug, currently known or developed in the future, having lipid-lowering and / or anti-inflammatory properties, can be used in the compositions described herein.

[0127] The provided compositions may further comprise one or more antihistamines, including, but not limited to, ethanolamines (such as diphenhydramine, carbinoxamine), ethoxidiamines (like tripelenamine, pyrrylamine), alkylamines (like chlorpheniramine, deoxylorfeniramine, brompeniramine, triprolidine), astemizole, latatatidine, fexofenadine, Bromfeniramine, Klemastine, Acetaminophen, Pseudoephedrine and Triprolidine.

[0128] Provided compositions may further comprise one or more antineoplastic agents, including, but not limited to, Acivicin, Aclubicin, Akodazole Hydrochloride, Acronin, Adozelesin, Aldesleukin, Altretamine, Ambomicin, Ametanthrone Acetate, Aminoglutethimide, Amsacrine, Anastrozole, Antramycin, Asparaginase Asperlin, Azactidine, Azetepe, Azotomycin, Batimastat, Benzodepe, Bikalutamide, Bisanthrine Hydrochloride, Bisnafide Dimesylate, Bizelesine, Bleomycin Sulfate, Sodium Brekwinar, Bropyrimine, Busulfan, Kaktynomycin, Kalusteron, Karacemide, Carbetimer, Carboplatinum, Karmustine, Carubicin hydrochloride, Karzelasin, Cedefingol, Chlorambucil, Cirolemycin, Cisplatin, Cladribine, Krisnatol Mesylate, Cyclophosphamide, Cytarabine, Dacarbazine, Daktinomycin, Daunorubicin Hydrochloride; decitabine; Dexormaplatin, Dezaguanine,

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Toremifene citrate, Trestolone acetate, Triticium, Trimetrexate, Trimetrexate glucuronate, Triptorelin, Tubulozole hydrochloride, Uracyl mustard, Uredepa, Wapreotide, Werteporfin, Vinblastine Sulfate, Vincristine sulfate, Windesin, Windesazine Sulfate, Winepidine Sulfate, Winglicinate Sulfate, Vinylsulfate Sulfate, Vinorelbine Sulfate , Winrozidine Sulfate, Vinzolidine Sulfate, Vorozol, Zeniplatin, Zinostatin and Zorubicin Hydrochloride.

[0129] The composition provided herein may further comprise one or more radiosensitisers, including, but not limited to, gemcitabine, 5-fluorouracil, pentoxifylline and vinorelbine.

5. Carriers [0130] The disclosed ECM mammal can be combined, conjugated or coupled to carriers and other compositions to aid delivery, delivery or other aspects of the ECM and their use. For convenience, such compositions are referred to herein as carriers. The carriers may be, for example, a small molecule, a pharmaceutical drug, a fatty acid, a detectable label, a coupling tag, a nanoparticle or an enzyme.

[0131] The disclosed compositions can be used therapeutically in combination with a pharmaceutically acceptable carrier. By "pharmaceutically acceptable" is meant a material that is not biologically or otherwise undesirable, i.e., the material may be administered to a subject together with the composition, without causing any undesirable biological effects or adversely interacting with any of the components of the pharmaceutical composition in which is included. The carrier will be naturally selected so as to minimize any degradation of the active ingredient and to minimize any undesirable side effects in the subject, as the person skilled in the art knows well.

[0132] Suitable carriers and formulations thereof are described in Remington: The Science and Practice of Pharmacy (19th ed) ed. AR Gennaro, Mack Publishing Company, Easton, PA 1995. Typically, a suitable amount of a pharmaceutically acceptable salt is used in the formulation, in order to obtain the isotonicity of the formulation. Examples of a pharmaceutically acceptable carrier include, but are not limited to, saline, Ringer's solution and dextrose solution. The pH of the solution is preferably from about 5 to about 8, and more preferably from about 7 to about 7.5. Additional carriers include sustained release formulations, such as semipermeable matrices of solid hydrophobic polymers containing the composition, which matrices are in the form of shaped articles, e.g. films, liposomes or microparticles. It will be obvious to those skilled in the art that

[0133] Pharmaceutical carriers are known to those skilled in the art. They will usually be standard carriers for administering drugs to people, including solutions such as sterile water, saline and buffered solutions at physiological pH.

[0134] The pharmaceutical compositions may contain carriers, thickeners, diluents, buffers, preservatives, surfactants and the like in addition to the selected molecule. The pharmaceutical compositions may also contain one or more active ingredients, such as antimicrobial agents, anti-inflammatories, anesthetics, and the like.

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[0135] Preparations for parenteral administration include sterile aqueous or non-aqueous solutions, suspensions and emulsions. Examples of non-aqueous solvents are propylene glycol, polyethylene glycol, vegetable oils such as olive oil and injectable organic esters such as ethyl oleate. Aqueous carriers include water, alcoholic / aqueous solutions, emulsions or suspensions, including physiological saline and buffered media. Parenteral carriers include sodium chloride solution, Ringer's dextrose, dextrose and sodium chloride, Ringer's lactate or non-volatile oils. Intravenous vehicles include fluid and nutrient replenishers, electrolyte replenishers (such as those based on Ringer's dextrose), and the like. Preservatives and other additives may also be present,

[0136] Formulations for topical administration may include ointments, lotions, creams, gels, drops, suppositories, sprays, liquids and powders. Conventional pharmaceutical carriers, aqueous, powder or oil bases, thickeners and the like may be necessary or desirable.

[0137] Compositions for oral administration include powders or granules, suspensions or solutions in water or non-aqueous vehicles, capsules, sachets or tablets. Thickeners, flavors, diluents, emulsifiers, dispersion-promoting aids or binders may be desirable.

[0138] Some of the compositions may potentially be administered as a pharmaceutically acceptable acidic or basic addition salt formed by reaction with inorganic acids such as hydrochloric acid, hydrobromic acid, perchloric acid, nitric acid, thiocyanic acid, sulfuric acid and phosphoric acid, and organic acids, such as formic acid, acetic acid, propionic acid, glycolic acid, lactic acid, pyruvic acid, oxalic acid, malonic acid, succinic acid, maleic acid and fumaric acid, or by reaction with an inorganic base such as sodium hydroxide , aluminum hydroxide, potassium hydroxide and organic bases such as mono-, di-, trialkyl and arylamines and substituted ethanolamines.

D. METHODS FOR MAKING COMPOSITIONS [0139] The compositions disclosed herein may be produced using any method known to those skilled in the art for that particular reagent or compound, unless specifically indicated otherwise. For example, U.S. Patent No. 5,275,826, U.S. Patent No. 5,554,389, U.S. Patent No. 6,099,567, and U.S. Patent No. 6,379,710, are disclosed herein by reference for methods of preparing compositions comprising intestinal submucosal mucosa (SIS), bladder wall submucosa (UBS), respectively, Stomach submucosal (SS) and hepatic submucosal (LS) or basal membrane (LBM).

E. EXAMPLES [0140] The following examples are set forth to provide those of ordinary skill in the art with full disclosure and description of how the compounds, compositions, articles and / or devices claimed herein are made and evaluated, and are intended to be exemplary only and are not intended to limit the disclosure. Efforts have been made to ensure accuracy with respect to numerical values ​​(e.g. quantities, temperature, etc.), but some errors and deviations should be taken into account. Unless otherwise indicated, the parts are parts by weight, the temperature is in ° C or the ambient temperature and the pressure is atmospheric or near atmospheric.

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1. Example 1: Retrospective assessment of new cases of post-operative atrial fibrillation in patients receiving CorMatrix® ECM ™.

[0141] A retrospective multicentre, two armed, analysis of disease card records using CorMatrix® ECM ™ was performed. The purpose of this retrospective study was to assess whether CorMatrix® ECM ™ for reconstructing the normal pericardium barrier could result in a lower rate of new post-operative atrial fibrillation compared to patients who were not subject to pericardium closure.

[0142] CorMatrix® ECM ™ can be used for the reconstruction and repair of the pericardium after open heart surgery. Undamaged, the pericardium provides a passive barrier to the heart preventing excessive expansion and helping to modulate sudden changes in volume. By reconstructing the pericardium with CorMatrix® ECM ™, the natural pericardium barrier can be restored. The aim of this retrospective clinical analysis was to assess whether a reduction in the number of new post-operative AF cases was observed, by analyzing patients who underwent reconstruction of their native pericardium with CorMatrix® ECM ™, compared to those who were not subject to pericarditis after the procedures isolated coronary artery bypass grafting (CABG).

[0143] CorMatrix® ECM ™ was provided in four-layered sheets with different dimensions, which can be cut to the size that the physician deems necessary to carry out the procedure.

[0144] The definition of new cases of post-operative atrial fibrillation used for this retrospective study is based on the definition used in the Society of Thoracic Surgeons (STS) Adult Cardiac Surgery Database 2007. The definition is: "Indication of whether or not the patient has had atrial fibrillation / atrial flutter ( AF) requiring treatment. It does not include a relapse of AF that occurred before surgery. It does NOT include patients who have had atrial fibrillation (treated or untreated) before surgery. The event must occur for the first time.

[0145] It was required that all patients meet the following inclusion criteria in order to be included as part of this retrospective clinical analysis: this heart surgery was the first or main heart surgery of the subject and the subject had to undergo the CABG isolated procedure.

[0146] Patients were not included as part of this retrospective clinical analysis if one or more of the following exclusion criteria were met: previous history of atrial fibrillation, previous history of open heart surgery, previous history of pericarditis, previous amiodarone intake in the last six months and at the same time planned operation of the valve.

[0147] Patients in whom their native pericardium was reconstructed with CorMatrix® ECM ™ had a statistically significant reduction in the incidence of atrial fibrillation (A-fib) compared to those who were not subject to pericardial closure following the CABG isolated procedures. The usual incidence of atrial fibrillation is approximately 25%. In these studies, the incidence of atrial fibrillation was between 4% and 8% (1/25 and 4/52).

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2. Reference Example 1: Modulation of cardiac remodeling with decellularised matrix emulsion is associated with myofibroblast proliferation and angiogenesis by recruitment of positive C-kit positive cells after myocardial infarction [0148] Degradation of the native extracellular matrix (ECM) is associated with non-adaptive remodeling heart after a heart attack. As shown herein, a xenogenic, cell-free matrix emulsion injected into the myocardium after myocardial infarction stimulated myofibroblast proliferation and angiogenesis by recruiting c-kit positive host cells.

[0149] Sixty-four rats were subjected to 45 minutes of local ischemia with the following 3, 7, 21 and 42 days of reperfusion. Histological examination was performed using immunohistological staining, and cardiac functions were analyzed by means of echocardiography. The ECM emulsion (30-50 μΐ) was injected into the affected area of ​​the myocardium after reperfusion, and the location of the emulsion was confirmed by Trichrome Masson staining. At 7 days after reperfusion, the population of c-kit positive cells in the emulsion region significantly increased relative to the control (32 ± 0.6 * vs. 15 ± 3/1000 testes), consistent with the significantly increased expression of 31 kDa stem cell factor detected using Western blot analysis. At the same time, with this change, myofibroblasts accumulated in the emulsion area to a significant degree compared to controls (59 ± 8 * vs 30 ± 3 / HPF). Strong immunoreactivity for VEGF was observed in the emulsion area, and angiogenesis was significantly increased compared to control, as demonstrated by the increased density of actin-positive smooth muscle α smooth muscle (70 ± 10 * to 20 ± 4 / HPF) and positive vWF positive dishes respectively. (95 ± 14 * versus 34 ± 8 / HPF). 42 days after reperfusion, echocardiography showed improvement in end-systolic volume (0.3 ± 0.1 * vs 0.6 ± 0.3 ml)), shortening fraction (33 ± 5 * vs 24 ± 6%) and ejection fraction ( 67 ± 6 * versus 53 ± 10%) in the emulsion group. The thickness of the medial wall of the anterior portion of the myocardial septum after the infarct in the emulsion group was also significantly greater than that in the Control (0.19 ± 0.02 * versus 0.15 ± 0.02 cm). as demonstrated by the increased density of actin-smooth smooth muscle α (70 ± 10 * to 20 ± 4 / HPF) and positive vWF positive dishes (95 ± 14 * versus 34 ± 8 / HPF), respectively. 42 days after reperfusion, echocardiography showed improvement in end-systolic volume (0.3 ± 0.1 * vs 0.6 ± 0.3 ml)), shortening fraction (33 ± 5 * vs 24 ± 6%) and ejection fraction ( 67 ± 6 * versus 53 ± 10%) in the emulsion group. The thickness of the medial wall of the anterior portion of the myocardial septum after the infarct in the emulsion group was also significantly greater than that in the Control (0.19 ± 0.02 * versus 0.15 ± 0.02 cm). as demonstrated by the increased density of actin-smooth smooth muscle α (70 ± 10 * to 20 ± 4 / HPF) and positive vWF positive dishes (95 ± 14 * versus 34 ± 8 / HPF), respectively. 42 days after reperfusion, echocardiography showed improvement in end-systolic volume (0.3 ± 0.1 * vs 0.6 ± 0.3 ml)), shortening fraction (33 ± 5 * vs 24 ± 6%) and ejection fraction ( 67 ± 6 * versus 53 ± 10%) in the emulsion group. The thickness of the medial wall of the anterior portion of the myocardial septum after the infarct in the emulsion group was also significantly greater than that in the Control (0.19 ± 0.02 * versus 0.15 ± 0.02 cm). shortening fraction (33 ± 5 * against 24 ± 6%) and ejection fraction (67 ± 6 * against 53 ± 10%) in the emulsion group. The thickness of the medial wall of the anterior portion of the myocardial septum after the infarct in the emulsion group was also significantly greater than that in the Control (0.19 ± 0.02 * versus 0.15 ± 0.02 cm). shortening fraction (33 ± 5 * against 24 ± 6%) and ejection fraction (67 ± 6 * against 53 ± 10%) in the emulsion group. The thickness of the medial wall of the anterior portion of the myocardial septum after the infarct in the emulsion group was also significantly greater than that in the Control (0.19 ± 0.02 * versus 0.15 ± 0.02 cm).

[0150] Injection into the myocardium of a decellularised extracellular matrix for ischemic / myocardial reperfusion weakened non-adaptive cardiac remodeling and maintained cardiac function, presumably mediated by increased myofibroblast proliferation and angiogenesis by recruitment of c-kit positive cells. * p <0.05 group with emulsion for control.

[0151] Preferred embodiments of the invention are listed below:

A method for treating or preventing a arrhythmia disorder in a subject, comprising administering to the myocardium a subject a therapeutically effective amount of a mammalian extracellular matrix composition, wherein the method does not comprise administering the small intestinal submucosa as a patch to the orifice of the cardiac heart.

2. A method according to embodiment 1, wherein the cardiac tissue is a myocardium, a spinal cord, an endocardium or an oculi.

3. The method according to embodiment 1, wherein the cardiac tissue is not pericardial.

4. The method of embodiment 1, wherein the subject is identified as being at risk of arrhythmias.

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5. The method of embodiment 1, wherein the mammalian extracellular matrix is ​​selected from the group consisting of small intestinal submucosa, bladder infiltrate, gastric submucosa, hepatic submucosal and hepatic basal membrane.

6. The method of embodiment 1, wherein the mammalian extracellular matrix is ​​not submucosa of the small intestine.

7. The method of embodiment 1, wherein the mammalian extracellular matrix is ​​produced in vitro from mammalian cell culture.

8. The method of embodiment 1, wherein the composition further comprises a synthetic extracellular matrix.

9. The method of embodiment 1, wherein the mammalian extracellular matrix is ​​selected from the group of tissues consisting of dermis, fascia, pericardium, parenchymal tissue and the extracellular matrix of the myocardium.

10. The method of embodiment 1, wherein the mammalian extracellular matrix is ​​a collagen scaffold from a mammalian tissue source.

11. The method of embodiment 1, wherein the arrhythmia is atrial fibrillation or ventricular fibrillation.

12. The method of embodiment 1, wherein the mammalian extracellular matrix is ​​a patch.

13. The method of embodiment 1, wherein the mammalian extracellular matrix is ​​suitable for injection.

14. The method of embodiment 1, wherein the mammalian extracellular matrix is ​​in liquefied form.

15. The method of embodiment 1, wherein the mammalian extracellular matrix is ​​emulsified.

16. The method of embodiment 1, wherein the mammalian extracellular matrix is ​​a powder or aerosol.

17. The method of embodiment 1, wherein the mammalian extracellular matrix is ​​sprayed onto cardiac tissue.

18. The method of embodiment 1, wherein the composition is administered to an orifice in the cardiac heart.

19. The method of embodiment 1, wherein the composition is injected into the myocardium of the heart.

20. The method of embodiment 1, wherein the composition is administered to the endocardial or cardiac surfaces.

21. The method of embodiment 1, wherein the subject has undergone heart surgery.

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22. The method of embodiment 1, wherein the subject has had a myocardial infarction.

23. The method of embodiment 1, wherein the composition further comprises a drug for rhythm disorders, a lipid lowering drug or an anti-inflammatory drug.

24. The method of embodiment 1, wherein the composition further comprises cells.

25. A method according to embodiment 24, wherein the cells are stem cells.

26. A method for treating or preventing a arrhythmia in a subject identified as being at risk of developing a arrhythmia, comprising administering to the myocardium a therapeutically effective amount of a composition comprising a mammalian extracellular matrix and further comprising an anti-arrhythmic agent, a lipid-lowering drug, cells, a protein or a combination thereof.

27. The method of embodiment 26, wherein the cells are stem cells.

28. The method of embodiment 26, wherein the protein is hyaluronic acid, a proteoglycan, an aminoglycate, or a combination thereof.

29. The method of embodiment 26, wherein the cardiac tissue is a myocardium, a spinal cord, an endocardium or a pericardium

30. The method of embodiment 26, wherein the mammalian extracellular matrix is ​​selected from the group consisting of small intestinal submucosa, bladder infiltrate, gastric submucosa, hepatic submucosal and hepatic basal membrane.

31. The method of embodiment 26, wherein the mammalian extracellular matrix is ​​produced in vitro from mammalian cell culture.

32. The method of embodiment 26, wherein the composition further comprises a synthetic extracellular matrix.

33. The method of embodiment 26, wherein the mammalian extracellular matrix is ​​selected from a group of tissues consisting of dermis, fascia, pericardium, parenchymal tissue and the extracellular matrix of the myocardium.

34. The method of embodiment 26, wherein the mammalian extracellular matrix is ​​a collagen scaffold from a mammalian tissue source.

35. The method of embodiment 26, wherein the arrhythmias are atrial fibrillation or ventricular fibrillation.

36. The method of embodiment 26, wherein the mammalian extracellular matrix is ​​a patch.

37. The method of embodiment 26, wherein the mammalian extracellular matrix is ​​suitable for injection.

38. The method of embodiment 26, wherein the mammalian extracellular matrix is ​​in liquefied form.

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39. The method of embodiment 26, wherein the mammalian extracellular matrix is ​​emulsified.

40. The method of embodiment 26, wherein the mammalian extracellular matrix is ​​a powder or aerosol.

41. The method of embodiment 26, wherein the mammalian extracellular matrix is ​​sprayed onto cardiac tissue.

42. A method according to embodiment 26, wherein the composition is administered to an orifice in the cardiac case.

43. The method of embodiment 26, wherein the composition is injected into the myocardium of the heart.

44. The method of embodiment 26, wherein the composition is administered to the endocardial or cardiac surfaces.

45. The method of embodiment 26, wherein the subject has undergone heart surgery.

46. ​​The method of embodiment 26, wherein the subject has had a myocardial infarction.

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