sign in sign up
<<
Home , Brofaromine, Cimoxatone, Minaprine, Nialamide, Phenelzine, Pheniprazine

US 2015 0011643A1 (19) United States (12) Patent Application Publication (10) Pub. No.: US 2015/0011643 A1 DiLisa et al. (43) Pub. Date: Jan. 8, 2015

(54) TREATMENT OF HEART FAILURE AND (60) Provisional application No. 61/038,230, filed on Mar. ASSOCATED CONDITIONS BY 20, 2008, provisional application No. 61/155,704, ADMINISTRATION OF MONOAMINE filed on Feb. 26, 2009. OXIDASE INHIBITORS (71) Applicants:Nazareno Paolocci, Baltimore, MD Publication Classification (US); Univeristy of Padua, Padova (IT) (51) Int. Cl. (72) Inventors: Fabio DiLisa, Padova (IT): Ning Feng, A63L/38 (2006.01) Baltimore, MD (US); Nina Kaludercic, (52) U.S. Cl. Baltimore, MD (US); Nazareno CPC ...... A61 K31/138 (2013.01) Paolocci, Baltimore, MD (US) USPC ...... S14/651 (21) Appl. No.: 14/332,234 (22) Filed: Jul. 15, 2014 (57) ABSTRACT Related U.S. Application Data Administration of inhibitors is useful in (63) Continuation of application No. 12/407,739, filed on the prevention and treatment of heart failure and incipient Mar. 19, 2009, now abandoned. heart failure. Patent Application Publication Jan. 8, 2015 Sheet 1 of 2 US 201S/0011643 A1

Figure 1 Sial 8. Sws-L)

Cleaved Š Caspase -3

)

Figure . Prevention of caspase-3 productief) fron cardiomyocytes Lapoi) reatment with clorgyi Be. Patent Application Publication Jan. 8, 2015 Sheet 2 of 2 US 201S/0011643 A1

Figure 2

8:38:8 ::::::::8:

US 2015/0011643 A1 Jan. 8, 2015

TREATMENT OF HEART FAILURE AND in the art is a variety of MAO inhibitors and their pharmaceu ASSOCATED CONDITIONS BY tically acceptable compositions for administration, in accor ADMINISTRATION OF MONOAMINE dance with the present invention, to including, but OXIDASE INHIBITORS not limited to, humans. CROSS-REFERENCE TO RELATED PATENT BRIEF DESCRIPTION OF THE DRAWINGS APPLICATIONS 0010 FIG. 1. Prevention of caspase-3 production from 0001. This application claims the benefit of U.S. Provi cardiomyocytes upon treatment with clorgyline. sional Application Ser. No. 61/038.230 filed 20 Mar. 2008, 0011 FIG. 2. Interaction of MAO-A and MAO-B iso and U.S. Provisional Application Ser. No. 61/155,704 filed 26 forms with /dietary amines in the central Feb. 2009, both of which applications are incorporated herein and peripheral nervous system. by reference in their entirety. DETAILED DESCRIPTION OF THE INVENTION STATEMENT REGARDING FEDERALLY SPONSORED RESEARCH ORDEVELOPMENT 0012. The detailed description of the various aspects and embodiments of the invention is divided into the following 0002 This invention was made with government support sections beginning with a definition of certain terms used under National Heart, Lung, and Blood Institute grant no. herein. R01 HL075265-01A2. The government has certain rights in the invention. Definitions BACKGROUND OF THE INVENTION 0013 The following definitions are provided to assist the reader. Unless otherwise defined, all terms of art, notations 0003 1. Field of the Invention and other scientific or medical terms or terminology used 0004. The present invention relates generally to the field of herein are intended to have the meanings commonly under medicine, pharmaceutical chemistry, biology and in particu stood by those of skill in the chemical and medical arts. In lar to methods for treating and inhibiting heart failure and Some cases, terms with commonly understood meanings are incipient heart failure. defined herein for clarity and/or for ready reference, and the 0005 2. State of the Art inclusion of such definitions herein should not necessarily be 0006. In spite of various known treatment and preventive construed to represent a substantial difference over the defi methods, heart failure continues to remain a major cause of nition of the term as generally understood in the art. As used worldwide mortality. There is a strong need for new effective herein, certain terms may have the following defined mean methods of therapy for prevention and treatment of heart ings. failure and incipient heart failure. The present invention pro 0014. As used herein, the singular form “a” “an and “the vides methods for preventing and treating heart failure and include singular and plural references unless the context incipient heart failure by administering monoamine oxidase clearly dictates otherwise. (MAO) inhibitors. 0015. As used herein, "compensated hypertrophy' refers to abnormal chamber function or enlargement of the heart SUMMARY OF THE INVENTION coincident with an increase in muscle mass. For example, in 0007. This invention is directed towards novel methods for cardiac disease, the compensation for the inefficiency of the preventing and treating heart failure, incipient heart failure, heart's pump action is by enlisting the various reserves of the and associated conditions, symptoms, signs, and disorders. In heart such as hypertrophy, enlargement, or increase in rate so one aspect, the invention provides a method of treating heart as to maintain circulatory equilibrium and prevent the appear failure in a in need thereof, which method comprises ance of the signs of congestive heart failure. administering to said mammal diagnosed with heart failure a 0016. As used herein, the term “comprising is intended to therapeutically effective amount of an MAO inhibitor. In mean that the compositions and methods include the recited another aspect, the invention provides a method of preventing elements, but not excluding others. “Consisting essentially heart failure in a mammal in need thereof, which method of when used to define compositions and methods, shall comprises administering to said mammal diagnosed with a mean excluding other elements of any essential significance susceptibility to heart failure a therapeutically effective to the composition or method. “Consisting of shall mean amount of a MAO inhibitor. excluding more than trace elements of other ingredients for 0008. In another aspect, the invention provides a method claimed compositions and Substantial method steps. Embodi of inhibiting incipient heart failure in a mammal in need ments defined by each of these transition terms are within the thereof, which method comprises administering to said mam Scope of this invention. Accordingly, it is intended that the mal diagnosed with incipient heart failure a therapeutically methods and compositions can include additional steps and effective amount of a MAO inhibitor. In another aspect, the components (comprising) or alternatively including steps and invention provides a method of treating incipient heart failure compositions of no significance (consisting essentially of) or in a mammal in need thereof, which method comprises alternatively, intending only the Stated method steps or com administering to said mammal diagnosed with incipient heart positions (consisting of). failure atherapeutically effective amount of a MAO inhibitor. 0017. As used herein, “cardiomyocytes' or “cardiac myo 0009. A variety of well known methods are useful for cytes' are specialized muscle cells which form the myocar diagnosing heart failure or incipient heart failure, their signs dium of the heart. Cardiomyocytes have five major compo and symptoms, and mammalian Subject Susceptible to Such nents: cell membrane (sarcolemma) and T-tubules for conditions. A variety of MAO inhibitors are useful for prac conduction; sarcoplasmic reticulum for contraction; ticing the methods of the present invention. Also well known contractile elements; mitochondria; and a nucleus. US 2015/0011643 A1 Jan. 8, 2015

0018. As used herein, “heart failure (HF) or “congestive 0023. As used herein, “mammals” include, but are not heart failure (CHF), refers to a condition in which the heart limited to, murines, rats, simians, humans, farm animals, can not pump enough blood to the body's other organs due to, sport animals and pets. for example, heart muscle malfunction, weakening of the 0024. As used herein, "monoamine oxidases” are heart muscle called cardiomyopathy, and other heart muscle that catalyze the oxidation of monoamines. Two related reasons. Congestive heart failure (CHF) is character monoamine oxidase (MAO) isoenzymes, MAO-A and MAO ized, among other effects, by left ventricle (LV) chamber B, are closely linked in opposite orientation on the X chro dilation, decreased LV contractility and elevated levels of mosome and are expressed in the outer mitochondrial mem circulating catecholamines. In another aspect, congestive brane. In vivo, MAO-A and MAO-B oxidize monoamine heart failure occurs due to ischemic and other reperfusion, neurotransmitters and dietary monoamines, the regulation of and other non-ischemic factors. Heart failure includes, but is which is important in maintaining normal mental states. not limited to, the following symptoms or signs: cardiac rep MAO-A prefers , , and as erfusion injury, compensated hypertrophy, human end stage substrates, whereas MAO-B prefers phenylethylamine and heart failure, hypertensive cardiomyopathy, left ventricular trace amines. These proteins have been sequenced and char , left or right ventricular dilation, left or right acterized, see for example, the National Center for Biotech Ventricular failure, maladaptive hypertrophy, myocardial nology Information (NCBI) GenBank Accession Nos. for structural disarrangement (apoptosis and loss of cardiomyo MAO-A gi572841 14|emb|CAI43.120.1||57284.114): cyte) and myocardial dysfunction (loss in contraction and/or gi57209563 emb|CAI42421.157209563: relaxation), and pressure overloaded heart. gi4557735|refNP 000231.1||4557735); 0019. As used herein, “incipient heart failure' refers to the gil 544023201gb|AAV34720.154402320): early and/or mild appearance of symptoms, precursors or gil 54402314|gb|AAV34717.154402314): signs of heart failure. For example, and without limitation, gil 544023021gb|AAV34711.154402302: humans and Such other mammals suffering from incipient gil 54402290gb|AAV34705.154402290; heart failure may show no symptoms of heart failure at rest gi57284213|emb|CAI43216.157284213) O and only mild symptoms of heart failure while exercising. gi57209566emb|CAI42424.157209566 and the Gen However, incipient heart failure can lead to full fledged or end Bank Accession NoS. for MAO-B stage heart failure. gi57209948 emb|CAI42522.157209948: 0020. As used herein, “left ventricular hypertension gil 187376Igb|AAA59551.1||187376): (LVH) is a condition wherein the cardiac muscle responds to gi38202207 refNP 000889.3 ||38202207): increased resistance in the circulation by becoming enlarged. gi57209564 emb|CAI42422.157209564: However, with time, the fibers of the hypertrophied heart gi57208148 emb|CAD92552.2157208148); muscle become thickened and shortened, and consequently gil 18490291 Igb|AAH22494.1 18490291; less able to relax. Hypertension makes the myocardium work gil 553527 Igb|AAB46386.1553527 O harder. The resulting hypertrophy is the product of the thick gil 187359gb|AAA59550.1||187359. ening and shortening of the muscle fibers of the heart. Under 0025. As used herein, "monoamine oxidase inhibitor” or these conditions, it becomes more difficult for the heart to "MAO inhibitor” refers to a compound that acts by inhibiting relax and go through the normal cycle of contraction and the activity of monoamine oxidase, including MAO-A and/or relaxation. Changes in the myocardium appearin the collagen MAO-B. In one aspect, MAO inhibitors prevent the break resulting in increased stiffness. The outcome of this process is down of monoamine neurotransmitters thereby increasing a heart that is less able to meet the output demands of normal their in vivo availability. In another aspect, MAO inhibitors circulation. There is an impaired diastolic relaxation, but also prevent the catabolism of dietary monoamines. In yet another heightened Vulnerability to ischemic events. aspect, MAO inhibitors prevent generation of reactive oxygen 0021. As used herein, “left ventricular dilation” refers to a species (ROS). MAO inhibitors are well known in the art and left ventricular enlargement, which can increase the Volume are used for the treatment of neurodegenerative diseases. of blood that is ejected from the ventricle, temporarily Examples of MAO-A inhibitors include, but are not limited improving cardiac output. This increase in size of the ven to, Clorgyline, , and the reversible MAO-A inhibi tricular cavity, however, also results in a reduction of the tors , , , , percentage of the left ventricular volume of blood that is , and . Examples of effected (called ejection fraction) and has significant physi MAO-B inhibitors include, but are not limited to, , ological implications. “Left ventricular dilation' is a well and . Examples of unselective MAO-A recognized precursor and sign of Ventricular dysfunction and and MAO-B inhibitors include, but are not limited to, Ipro congestive heart failure after myocardial infarction. Simi clozide (Sursum), (Marsilid, Iprozid, Ipronid, larly, “right ventricular dilation” refers to a right ventricular Rivivol, Propilniazida), (Marplan), Mebana enlargement and associated signs or disorder. zine (Actomol), Metfendrazine (H.M.-11), (Nia 0022. As used herein, “left ventricular failure” refers to a mid), (Nardil), (Catron), Phenox disorder where the left side of the heart fails to pump blood ypropazine (Drazine), Pivalylbenzhydrazine (Tersavid, effectively. This results in a back flow, pressure and/or con Neomarsilid), (Safra) and (Par gestion of blood into the lungs. Signs indicating "left ven nate). See, for example, Remington: The Science and Practice tricular failure' include a laterally displaced apex beat (which of Pharmacy, 21 Edition, (2005, Lippincott Williams & occurs if the heart is enlarged). A gallop rhythm (additional Wilkins), pages 1517-1523, and Physicians’ Desk Reference, heart sounds) may be heard as a marker of increased blood Edition 60 (2006, Thomson PDR) page 1499 (each of which flow, or increased intra-cardiac pressure. Similarly, "right is incorporated herein by reference). ventricular failure' refers to a sign or disorder where the right 0026. As used herein, "preventing or “prevention' of a side of the heart fails to pump blood effectively. disease, disorder, symptom or condition means that the onset US 2015/0011643 A1 Jan. 8, 2015

of the disease, disorder, symptom or condition in a mammal inhibitors have been used for treating patients with Parkin predisposed thereto is prevented Such that the mammal does Sons and Alzheimer's disease. not manifest the disease, disorder, symptom or condition. 0033. In contrast, less attention has been given to the 0027. As used herein, a “therapeutically effective amount byproducts of MAO’s monoamine catabolism. The monoam or an “effective amount” is used synonymously with and ine catabolism byproducts generated by MAO are aldehydes, intends an amount sufficient to effect beneficial or desired ammonia and H2O. H.O. is a toxic, reactive oxygen species, results. An effective amount can be administered in one or '' or can generate toxic hydroxyl radicals in the presence of more administrations, applications, or dosages. Fe2. 0028. As used herein, “treating or “treatment of a dis 0034. In the heart, MAO-A is the prevalent isoform, and ease, disorder, symptom or condition will depend on the the majority of the is situated in the cardiomyo disease, disorder, symptom or condition to be treated, and the cytes'. In the peripheral tissues, MAO protects the body by mammal to be treated. In general, treatment intends one or oxidizing blood-derived amines or by preventing their entry more of inhibiting the progression of the manifested disease, into the bloodstream (FIG. 2). disorder, symptom or condition as measured by clinical or sub-clinical parameters (where the term “inhibiting or “inhi Therapies bition' is intended to be a subset of “treating or “treatment'), 0035. The present invention relates in part to the discovery arresting the development of the disease, disorder, symptom that MAO-A and MAO-B have major implications in the or condition as measured by clinical or sub-clinical param progression of compensated hypertrophy to left ventricular eters, ameliorating or causing regression of the disease, dis dilation and failure. Accordingly, the present invention pro order, symptom or condition as measured by clinical or Sub vides that inhibition of MAOs, such as MAO-A, effectively clinical parameters, or reducing pain or discomfort for the prevents or retards this progression, improves myocardial mammal treated as measured by clinical and/or pharmaco function, and may also prevent loss of viable myocardial cells logical parameters. "Treating does not include preventing due to increased oxidative stress and pro-apoptotic signaling. the onset of the disease or condition. 0036. In one aspect, the invention provides a method of treating heart failure in a mammal in need thereof, which Monoamine Oxidase (MAO) and Its Role In Heart Failure. method comprises administering to said mammal diagnosed 0029 Monoamine oxidases (MAO) are flavoenzymes with heart failure a therapeutically effective amount of an located within the outer mitochondrial membrane. MAOs are MAO inhibitor. In another aspect, the invention provides a responsible for oxidative deamination of monoamino neu method of preventing heart failure in a mammal in need rotransmitters and dietary monoamines'. These exist in two thereof, which method comprises administering to said mam different isoforms, MAO-A (FIG. 2) and MAO-B', sharing mal diagnosed with a Susceptibility to heart failure a thera 70% sequence homology. These are distinguished by differ peutically effective amount of a MAO inhibitor. ent substrate affinity and inhibitor sensitivity. 0037. In another aspect, the invention provides a method 0030. MAO-A preferentially catalyses the oxidative of inhibiting incipient heart failure in a mammal in need deamination of norepinephrine (NE) and serotonin (5-HT) thereof, which method comprises administering to said mam and is inhibited by low concentrations of clorgyline. MAO-B mal diagnosed with incipient heart failure a therapeutically has affinity for phenylethylamine and benzylamine, and is effective amount of a MAO inhibitor. In another aspect, the inhibited by selegiline. Both isoforms catalyze the deamina invention provides a method of treating incipient heart failure tion of dopamine, , and and in a mammal in need thereof, which method comprises are inhibited by pargyline. Deletion of MAO-A and MAO-B administering to said mammal diagnosed with incipient heart genes has proven their role in failure atherapeutically effective amount of a MAO inhibitor. and behavior. 0038. In certain embodiments, the MAO-A (or-B) inhibi 0031 MAO-A null mice have elevated brain levels of tors administered in accordance with the invention are those, 5-HT, NE, and, to a lesser extent, dopamine. 2-Phenylethy which, when administered, do not give rise to the potentially lamine is increased in MAO-B knockout (KO) mice". Com harmful side-reaction called "cheese-effect.” Cheese-effect pulsive-aggressive behavior results from lack of MAO-A refers to hypertensive crises in patients under treatment with function both in humans and mice". It appears that MAO-A MAO inhibitors following the ingestion of food rich in is active during development since this effect can be mim tyramine (chocolate, cheese, etc)' decompensated, mal icked by the administration of MAO inhibitors during the adaptive cardiac hypertrophy and functional failure. Thus, in early postnatal period. However, these studies report that certain embodiment, the MAO inhibitors administered in MAO may not be essential for survival". accordance with the present invention exclude irreversible 0032. The role of MAO in terminating the actions of neu MAO inhibitors that cause "cheese-effect.” In certain other rotransmitters/dietary amines in central and peripheral ner embodiments, the MAO inhibitors administered in accor Vous system (and in the extraneuronal tissue) has been dance with the present invention are the reversible MAO-A or reported. MAO inhibitors are used for treating neurodegen RIMA inhibitors. Such MAO inhibitors are well known to erative diseases and affective disorders. MAO-A inhibitors one of skill in the art. are particularly effective in the treatment of depression by 0039. In certain embodiments, the methods of the inven increasing brain levels of dopamine, NE and 5-HT. MAO-B tion exclude methods of preventing postischemic oxidative US 2015/0011643 A1 Jan. 8, 2015

stress and myocyte hypertrophy. In certain other embodi dilation, reduced basal and beta-stimulated contractility and ments, the methods of the invention include methods of pre depressed overall LV function. venting postischemic oxidative stress and myocyte hypertro phy. TABLE 1 0040. The invention having been described in summary, in MAO-A gene expression in mouse detail, and by the accompanying figure, is exemplified, and hearts after 6 weeks of TAC. not limited, by the following examples which demonstrate the MAO-AGAPDH usefulness of administering MAO inhibitors for the preven Sham tion and treatment of heart failure, incipient heart failure, and TAC 6 weeks one or more of their signs and symptoms.

EXAMPLES 0043. The effects of MAO-A inhibition under pharmaco 0041. The practice of the present invention employs, logical conditions on the left ventricular hypertension (LVH) unless otherwise indicated, conventional techniques of development were also tested. As shown in Table 2, after 3 molecular biology (including recombinant techniques), weeks of TAC, wall thickness was increased in Saline- and microbiology, cell biology, biochemistry, , clorgyline-treated mice (CLO, 1 mg/kg/day, n=6) when com immunology, and chemistry, which are well within the skill of pared to sham operated animals, likely reflecting a compen on of art. Such techniques are explained fully in the literature sation for the increase in pressure. However, LV function was for example in the following publications. See, e.g., Sam significantly better in CLO-treated group (CLO-mice): both brook and Russell eds. MOLECULAR CLONING: A fractional shortening and ejection fraction were comparable LABORATORY MANUAL, 3' edition (2001); the series to values reported for control mice (sham). At 6 weeks of CURRENT PROTOCOLS IN MOLECULARBIOLOGY (F. TAC, differences between the two groups became even more M. Ausubel et al. eds. (2007)); the series METHODS IN pronounced. The CLO-mice displayed improved LV func ENZYMOLOGY (Academic Press, Inc., N.Y.); PCR 1: A tion: fractional shortening was 30+5.4 61+0.8% , and PRACTICAL APPROACH (M. MacPherson et al. IRL Press ejection fraction 60.2+8.2 vs 93.6+0.4% (Table 2). TABLE 2 Time-dependent changes in cardiac morphology and in vivo Ventricular function induced by TAC in control and clorgyline treated mice. IVS: interventricular septum, LVEDD: left ventricular end-diastolic dimension, LVESD: left ventricular end-systolic dimension, LVPW: left ventricular posterior wall, FS: fractional shortening, EF: ejection fraction, BW: body weight. TSW. TAC 3 weeks, TOW: TAC 6 weeks, CLO. clorgyline. Sham T3w T3w - CLO T6w T6w - CLO BW (g) 27.21.3 25.4 - 0.6 25.60.6 26.6 0.7 27.10.4 Heart rate (bpm) 738 + 11.2 670 + 11. 702 + 12 627.5 +24. 670 13.7 IVS (mm) O.8 O.O2 1.3 + 0.04 1.3 + 0.02 1.2 : 0.04 1.4 + 0.03 LVEDD (mm) 3.2, O.O2 2.90.2 2.9 OO6 4.O. O.2 3.0 + 0.08° LVESD (mm) 12 O.O2 17 O.2 12 OO6 2.9 + 0.4 1.2 + 0.04? LVPW (mm) O.8 O.O2 1.3 + 0.04 1.2 + 0.03 1.1 + 0.05 13 O.O3. P FS (%) 63.5 + 0.9 43.9 +3.2 56.8 + 1.9, 30.6+ 5.3 61.6+ 0.9 EF (%) 94.70.4 81 + 7.9- 91...6+ 0.4 62.4 + 7.9 94.3 + 0.4° LV mass/BW (mg/g) 28 0.1 6.9 + 0.4 6.4 + 0.3 9.3 + 0.5 7.2 - 0.48. P “p < 0.05 vs sham, p <0.001 vs sham, p < 0.05 vs TAC 3 weeks, Pp < 0.05 ws TAC 6 weeks, at Oxford University Press (1991)); CULTURE OF ANIMAL 0044) Moreover, the CLO-mice showed reduced levels of CELLS: A MANUAL OF BASICTECHNIQUE (R.I. Fresh hypertrophy: LV mass, left ventricular end diastolic dimen ney 5' edition (2005)); and GOOMAN AND GILLMAN'S sion (LVEDD), and end systolic dimension (LVESD) were all THE PHARMACOLOGICAL BASIS OF THERAPEU significantly reduced (Table 2). These results demonstrate (Brunton et al. McGraw Hill Publishing (2005)). that inhibiting MAO-A by administering a therapeutically 1. In vivo Administration of an MAO Inhibitor Prevents Car effective amount of an MAO inhibitor prevents or reduces diac Decompensation after 6 Weeks of Transverse Aortic maladaptive LV hypertrophy, chamber dilation, and LV dys Constriction (TAC). function in pressure-overloaded hearts, and is thus useful in 0042. MAO as a source of ROS in cardiac reperfusion practicing the methods of the invention. injury' and its role in in vitro pro-hypertrophic effects of 2. Administration of the MAO-A Inhibitor Clorgyline Results serotonin has been explored'. In this experiment, MAO's in Reduced Levels of Apoptosis in Heart Cells after 6 Weeks expression, activity, and role in congestive heart failure of TAC. (CHF) was tested in vivo. The MAO-A gene expression was 0045. The MAO-A inhibitor clorgyline impacted the pro 3.5-fold higher in C57B16 mice after 6 weeks of TAC-in apoptotic signaling cascade as demonstrated below. Levels of duced pressure overload (T6W. n=6, Table 1). TAC-induced cleaved caspase-3 were markedly elevated after 6 weeks in pressure overload is a condition associated with chamber TAC mice vs shams. Clorgyline treatment prevented US 2015/0011643 A1 Jan. 8, 2015 caspase-3 activation demonstrating that loss of viable cardi 0048 Preincubation with clorgyline prior to tyramine omyocytes is a major contributor to impaired ventricular treatment reduced this effect significantly. function and onset of chamber dilation (FIG. 1). 0049. These results demonstrate that MAO activity in the heart can trigger the pro-hypertrophic response via activation 3. MAO-A Expression is Induced by Pro-Hypertrophic of pathways promoting maladaptive hypertrophy and cause Stimuli and the Up-Regulation of MAO-A Triggers heart failure; administration of an MAO-A inhibitor reduces Hypertrophic Signaling. these symptoms, and is therapeutically beneficial. 0046) 4. MAO-A is a Major Source of Reactive Oxygen Species (ROS) and Inhibition of MAO-A Prevents Oxidative Stress in TABLE 3 Cardiomyocytes. MAO-A and BNP gene expression after treatment with NE in the absence or presence of 0050. This example demonstrates that the administration clorgyline. of MAO inhibitors prevents oxidative stress in cardiomyo cytes and is useful in preventing and treating heart failure or MAO-A BNP an associated symptom or condition in accordance with the (vs. GAPDH) (vs. GAPDH) invention. In vivo, mitochondria and the respiratory chain, in control O.S8 O.OS 1 - O particular, are considered a major intracellular ROS source'. NE 1.85 O.O4* 8.9 1.4 NE- 4.85 + 0.9*" MAOs are located in the outer membrane of these organelles clorgyline and MAO-A can increase oxidative burden. When HL-1 car diomyocytes were incubated with 100 uM HO or 5 uM *p < 0.05 vs control, arachidonic acid, a rise in ROS production at mitochondrial "p < 0.05 ws NE. level was observed (Table 6). 0047. This experiment demonstrates a causative link between pro-hypertrophic agents and MAO-A. Incubation of TABLE 6 neonatal rat cardiomyocytes with NE (10 uM) for 24 hrs Levels of oxidative stress upon stimulation with H2O2 or resulted in up-regulation of MAO-A gene expression and arachidonic acid in the absence or presence of deprenyl, induction of hypertrophy, as measured by an increase in brain clorgyline or in siRNA treated cells. natriuretic peptide (BNP) expression (Table 3). MAO-A inhi H2O2 Arachidonic acid bition partially reduced NE induced hypertrophy, suggesting (% increase vs (% increase vs that this hormone is transported into the cell and degraded by control) control) MAO-A while H2O is produced. This process may contrib No treatment 156.24.3 145.638 ute to the development of +deprenyl 150.7 - 4.9 1424 - 0.3 +clorgyline 122.63* 98.3 1.3* TABLE 4 +MAO-A 1223 O.4* 99 2.7 siRNA EMT gene expression in mouse hearts upon stimulation with tyramine. *p < 0.05 vs no treatment, EMT, GAPDH 0051. The rise in ROS production was completely pre vented, or reduced, by cell pretreatment with clorgyline (1 control O.S. O.04 uM). No protection was observed with MAO-B inhibitor tyramine O.97 O.2* selegiline. These results were also confirmed in siRNA *p < 0.05 vs control. treated cells in which MAO-A expression was reduced by 90%. hypertrophy. Exposing myocytes to another MAO Substrate, tyramine (10 uM), resulted in a 2-fold increase in EMT gene REFERENCE LIST expression (Table 4), the main monoamine transporter present at the cardiomyocyte level. A direct link between 0052 (1) Kumar M J, Nicholls DG, Andersen J. K. Oxi MAO-A activity and myocyte hypertrophy was further cor dative alpha-ketoglutarate dehydrogenase inhibition via roborated by the fact that tyramine incubation of cardiomyo subtle elevations in levels results in cytes resulted in 2-fold increase of NFAT4 expression, a loss of spare respiratory capacity: implications for Parkin transcription factor involved in maladaptive hypertrophy son's disease. J Biol Chem 2003 Nov. 21; 278(47):46432 (Table 5). 9 TABLE 5 0053 (2) Shih J C, Chen K. Regulation of MAO-A and MAO-B gene expression. Curr Med Chem 2004 August; NFAT4 gene expression (normalized to GAPDH) upon stimulation with tyramine in the 11(15): 1995-2005. absence or presence of clorgyline. 0054 (3) Youdim M. B. Finberg J. P. New directions in and B selective inhibitors and sub -clorgyline +clorgyline strates. Biochem Pharmacol 1991 Jan. 15:41(2):155-62. Control O.24 O.O3 O.23 O.O2 0055 (4) Youdim M B, Edmondson D, Tipton K F. The Tyramine O.38 O.O1* 0.32 + 0.02" therapeutic potential of monoamine oxidase inhibitors. Nat *p < 0.05 vs control, Rev Neurosci 2006 April; 7(4):295-309. "p < 0.05 vs tyramine. 0056 (5) Brunner H.G., Nelen M. Breakefield XO, Ropers H H, van Oost BA. Abnormal behavior associated with a US 2015/0011643 A1 Jan. 8, 2015

point mutation in the structural gene for monoamine oxi 0065 (14) Bianchi P. Pimentel DR, Murphy MP, Colucci dase A. Science 1993 Oct. 22:262(5133):578-80. WS, Parini A. A new hypertrophic mechanism of serotonin 0057 (6) Shih.J.C. Cloning, after cloning, knock-out mice, in cardiac myocytes: receptor-independent ROS genera and physiological functions of MAO-A and B. Neurotoxi tion. FASEBJ 2005 April; 19(6):641-3. cology 2004 January; 25(1-2):21-30. 0066 (15) Di Lisa, Canton M, Menabo R, Kaludercic N, 0058 (7) Lenders J. W. Eisenhofer G, Abeling NG et al. Bernardi P. Mitochondria and cardioprotection. Heart Fail Specific genetic deficiencies of the A and B isoenzymes of Rev 2007 December; 12(3-4):249-60. monoamine oxidase are characterized by distinct neuro 0067 (16)Youdim M. B. Weinstock M Therapeutic appli chemical and clinical phenotypes. J Clin Invest 1996 Feb. cations of selective and non-selective inhibitors of 15;97(4): 1010-9. monoamine oxidase A and B that do not cause significant 0059 (8) Billett E. E. Monoamine oxidase (MAO) in tyramine potentiation. Neurotoxicology. 2004 January; human peripheral tissues. Neurotoxicology 2004 January; 25(1-2):243-50. 25(1-2): 139-48. 0068 Throughout this disclosure, various publications, 0060 (9)Yang L, Omori K. Suzukawa J., Inagaki C. Cal patents and/or published patent specifications are referenced cineurin-mediated BAD Ser155 dephosphorylation in by an identifying citation. The disclosures of these publica ammonia-induced apoptosis of cultured rat hippocampal tions, patents and published patent specifications are hereby neurons. Neurosci Lett 2004 Feb. 26:357(1):73-5. incorporated by reference into the present disclosure to more 0061 (10) Halliwell B. Reactive oxygen species and the fully describe the state of the art to which this invention central nervous system. J Neurochem 1992 November; pertains. 59(5):1609-23. What is claimed is: 0062 (11) Lamensdorf I, Eisenhofer G, Harvey-White J, 1. A method of treating heart failure in a mammal in need Hayakawa Y. Kirk K. Kopin I J. Metabolic stress in PC12 thereof, which method comprises administering to said mam cells induces the formation of the endogenous dopaminer mal diagnosed with heart failure a therapeutically effective gic neurotoxin, 3,4-dihydroxyphenylacetaldehyde. J Neu amount of a monoamine oxidase inhibitor. rosci Res 2000 May 15; 60(4):552-8. 2. A method of inhibiting incipient heart failure in a mam 0063 (12) Partanen S, Penttila O, Kyosola K, Merikallio mal in need thereof, which method comprises administering E. Siltanen P. Histochemically demonstrable monoamine to said mammal diagnosed with incipient heart failure athera oxidase activity in the adult human heart in various cardiac peutically effective amount of a monoamine oxidase inhibi diseases. Virchows Arch A Pathol Anat Histol 1976 Jul. 21; tOr. 370(4):291-6. 3. A method of treating incipient heart failure in a mammal 0064 (13) Bianchi P. Kunduzova O, Masini E et al. Oxi in need thereof, which method comprises administering to dative stress by monoamine oxidase mediates receptor said mammal diagnosed with incipient heart failure a thera independent cardiomyocyte apoptosis by serotonin and peutically effective amount of a monoamine oxidase inhibi postischemic myocardial injury. Circulation 2005 Nov. 22: tOr. 112(21):3297-305.