WO2019068326A1 - Inhibiteurs de la lsd1 pour le traitement et la prévention de cardiomyopathies - Google Patents

Inhibiteurs de la lsd1 pour le traitement et la prévention de cardiomyopathies Download PDF

Info

Publication number
WO2019068326A1
WO2019068326A1 PCT/EP2017/075318 EP2017075318W WO2019068326A1 WO 2019068326 A1 WO2019068326 A1 WO 2019068326A1 EP 2017075318 W EP2017075318 W EP 2017075318W WO 2019068326 A1 WO2019068326 A1 WO 2019068326A1
Authority
WO
WIPO (PCT)
Prior art keywords
cardiomyopathy
lmna
lsdl
inhibitor
lsdl inhibitor
Prior art date
Application number
PCT/EP2017/075318
Other languages
English (en)
Inventor
Michel Puceat
Imen JEBENIANI
Gisèle Jeanne Liliane BONNE
Anne-Claire Irène Marie GUENANTIN
Nicolas VIGNIER
Original Assignee
Université D'aix-Marseille
Centre National De La Recherche Scientifique - Cnrs -
INSERM (Institut National de la Santé et de la Recherche Médicale)
Universite Pierre Et Marie Curie (Paris 6)
Association Institut De Myologie
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Université D'aix-Marseille, Centre National De La Recherche Scientifique - Cnrs -, INSERM (Institut National de la Santé et de la Recherche Médicale), Universite Pierre Et Marie Curie (Paris 6), Association Institut De Myologie filed Critical Université D'aix-Marseille
Priority to PCT/EP2017/075318 priority Critical patent/WO2019068326A1/fr
Publication of WO2019068326A1 publication Critical patent/WO2019068326A1/fr

Links

Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/13—Amines
    • A61K31/135—Amines having aromatic rings, e.g. ketamine, nortriptyline
    • A61K31/137—Arylalkylamines, e.g. amphetamine, epinephrine, salbutamol, ephedrine or methadone
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A61K31/44—Non condensed pyridines; Hydrogenated derivatives thereof
    • A61K31/445—Non condensed piperidines, e.g. piperocaine
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A61K31/44—Non condensed pyridines; Hydrogenated derivatives thereof
    • A61K31/445—Non condensed piperidines, e.g. piperocaine
    • A61K31/4468—Non condensed piperidines, e.g. piperocaine having a nitrogen directly attached in position 4, e.g. clebopride, fentanyl
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
    • A61K45/06—Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P9/00—Drugs for disorders of the cardiovascular system
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
    • C12N15/113—Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing
    • C12N15/1137—Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing against enzymes
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2310/00—Structure or type of the nucleic acid
    • C12N2310/10—Type of nucleic acid
    • C12N2310/14—Type of nucleic acid interfering N.A.
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Y—ENZYMES
    • C12Y114/00—Oxidoreductases acting on paired donors, with incorporation or reduction of molecular oxygen (1.14)
    • C12Y114/11—Oxidoreductases acting on paired donors, with incorporation or reduction of molecular oxygen (1.14) with 2-oxoglutarate as one donor, and incorporation of one atom each of oxygen into both donors (1.14.11)

Definitions

  • Cardiomyopathy is a group of anatomic and pathologic conditions associated with heart muscle dysfunction of the heart, damaging the muscle tone of the heart and thus reducing its ability to pump blood to the rest of the body. Cardiomyopathy is the leading cause of heart failure and at the late stage requires heart transplant. It differs from other heart problems in that it frequently affects younger people. Cardiomyopathies, which can be acquired or congenital, have many causes, symptoms, and treatments. The main types of cardiomyopathy are dilated cardiomyopathy, hypertrophic cardiomyopathy, restrictive cardiomyopathy, arrhythmogenic right ventricular dysplasia and unclassified cardiomyopathy.
  • Dilated cardiomyopathy is the most common form of cardiomyopathy. It is characterized by an increase in both myocardial mass and volume, which compromises cardiac contractibility and ultimately results in reduced left ventricular function. Dilated cardiomyopathy can lead to heart failure, heart valve disease, arrhythmia (irregular heart rate), and blood clots in the heart. Dilated cardiomyopathy may result from excess alcohol consumption, heavy metals exposure, ***e use, inflammation or viral infections. However, genetically inherited forms of dilated cardiomyopathy have been identified in 25-35% of patients presenting with this disease.
  • Inherited dilated cardiomyopathies are caused by mutations in genes that encode components of a wide variety of cellular components and pathways, including the nuclear envelope ⁇ e.g., LMNA, EMD), contractile apparatus ⁇ e.g., MYH7, ACTC1, TPM1, TIN), force transduction apparatus e.g., MLP, DES, TNNT2), and calcium handling ⁇ e.g., SERCA) (Bonne and Muchir, Chapter 7, in "Cardiology and Cardiovascular Medicine - Cardiomyopathies: Types and Treatments” , Kaan Kirali, 2017).
  • LMNA nuclear envelope
  • EMD contractile apparatus
  • MYH7 e.g., MYH7, ACTC1, TPM1, TIN
  • force transduction apparatus e.g., MLP, DES, TNNT2
  • SERCA calcium handling
  • the LMNA gene which encodes A-type nuclear lamins, may be the most prevalent dilated cardiomyopathy causative gene, as LMNA mutations appear to account for about 5-10% of inherited dilated cardiomyopathy.
  • mutations of the LMNA gene are also known to be implicated in a wide spectrum of disorders, referred to as laminopathies, which include skeletal muscle dystrophies ⁇ e.g., Autosomal Emery- Dreifuss muscular dystrophy), - - lipodystrophies (e.g., Dunnigan-type familial partial lipodystrophy), peripheral neuropathy (e.g., Charcot-Marie-Tooth disorder type 2B1) and premature ageing (progerias).
  • laminopathies which include skeletal muscle dystrophies ⁇ e.g., Autosomal Emery- Dreifuss muscular dystrophy), - - lipodystrophies (e.g., Dunnigan-type familial partial lipodystrophy), peripheral neuropathy (e.g
  • LMNA dilated cardiomyopathy is characterized by cardiac dilatation and impaired systolic function. It usually presents in early to mid-adulthood with symptomatic conduction system disease generally accompanied with arrhythmias or with symptomatic dilated cardiomyopathy. In LMNA dilated cardiomyopathy, which is known to have a more aggressive course than other inherited dilated cardiomyopathies, arrhythmias gradually become more frequent with age, potentially leading to sudden death.
  • the present invention identifies the lysine (K)-specific demethylase, LSD1, as a new therapeutic target in cardiomyopathies, in particular in cardiomyopathies associated with mutations of lamins A/C genes. More specifically, as described in the Examples section below, in order to better understand the origin of the disease and to identify potential pharmacological targets, the present Inventors have used mouse embryonic stem cells harboring the p.H222P LMNA mutation, which faithfully recapitulate early embryonic development, including cardiogenesis, and constitute a valuable model to study lamin- associated development defects. The cardiac differentiation process was observed to be blocked at an early stage of development pointing to a defect in the process of epithelial-to-mesenchymal transition (EMT).
  • EMT epithelial-to-mesenchymal transition
  • the Inventors used the - - homozygous H222P LM/VA mouse model, which recapitulates the dilated cardiomyopathy observed in patients, and found that cardiomyopathy was already detectable at the foetal stage pointing to an early onset of the disease. Indeed, echocardiography carried out in utero, revealed a defect in contractibility of the left ventricle in E13.5 mouse embryos. Using cardiac cell derivatives of pluripotent stem cells and a chomatin immunoprecipitation assay, the Inventors found a defect in histone mark H3K4mel in regulatory regions of genes playing a role in EMT during cardiogenesis, which accounted for the defect in EMT in mutant cells.
  • the present invention relates to an LSDl inhibitor for use in a method of treatment or prevention of a cardiomyopathy.
  • the present invention relates to the use of an LSDl inhibitor in the manufacture of a medicament for the treatment or prevention of a cardiomyopathy.
  • the present invention relates to a method for treating or preventing a cardiomyopathy in a subject in need thereof, the method comprising a step of administering to said subject, an effective amount of an LSDl inhibitor, or of a pharmaceutically acceptable composition thereof.
  • the cardiomyopathy is a dilated cardiomyopathy.
  • the cardiomyopathy is an LM/VA-related cardiomyopathy.
  • the LM/VA-related cardiomyopathy may be an LM/VA-related dilated cardiomyopathy.
  • the LMVA-related cardiomyopathy may alternatively be selected from the group consisting of arrhythmo genie right ventricular dysplasia/cardiomyopathy, an inherited form of early onset of myocardial fibrosis, and left ventricular non-compaction.
  • the LM/VA-related cardiomyopathy may be selected from the group consisting of cardiomyopathies secondary to a skeletal muscle dystrophy or - - lipodystrophy in a laminopathy.
  • the laminopathy may be Emery-Dreifuss muscular dystrophy (EDMD), limb-girdle muscular dystrophy type IB (LGMD1B), Dunnigan- type familial partial lipodystrophy 2, and progeria.
  • the LSDl inhibitor is an irreversible LSDl inhibitor or a reversible LSDl inhibitor.
  • the LSDl inhibitor is a small molecule, or a physiologically acceptable salt thereof.
  • the small molecule LSDl inhibitor may be selected from the group consisting of GSK-LSD1, ORY-1001, ORY-2001, ORY-3001, GSK-2879552, INCB-059872, IMG-7289, CC-90011, RN-1, RN-7, GSK-2699537, GSK-2879552, 4SC-202 and T-3775440.
  • the small molecule LSDl inhibitor is selected from GSK-LSD1, GSK-2879552, and ORY-1001.
  • the LSDl inhibitor is an SiRNA.
  • the present invention provides a pharmaceutical composition for use in the treatment or prevention of a cardiomyopathy, wherein said pharmaceutical composition comprises an effective amount of an LSDl inhibitor and a pharmaceutically acceptable carrier or excipient.
  • the LSDl inhibitor comprised in a pharmaceutical composition according to the present invention may be as described above, an irreversible LSDl inhibitor or a reversible LSDl inhibitor. It may be a small molecule (such as GSK-LSD1, ORY- 1001, ORY-2001, ORY-3001, GSK-2879552, INCB-059872, IMG-7289, CC-90011, RN-1, RN-7, GSK-2699537, 4SC-202 or T-3775440) or an SiRNA.
  • a pharmaceutical composition according to the present invention further comprises at least one additional therapeutic agent.
  • the additional therapeutic agent may be selected from the group consisting of angiotensin-converting enzyme (ACE) inhibitors, angiotensin II receptor blockers (ARBs), beta blockers, cardiac glycosides, diuretics, antiarrhythmics, inotropic agents, and anticoagulants.
  • ACE angiotensin-converting enzyme
  • ARBs angiotensin II receptor blockers
  • beta blockers beta blockers
  • cardiac glycosides diuretics
  • antiarrhythmics inotropic agents
  • anticoagulants anticoagulants
  • the present invention relates to a kit for use in the treatment or prevention of a cardiomyopathy, comprising at least one LSDl inhibitor, or a pharmaceutical composition thereof, and at least one additional therapeutic agent, wherein the at least one LSDl inhibitor and at least one additional therapeutic agent are comprised in separate containers.
  • the LSD1 inhibitor, or pharmaceutical composition thereof, and the at least one additional therapeutic agent may be as described herein.
  • Figure 1 Lmna +/H222V mutation results in severe structural defects in vivo.
  • A Echocardiography analysis of dilated embryonic hearts presenting a weak contractility, depending of the genotypes of the embryos. The presented values are expressed as percentages.
  • B Echocardiography analysis of different parameters: ejection fraction (EF), Left Ventricule volume during diastole or systole (LV vol D or LV vol S, respectively), Left Ventricule Internal Diameter, diastole or systole (LVIDD or LVIDs).
  • C Hematoxilin and Eosin (H&E) staining for wild-type (WT) (top panel) and H222P homozygote (HMZ) embryonic hearts at El 3.5 (left panel and middle panel). Immunohistochemistery with an antibody against ALPHA ACTININE (right panel) on WT and H222P homozygote (HMZ) embryonic hearts at E13.5. *p ⁇ 0.05 data are represented + SEM.
  • FIG. 2 mESC carrying Lmna +/H222V mutation shows delayed cardiogenesis program and functionnal defects during differentiation in vitro.
  • A Time course expression analysis of the cardiac mesodermal genes MesPl, l, Tbx5, Nkx2.5 and Mef2c. *p ⁇ 0.05 data are represented + SEM.
  • C Assessement of cardiac functionality in vitro by scoring the number of beating EBs ***p ⁇ 0.005 data are represented + SEM.
  • Figure 3 Lmna +/m22V mutation affects mesodermal differentiation throught
  • FIG. 4 Inhibition of LSD1 is sufficient to correct epigenetic and genetic defects in differentiation ES Lmna +/H222P cells in vitro.
  • A Transfection of embryonic stem cells (ESCs) prior to differentiation with the LSD1 siRNA was efficient to downregulate the demethylase. ChIP was used to interrogate the H3K4mel epigenetic mark sitting on both twist and MesPl regulatory regions. Downregulation of LSD1 in ESCs allowed for a loss of H3K4mel on both genes surrounded by chromatin extracted from Lmna +/H222P mutated embryoid bodies (EBs) when compared to wild-type (wt) EBs, both collected at Day 4.
  • EBs Lmna +/H222P mutated embryoid bodies
  • C LSD1 inhibition allows for a recovery in expression of both atrial and ventricular myosin light chain, MLC2a and ML2v respectively, in differentiating Lmna +/H222P mutated EBs cultured in differentiation medium for twelve days.
  • D Beating index of EBs wt or Lmna m22PI+ used as an index of cardiac differentiation, treated or not with GSK1-LSD1 (LSD1 inhibitor). p ⁇ 0.005 data are represented + SEM: * comparison of wt EBs with wt EBs + inhibitor; ⁇ comparison of Lmna m22FI+ EBs treated or not by GSK-LSD 1.
  • the term "subject” refers to a human or another mammal (e.g., primate, dog, cat, goat, horse, pig, mouse, rat, rabbit, and the like), that can develop a cardiomyopathy, but may or may not be suffering from the disease.
  • Non- human subjects may be transgenic or otherwise modified animals.
  • non- human subjects may be domesticated animals and/or pets.
  • the subject is a human being.
  • the subject is often referred to as an "individual" or a "patient”. These terms do not denote a particular age, and thus encompass adults, teenagers, children, babies, newborns, and foetuses.
  • a “cardiomyopathy patient” is an individual suffering from a cardiomyopathy.
  • the term “cardiomyopathy” refers to a group of diseases that adversely affect cardiac cell tissue leading to measurable deterioration in myocardial function (e.g., systolic function, diastolic function), and that are unexplained by - - abnormal loading conditions (hypertension, ischemic heart disease, valve disease, etc .), congenital cardiac abnormalities, and ischemic heart disease.
  • cardiomyopathy is used herein to refer to any disease that is encompassed by the 2006 American Heart Association definition and classification of cardiomyopathies (Maron et ah, Circulation, 20016, 113: 1807) and/or by the 2008 European Society of Cardiology definition and classification of cardiomyopathies (Elliott et ah, Eur. Heart J., 2008, 29: 270).
  • the main cardiomyopathy phenotypes are: dilated, hypertrophic, restrictive, arrhythmo genie right ventricular, and unclassified cardiomyopathies. Each cardiomyopathy phenotype is caused by a variety of familial and non-familial disorders. Identification of various cardiomyopathy phenotypes relies primarily upon echocardio graphic evaluation. In many embodiments of the present invention, the cardiomyopathy is a dilated cardiomyopathy.
  • dilatation refers to a disease that is characterized by dilatation and impaired contraction of one or both ventricles.
  • the dilatation can be severe and is invariably accompanied by an increase in total cardiac mass (hypertrophy).
  • Affected patients have impaired systolic function and may or may not develop overt heart failure.
  • Methods for diagnosing dilated cardiomyopathy include, but are not limited to, echocardiography, a magnetic resonance image (MRI) to determine whether the subject's heart is enlarged, an electrocardiogram to detect abnormal electrical activity of the heart, an echocardiogram to determine the size and shape of the heart, a radionuclide ventriculogram to assess heart function, or cardiac catheterization of dyes coupled with the use of X-ray examination to determine cardiac structure and function.
  • a diagnosis of dilated cardiomyopathy can be made when dilatation and increase sphericity of, typically, at least the left ventricle and reduced systolic function are observed.
  • Subjects with dilated cardiomyopathy can be asymptomatic or can exhibit one or more symptoms associated with dilated cardiomyopathy, which include, but are not limited to, shortness of breath, dyspnea, fatigue, peripheral edema, muscle weakness, dystrophy, syncope, arrhythmia, thromboembolism, and/or congestive heart failure.
  • symptoms associated with dilated cardiomyopathy include, but are not limited to, shortness of breath, dyspnea, fatigue, peripheral edema, muscle weakness, dystrophy, syncope, arrhythmia, thromboembolism, and/or congestive heart failure.
  • dilated cardiomyopathy can also have a familial or genetic predisposition, and has been associated with mutations in genes that encode components of a wide variety of cellular components and pathways.
  • the dilated cardiomyopathy is an LMNA- associated dilated cardiomyopathy.
  • the terms "LMNA -associated dilated cardiomyopathy”, “LMNA-related dilated cardiomyopathy” and “LMNA dilated cardiomyopathy” are used herein interchangeably. They refer to a dilated cardiomyopathy that is caused by a mutation of the LMNA gene (also called dilated cardiomyopathy 1A).
  • LMNA cardiomyopathy there are currently no clinical criteria that can reliably distinguish LMNA cardiomyopathy from other forms of idiopathic dilated cardiomyopathy. Similar to other dilated cardiomyopathies, LMNA- associated dilated cardiomyopathy is characterized by chamber enlargement and systolic dysfunction of one or both ventricles. However, compared to other dilated cardiomyopathies, the natural course of LMNA dilated cardiomyopathy is aggressive, often leading to premature death or cardiac transplant. LMNA gene testing is utilized to confirm a diagnosis of dilated cardiomyopathy in patients with clinically evident disease. Genetic testing allows for early identification and diagnosis of individuals at greater risk prior to the expression of typical clinical manifestations.
  • LMNA -associated cardiomyopathy refers (1) to a cardiomyopathy that is caused by a mutation of the LMNA gene or to a cardiomyopathy that is secondary to skeletal muscle dystrophy or lipodystrophy in a laminopathy ⁇ i.e., a disease caused by a LMNA mutation).
  • cardiomyopathies caused by a mutation of the LMNA gene include the LMNA dilated cardiomyopathy or dilated cardiomyopathy 1A (see above), arrhythmogenic right ventricular dysplasia/cardiomyopathy (Quarta et al., Eur.
  • Heart J., 2012, 33: 1128-1136 inherited form of early onset of myocardial fibrosis (van Tintelen et al., J. Am. Coll. Cardiol., 2007, 49: 2430-2439), left ventricular non-compaction (Hermida-Prieto et al., Am. J. Cardiol., 2004, 94: 50-54).
  • laminopathies in which cardiomyopathy is secondary to skeletal muscle dystrophy, or lipodystrophy include, without limitation, Emery- Dreifuss muscular dystrophy (EDMD), limb-girdle muscular dystrophy type IB (LGMD1B) (Antoniades et al, J. Interv. Card.
  • treatment is used herein to characterize a method or process that is aimed at (1) slowing down or stopping the progression, aggravation, or deterioration of at least one symptom of the disease or condition; (2) bringing about amelioration of at least one symptom of the disease or condition; and/or (3) curing the disease or condition.
  • a treatment is administered after onset of the disease or condition, for a therapeutic action.
  • prevention or “preventive treatment” are used herein interchangeably. They refer to a method or process that is aimed at (1) delaying or preventing the onset of a disease or disorder; and/or (2) delaying or preventing the onset at least one symptom of the disease or disorder.
  • a preventive treatment is administered prior to the onset of the disease or condition, and/or prior to the onset of at least one symptom of the disease or condition, for a prophylactic action.
  • a “pharmaceutical composition” is defined herein as comprising an effective amount of at least one LSD1 inhibitor of the invention, or a physiologically tolerable salt thereof, and at least one pharmaceutically acceptable carrier or excipient.
  • physiologically tolerable salt refers to any acid addition or base addition salt that retains the biological activity and properties of the free base or free acid, respectively, and that is not biologically or otherwise undesirable.
  • Acid addition salts are formed with inorganic acids (e.g., hydrochloric, hydrobromic, sulfuric, nitric, phosphoric acids, and the like); or organic acids (e.g., acetic, propionic, pyruvic, maleic, malonic, succinic, fumaric, tartaric, citric, benzoic, mandelic, methanesulfonic, ethanesulfonic, p-toluenesulfonic, salicylic acids, and the like).
  • inorganic acids e.g., hydrochloric, hydrobromic, sulfuric, nitric, phosphoric acids, and the like
  • organic acids e.g., acetic, propionic, pyruvic, maleic, malonic, succinic, fumaric, tart
  • Base addition salts can be formed with inorganic bases (e.g. , sodium, potassium, lithium, ammonium, calcium, magnesium, zinc, aluminum salts, and the like) or organic bases (e.g. , salts of primary, secondary, and tertiary amines, substituted amines including naturally occurring substituted amines, cyclic amines and basic ion exchange resins, such as isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, ethanolamine, 2-dimethyl-aminoethanol, 2-diethylaminoethanol, trimethamine, dicyclohexylamine, lysine, arginine, histidine, caffeine, procaine, hydrabamine, choline, - - betaine, ethylenediamine, glucosamine, methylglucamine, theobromine, purines, piperazine, piperidine, N-ethylpiperidine, polyamine resins,
  • the term "effective amount” refers to any amount of a molecule, compound, agent, or composition that is sufficient to fulfil its intended purpose(s), e.g., a desired biological or medicinal response in a cell, tissue, system or subject, such as, for example, the effect for which it is administered, to treat or prevent a disease, reduce an enzymatic activity, reduce one or more symptoms of the disease or condition, etc .).
  • An example of an “effective amount” is a “therapeutically effective amount", which is an amount sufficient to contribute to the treatment or reduction of a symptom or symptoms of a disease.
  • a "reduction" of a symptoms or symptoms means decreasing of the severity or frequency of the symptom(s), or elimination of the symptom(s).
  • an “effective amount” is a “prophylactic effective amount”, which is an amount sufficient to prevent or delay the onset (or reoccurrence) of a disease, pathology or condition, or reducing the likelihood of the onset (or reoccurrence) of the disease, pathology or condition, or their symptoms.
  • the full therapeutic or prophylactic effect does not necessarily occur by administration of one dose, and may occur only after administration of a series of doses.
  • a therapeutic or prophylactic effective amount may be administered in one or more administrations. The exact amounts will depend on the purpose of the treatment, and will be ascertainable by one skilled in the art using known techniques (see, e.g., Lieberman, "Pharmaceutical Dosage Forms", vols.
  • pharmaceutically acceptable carrier or excipient refers to a carrier medium which does not interfere with the effectiveness of the biological and/or therapeutic activity of the active ingredient(s) and which is not excessively toxic to the host at the concentration at which it is administered.
  • carrier medium includes solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic agents, and adsorption delaying agents, and the like.
  • the present invention identifies LSDl as a new therapeutic target in cardiomyopathies, and provides for the use of LSDl inhibitors in the treatment and/or prevention of cardiomyopathies, in particular dilated cardiomyopathies, such as L VA-related dilated cardiomyopathies.
  • LSDl belongs to the flavoenzyme-dependent demethylases. There are two members in this family: LSDl (also called KDM1A) and LSD2 (also known as KDM1B). Cry stallo graphic studies have shown that LSDl and LSD2 share a conserved C-terminal amino oxidase domain (AOD) and a SWIRM domain. LSDl functions to demethylate mono- and di-methylated lysines, in particular H3K4mel/2, H3K9mel/2, and non-histone proteins including p53, E2F1 andDNMTl. LSDl is present in different protein complexes, such as the HDAC/CoREST/REST complex and the Mi- 2/nucleosome remodeling and deacetylase (NuRD) complex, and displays diverse functions.
  • NuRD Mi- 2/nucleosome remodeling and deacetylase
  • LSDl refers to a protein called lysine (K)-specific demethylase 1A (LSDl) or lysine-specific histone demethylase 1A (KDM1A) (UniProtKB/Swiss-Prot Number: 060341 in humans), that is also known as: AOF2, BHC110, KDM1, LSDl, CPRF, and lysine demethylase 1A.
  • this enzyme is encoded by the KDM1A gene, which is situated on the short (p) arm of human chromosome 1 at position 36.12 (Gene ID: 23028).
  • LSDl is involved in a wide variety of normal physiological processes, including stem cell maintenance and differentiation, it is also a key player in oncogenic processes, including compromised differentiation, enhanced cell motility and metabolic reprogramming. Therefore, the inhibition of LSDl is considered as a potential therapeutic approach for the treatment for cancer, and a large number of LSDl inhibitors have been reported and are currently undergoing clinical assessment for the treatment of acute myeloid leukemia, small-cell - - lung cancer, etc... (Lynch et al., Expert. Opin. Ther. Targets, 2012, 13: 1239-1249; Hino et al, Cancer Sci., 2016, 107: 1187-1192; Fu et al, Future Med. Chem., 2017, 9: 1227-1242).
  • the present Inventors have observed that the lysine demethylase LSDl is more active in Lmna +/H222P mutated embryonic stem cells than in wild-type embryonic stem cells (see Examples section below). They have also demonstrated that blunting LSDl activity by a siRNA or a pharmacological LSDl inhibitor rescued the epigenetic landscape in mutated embryonic stem cells and that treating H222P homozygous pregnant mice with a pharmacological LSDl inhibitor at an early stage of pregnancy prevented cardiomyopathy in the offspring. Thus, the present invention proposes the use of LSDl inhibitors in the treatment and/or prevention of cardiomyopathies.
  • LSDl inhibitor includes and encompasses any active agent or compound that, irrespective of its mechanism(s) of action, reduces or inhibits the accumulation, function or stability of the histone demethylase LSDl, or that decreases or prevents expression of the KDM1A gene.
  • reduce reduce
  • inhibit decrease
  • prevent are used relative to a control.
  • One skilled in the art would readily identify the appropriate control to be used for comparison. For example, reduction or inhibition of the accumulation, function or stability of LSDl in a subject or cell that is treated with a LSDl inhibitor is assessed by comparison to the accumulation, function or stability of LSDl in a subject or cell that is not treated with the LSDl inhibitor.
  • LSDl inhibitors that are suitable for use in the present invention may be reversible or irreversible, direct or indirect.
  • Suitable LSDl inhibitors may be nucleic acids, peptides, polypeptides, peptidomimetics, carbohydrates, polysaccharides, lipopolysaccharides, lipids, and other organic or inorganic molecules.
  • a LSDl inhibitor may be any member of a class of compounds (e.g., a small molecule, or an antibody or fragment or derivative of such antibody) that binds LSDl and inhibits a biological activity (e.g., demethylase activity) of LSDl or a protein complex in which - -
  • LSDl exerts its function (e.g. , LSDl complexed to co-REST and/or other protein members of the nucleosome).
  • a LSDl inhibitor may also be any member of a class of compounds that decreases the expression of a nucleic acid encoding a LSDl protein (e.g. , a nucleic acid that inhibits the transcription or translation of the KDM1A gene or that mediates RNA interference, RNAi).
  • small molecule refers to a molecule having a molecular weight of less than 1,000 daltons, preferably of less than 700 daltons.
  • the LSDl inhibitor is a selective LSDl inhibitor, while in other embodiments of the present invention, the LSDl inhibitor is dual LSD1/MAO inhibitor.
  • selective LSDl inhibitor refers to an LSDl inhibitor, which has a half maximal inhibitory concentration (i.e. , IC 50 ) value for LSDl that at least 2-fold lower than its IC 50 values for other FAS utilizing enzymes (i.e., LSD2, MAO- A and MAO-B).
  • IC 50 half maximal inhibitory concentration
  • a selective LSDl inhibitor has an IC 50 value for LSDl which is at least 5-fold, at least 10-fold, or at least 20-fold, lower than its IC 50 values for LSD2, MAO- A and MAO-B.
  • a selective LSDl inhibitor has an IC 50 value that is at least 50-fold, or at least 100-fold, lower than its IC 50 values for LSD2, MAO- A and MAO-B. Even more preferably, a selective LSDl inhibitor has an IC 50 value that is more than 100-fold lower than its IC 50 values for LSD2, MOA-A and MOA-B.
  • IC 50 values are more than 100-fold lower than its IC 50 values for LSD2, MOA-A and MOA-B.
  • LSDIZMAO-A(B) inhibitor selective inhibitor of LSDl and MAO-A(B)
  • LSDIZMAO-A(B) inhibitor selective inhibitor of LSDl and MAO-A(B)
  • LSDIZMAO-A(B) inhibitor selective inhibitor of LSDl and MAO-A(B)
  • LSDIZMAO-A(B) selective inhibitor refers to an LSDl inhibitor which has IC 50 values for LSDl and MAO- A (or MAO-B) that are at least 2-fold lower than its IC 50 value for MAO-B (or MAO-A, respectively).
  • LSDl inhibitor which has IC 50 values for LSDl and MAO-A (or MAO-B) that are at least 5- fold, at least 10-fold, or at least 20-fold, lower than its IC 50 value for MAO-B (or MAO- - - -
  • LSDl inhibitor which has IC 50 values for LSDl and MAO-A (or MAO-B) that are at least 50-fold, or at least 100-fold, lower than its IC 50 value for MAO-B (or MAO-A, respectively). Even more preferably, they refer to an LSDl inhibitor which has IC 50 values for LSDl and MAO-A (or MAO- B) that are at more than 100-fold lower than its IC 50 value for MAO-B (or MAO-A, respectively).
  • the LSDl inhibitor is an irreversible small molecule LSDl inhibitor.
  • irreversible small molecule inhibitors include, without limitation, pargyline (propargylamine) derivatives or analogs (Culhane et al, J. Am. Chem. Soc, 2006, 128: 4536-4537; Schmitt et al, J. Med. Chem., 2013, 56: 7334-7342), phenelzine derivatives or analogs (Prusevich et al, ACS Chem. Biol., 2014, 9: 1284-1293), and cyclopropylamine derivatives or analogs.
  • the LSDl inhibitor is selected among cyclopropylamine derivatives or analogs, in particular among tranylcypromine ((+)-iraw5'-2-phenylcyclopropylamine) derivatives.
  • cyclopropylamine and arylcyclopropylamine derivatives and analogues that can be used as LSDl inhibitors in the practice of the present invention include, without limitation, the ones described in US 2010/0324147; WO 2011/131576; Gooden et al, Bioorg. Med. Chem. Let., 2008 18: 3047-3051; 6827-6833; Ueda et al, J. Am. Chem. Soc, 2009, 131: 17536-17537; Mimasu et al, Biochemistry, 2010, 49: 6494-6503; - -
  • N-alkylated tranylcypromine derivatives including ORY-1001 (Oryzon Genomics, Roche), ORY-2001 (Oryzon Genomics), ORY-3001 (Oryzon Genomics), GSK-2879552 (GlaxoSmithKline), INCB-059872 (Incyte), IMG-7289 (Imago Biosciences), and CC-90011 (Celgene), are currently in clinical trials for treatments of cancers, such as acute myeloid leukemia (AML), small cell lung cancer (SCLC), myelodysplasia syndrome (MDS), and non-Hodgkin's lymphoma (NHL), and for treatments of neurodegenerative diseases such as Alzheimer's disease, Parkinson's disease, Huntington's disease.
  • AML acute myeloid leukemia
  • SCLC small cell lung cancer
  • MDS myelodysplasia syndrome
  • NHL non-Hodgkin's lymphoma
  • the LSD1 inhibitor to be used in the practice of the present invention is selected from ORY-1001, ORY-2001, ORY-3001, GSK-2879552, INCB-059872, IMG-7289, and CC-90011.
  • tranylcypromine derivatives that can be used as LSD1 inhibitors to be used in the practice of the present invention include, for example, RN-1 and RN-7 (Neelamegam et al, ACS Chem. Neurosci., 2012, 3: 120-128; Cui et al, Blood, 2015, 126: 386-396; McGrath et al, Cancer Res., 2016, 76: 1975-1988), GSK-2699537 (developed by GlaxoSmithKline) and T-3775440 (developed by Takeda; Ishikawa et al, Mol. Cancer Ther., 2017, 16(2): 273-284).
  • RN-1 and RN-7 Neelamegam et al, ACS Chem. Neurosci., 2012, 3: 120-128; Cui et al, Blood, 2015, 126: 386-396; McGrath et al, Cancer Res., 2016, 76: 1975-1988
  • GSK-2699537 developed by GlaxoSmithK
  • the selective LSD1 inhibitor is GSK-LSDI (N-[(lR,2S)-2-phenylcyclopropyl]piperidin-4-amine dihydrochloride; Cas Number: 1431368-48-7 (for the free base)).
  • GSK-LSDI which is a selective and irreversible inhibitor of LSD1, has the following chemical structure:
  • GSK-LSDI inhibits LSD1 with an IC 50 of 16 nM and is more than 1000 fold selective over other closely related FAD utilizing enzymes ⁇ i.e., LSD2, MAO-A, MAO-B).
  • the selective LSD1 inhibitor is GSK-2879552 (4-[[4-[[((lR,2S)-2-Phenylcyclopropyl)amino]- - - methyl]piperidin-l-yl]methyl]benzoic acid; Cas Number: Cas 1401966-69-5).
  • GSK- 2879552 which is a selective and irreversible inhibitor of LSD1, has the following chemical structure:
  • the selective LSD1 inhibitor is ORY-1001 (re/-N 1 -[(lR,2S)-2-phenylcyclopropyl]-l,4-cyclohexanediamine, dihydrochloride; CAS Number: 1431303-72-8 for the 2 HC1 salt; CAS Number: 1431303-71-7 for the xHCl salt; and CAS Number: 1431304-21-0 for the free base).
  • ORY-1001 which is a selective inhibitor of LSD1 (IC 50 ⁇ 20 nM), has the following chemical structure:
  • LSD1 inhibitor is 4SC-202 (4SC AG).
  • the LSD1 inhibitor is an irreversible small molecule LSD1 inhibitor.
  • LSD1 inhibitors examples include, without limitation, LSD1 inhibitors containing a benzohydrazide scaffold, such as the ones described in Soma et al, J. Med.Chem., 2013, 56: 9496-9508; Zhou et al, Bioorg. Med. Chem. Lett, 2016, 26: 4552-4557; WO 2013/025805, WO 2014/205213; WO 2017/004519), for example HCI-2509 (also named SP2059 or LSD1-C12 - CAS Number: 1423715-09-6) (Sankar et al, Clin.
  • reversible LSD1 inhibitors suitable for use in the context of the present invention include dithiocarbamates-based compounds, such as the ones described by Zheng et al, J. Med. Chem., 2013, 56: 8543-8560; Ye et al, Med. Chem. Commun., 2014, 5: 650-654. - -
  • reversible LSD1 inhibitors suitable for use in the present invention include pyridine, pyrazine and pyrimidine derivatives and analogs, such as the ones developed by Incyte Corporation and described in WO 2016/007722, WO 2016/007727, WO 2016/007731 and WO 2016/007736; and the ones described by Li et al, ACS Med. Chem. Lett., 2017; 8: 384-389 and Wang et al, Eur. J. Med. Chem., 2017, 125: 940-951.
  • reversible LSD1 inhibitors suitable for use in the present invention include diverse triazole- and tetrazole-based compounds, such as the ones described by Kutz et al, Med. Chem. Commun., 2014, 5: 1863-1870; WO 2015/120281, and the reversible LSD1 inhibitors with a 5-hydroxypyrazole scaffold, such as the ones described by Mould et al, Bioorg. Med. Chem. Lett., 2017, 27: 3190-3195.
  • reversible LSD1 inhibitors include polyamine analogs such as bisguanidine and biguanide (Huang et al, PNAS, 2007, 104: 8023-8028; Zhu et al, Amino Acids, 2012, 42, 42: 887-898); bisurea and bisthiourea derivatives (WO 2007/021839; WO 2008/127734; WO 2011/022489; Sharma et al, J. Med. Chem., 2010, 43: 5189-5212; Nowotarski et al, Bioorg. Med.
  • polyamine analogs such as bisguanidine and biguanide (Huang et al, PNAS, 2007, 104: 8023-8028; Zhu et al, Amino Acids, 2012, 42, 42: 887-898); bisurea and bisthiourea derivatives (WO 2007/021839; WO 2008/127734; WO 2011/022489; Sharma et al, J. Med. Chem., 2010, 43
  • guanidinium-based compounds Wang et al, Cancer Res., 2011, 71: 7238-7249, such as CBB1007; lysine-based compounds (WO 2014/084298); and oligoamine analogues ⁇ i.e., long chain polyamine analogues) (Huang et al, Clin. Cancer Res., 2009, 15: 7217- 7228).
  • Linear and cyclic peptides have also been developed as irreversible small molecule LSD1 inhibitors.
  • Examples of such peptides include, without limitation, the ones described in Kumarasinghe et al, ACS Med. Chem. Lett., 2013, 5: 29-33; Tortorici et al, ACS Chem. Biol., 2013, 8: 1677-1682; Kakizawa et al, Bioorg. Med. - -
  • any irreversible small molecule LSD1 inhibitor may find application in the present invention, including the irreversible small molecule LSD1 inhibitors currently in development, including, without limitation, the carboxamides derivatives (Sartori et al., J. Med. Chem., 2017, 61: 1673-1692; Vianello et al., J. Med. Chem. 2017, 60: 1693-1715), benzesulfonamide derivatives (Xi et al., Bioorg. Chem., 2017, 72: 182-189), etc...
  • LSD1 inhibitors include RNA interfering agents.
  • RNA interference As used herein, the term “RNA interference” (or “RNAi”) has its art understood meaning and refers to a biological process in which RNA molecules silence, inhibit or down-regulate gene expression by causing the destruction, degradation and/or cleavage of specific mRNA molecules or by blocking the translation thereof. As known in the art, RNA interference is now exploited in therapy. Indeed, RNAi can be initiated by the hand of man, for example, to silence the expression of target genes.
  • RNAi agent and “RNA interfering agent” are used herein interchangeably. They refer to any RNA molecule that is capable of specifically inhibiting or down-regulating the expression of a target gene (here the KDM1A gene).
  • RNAi agent By “silencing, inhibiting or down- regulating expression of a target gene”, it is meant that the expression of the target gene, or level of RNA molecules or equivalent RNA molecules encoding one or more proteins or protein subunits, or activity of one or more proteins or protein subunits, is reduced below that observed in the absence of the RNAi agent.
  • an RNA interfering agent reduces the expression of the KDM1A gene or transcript to less than about 90%, 80%, 70%, 60%, 50%, 25%, 10%, 5%, 1%, or to less than 0.1% (for example less than about 10 "4 , 10 "5 , 10 “6 , 10 “7 , 10 “8 , 10 “9 , 10 “10 , 10 “11 , 10 “12 , 10 “13 , 10 “14 , or 10 "15 ) of the expression of the KDM1A gene or transcript in the absence of the RNA interfering agent.
  • RNAi agent may be any single-stranded RNA (e.g., mature miRNA, ssRNAi oligonucleotides, ssDNAi oligonucleotides) or double-stranded RNA (i.e., duplex RNA such as siRNA, Dicer-substrate dsRNA, shRNA, aiRNA, or pre-miRNA) that is capable of reducing or inhibiting the expression of a target gene or sequence (e.g., by mediating - - the degradation or inhibiting the translation of mRNAs which are complementary to the interfering RNA sequence) when the RNAi agent is in the same cell as the target gene or sequence.
  • RNAi agent may be any single-stranded RNA (e.g., mature miRNA, ssRNAi oligonucleotides, ssDNAi oligonucleotides) or double-stranded RNA (i.e., duplex RNA such as siRNA, Dicer
  • RNAi agent thus refers to the single- stranded RNA that is complementary to a target mRNA sequence (or circRNA sequence) or to the double- stranded RNA formed by two complementary strands or by a single, self- complementary strand.
  • An RNAi agent may have substantial or complete identity to the target gene mRNA (or circRNA) or sequence, or may comprise a region of mismatch (i.e., a mismatch motif). Consequently, the term "RNAi agent” refers to a RNA molecule comprising a strand having a sequence sufficiently complementary to a target mRNA (or circRNA) sequence to direct target- specific RNA interference (RNAi) thereby inhibiting or down-regulating the expression of the target gene.
  • RNAi agent target-specific RNA interference
  • RNAi agent can comprise naturally occurring RNA, synthetic RNA, or recombinantly produced RNA, as well as altered RNA that differs from naturally- occurring RNA by the addition, deletion, substitution and/or alteration of one or more nucleotides. Such alterations can include addition of non-nucleotide material, such as to the end of the molecule or to one or more internal nucleotides of the RNAi, including modifications that make the RNAi agent resistant to nuclease digestion.
  • An RNAi agent may be administered in free (naked) form or by the use of delivery systems that enhance stability and/or targeting, or may be administered in the form of its precursor or encoding DNA.
  • an RNAi agent is a siRNA (small interfering RNA), a shRNA (short hairpin RNA), a micro-RNA (micro RNA), or an aiRNA (asymmetric interfering RNA).
  • siRNA small interfering RNA
  • shRNA short hairpin RNA
  • micro-RNA micro-RNA
  • aiRNA asymmetric interfering RNA
  • the LSD1 RNA interfering agent used in the practice of the present invention is a small interfering RNA.
  • small interfering RNA and “siRNA” are used herein interchangeably. They refer to an RNA (or RNA analog) comprising between about 10-50 nucleotides (or nucleotide analogs) which is capable of directing or mediating RNA interference.
  • a siRNA comprises between about 15-30 nucleotides or nucleotide analogs, more preferably between about 16-25 nucleotides (or - - nucleotide analogs), even more preferably between about 18-23 nucleotides (or nucleotide analogs), and even more preferably between about 19-22 nucleotides (or nucleotide analogs) (e.g., 19, 20, 21 or 22 nucleotides or nucleotide analogs).
  • KDM1A transcripts include nucleotide sequences corresponding to any one the following sequences: (1) human LSD1 nucleotide sequences as set forth for example in GenBank Accession Nos.
  • nucleotide sequences that share at least 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99% sequence identity with any one of the sequences referred to in (1); (3) nucleotide sequences that hybridize under at least low, medium or high stringency conditions to the sequences referred to in (1); (4) nucleotide sequences that encode any one of the following amino acid sequences: human LSD1 amino acid sequences as set forth for example in GenPept Accession Nos.
  • siRNAs molecules may be prepared by any suitable method known in the art (see for example WO 02/44321). Sequence specific gene silencing can be achieved in mammalian cells using synthetic, short double- stranded RNAs that mimic the siRNAs produced by the enzyme dicer. siRNAs can be chemically or in viiro-synthesized or can be the result of short double- stranded hairpin-like RNAs (shRNAs) that are processed into siRNAs inside the cell. Synthetic siRNAs are generally designed using algorithms and a conventional DNA/RNA synthesizer.
  • siRNAs can also be synthesized in vitro using kits such as Ambion's SILENCERTM siRNA Construction Kit. - -
  • LSD1 siRNA molecules have been described, for example, in Shi et al, Cell, 2004, 119: 941-953; Metzger et al, Nature, 2005, 437: 436-439; Zhu et al., PLoS ONE, 2008, 3: el446; Wang et al., Cell, 2009, 138: 660-672; Sun et al., Mol. Cell Biol, 2010, 30: 1997-2005; Hino et al, Nature Comm. 2012, 3: article number 758; and Xu et al., Cell Physiol. Biochem., 2013, 31: 854-862; and are commercially available from Santa Cruz Biotechnology, Inc. (Santa Cruz, CA, USA) and OriGene Technologies, Inc. (Rockville, MD, USA).
  • the present invention provides for the use of an LSD1 inhibitor in a method of treatment and/or prevention of a cardiomyopathy, in particular a dilated cardiomyopathy, in a subject.
  • the cardiomyopathy is a LMNA- associated cardiomyopathy, for example a LMVA-associated dilated cardiomyopathy.
  • a method of treatment and/or prevention according to the present invention may be accomplished using an LSD1 inhibitor (as defined herein), or a pharmaceutical composition thereof.
  • the method generally comprises a step of administering, directly or indirectly, to the subject in need thereof, an effective amount of an LSD1 inhibitor, or a pharmaceutical composition thereof.
  • the LSD1 inhibitors and methods of the present invention are used for treating a cardiomyopathy in a subject in need thereof.
  • treatment methods are administered after onset of the disease to be treated.
  • the effective amount of LSD1 inhibitor to be administered is a therapeutically effective amount of LSD 1 inhibitor.
  • cardiomyopathies examples include, without limitation, the primary cardiomyopathies that predominantly involve the heart: dilated cardiomyopathy (see below), hypertrophic cardiomyopathy, restrictive cardiomyopathy, arrhythmo genie right ventricular cardiomyopathy, unclassified cardiomyopathy, and acquired cardiomyopathies (e.g., stress induced cardiomyopathies, cardiomyopathy in peripartum, tachycardia-induced cardiomyopathy,...); and secondary cardiomyopathies that can have multi-organ involvement: cardiomyopathies from infiltrative conditions - -
  • cardiomyopathies from storage conditions e.g. , haemochromatosis, Fabry' s disease
  • cardiomyopathies from toxic agents e.g. , endomyocardial fibrosis, Loeffler endocarditis
  • cardiomyopathies from endocrine conditions e.g. , diabetes mellitus, hyperthyroidism, acromegaly
  • cardiomyopathies from underlying systemic autoimmune conditions e.g.
  • cardiomyopathies from neuromuscular conditions e.g. , myotonic dystrophy, Friedreich ataxia
  • cardiofacial cardiomyopathies e.g. , Noonan syndrome
  • Other types such as obesity-associated cardiomyopathy.
  • the cardiomyopathy to be treated using a method according to the invention may also be an LMNA cardiomyopathy, as defined above.
  • the LMNA cardiomyopathy may be a disease caused by an LMNA mutation and that mainly affects the heart, or may be a cardiomyopathy that is secondary to skeletal muscle dystrophy or lipodystrophy in a laminopathy (i.e. , a disease caused by a LMNA mutation).
  • the subject in need of treatment is a cardiomyopathy patient.
  • the patient may be a person having subjective symptoms, such as exertional or non-exertional shortness of breath, dyspnea, orthopnea, malaise, easy fatigue, chest discomfort, chest pain, etc., or may be an asymptomatic person.
  • the subject may include an individual who is suspected of having a cardiomyopathy according to known diagnostic criteria by medical interview, blood pressure examination, physical examination such as heart sound examination and electrocardiogram examination, chest X-ray examination, chest CT examination, cardiac MR examination, standard 12 induction electrocardiogram examination, echocardiographic examination, biochemical examination of serum or plasma for measuring values of creatinine kinase (CK), aspartate aminotransferase (AST), alanine aminotransferase (ALT), lactate dehydrogenase (LDH), BNP, NT-proBNP, etc.
  • CK creatinine kinase
  • AST aspartate aminotransferase
  • ALT alanine aminotransferase
  • LDH lactate dehydrogenase
  • BNP NT-proBNP, etc.
  • the cardiomyopathy to be treated with a method according to the present invention is a dilated cardiomyopathy.
  • the subject in need of treatment is a dilated cardiomyopathy patient.
  • the subject can be one who has been previously diagnosed with or identified as suffering from, or having, dilated cardiomyopathy or one or more symptoms or complications related to dilated cardiomyopathy, and may or may not have already undergone treatment for dilated - - cardiomyopathy or the one or more symptoms or complications related to dilated cardiomyopathy.
  • the subject can be one who has not been previously diagnosed as having dilated cardiomyopathy or one or more symptoms or complications related to dilated cardiomyopathy.
  • dilated cardiomyopathy The most common symptoms of dilated cardiomyopathy include; shortness of breath (dyspnea) with exertion, which may progress to shortness of breath at rest, shortness of breath when lying down (orthopnea), impaired ability to be active or exercise, swelling of the lower legs, ankles and feet (peripheral edema). Symptoms of heart failure do not occur until late in the disease. Symptoms may worsen gradually or abruptly appear or worsen. Often, there are no early symptoms of dilated cardiomyopathy and the heart is discovered to be enlarged as a result of a test done for another reason (e.g., a chest x-ray).
  • Dilated cardiomyopathy can easily be diagnosed using echocardiography Echocardiography, however, does not give information on the cause underlying cardiomyopathy.
  • Conduction system disease is detected by a 12-lead electrocardiogram (ECG); arrhythmias are detected by an ECG, 24-hour rhythm recording, or event monitor.
  • ECG electrocardiogram
  • Left ventricular enlargement is diagnosed with cardiac imaging; reduced systolic function is assessed by two-dimensional echocardiography, angiography, radioisotope scanning, or magnetic resonance imaging.
  • the dilated cardiomyopathy is an LMVA-related dilated cardiomyopathy (as defined above).
  • the subject in need or treatment is an LM/VA-related dilated cardiomyopathy.
  • LMNA sequence analysis identifies pathogenic variants in most individuals with LMNA related dilated cardiomyopathy. 2.
  • LM/VA-related dilated cardiomyopathy Today, pregnancy is contraindicated in most women with dilated cardiomyopathy, and pregnant women with idiopathic or familial dilated cardiomyopathy are generally followed with high-risk obstetrician. Each child of an individual with LM/VA-related dilated cardiomyopathy has a 50% chance of inheriting the parent's pathogenic variant. Prenatal diagnosis for LM/VA-related dilated cardiomyopathy is technically possible when the pathogenic variant in the family is known. Once an LMNA pathogenic variant has been identified in an affected family member, prenatal testing and preimplantation - - genetic diagnosis for a pregnancy at increased risk for L NA-related dilated cardiomyopathy are possible options.
  • the LSDl inhibitors and methods of the present invention are used in a method for preventing a cardiomyopathy, preferably an LMVA-related cardiomyopathy, such as an L VA-related dilated cardiomyopathy, in an embryo.
  • the method comprises administering to the woman pregnant with said embryo an effective amount of an LSDl inhibitor, preferably a prophylactic ally effective amount of an LSDl inhibitor.
  • the pregnant woman and/or the embryo's father are known to carry a cardiomyopathy-causing LMNA variant.
  • the method is carried out to prevent the presence, in the newborn, of the cardiomyopathy- causing LMNA variant.
  • the method of prevention of the present invention is carried out to prevent or delay the onset of an L NA-associated cardiomyopathy (such as an LMNA -related dilated cardiomyopathy) or of one or more symptoms or complications of LMVA-associated cardiomyopathy.
  • the method comprises administering to the subject in need thereof, an effective amount of an LSDl inhibitor, preferably a prophylactically effective amount of an LSDl inhibitor.
  • the subject to be treated is preferably an individual carrying a cardiomyopathy-causing LMNA variant.
  • a prophylactic method according to the present invention may be administered to asymptomatic individuals, in order to prevent or delay onset of the disease or of one or more symptoms associated with the disease.
  • the subject may be a family member of an individual who has been diagnosed with an LMVA-associated cardiomyopathy.
  • LMNA gene testing may be utilized to confirm the presence of the cardiomyopathy- causing LMNA variant, before administration of a preventive treatment according to the present invention.
  • a prophylactic method may be administered to at-risk children or teenagers. Indeed, although the likelihood of clinically detectable LMNA -related cardiomyopathy in pediatric cases is low, genetic testing should be offered as it can facilitate identification of at-risk children who may benefit from early treatment, potentially forestalling the development of advanced disease. - -
  • a prophylactic method according to the present invention may also be administered to a patient suffering from a laminopathy in which cardiomyopathy is secondary to skeletal muscle dystrophy, or lipodystrophy.
  • the subject has been diagnosed with such a laminopathy and is at-risk of developing a cardiomyopathy but has not developed the associated cardiomyopathy or any of the symptoms.
  • the subject may have been diagnosed with Emery- Dreifuss muscular dystrophy (EDMD), limb-girdle muscular dystrophy type IB (LGMD1B), Dunnigan-type familial partial lipodystrophy 2, or progeria, who are at risk of developing a cardiomyopathy.
  • EDMD Emery- Dreifuss muscular dystrophy
  • LGMD1B limb-girdle muscular dystrophy type IB
  • Dunnigan-type familial partial lipodystrophy 2 or progeria
  • a prophylactic method according to the present invention may also be administered to a patient presenting a risk of developing a cardiomyopathy.
  • Risk factors for cardiomyopathy include, but are not limited to, prior instances of myocardial infarction, infection with coxsacki B virus or enteroviruses, autoimmune disease, alcohol abuse, chemotherapy (particularly doxorubicin and cobalt), thyroid disease, tachycardia, stimulant use, extrasystole, and pregnancy.
  • an LSD1 inhibitor of the present invention, or a pharmaceutical composition thereof is administered alone according to a method of treatment or prevention described herein. In other embodiments, an LSD1 of the present invention, or a pharmaceutical composition thereof, is administered in combination with at last one additional therapeutic agent or therapeutic procedure.
  • the LSD1 inhibitor, or pharmaceutical composition thereof may be administered prior to administration of the therapeutic agent or therapeutic procedure, concurrently with the therapeutic agent or procedure, and/or following administration of the therapeutic agent or procedure.
  • Therapeutic procedures that may be performed in combination with administration of an LSD1 inhibitor of the invention, or of a pharmaceutical composition thereof, include, but are not limited to, implantations of left ventricular assist devices, cardiac resynchronization therapy (biventricular pacing), implantations of automatic cardioverter-defibrillators, and ventricular restoration surgery. - -
  • Therapeutic agents that may be administered in combination with one of the LSD1 inhibitors of the invention, or with a pharmaceutical composition thereof, may be selected among a large variety of biologically active compounds that are known to have a beneficial effect in the treatment of a cardiomyopathy or that are known to be beneficial to a patient in general (e.g., anti-inflammatory agents, immunomodulatory agents, analgesics, antimicrobial agents, antibacterial agents, antibiotics, antioxidants, antiseptic agents, and combinations thereof); or that may not be effective alone, but my contribute to the efficacy of the active agent(s) (called adjunctive agents).
  • biologically active compounds that are known to have a beneficial effect in the treatment of a cardiomyopathy or that are known to be beneficial to a patient in general (e.g., anti-inflammatory agents, immunomodulatory agents, analgesics, antimicrobial agents, antibacterial agents, antibiotics, antioxidants, antiseptic agents, and combinations thereof); or that may not be effective alone, but my contribute to the efficacy of the active agent(s)
  • Therapeutic agents known to be useful in the treatment of a cardiomyopathy include, angiotensin-converting enzyme (ACE) inhibitors (e.g., enalipril, lisinopril); angiotensin II receptor blockers (ARBs) (e.g. , losartan, alsartan); beta blockers (e.g. , metoprolol, such as Lopressor ® or Toprol-XL ® ); cardiac glycosides (e.g. , digoxin, digitoxin), diuretics such as aldosterone antagonists (e.g. , spironolactone, eplerenone), loop diuretics (e.g.
  • ACE angiotensin-converting enzyme
  • ARBs angiotensin II receptor blockers
  • beta blockers e.g. , metoprolol, such as Lopressor ® or Toprol-XL ®
  • cardiac glycosides e.g. , digoxin, digitoxin
  • Anticoagulants may be used in selected cardiomyopathy patients.
  • the LSD1 inhibitor and the additional therapeutic agent may be administered concurrently (i.e. , together or separately but at about the same time, e.g. , within 5 minutes, 15 minutes or 30 minutes of each other), or alternatively, they may be administered sequentially (i.e., separately and at different times, e.g., different times of the same day (for instance within 1, 2, 4, 5, 8, 10, 12, 20, or 24 hours of each other) or different times of the same week or different times of the same month, etc).
  • Co-administration can be accomplished by co-formulation, i.e. , by preparing a single pharmaceutical composition including both active agents. Alternatively, the active agents can be formulated separately.
  • An LSD1 inhibitor according to the present invention (optionally after formulation with one or more appropriate pharmaceutically acceptable carriers or excipients), in a desired dosage, can be administered to a subject in need thereof by any suitable route. - -
  • an LSD1 inhibitor of the present invention including tablets, capsules, injectable solutions, encapsulation in liposomes, microparticles, microcapsules, etc.
  • Methods of administration include, but are not limited to, dermal, intradermal, intramuscular, intraperitoneal, intralesional, intravenous, subcutaneous, intranasal, pulmonary, epidural, ocular, and oral routes.
  • An LSD1 inhibitor of the present invention, or a pharmaceutical composition thereof may be administered by any convenient or other appropriate route, for example, by infusion or bolus injection, by adsorption through epithelial or mucocutaneous linings (e.g.
  • LSD1 inhibitor in embodiments where an LSD1 inhibitor is administered along with an additional therapeutic agent, the LSD1 inhibitor and the therapeutic agent may be administered by the same route (e.g. , orally) or by different routes (e.g., orally and intravenously).
  • an LSD1 inhibitor (or a pharmaceutical composition thereof) according to the present invention will be in a dosage such that the amount delivered is effective for the intended purpose.
  • the route of administration, formulation, and dosage administered will depend upon the therapeutic/prophylactic effect desired, the severity of the disorder being treated, the presence of any infection, the age, sex, weight and general health condition of the patient as well as upon the potency, bioavailability and in vivo half-life of the LSD1 inhibitor used, the use (or not) of concomitant therapies, and other clinical factors. These factors are readily determinable by the attending physician in the course of the therapy.
  • the dosage to be administered can be determined from studies using animal models and/or human clinical trials. Adjusting the dose to achieve maximal efficacy based on these and other methods are well known in the art and are within the capabilities of trained physicians.
  • a treatment according to the present invention may consist of a single dose or multiple doses.
  • administration of an LSD1 inhibitor, or of a pharmaceutical composition thereof may be constant for a certain period of time or periodic and at specific intervals, e.g. , hourly, daily, weekly (or at some other multiple day interval); monthly, yearly (e.g., in a time release form).
  • the delivery may occur at - - multiple times during a given time period, e.g., two or more times per week, two or more times per month, and the like.
  • the delivery may be continuous delivery for a period of time, e.g., intravenous delivery.
  • the amount of the LSD1 inhibitor (or a pharmaceutical composition thereof) administered will preferably be in the range of about 1 ng/kg to about 1000 mg/kg body weight of the subject, for example, between about 100 ng/kg and about 500 mg/kg body weight of the subject; or between about 1 g/kg and about 100 mg/kg body weight of the subject; or between about 100 g/kg and about 10 mg/kg body weight of the subject; or between about 500 g/kg and about 1 mg/kg body weight of the subject.
  • the effects of a therapeutic or prophylactic treatment according to the present invention may be assessed using any technique used to diagnose cardiomyopathy, including, but not limited to, physical examination, echocardiodiagram, electrocardiogram, magnetic resonance image; radionuclide ventriculogram, etc.
  • an LSD1 inhibitor of the present invention may be administered per se or as a pharmaceutical composition.
  • the present invention provides pharmaceutical compositions comprising an effective amount of a LSD1 inhibitor and at least one pharmaceutically acceptable carrier or excipient for use in the treatment or prevention of a cardiomyopathy.
  • the composition further comprises one or more additional biologically active agents.
  • the optimal pharmaceutical formulation of an LSD1 inhibitor according to the present invention can be varied depending upon the route of administration and desired dosage. Such formulations may influence the physical state, stability, rate of in vivo release, and rate of in vivo clearance of the administered active ingredient.
  • compositions of the present invention may be formulated in dosage unit form for ease of administration and uniformity of dosage.
  • unit dosage form refers to a physically discrete unit for the patient to be treated. It will be understood, however, that the total daily dosage of the compositions will be decided by the attending physician within the scope of sound medical judgement. - -
  • sterile injectable preparations for example, sterile injectable aqueous or oleaginous suspensions may be formulated according to the known art using suitable dispersing or wetting agents, and suspending agents.
  • the sterile injectable preparation may also be a sterile injectable solution, suspension or emulsion in a non-toxic parenterally acceptable diluent or solvent, for example, as a solution in 2,3-butanediol.
  • acceptable vehicles and solvents that may be employed are water, Ringer's solution, U.S.P. and isotonic sodium chloride solution.
  • sterile, fixed oils are conventionally employed as a solution or suspending medium.
  • any bland fixed oil can be employed including synthetic mono- or di-glycerides.
  • Fatty acids such as oleic acid may also be used in the preparation of injectable formulations.
  • Sterile liquid carriers are useful in sterile liquid form compositions for parenteral administration.
  • Injectable formulations can be sterilized, for example, by filtration through a bacterial-retaining filter, or by incorporating sterilizing agents in the form of sterile solid compositions which can be dissolved or dispersed in sterile water or other sterile injectable medium prior to use.
  • Liquid pharmaceutical compositions which are sterile solutions or suspensions can be administered by, for example, intravenous, intramuscular, intraperitoneal or subcutaneous injection. Injection may be via single push or by gradual infusion. Where necessary or desired, the composition may include a local anesthetic to ease pain at the site of injection.
  • Injectable depot forms are made by forming micro-encapsulated matrices of the active ingredient in biodegradable polymers such as polylactide-polyglycolide. Depending upon the ratio of active ingredient to polymer and the nature of the particular polymer employed, the rate of ingredient release can be controlled. Examples of other biodegradable polymers include poly(orthoesters) and poly(anhydrides). Depot injectable formulations can also be prepared by entrapping the active ingredient in liposomes or microemulsions which are compatible with body tissues.
  • Liquid dosage forms for oral administration include, but are not limited to, pharmaceutically acceptable emulsions, microemulsions, solutions, suspensions, syrups, - - elixirs, and pressurized compositions.
  • the liquid dosage form may contain inert diluents commonly used in the art such as, for example, water or other solvent, solubilising agents and emulsifiers such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, dimethylformamide, oils (in particular, cotton seed, ground nut, corn, germ, olive, castor, and sesame oils), glycerol, tetrahydrofurfuryl alcohol, polyethylene glycols, and fatty acid esters of sorbitan and mixtures thereof.
  • inert diluents commonly used in the art such as
  • the oral compositions can also include adjuvants such as wetting agents, suspending agents, preservatives, sweetening, flavouring, and perfuming agents, thickening agents, colors, viscosity regulators, stabilizes or osmo-regulators.
  • suitable liquid carriers for oral administration include water (potentially containing additives as above, e.g. , cellulose derivatives, such as sodium carboxymethyl cellulose solution), alcohols (including monohydric alcohols and polyhydric alcohols such as glycols) and their derivatives, and oils (e.g., fractionated coconut oil and arachis oil).
  • the liquid carrier can be halogenated hydrocarbon or other pharmaceutically acceptable propellant.
  • Solid dosage forms for oral administration include, for example, capsules, tablets, pills, powders, and granules.
  • the LSD1 inhibitor may be mixed with at least one inert, pharmaceutically acceptable excipient or carrier such as sodium citrate or dicalcium phosphate and one or more of: (a) fillers or extenders such as starches, lactose, sucrose, glucose, mannital, and silicic acid; (b) binders such as, for example, carboxymethylcellulose, alginates, gelatine, polyvinylpyrrolidone, sucrose, and acacia; (c) humectants such as glycerol; (d) disintegrating agents such as agar-agar, calcium carbonate, potato or tapioca starch, alginic acid, certain silicates, and sodium carbonate; (e) solution retarding agents such as paraffin; absorption accelerators such as quaternary ammonium compounds; (g) wetting agents such as, for example, cetylene glycol
  • excipients suitable for solid formulations include surface modifying agents such as non-ionic and anionic surface modifying agents.
  • surface modifying agents include, but are not limited to, poloxamer 188, benzalkonium chloride, calcium stearate, cetostearyl - - alcohol, cetomacrogol emulsifying wax, sorbitan esters, colloidal silicon dioxide, phosphates, sodium dodecylsulfate, magnesium aluminum silicate, and triethanolamine.
  • the dosage form may also comprise buffering agents.
  • Solid compositions of a similar type may also be employed as fillers in soft and hard-filled gelatine capsules using such excipients as lactose or milk sugar as well as high molecular weight polyethylene glycols and the like.
  • the solid dosage forms of tablets, dragees, capsules, pills, and granules can be prepared with coatings and shells such as enteric coatings, release controlling coatings and other coatings well known in the pharmaceutical formulating art. They may optionally contain opacifying agents and can also be of a composition such that they release the active ingredient(s) only, or preferably, in a certain part of the intestinal tract, optionally, in a delaying manner.
  • Examples of embedding compositions which can be used include polymeric substances and waxes.
  • an inventive composition may be desirable to administer an inventive composition locally to a specific area. This may be achieved, for example, and not by way of limitation, by local infusion during surgery, topical application, by injection, by means of a catheter, by means of suppository, or by means of a skin patch or stent or other implant.
  • the composition is preferably formulated as a gel, an ointment, a lotion, or a cream which can include carriers such as water, glycerol, alcohol, propylene glycol, fatty alcohols, triglycerides, fatty acid esters, or mineral oil.
  • Topical carriers include liquid petroleum, isopropyl palmitate, polyethylene glycol, ethanol (95%), polyoxyethylenemonolaurat (5%) in water, or sodium lauryl sulfate (5%) in water.
  • Other materials such as antioxidants, humectants, viscosity stabilizers, and similar agents may be added as necessary.
  • the inventive compositions may be disposed within transdermal devices placed upon, in, or under the skin.
  • transdermal devices include patches, implants, and injections which release the active ingredient by either passive or active release mechanisms.
  • Transdermal administrations include all administrations across the surface of the body and the inner linings of bodily passage including epithelial and mucosal tissues. Such administrations may be carried out using - - the present compositions in lotions, creams, foams, patches, suspensions, solutions, and suppositories (rectal and vaginal).
  • Transdermal administration may be accomplished through the use of a transdermal patch containing an active ingredient (i.e., a LSD1 inhibitor according to the present invention) and a carrier that is non-toxic to the skin, and allows the delivery of the ingredient for systemic absorption into the bloodstream via the skin.
  • the carrier may take any number of forms such as creams and ointments, pastes, gels, and occlusive devices.
  • the creams and ointments may be viscous liquid or semisolid emulsions of either the oil-in-water or water-in-oil type. Pastes comprised of absorptive powders dispersed in petroleum or hydrophilic petroleum containing the active ingredient may be suitable.
  • occlusive devices may be used to release the active ingredient into the bloodstream such as a semi-permeable membrane covering a reservoir containing the active ingredient with or without a carrier, or a matrix containing the active ingredient.
  • Suppository formulations may be made from traditional materials, including cocoa butter, with or without the addition of waxes to alter the suppository's melting point, and glycerine.
  • Water soluble suppository bases such as polyethylene glycols of various molecular weights, may also be used.
  • RNAi agent may be administered in free (naked) form or by the use of delivery systems that enhance stability and/or targeting, e.g., liposomes, or incorporated into other vehicles, such as hydrogels, cyclodextrins, biodegradable nanocapsules, bioadhesive microspheres, or proteinaceous vectors (WO 00/53722), or in combination with a cationic peptide (US 2007/275923).
  • RNAi agents may also be administered in the form of their precursors or encoding DNAs. Materials and methods for producing various formulations are known in the art and may be adapted for practicing the subject invention. Suitable formulations for the delivery of antibodies can be found, for example, in "Remington's Pharmaceutical Sciences", E.W. Martin, 18 th Ed., 1990, Mack Publishing Co.: Easton, PA.
  • an LSD1 inhibitor of the invention is the only active ingredient in a pharmaceutical composition of the present invention.
  • the pharmaceutical composition further comprises one or more additional - - biologically active agents.
  • suitable biologically active agents include, but are not limited to, biologically active agents generally used in the treatment of cardiomyopathy (such as those listed above), anti-inflammatory agents, immunomodulatory agents, analgesics, antimicrobial agents, antibacterial agents, antibiotics, antioxidants, antiseptic agents, and combinations thereof.
  • the LSD1 inhibitor and the at least one additional therapeutic agent may be combined in one or more preparations for simultaneous, separate or sequential administration of the LSD1 inhibitor and therapeutic agent(s). More specifically, an inventive composition may be formulated in such a way that the LSD1 inhibitor and therapeutic agent(s) can be administered together or independently from each other. For example, the LSD1 inhibitor and a therapeutic agent can be formulated together in a single composition. Alternatively, they may be maintained (e.g., in different compositions and/or containers, for example under the form of a pharmaceutical pack or kit) and administered separately.
  • the present invention provides a pharmaceutical pack or kit comprising one or more containers (e.g., vials, ampoules, test tubes, flasks or bottles) containing one or more ingredients allowing administration of an LSD1 inhibitor of the present invention.
  • containers e.g., vials, ampoules, test tubes, flasks or bottles
  • Different ingredients of a pharmaceutical pack or kit may be supplied in a solid
  • kits may include media for the reconstitution of lyophilized ingredients. Individual containers of the kits will preferably be maintained in close confinement for commercial sale.
  • a pack or kit includes one or more additional therapeutic agent(s), such as those listed above.
  • additional therapeutic agent(s) such as those listed above.
  • Optionally associated with the container(s) can be a notice or package insert in the form prescribed by a governmental agency regulating the manufacture, use or sale of pharmaceutical or biological products, which notice reflects approval by the agency of manufacture, use or sale for human administration.
  • the notice of package insert may contain instructions for use of a pharmaceutical composition according to methods of treatment disclosed herein. - -
  • An identifier e.g., a bar code, radio frequency, ID tags, etc.
  • the identifier can be used, for example, to uniquely identify the kit for purposes of quality control, inventory control, tracking movement between workstations, etc. Examples
  • A-type lamins are ubiquitous proteins encoded by the LMNA gene via alternative splicing. They constitute, together with B-type lamins, the nuclear lamina, which is essential for nuclear architecture.
  • A-type laminas play critical roles in cell and organ physiology, including cell mechanotransduction, intracellular signal transduction, chromatin organization through the lamin- associated domains (LADs) (Solovei et al., Cell, 2013, 152: 584-598) mostly, but not exclusively (Gesson et al, Genome Res., 2016, 26: 462-473), repressing gene transcription , DNA replication and repair (Gruenbaum and Foisner, Annu. Rev. Biochem. 2015, 84: 131-164; Muchir et al., Biochem. Biophys. Res. Commun., 2014, 592: 958-961; Muchir et al., Trends Cardiovasc, 2010, 20: 217-221).
  • Laminopathies include skeletal muscle dystrophies, cardiomyopathies, lipodystrophies, and premature ageing (progeria).
  • LMNA mutations are mostly missense mutations that cause either a haploinsufficiency and/or lead to non-functional proteins with potential toxicity (Dittmer and Misteli, Genome Biol., 2011, 12: 222; Cattin et al, Curr. Opin. Cardiol., 2013, 28: 297-304).
  • A/C lamins as scaffolding proteins for the nucleoskeleton, as signal transduction mediators and as mechanotransduction sensors have been described during the last decade.
  • LADs lamin-associated domains
  • the mouse anti- SSEA1 antibody was obtained from Hybridoma Bank (University of Iowa, Iowa City, IA), the rabbit anti-Oct-3/4 antibody, the anti-Sox2 from Santa-Cruz (CA, USA), the anti-nanog from RID System, the mouse anti-sarcomeric a-actinin antibody from Sigma (23) (Meyer et al, FEBS Lett., 2000, 478: 151-158), the anti-Nkx2.5 antibody from Santa Cruz (HI 14), the anti Isll antibody from Hybridoma Bank, and the anti-Mef2c antibody from Aviva Systems Biology.
  • the anti-LSD 1 antibody was purchased from abeam and the anti- PMHC antibody was obtained from the Hybridoma Bank Iowa.
  • Lmna +/H222P mutant were propagated on a feeder layer as previously described, in a medium containing recombinant LIF (1/1000 to 1/5000) (Meyer et ah, FEBS Lett., 2000, 478: 151-158).
  • the cells were trypsinized and replated every other day. Differentiation was carried out using the hanging drop method, as previously described (Meyer et al, FEBS Lett., 2000, 478: 151-158).
  • the cells were phenotyped every 10 passages using anti-OCTA (Santa Cruz) and NANOG (R&D Systems) antibodies, which were used at 1/100. The cell karyotype was found normal and stable in the course of the experiments.
  • Embryoid Bodies Beating Calculation. After 5 days of differentiation, EBs were plated on gelatin-coated 48-well-plates. To assess differentiation efficiently, the percentage of beating EBs was evaluated every day from Day 7 to Day 8.
  • RNAs were extracted from embryonic stem cells using a Zymo Research kit (Proteingene, France). One microgram of RNA was reverse-transcribed using the SuperscriptTM II reverse transcriptase (Invitrogen, France) and oligo(16)dT. Quantitative PCR was performed using a LightCycler LC480 instrument (Roche). The amplification reactions were carried out as recommended by the manufacturer.
  • reaction mixture contained 10 ⁇ of LightCycler ® -DNA Master SYBR Green I mix (FAST Start KIT, containing Taq DNA polymerase, reaction buffer, deoxynucleoside trisphosphate mix, and SYBR Green I dye), 3 mM MgCl2, 0.5 ⁇ of appropriate primer and 2 ⁇ of cDNA.
  • LightCycler ® -DNA Master SYBR Green I mix FAST Start KIT, containing Taq DNA polymerase, reaction buffer, deoxynucleoside trisphosphate mix, and SYBR Green I dye
  • the data were normalized by PCR analysis of ATP50 expression.
  • the results were expressed as a function of the level of expression of the gene of interest in control undifferentiated stem cells using a previously described mathematical model (Pfaffl, Nucleic Acids Res., 2001, 29: e45).
  • the amplification program included an initial denaturation step at 95°C for 8 minutes, and 40 cycles of denaturation at 95°C for 3 seconds, annealing at 65°C for 10 seconds, and extension at 72°C for 10 seconds.
  • the temperature transition rate was 20°C/s. Fluorescence was measured at the end of each - - extension step.
  • a melting curve was acquired by heating the product at 20°C/s to 95°C, cooling it at 20°C/s to 70°C, keeping it at 70°C for 20 seconds, and then slowly heating it at 0.1°C/s to 95°C. Fluorescence was measured through the slow heating phase. Melting curves were used to determine the specificity of PCR products, which were confirmed using conventional gel electrophoresis.
  • the primers (and their sequences) used in this study are listed in Table 1.
  • Embryonic stem (ES) cells and embryoid bodies (EBs) were fixed in paraformaldehyde for 15 minutes at room temperature. After blocking in a mixture of PBS, 3% BSA, 0.1 % Triton X100, cells were incubated in PBS, 0.01 % Triton X100, 3% BSA with a mouse anti-SSEAl, rabbit anti-Oct-3/4 antibody, or an - - mouse anti-sarcomeric a-actinin antibody (1/500), or an anti-Nkx2.5 antibody, or an anti Isll antibody, or an anti-Mef2c antibody, or an anti-LSD 1 antibody, or an anti- PMHC antibody.
  • ES Embryonic stem
  • EBs embryoid bodies
  • the cells were then incubated with the appropriate secondary antibody, anti-rabbit or mouse IgG coupled to Alexa 488 or Alexa 546 (Molecular Probes Life technologies) diluted to 1/500 in PBS, 0.01% triton X100, 3% BSA.
  • the cells were incubated for 5 minutes in DAPI (Sigma-Aldrich) before mounting (Fluoromount-G, Southern Biotech, Birmingham, AL, USA). Images were acquired using an apotome (Zeiss) and deconvoluted using autoquant software (Bitplane, Switzerland) Plasmid Construction, Embryonic Stem Cell Clone Lmna +/H222PR .
  • the pEGFPCl/hLMNA plasmid containing prelamin A under the control of the SV40 promotor was a kind gift from Dr. Buendia (Favreau et ah, Mol. Cell Biol., 2004, 24: 1481-1492).
  • the CMV promoter was digested using Nhel and Ndel.
  • the cDNA encoding the cardiac alpha actinin (Grey et ah, Hum. Mol. Genet., 2005, 14: 1367- 1377) was then modified with the following primers, in order to integrate Ndel and Nhel restriction sites:
  • Lmna +/H222PR embryonic stem cells clone were propagated in the presence of LIF, selected for ten days by incubation with neomycin (200 ⁇ g/mL) and further screened by PCR.
  • Chromatin Immunoprecipitation Assay Chromatin Immunoprecipitation assays were performed according to previously reported protocols (Jebeniani et ah, J. Vis. Exp., 2016 Jun 3;(112). doi: 10.3791/538742016).
  • the ChIP grade antibodies anti H3K4mel and anti H3K4ac27 used were from Diagenode.
  • Embryos Whole-mount Staining Whole-mount staining of heart sections was performed as previously described (Abboud et ah, Nature Commun., 2015, 6: 6749). Briefly, embryos were fixed overnight at 4°C with 4% paraformaldehyde, and then - - washed with PBS . The samples were dehydrated using ethanol 50%, 70% and 100% and then xylene prior to paraffin embedding. The embryo samples were cut with a microtome, and the obtained heart sections were stained with eosin hematoxylin or permeabilised with PBS Triton 0.1% and blocked with PBS 0.1% Triton X100 added with 10% donkey serum (DS). Mouse anti-sarcomeric a-actinin antibody (1/500) was then added in PBS 10% DS and 1% Triton X100. The secondary antibody Alexa fluor 488 was added in PBS 0.1% Triton X100.
  • the Lmna +/H222P mutation is responsible for severe cardiac defects at early embryonic stage.
  • the cardiac function of E13.5 embryos was monitored by ultrasound imaging of the live fetus.
  • the present Inventors investigated the cardiac function of embryos carrying Lmna H222P/H222P , Lmna +/H222 P mutation or Lmna +/+ siblings.
  • ESCs Lmna +I+ and Lmna +/m22P embryonic stem cells (ESCs) to differentiate into cardiac mesoderm and cardiac myocytes.
  • ESCs were - - differentiated within embryoid bodies (EBs) using the hanging drop method (Grey et al, Hum. Mol. Genet., 2005, 14: 1367-1377; Maltsev et al, Circ. Res., 1994, 75: 233-
  • the present Inventors investigated the functionality of Lmna +/H222P cells during cardiac differentiation.
  • Daily counting of beating EBs from day 7 to day 32 showed a reduced beating activity of embryoid bodies (EBs) and a significant delay in cardiac differentiation, as Lmna +/H222P cardiomyocytes started to beat at day 14, whereas wild-type cardiomyocytes started beating at day 7 ( Figure 2(C)).
  • EBs embryoid bodies
  • Lmna +/H222P mutated mouse embryonic stem cells recapitulated developmental cardiac defects that may account for dilated cardiomyopathy in mouse embryos.
  • the Lmna +/H222P mutation affects mesodermal differentiation through Epithelial-to-mesenchymal transition (EMT) defects.
  • EMT Epithelial-to-mesenchymal transition
  • the Inventors hypothesized that as MesPl regulates the epithelial-to-mesenchymal transition (EMT) of mesendodermal cells and allows the cells to migrate from the posterior to the anterior region of the embryo (Lindsey et al, Cell Stem Cell, 2008, 3: 55-68), the mutant may feature a downregulation of EMT genes. Expression of E-cadherin, Snail and Twist were monitored in the course of differentiation of both wild-type and Lmna +/H222P mutated mouse embryonic stem cells.
  • Figure 3 A shows that E-cadherin was upregulated at Day 4 of differentiation in mutated cells when compared to wild-type - - cells.
  • both Twist, an early player in the EMT process, and Snail were first induced in embryoid bodies at Day 4 of differentiation in both wild-type and mutated cells. Expression of both genes still increased at differentiation Days 5 and 6 in wild-
  • Type A-lamins are associated with chromatin and are likely to regulate chromatin- modifiers.
  • EMT is a process accompanied with chromatin remodeling dependent upon the demethylase LSDl (McDonald et al , Nat. Struct. Mol. Biol., 2001, 18: 867-874).
  • LSDl demethylase LSDl
  • FIG. 3B illustrates the results obtained in these experiments.
  • the left panel indicates the genomic regulatory region investigated on both Twist and MesplXoci.
  • H3K4mel occupied the 3' and 5' regulatory regions of both genes (data not shown). However, less mark was deposited on regulatory region E of Twist and on regions El,
  • LSDl Inhibition of LSDl is sufficient to correct epigenetic and genetic defects in differentiation Lmna +/H222P mutated embryonic stem cells in vitro. Since LSDl was
  • +/H222P found to potentially be more active in embryonic stem cells carrying the Lmna mutation than in wild-type embryonic stem cells, experiments were then designed in order to inhibit LSDl using both a siRNA approach and a selective pharmacological inhibitor, GSK-LSD1.
  • Figure 4A shows that transfection of embryonic stem cells prior to differentiation with the LSDl siRNA was efficient to downregulate the demethylase.
  • H3K4mel +/H222P of H3K4mel on both genes surrounded by chromatin extracted from Lmna - - mutated embryoid bodies when compared to wild-type embryoid bodies, both collected at Day 4.
  • Lmna +/H222P mutated embryoid bodies derived from LSDl-downregulated embryonic stem cells was above that observed in chromatin extracted from wild-type embryoid bodies.
  • LSD1 inhibition was associated with a recovery in expression of both MLC2a (atrial myosin light chain) and ML2v (ventricular myosin light chain), in differentiating
  • the level of expression of these genes reached, or even exceeded, the level of genes expression in wild-type cells ( Figure 4B).
  • blunting LSD1 expression in embryonic stem cells prior to differentiation of wild-type cells did not significantly affect MLC2a or MLC2v expression.
  • FIG. 4(C) shows that treatment of Lmna +/H222P mutated embryoid bodies with GSK-LSDl, a pharmacological inhibitor of LSD1, rescued cardiac differentiation as indexed by sarcomeric cc-actinin immunostaing, which revealed well-organized sarcomeric structures.
  • GSK-LSDl treatment of Lmna H222P/H222P mice prevents cardiomyopathy. Since, GSK-LSDl treatment of Lmna H222P/+ mutated embryoid bodies was found to rescue the cardiac phenotype, the Inventors designed experiments to test the potential therapeutic strategy on Lmna H222P /H222P mice. Lmna H222P /+ mice were bred together. - -
  • Pregnant female were gavaged with 1.5 mg/kg of GSK-LSD1 at E7.5.
  • In utero E13.5 embryonic heart function was monitored using echocardiography. The embryos were then collected for histological analysis.
  • Echocardiography revealed the absence of a cardiomyopathy.
  • the functional parameters of treated homozygous mice were indistinguishable of those of heterozygous mice (see Table 2 below).
  • the histological analysis of treated homozygous mice also failed to reveal any cardiac defects.
  • IVS intraventricular septum d: diameter; LVID: left ventricular internal diameter;
  • LVPDW left ventriclular parietal wall d: diameter
  • LVIDs left ventricular internal diameter in systole
  • LVPDWs left ventriclular parietal wall in sytole d: diameter;
  • LV Vol, s left ventricular volume in systole
  • Laminopathies that affect cardiac and skeletal muscle represent more than 79% of identified mutations in the LMNA gene so far.
  • p.H222P is a missense mutation that is responsible for the autosomal-Dominant Emery-dreifuss muscular dystrophy (AD-EDMD).
  • AD-EDMD patients suffer from a muscular - - dystrophy and a dilated cardiomyopathy. How a specific mutation on a protein that is ubiquitously expressed protein can feature such as specific phenotype is still unknown.
  • This cardiomyopathy likely originates from a defect in ventricular trabeculation and/or compaction - a severe cardiac defect, which impacts ventricular function as monitored by echocardiography.
  • laminopathy is an early onset disease with an embryonic origin.
  • a recently published scientific paper (Meharini et ah, Nature Commun., 2014, 5: 5515) reported that familial cardiac hypertrophy displays an embryonic cardiac origin. Altogether, this suggest that genetic diseases often diagnosed at the adult stage feature an embryonic origin so far ignored and that may be used as a therapeutic window.
  • MesPl is a key gene that is responsible for the epithelial-to-mesenchymal transition (EMT) process of epiblast cells, allowing them to migrate from the posterior to the anterior pole of E7.5 embryos (Kitakima et al., Development, 2000, 127: 3215- 3226; Lindsley et al, Cell Stem Cell, 2008, 3(1): 55-68). Together with the observation that expression of both Twist and snail was slowed down in Lmna H222P/+ embryoid bodies, the present Inventors reasoned that a defect in EMT was, in fact, a key event in the cardiac cell differentiation impairment of embryonic stem cells.
  • EMT epithelial-to-mesenchymal transition
  • chromatin immunoprecipitation (ChIP) assays revealed that the H3K4mel epigentic mark was significantly lost in EMT genes, including Mespl, in Lmna H222P /+ embryoid bodies.
  • MesPl is expressed in bi-potent skeletal and cardiac progenitor cells (Chan et al., Stem Cell Reports, 2016, 12: 26-34).
  • a defect in MesPl expression in cells carrying the H222P Lmna mutation could also play a role in development of skeletal muscle and further in skeletal muscle dystrophy observed in AD-EMD patients.
  • H222P LMNA mutation leads to an hyperactivity of LSD1, either by sequestering LSD1 and its co-REST complex in a lamin- associated domain (LAD) including MesPl and twist, both located on chromosome 7 and distanced from each other by 3 megabases.
  • LAD lamin- associated domain
  • the H222P mutation on LMNA could induce changes in chromatin 3D configuration including the histone H3 that regulates LSDl/CoREST conformational dynamics (Baron et al., Proc. Natl. Acad. Sci .USA, 2012, 109: 12509-12514).
  • the H3K4mel turns out to be crucial in the mesodermal cardiogenic transcriptional scenario.
  • This epigenetic mark, and in turn LSDl that regulates the histone modification, is central to another genetic disease such as the Di-Georges syndrome (Fulcoli et al., Nat. Commun., 2016, 7: 11688).
  • the data obtained in the present study identify LSDl inhibitors, which are currently used in cancer therapy, as therapeutic molecules in cardiomyopathy, in particular dilated cardiomyopathy associated to a laminopathy.

Abstract

La présente invention identifie la lysine déméthylase LSD1 comme nouvelle cible thérapeutique dans des cardiomyopathies, et concerne l'utilisation d'inhibiteurs de la LSD1 dans le traitement et/ou la prévention de cardiomyopathies, en particulier de cardiomyopathies associées à LMNA, telles que des cardiomyopathies dilatées associées à LMNA.
PCT/EP2017/075318 2017-10-05 2017-10-05 Inhibiteurs de la lsd1 pour le traitement et la prévention de cardiomyopathies WO2019068326A1 (fr)

Priority Applications (1)

Application Number Priority Date Filing Date Title
PCT/EP2017/075318 WO2019068326A1 (fr) 2017-10-05 2017-10-05 Inhibiteurs de la lsd1 pour le traitement et la prévention de cardiomyopathies

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
PCT/EP2017/075318 WO2019068326A1 (fr) 2017-10-05 2017-10-05 Inhibiteurs de la lsd1 pour le traitement et la prévention de cardiomyopathies

Publications (1)

Publication Number Publication Date
WO2019068326A1 true WO2019068326A1 (fr) 2019-04-11

Family

ID=60138348

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/EP2017/075318 WO2019068326A1 (fr) 2017-10-05 2017-10-05 Inhibiteurs de la lsd1 pour le traitement et la prévention de cardiomyopathies

Country Status (1)

Country Link
WO (1) WO2019068326A1 (fr)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP3964204A1 (fr) 2020-09-08 2022-03-09 Université d'Aix-Marseille Composés destinés à être utilisés dans le traitement et la prévention de la fibrose de tissus

Citations (35)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2000053722A2 (fr) 1999-03-10 2000-09-14 Phogen Limited Administration de substances a des cellules
WO2002044321A2 (fr) 2000-12-01 2002-06-06 MAX-PLANCK-Gesellschaft zur Förderung der Wissenschaften e.V. Petites molecules d'arn mediant l'interference arn
WO2007021839A2 (fr) 2005-08-10 2007-02-22 Johns Hopkins University Polyamines utiles en tant q'agents therapeutiques antiparasites et anticancereux et en tant qu'inhibiteurs de demethylase specifiques a la lysine
US20070275923A1 (en) 2006-05-25 2007-11-29 Nastech Pharmaceutical Company Inc. CATIONIC PEPTIDES FOR siRNA INTRACELLULAR DELIVERY
WO2008127734A2 (fr) 2007-04-13 2008-10-23 The Johns Hopkins University Inhibiteurs de la déméthylase spécifique de la lysine
WO2010043721A1 (fr) 2008-10-17 2010-04-22 Oryzon Genomics, S.A. Inhibiteurs d’oxydases et leur utilisation
WO2010084160A1 (fr) 2009-01-21 2010-07-29 Oryzon Genomics S.A. Dérivés de phénylcyclopropylamine et leur utilisation médicale
US20100324147A1 (en) 2009-06-02 2010-12-23 Mccafferty Dewey G Arylcyclopropylamines and methods of use
WO2011022489A2 (fr) 2009-08-18 2011-02-24 The Johns Hopkins University Composés (bis)urée et (bis)thiourée en tant que modulateurs épigéniques de la déméthylase 1 spécifique à la lysine et méthodes de traitement de troubles
WO2011035941A1 (fr) 2009-09-25 2011-03-31 Oryzon Genomics S.A. Inhibiteurs de déméthylase-1 spécifique de la lysine et leur utilisation
WO2011042217A1 (fr) 2009-10-09 2011-04-14 Oryzon Genomics S.A. Acétamides d'hétéroaryl- et aryl-cyclopropylamine substitués et leur utilisation
US20110110916A1 (en) 2008-05-01 2011-05-12 Worman Howard J Methods for treating and/or preventing cardiomyopathies by erk or jnk inhibition
WO2011131697A1 (fr) 2010-04-19 2011-10-27 Oryzon Genomics S.A. Inhibiteurs de la déméthylase-1 spécifiques de la lysine, et leur utilisation
WO2011131576A1 (fr) 2010-04-20 2011-10-27 Università Degli Studi Di Roma "La Sapienza" Dérivés de tranylcypromine comme inhibiteurs de l'histone déméthylase lsd1 et/ou lsd2
WO2012013728A1 (fr) 2010-07-29 2012-02-02 Oryzon Genomics S.A. Inhibiteurs de déméthylase lsd1 base d'aryclcyclopropylamine et leur utilisation médicale
WO2012013727A1 (fr) 2010-07-29 2012-02-02 Oryzon Genomics S.A. Dérivés de cyclopropylamine utiles en tant qu'inhibiteurs de lsd1
WO2012045883A1 (fr) 2010-10-08 2012-04-12 Oryzon Genomics S.A. Inhibiteurs d'oxydases de cyclopropylamine
WO2012135113A2 (fr) 2011-03-25 2012-10-04 Glaxosmithkline Llc Cyclopropylamines en tant qu'inhibiteurs de lsd1
WO2012156537A2 (fr) * 2011-05-19 2012-11-22 Oryzon Genomics, S.A. Inhibiteurs de la lysine déméthylase destinés au traitement de la thrombose et de maladies cardiovasculaires
WO2013022047A1 (fr) 2011-08-09 2013-02-14 武田薬品工業株式会社 Composés de cyclopropanamine
WO2013025805A1 (fr) 2011-08-15 2013-02-21 University Of Utah Research Foundation Analogues de (e)-n'-(1-phényléthylidène)benzohydrazide substitués en tant qu'inhibiteurs de l'histone déméthylase
WO2013057320A1 (fr) 2011-10-20 2013-04-25 Oryzon Genomics, S.A. Composés (hétéro)aryle cyclopropylamines en tant qu'inhibiteurs de lsd1
WO2013057322A1 (fr) 2011-10-20 2013-04-25 Oryzon Genomics, S.A. Composés d'(hétéro)aryl-cyclopropylamine à titre d'inhibiteurs de lsd1
WO2014084298A1 (fr) 2012-11-28 2014-06-05 京都府公立大学法人 Inhibiteur sélectif de lsd1 à structure lysine
WO2014164867A1 (fr) * 2013-03-11 2014-10-09 Imago Biosciences Inhibiteurs de kdm1a pour le traitement d'une maladie
WO2014205213A1 (fr) 2013-06-19 2014-12-24 University Of Utah Research Foundation Analogues de (e)-n'-(1-phényléthylidène)benzohydrazide substitués servant d'inhibiteurs de l'histone déméthylase
WO2015120281A1 (fr) 2014-02-07 2015-08-13 Musc Foundation For Research Development Inhibiteurs de kdm1a à base d'aminotroazole et d'aminotétrazole comme modulateurs épigénétiques
WO2015123408A1 (fr) 2014-02-13 2015-08-20 Incyte Corporation Cyclopropylamines en tant qu'inhibiteurs de lsd1
WO2015123437A1 (fr) 2014-02-13 2015-08-20 Incyte Corporation Cyclopropylamines en tant qu'inhibiteurs de lsd1
WO2015123424A1 (fr) 2014-02-13 2015-08-20 Incyte Corporation Cyclopropylamines en tant qu'inhibiteurs de lsd1
WO2016007722A1 (fr) 2014-07-10 2016-01-14 Incyte Corporation Triazolopyridines et triazolopyrazines utilisables comme inhibiteurs de lsd1
WO2016007731A1 (fr) 2014-07-10 2016-01-14 Incyte Corporation Imidazopyridines et imidazopyrazines à utiliser en tant qu'inhibiteurs de lsd1
WO2016007727A1 (fr) 2014-07-10 2016-01-14 Incyte Corporation Triazolopyridines et triazolopyrazines utilisables comme inhibiteurs de lsd1
WO2016007736A1 (fr) 2014-07-10 2016-01-14 Incyte Corporation Imidazopyrazines en tant qu'inhibiteurs de lsd1
WO2017004519A1 (fr) 2015-07-02 2017-01-05 University Of Utah Research Foundation Analogues de benzohydrazide substitués utiles en tant qu'inhibiteurs de l'histone déméthylase

Patent Citations (35)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2000053722A2 (fr) 1999-03-10 2000-09-14 Phogen Limited Administration de substances a des cellules
WO2002044321A2 (fr) 2000-12-01 2002-06-06 MAX-PLANCK-Gesellschaft zur Förderung der Wissenschaften e.V. Petites molecules d'arn mediant l'interference arn
WO2007021839A2 (fr) 2005-08-10 2007-02-22 Johns Hopkins University Polyamines utiles en tant q'agents therapeutiques antiparasites et anticancereux et en tant qu'inhibiteurs de demethylase specifiques a la lysine
US20070275923A1 (en) 2006-05-25 2007-11-29 Nastech Pharmaceutical Company Inc. CATIONIC PEPTIDES FOR siRNA INTRACELLULAR DELIVERY
WO2008127734A2 (fr) 2007-04-13 2008-10-23 The Johns Hopkins University Inhibiteurs de la déméthylase spécifique de la lysine
US20110110916A1 (en) 2008-05-01 2011-05-12 Worman Howard J Methods for treating and/or preventing cardiomyopathies by erk or jnk inhibition
WO2010043721A1 (fr) 2008-10-17 2010-04-22 Oryzon Genomics, S.A. Inhibiteurs d’oxydases et leur utilisation
WO2010084160A1 (fr) 2009-01-21 2010-07-29 Oryzon Genomics S.A. Dérivés de phénylcyclopropylamine et leur utilisation médicale
US20100324147A1 (en) 2009-06-02 2010-12-23 Mccafferty Dewey G Arylcyclopropylamines and methods of use
WO2011022489A2 (fr) 2009-08-18 2011-02-24 The Johns Hopkins University Composés (bis)urée et (bis)thiourée en tant que modulateurs épigéniques de la déméthylase 1 spécifique à la lysine et méthodes de traitement de troubles
WO2011035941A1 (fr) 2009-09-25 2011-03-31 Oryzon Genomics S.A. Inhibiteurs de déméthylase-1 spécifique de la lysine et leur utilisation
WO2011042217A1 (fr) 2009-10-09 2011-04-14 Oryzon Genomics S.A. Acétamides d'hétéroaryl- et aryl-cyclopropylamine substitués et leur utilisation
WO2011131697A1 (fr) 2010-04-19 2011-10-27 Oryzon Genomics S.A. Inhibiteurs de la déméthylase-1 spécifiques de la lysine, et leur utilisation
WO2011131576A1 (fr) 2010-04-20 2011-10-27 Università Degli Studi Di Roma "La Sapienza" Dérivés de tranylcypromine comme inhibiteurs de l'histone déméthylase lsd1 et/ou lsd2
WO2012013728A1 (fr) 2010-07-29 2012-02-02 Oryzon Genomics S.A. Inhibiteurs de déméthylase lsd1 base d'aryclcyclopropylamine et leur utilisation médicale
WO2012013727A1 (fr) 2010-07-29 2012-02-02 Oryzon Genomics S.A. Dérivés de cyclopropylamine utiles en tant qu'inhibiteurs de lsd1
WO2012045883A1 (fr) 2010-10-08 2012-04-12 Oryzon Genomics S.A. Inhibiteurs d'oxydases de cyclopropylamine
WO2012135113A2 (fr) 2011-03-25 2012-10-04 Glaxosmithkline Llc Cyclopropylamines en tant qu'inhibiteurs de lsd1
WO2012156537A2 (fr) * 2011-05-19 2012-11-22 Oryzon Genomics, S.A. Inhibiteurs de la lysine déméthylase destinés au traitement de la thrombose et de maladies cardiovasculaires
WO2013022047A1 (fr) 2011-08-09 2013-02-14 武田薬品工業株式会社 Composés de cyclopropanamine
WO2013025805A1 (fr) 2011-08-15 2013-02-21 University Of Utah Research Foundation Analogues de (e)-n'-(1-phényléthylidène)benzohydrazide substitués en tant qu'inhibiteurs de l'histone déméthylase
WO2013057320A1 (fr) 2011-10-20 2013-04-25 Oryzon Genomics, S.A. Composés (hétéro)aryle cyclopropylamines en tant qu'inhibiteurs de lsd1
WO2013057322A1 (fr) 2011-10-20 2013-04-25 Oryzon Genomics, S.A. Composés d'(hétéro)aryl-cyclopropylamine à titre d'inhibiteurs de lsd1
WO2014084298A1 (fr) 2012-11-28 2014-06-05 京都府公立大学法人 Inhibiteur sélectif de lsd1 à structure lysine
WO2014164867A1 (fr) * 2013-03-11 2014-10-09 Imago Biosciences Inhibiteurs de kdm1a pour le traitement d'une maladie
WO2014205213A1 (fr) 2013-06-19 2014-12-24 University Of Utah Research Foundation Analogues de (e)-n'-(1-phényléthylidène)benzohydrazide substitués servant d'inhibiteurs de l'histone déméthylase
WO2015120281A1 (fr) 2014-02-07 2015-08-13 Musc Foundation For Research Development Inhibiteurs de kdm1a à base d'aminotroazole et d'aminotétrazole comme modulateurs épigénétiques
WO2015123408A1 (fr) 2014-02-13 2015-08-20 Incyte Corporation Cyclopropylamines en tant qu'inhibiteurs de lsd1
WO2015123437A1 (fr) 2014-02-13 2015-08-20 Incyte Corporation Cyclopropylamines en tant qu'inhibiteurs de lsd1
WO2015123424A1 (fr) 2014-02-13 2015-08-20 Incyte Corporation Cyclopropylamines en tant qu'inhibiteurs de lsd1
WO2016007722A1 (fr) 2014-07-10 2016-01-14 Incyte Corporation Triazolopyridines et triazolopyrazines utilisables comme inhibiteurs de lsd1
WO2016007731A1 (fr) 2014-07-10 2016-01-14 Incyte Corporation Imidazopyridines et imidazopyrazines à utiliser en tant qu'inhibiteurs de lsd1
WO2016007727A1 (fr) 2014-07-10 2016-01-14 Incyte Corporation Triazolopyridines et triazolopyrazines utilisables comme inhibiteurs de lsd1
WO2016007736A1 (fr) 2014-07-10 2016-01-14 Incyte Corporation Imidazopyrazines en tant qu'inhibiteurs de lsd1
WO2017004519A1 (fr) 2015-07-02 2017-01-05 University Of Utah Research Foundation Analogues de benzohydrazide substitués utiles en tant qu'inhibiteurs de l'histone déméthylase

Non-Patent Citations (114)

* Cited by examiner, † Cited by third party
Title
"Remington: The Science and Practice of Pharmacy", 2003, LIPPINCOTT, WILLIAMS & WILKINS, article 20TH ED.
ABBOUD ET AL., NATURE COMMUN., vol. 6, 2015, pages 6749
ALBA MAIQUES-DIAZ ET AL: "LSD1: biologic roles and therapeutic targeting", EPIGENOMICS, vol. 8, no. 8, 1 August 2016 (2016-08-01), United Kingdom, pages 1103 - 1116, XP055438086, ISSN: 1750-1911, DOI: 10.2217/epi-2016-0009 *
AMANO ET AL., BIOORG. MED. CHEM., vol. 25, 2017, pages 2617 - 2624
ANTONIADES ET AL., J. INTERV. CARD. ELECTROPHYSIOL., vol. 19, 2007, pages 1 - 7
ARIMURA ET AL., HUM. MOL. GENET., vol. 14, 2005, pages 155 - 169
BARON ET AL., PROC. NATL. ACAD. SCI .USA, vol. 109, 2012, pages 12509 - 12514
BEN YAOU ET AL., REV. NEUROL., vol. 161, 2005, pages 42 - 54
BENELKEBIR ET AL., BIOORG. MED. CHEM., vol. 19, 2011, pages 3709 - 3716
BINDA ET AL., J. AM. CHEM. SOC., vol. 132, 2010, pages 6827 - 6833
BONN ET AL., NATURE GENET., vol. 44, 2012, pages 148 - 156
BONNE; MUCHIR: "Kaan Kirali", 2017, article CHAPTER 7: "Cardiology and Cardiovascular Medicine - Cardiomyopathies: Types and Treatments"
BORRELLO ET AL., BIOORG. MED. CHEM. LETT., vol. 27, 2017, pages 2099 - 2101
CATTIN ET AL., CURR. OPIN. CARDIOL., vol. 28, 2013, pages 297 - 304
CHAN ET AL., STEM CELL REPORTS, vol. 12, 2016, pages 26 - 34
CHEMICAL ABSTRACTS, Columbus, Ohio, US; abstract no. 1423715-09-6
CHEMICAL ABSTRACTS, Columbus, Ohio, US; abstract no. 1431368-48-7
CUI ET AL., BLOOD, vol. 126, 2015, pages 386 - 396
CULHANE ET AL., J. AM. CHEM. SOC., vol. 128, 2006, pages 4536 - 4537
DATABASE BIOSIS [online] BIOSCIENCES INFORMATION SERVICE, PHILADELPHIA, PA, US; 1994, KOGA YOSHINORI ET AL: "Therapeutic management of dilated cardiomyopathy", XP002781820, Database accession no. PREV199497362984 *
DHANAK: "Proceedings of the 104th Annual Meeting of the American Association for Cancer Research", 2013, article 104TH ED.: "Drugging the cancer genome"
DITTMER; MISTELI, GENOME BIOL., vol. 12, 2011, pages 222
E.W. MARTIN: "Remington's Pharmaceutical Sciences", 1990, MACK PUBLISHING CO., article 18TH ED.
ECKERSLEY-MASLIN ET AL., NUCLEUC, vol. 4, 2013, pages 53 - 60
ELLIOTT ET AL., EUR. HEART J., vol. 29, 2008, pages 270
FAVREAU ET AL., MOL. CELL BIOL., vol. 24, 2004, pages 1481 - 1492
FOSTER ET AL., MOL. CELL. BIOL., vol. 30, no. 20, pages 4851 - 4863
FU ET AL., FUTURE MED. CHEM., vol. 9, 2017, pages 1227 - 1242
FULCOLI ET AL., NAT. COMMUN., vol. 7, 2016, pages 11688
GESSON ET AL., GENOME RES., vol. 26, 2016, pages 462 - 473
GOLZIO ET AL., EUROPACE, vol. 9, no. 12, 2007, pages 1158 - 1160
GOODEN ET AL., BIOORG. MED. CHEM. LET., vol. 18, 2008, pages 3047 - 3051,6827-6833
GREY ET AL., HUM. MOL. GENET., vol. 14, 2005, pages 1367 - 1377
GREY ET AL., HUM. MOL. GENET., vol. 14, no. 10, 2005, pages 1367 - 1377
GRUENBAUM; FOISNER, ANNU. REV. BIOCHEM., vol. 84, 2015, pages 131 - 164
HAYWARD; COLE, METHODS ENZYMOL., vol. 573, 2016, pages 261 - 278
HERMIDA-PRIETO ET AL., AM. J. CARDIOL., vol. 94, 2004, pages 50 - 54
HINO ET AL., CANCER SCI., vol. 107, 2016, pages 1187 - 1192
HINO ET AL., NATURE COMM., vol. 3, 2012
HUANG ET AL., CLIN. CANCER RES., vol. 15, 2009, pages 7217 - 7228
HUANG ET AL., PNAS, vol. 104, 2007, pages 8023 - 8028
IMEN JEBENIANI: "Etude in vitro et in vivo d'une cardiomyopathie secondaire à une laminopathie", THESES.FR, 27 January 2017 (2017-01-27), pages 1 - 2, XP055482864, Retrieved from the Internet <URL:http://www.theses.fr/2017AIXM0025> [retrieved on 20180611] *
ISHIKAWA ET AL., MOL. CANCER THER., vol. 16, no. 2, 2017, pages 273 - 284
JEBENIANI ET AL., J. VIS. EXP., vol. 112, 3 June 2016 (2016-06-03)
KAKIZAWA ET AL., BIOORG. MED. CHEM. LETT., vol. 25, 2015, pages 1925 - 1928
KHAN ET AL., MED. RES. REV., vol. 33, 2013, pages 873 - 910
KISKUS ET AL., LEUKEMIA, vol. 28, 2014, pages 2155 - 2164
KITAKIMA ET AL., DEVELOPMENT, vol. 127, 2000, pages 3215 - 3226
KOGA YOSHINORI ET AL: "Therapeutic management of dilated cardiomyopathy", CARDIOVASCULAR DRUGS AND THERAPY, vol. 8, no. 1, 1994, pages 83 - 88, ISSN: 0920-3206 *
KRESS ET AL., MAMM. GENOME, vol. 9, 1998, pages 998 - 1001
KUMARASINGHE ET AL., ACS MED. CHEM. LETT., vol. 5, 2013, pages 29 - 33
KUTZ ET AL., MED. CHEM. COMMUN., vol. 5, 2014, pages 1863 - 1870
LEE ET AL., CHEM. BIOL., vol. 13, no. 6, 2006, pages 563 - 567
LI ET AL., ACS MED. CHEM. LETT., vol. 8, 2017, pages 384 - 389
LIEBERMAN, PHARMACEUTICAL DOSAGE FORMS, vol. 1-3, 1992
LINDSEY ET AL., CELL STEM CELL, vol. 3, 2008, pages 55 - 68
LINDSLEY ET AL., CELL STEM CELL, vol. 3, no. 1, 2008, pages 55 - 68
LLOYD, THE ART, SCIENCE AND TECHNOLOGY OF PHARMACEUTICAL COMPOUNDING, 1999
LYNCH ET AL., EXPERT. OPIN. THER. TARGETS, vol. 13, 2012, pages 1239 - 1249
MALCOMSON ET AL., FEBS J., vol. 282, 2015, pages 3190 - 3198
MALTSEV ET AL., CIRC. RES., vol. 75, 1994, pages 233 - 244
MARON ET AL., CIRCULATION, vol. 113, pages 1807
MCDONALD ET AL., NAT STRUCT MOL BIOL., vol. 18, 2011, pages 867 - 874
MCDONALD ET AL., NAT. STRUCT. MOL. BIOL., vol. 18, 2001, pages 867 - 874
MCGRATH ET AL., CANCER RES., vol. 76, 2016, pages 1975 - 1988
MEHARINI ET AL., NATURE COMMUN., vol. 5, 2014, pages 5515
METZGER ET AL., NATURE, vol. 437, 2005, pages 436 - 439
MEUNE ET AL., NEW ENGL. J. MED., vol. 354, no. 2, 2006, pages 209 - 210
MEYER ET AL., FEBS LETT., vol. 478, 2000, pages 151 - 158
MIMASU ET AL., BIOCHEMISTRY, vol. 49, 2010, pages 6494 - 6503
MOULD ET AL., BIOORG. MED. CHEM. LETT., vol. 27, 2017, pages 3190 - 3195
MUCHIR ET AL., BIOCHEM. BIOPHYS. RES. COMMUN., vol. 592, 2014, pages 958 - 961
MUCHIR ET AL., TRENDS CARDIOVASC., vol. 20, 2010, pages 217 - 221
NEELAMEGAM ET AL., ACS CHEM. NEUROSCI., vol. 3, 2012, pages 120 - 128
NOWOTARSKI ET AL., BIOORG. MED. CHEM., vol. 23, 2015, pages 1601 - 1612
OLIVE ET AL., ARTERIOSCLER. THROMB. VASE. BIOL., vol. 11, 2010, pages 2301 - 2309
PERIC-HUPKES ET AL., MOL. CELL, vol. 38, 2010, pages 603 - 613
PEROVANOVIC ET AL., SCI. TRANSL. MED., vol. 8, 2016, pages 335ra58
PFAFFL, NUCLEIC ACIDS RES., vol. 29, 2001, pages e45
PICKAR, DOSAGE CALCULATIONS, 1999
PRUSEVICH ET AL., ACS CHEM. BIOL., vol. 9, 2014, pages 1284 - 1293
QUARTA ET AL., EUR. HEART J., vol. 33, 2012, pages 1128 - 1136
SAKANE ET AL., BIOCHEM. BIOPHYS. RES. COMMUN., vol. 444, 2014, pages 24 - 29
SANKAR ET AL., CLIN. CANCER RES., vol. 20, 2014, pages 4584 - 4597
SARTORI ET AL., J. MED. CHEM., vol. 61, 2017, pages 1673 - 1692
SCHMITT ET AL., J. MED. CHEM., vol. 56, 2013, pages 7334 - 7342
SCHULZ-FINCKE ET AL., MOL. CANCER THER., vol. 14, no. 12, 2015
SHARMA ET AL., J. MED. CHEM., vol. 43, 2010, pages 5189 - 5212
SHI ET AL., CELL, vol. 119, 2004, pages 941 - 953
SOLOVEI ET AL., CELL, vol. 152, 2013, pages 584 - 598
SOMA ET AL., J. MED.CHEM., vol. 56, 2013, pages 9496 - 9508
SUN ET AL., MOL. CELL BIOL., vol. 30, 2010, pages 1997 - 2005
TORTORICI ET AL., ACS CHEM. BIOL., vol. 8, 2013, pages 1677 - 1682
TRIFIRO ET AL., FUTURE MED. CHEM., vol. 9, 2017, pages 1161 - 1174
UEDA ET AL., J. AM. CHEM. SOC., vol. 131, 2009, pages 17536 - 17537
VALENTE ET AL., ACS MED. CHEM. LETT., vol. 6, 2014, pages 173 - 177
VALENTE ET AL., EUR. J. MED. CHEM., vol. 94, 2015, pages 163 - 174
VAN BERLO ET AL., J. MOD. MED., vol. 83, no. 1, 2005, pages 79 - 83
VAN TINTELEN ET AL., J. AM. COLL. CARDIOL., vol. 49, 2007, pages 2430 - 2439
VAN VLIET ET AL., CARDIOVASC. RES., vol. 96, 2012, pages 352 - 362
VIANELLO ET AL., EUR. J. MED. CHEM., vol. 86, 2014, pages 352 - 363
VIANELLO ET AL., J. MED. CHEM., vol. 60, 2017, pages 1693 - 1715
WANG ET AL., CANCER RES., vol. 71, 2011, pages 7238 - 7249
WANG ET AL., CELL, vol. 138, 2009, pages 660 - 672
WANG ET AL., EUR. J. MED. CHEM., vol. 125, 2017, pages 940 - 951
WILLMANN ET AL., INT. J. CANCER, vol. 131, 2012, pages 2704 - 2709
XI ET AL., BIOORG. CHEM., vol. 72, 2017, pages 182 - 189
XU ET AL., CELL PHYSIOL. BIOCHEM., vol. 31, 2013, pages 854 - 862
YANG KE (CORRESPONDENCE) ET AL: "Attenuation of LSD1 inhibitor on ISO-induced cardiac contractile response", CARDIOLOGY, KARGER, BASEL, CH, vol. 129, no. Suppl.1, 1 August 2014 (2014-08-01), pages 52 - 53, XP009505859, ISSN: 0008-6312 *
YE ET AL., MED. CHEM. COMMUN., vol. 5, 2014, pages 650 - 654
ZHENG ET AL., J. MED. CHEM., vol. 56, 2013, pages 8543 - 8560
ZHOU ET AL., BIOORG. MED. CHEM. LETT., vol. 26, 2016, pages 4552 - 4557
ZHU ET AL., AMINO ACIDS, vol. 42, no. 42, 2012, pages 887 - 898
ZHU ET AL., PLOS ONE, vol. 3, 2008, pages e1446

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP3964204A1 (fr) 2020-09-08 2022-03-09 Université d'Aix-Marseille Composés destinés à être utilisés dans le traitement et la prévention de la fibrose de tissus
WO2022053520A1 (fr) 2020-09-08 2022-03-17 Université D'aix Marseille Inhibiteurs de lsd1 destinés à être utilisés dans le traitement et la prévention de la fibrose tissulaire

Similar Documents

Publication Publication Date Title
US11458137B2 (en) Compositions and methods of using tyrosine kinase inhibitors
US11931402B2 (en) Compositions for treating heart disease by inhibiting the action of mAKAP-β
JP2019506459A (ja) アルファ−１−アドレナリン受容体アゴニスト療法
US20190381047A1 (en) Methods for treatment of retinal disease by photoreceptor gene expression modulation
AU2018234632A1 (en) Dominant active yap, a hippo effector, induces chromatin accessibility and cardiomyocyte renewal
WO2019037222A1 (fr) Utilisation d&#39;un inhibiteur de gpr31 dans la préparation de médicaments
JP5686730B2 (ja) 心臓肥大を抑制、遅延、および／または予防するための方法、および医薬組成物
US11077110B2 (en) Compositions and methods for treating and preventing metabolic disorders
WO2019068326A1 (fr) Inhibiteurs de la lsd1 pour le traitement et la prévention de cardiomyopathies
WO2017182580A1 (fr) Procédés de diagnostic et de traitement de troubles liés à la reproduction et procédés de contraception
EP3658157B1 (fr) Traitement de cardiopathies par inhibition de l&#39;action des protéines d&#39;ancrage aux protéines kinases a (makap) du muscle
WO2013020372A1 (fr) Méthodes et réactifs pour la prévention et la guérison d&#39;une insulinorésistance et du diabète sucré
US20210353585A1 (en) Methods and compositions for treating laminopathies affecting skeletal or cardiac muscle
US10973912B2 (en) Treatment for myopathy
US20150031622A1 (en) Rhogef12 is a therapeutic target for the treatment of heart failure
JP2021509914A (ja) Ｄｌｋ１−Ｄｉｏ３クラスターに位置するｍｉＲＮＡまたはそのバリアントを活性成分として含む、筋疾患またはカヘキシーの予防または処置のための医薬組成物
US9309514B2 (en) G protein-coupled purinergic receptor Gpr17 mediates orexigenic effects of FoxO1 in AgRP neurons
KR101436684B1 (ko) 저산소성 허혈 예방 또는 치료용 조성물
TW201138777A (en) Involvement of androgen/androgen receptor pathway in Fabry disease
US20110293599A1 (en) Method for modulating muscle differentiation or regeneration
US20210261966A1 (en) TREATMENT OF CARDIOMYOPATHY THROUGH MODULATION OF HYPOXIA-INDUCED eRNA ACTIVITY

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 17786857

Country of ref document: EP

Kind code of ref document: A1

NENP Non-entry into the national phase

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 17786857

Country of ref document: EP

Kind code of ref document: A1