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WO2018130840A1 - Inhibiteur de méthylation de l'arginine pour le traitement ou la prévention d'une maladie cardiovasculaire - Google Patents

Inhibiteur de méthylation de l'arginine pour le traitement ou la prévention d'une maladie cardiovasculaire Download PDF

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WO2018130840A1
WO2018130840A1 PCT/GB2018/050080 GB2018050080W WO2018130840A1 WO 2018130840 A1 WO2018130840 A1 WO 2018130840A1 GB 2018050080 W GB2018050080 W GB 2018050080W WO 2018130840 A1 WO2018130840 A1 WO 2018130840A1
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methyl
arginine methylation
methylation inhibitor
pharmaceutically acceptable
solvate
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Pedro BELTRAN-ALVAREZ
Francisco RIVERO
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University of Hull
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University of Hull
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    • 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
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/16—Amides, e.g. hydroxamic acids
    • A61K31/165—Amides, e.g. hydroxamic acids having aromatic rings, e.g. colchicine, atenolol, progabide
    • A61K31/167—Amides, e.g. hydroxamic acids having aromatic rings, e.g. colchicine, atenolol, progabide having the nitrogen of a carboxamide group directly attached to the aromatic ring, e.g. lidocaine, paracetamol
    • 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
    • A61K31/16—Amides, e.g. hydroxamic acids
    • A61K31/17—Amides, e.g. hydroxamic acids having the group >N—C(O)—N< or >N—C(S)—N<, e.g. urea, thiourea, carmustine
    • 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
    • A61K31/335—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin
    • A61K31/34—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having five-membered rings with one oxygen as the only ring hetero atom, e.g. isosorbide
    • 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
    • A61K31/335—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin
    • A61K31/35—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having six-membered rings with one oxygen as the only ring hetero atom
    • A61K31/352—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having six-membered rings with one oxygen as the only ring hetero atom condensed with carbocyclic rings, e.g. methantheline 
    • 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
    • A61K31/41—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with two or more ring hetero atoms, at least one of which being nitrogen, e.g. tetrazole
    • A61K31/4245—Oxadiazoles
    • 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
    • A61K31/41—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with two or more ring hetero atoms, at least one of which being nitrogen, e.g. tetrazole
    • A61K31/433—Thidiazoles
    • 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
    • A61K31/4353—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 ortho- or peri-condensed with heterocyclic ring systems
    • A61K31/4355—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 ortho- or peri-condensed with heterocyclic ring systems the heterocyclic ring system containing a five-membered ring having oxygen as a ring hetero atom
    • 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
    • A61K31/47—Quinolines; Isoquinolines
    • A61K31/473—Quinolines; Isoquinolines ortho- or peri-condensed with carbocyclic ring systems, e.g. acridines, phenanthridines
    • A—HUMAN NECESSITIES
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    • 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
    • A61K31/53—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with three nitrogens as the only ring hetero atoms, e.g. chlorazanil, melamine
    • 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
    • A61K31/70—Carbohydrates; Sugars; Derivatives thereof
    • A61K31/7042—Compounds having saccharide radicals and heterocyclic rings
    • A61K31/7052—Compounds having saccharide radicals and heterocyclic rings having nitrogen as a ring hetero atom, e.g. nucleosides, nucleotides
    • A61K31/706—Compounds having saccharide radicals and heterocyclic rings having nitrogen as a ring hetero atom, e.g. nucleosides, nucleotides containing six-membered rings with nitrogen as a ring hetero atom
    • A61K31/7064—Compounds having saccharide radicals and heterocyclic rings having nitrogen as a ring hetero atom, e.g. nucleosides, nucleotides containing six-membered rings with nitrogen as a ring hetero atom containing condensed or non-condensed pyrimidines
    • A61K31/7076—Compounds having saccharide radicals and heterocyclic rings having nitrogen as a ring hetero atom, e.g. nucleosides, nucleotides containing six-membered rings with nitrogen as a ring hetero atom containing condensed or non-condensed pyrimidines containing purines, e.g. adenosine, adenylic acid
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P7/00—Drugs for disorders of the blood or the extracellular fluid
    • A61P7/02—Antithrombotic agents; Anticoagulants; Platelet aggregation inhibitors
    • 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

Definitions

  • aspects and embodiments of the present invention relate to the treatment and/or prevention of cardiovascular disease, for example thromboembolic conditions, such as atherothrombotic disease.
  • cardiovascular disease for example thromboembolic conditions, such as atherothrombotic disease.
  • the present invention relates at least in part to an arginine methylation inhibitor ('ArgMe' inhibitor) for use in the treatment and/or prevention of cardiovascular disease such as thromboembolic conditions (for example atherothrombotic disease).
  • 'ArgMe' inhibitor arginine methylation inhibitor
  • the invention also provides inter alia pharmaceutical compositions comprising an arginine methylation inhibitor, together with methods of treating and/or preventing such conditions, diseases and/or disorders as well as other subject matter.
  • Atherosclerosis is characterised by the hardening and narrowing of arteries due to the deposition of fatty deposits (known as atheromas or atherosclerotic plaques) on their walls. In an advanced state, atherosclerosis increases the risk of thrombi that can potentially block the blood flow to organs like the heart or brain, leading to myocardial infarction or stroke, respectively.
  • integrins facilitates the binding of the plasma protein fibrinogen, which subsequently supports platelet aggregation and consolidation of the clot.
  • Aggregation is initially a reversible process but is consolidated by the release of soluble agonists such as adenosine diphosphate ('ADP') and thromboxane A2 (TXA2 ), ensuring rapid entrapment of platelets (Jurk et al., Semin. Thromb. Hemost. 2005, 31, 381-392). While this process is critical to haemostatic protection of the vasculature after injury, the rupture of atherosclerotic plaques drives uncontrolled platelet activation that leads to arterial thrombosis and the clinical events mentioned above.
  • soluble agonists such as adenosine diphosphate ('ADP') and thromboxane A2 (TXA2 )
  • the inventors have developed such an understanding and, in particular, have determined that the post-translational modification of arginine residues in proteins by methylation is very likely to play a critical role in the signalling pathways that regulate platelet physiology.
  • the inventors have found that inhibition of arginine methylation significantly delays platelet aggregation in response to agonists.
  • activatory signalling pathways triggered by agonists are counterbalanced by inhibitory signalling pathways triggered by prostacyclin ('PGI2') and nitric oxide ('NO') released by the endothelium.
  • 'PGI2' prostacyclin
  • 'NO' nitric oxide
  • PGI2 and NO for example, trigger the formation of cyclic nucleotides, which in turn activate protein kinases A and G, respectively, which phosphorylate numerous substrates.
  • Arginine methylation consists of the introduction of one ('monoArgMe') or two ('diArgMe') methyl groups on the ⁇ -nitrogen of arginine residues.
  • Arginine methylation has been implicated in a number of cellular processes including signal transduction, RNA processing, and protein- protein interactions (Bedford etal., Mol. Cell. 2009, 33, 1-13).
  • the effect of arginine methylation in the cardiovascular system, in particular of cardiac ion channels in the context of sudden cardiac death and heart failure has already been investigated (Beltran-Alvarez et al., Proteome Res. 2011, 10, 3712-3719; Beltran-Alvarez et al., FEBS Lett. 2013, 587, 3159-3165; Beltran-Alvarez et al., J. Mol. Cell. Cardiol.
  • Arginine methylation is catalysed by protein arginine methyl transferases ('PRMTs'). There are at least 9 enzymes encoded in the human genome ('PRMT1-9'), and proteomics analysis indicates the presence in platelets of at least PRMT1 and -5 (Kim et al., Nature 2014, 509, 575-581).
  • an arginine methylation inhibitor for example, a PRMT inhibitor such as AMI-1 (Cheng et al., J. Biol. Chem. 2004, 279, 23892-23899)
  • a PRMT inhibitor such as AMI-1
  • TXA2 pathway e.g. aspirin
  • ADP pathway e.g. clopidogrel
  • GPIIb/llla e.g. abciximab
  • thrombin pathway e.g. vorapaxar
  • arginine methylation inhibitors are effective as antiplatelet agents by a mechanism that targets the inhibitory pathways, and at least the cAMP pathway. It is therefore believed that arginine methylation inhibitors are able to produce potent antiplatelet effects without compromising the ability of thrombin to generate fibrin, and without completely inhibiting platelet activation. This could result in antiplatelet activity in the event of a blood clot, while maintaining protective haemostatic platelet functions, thus minimising risk of bleeding.
  • the present invention therefore provides for the use of arginine methylation inhibitor(s) as antiplatelet agent(s) for the treatment of cardiovascular disease and, in particular, thromboembolic conditions (for example atherothrombotic disease).
  • VASP vasodilator stimulated phosphoprotein
  • VASP vasodilator stimulated phosphoprotein
  • Ena-VASP family members contain an EHV1 N-terminal domain that binds proteins containing E/DFPPPPXD/E motifs and targets Ena-VASP proteins to focal adhesions.
  • the NCBI accession number of VASP (homo sapiens) is NP_003361 (version 1).
  • VASP proteins are actin-associated proteins involved in a range of processes dependent on cytoskeleton remodelling and cell polarity such as axon guidance, lamellipodial and filopodial dynamics, platelet activation and cell migration. VASP promotes actin filament elongation.
  • VASP stimulates actin filament elongation by promoting the transfer of profilin-bound actin monomers onto the barbed end of growing actin filaments. VASP thus regulates actin dynamics in platelets and plays an important role in regulating platelet aggregation. In platelets, VASP is modified by phosphorylation e.g. at Serine 157 and platelet reactivity inversely correlates with VASP S157 phosphorylation levels (Horstrup K, et al. 1994 Eur J Biochem 225, 21-7).
  • VASP phosphorylation index has been developed and kits to measure VASP phosphorylation index are commercially available (Diagnostica Stago, Asnieres, France) and used in research studies of cardiovascular disease (Laine M, et al. 2013 Thromb Res. 132(1):e15-8.
  • an arginine methylation inhibitor for use as an antiplatelet agent in the event of a blood clot wherein the use of the arginine methylation inhibitor has a reduced risk of bleeding. It is an aim of certain embodiments of the present invention to provide an arginine methylation inhibitor for use in the treatment and/or prevention of atherothrombotic disease wherein the use of the arginine methylation inhibitor maintains protective haemostatic platelet functions whilst reducing risk of bleeding.
  • an arginine methylation inhibitor for use in the treatment and/or prevention of thrombosis (for example the prevention of thrombosis) in a subject undergoing a surgical procedure, for example an angioplasty procedure. It is an aim of certain embodiments of the present invention to identify target proteins important to platelet physiology wherein arginine methylation may be inhibited. It is envisaged that such inhibition may result in the provision of an arginine methylation inhibitor for use in the treatment and/or prevention of cardiovascular disease and, in particular, thromboembolic conditions, such as atherothrombotic disease.
  • an arginine methylation inhibitor for use in the treatment and/or prevention of cardiovascular disease.
  • the arginine methylation inhibitor is for use in the treatment and/or prevention of a thromboembolic condition.
  • the arginine methylation inhibitor is for use in the treatment and/or prevention of atherothrombotic disease.
  • the arginine methylation inhibitor is for use in the treatment and/or prevention of a disease selected from coronary artery disease ('CAD'), ischemic heart disease ('IHD' also known as coronary heart disease ('CHD')), acute coronary syndrome ('ACS') (for example myocardial infarction and/or angina), left ventricular thrombus ('LVT'), cerebral ischemia, critical limb ischemia, peripheral artery disease ('PAD'), deep vein thrombosis ('DVT') and pulmonary embolism.
  • a disease selected from coronary artery disease ('CAD'), ischemic heart disease ('IHD' also known as coronary heart disease ('CHD')), acute coronary syndrome ('ACS') (for example myocardial infarction and/or angina), left ventricular thrombus ('LVT'), cerebral ischemia, critical limb ischemia, peripheral artery disease ('PAD'), deep vein thrombosis ('DVT') and
  • the arginine methylation inhibitor is for use as an antiplatelet agent and/or antithrombotic agent.
  • the arginine methylation inhibitor is for use in the treatment and/or prevention of thrombosis (for example the prevention of thrombosis) in a subject undergoing a surgical procedure, for example an angioplasty procedure.
  • a pharmaceutical composition for use in the treatment and/or prevention of cardiovascular disease wherein the pharmaceutical composition comprises an arginine methylation inhibitor and a pharmaceutically acceptable carrier.
  • the pharmaceutical composition is for use in the treatment and/or prevention of a thromboembolic condition.
  • the pharmaceutical composition is for use in the treatment and/or prevention of atherothrombotic disease.
  • the pharmaceutical composition is for use in the treatment and/or prevention of a disease selected from coronary artery disease ('CAD'), ischemic heart disease ('IHD' also known as coronary heart disease ('CHD')), acute coronary syndrome ('ACS') (for example myocardial infarction and/or angina), left ventricular thrombus ('LVT), cerebral ischemia, critical limb ischemia, peripheral artery disease ('PAD'), deep vein thrombosis ('DVT) and pulmonary embolism.
  • a disease selected from coronary artery disease ('CAD'), ischemic heart disease ('IHD' also known as coronary heart disease ('CHD')), acute coronary syndrome ('ACS') (for example myocardial infarction and/or angina), left ventricular thrombus ('LVT), cerebral ischemia, critical limb ischemia, peripheral artery disease ('PAD'
  • the pharmaceutical composition is for use as an antiplatelet agent and/or antithrombotic agent.
  • the pharmaceutical composition is for use in the treatment and/or prevention of thrombosis (for example the prevention of thrombosis) in a subject undergoing a surgical procedure, for example an angioplasty procedure.
  • kits including such a pharmaceutical composition.
  • a method of treating and/or preventing cardiovascular disease comprising administering a pharmaceutically effective amount of an arginine methylation inhibitor as described herein to a subject in need thereof.
  • the method is for treating and/or preventing a thromboembolic condition.
  • the method is for treating and/or preventing an atherothrombotic disease.
  • the method is for treating and/or preventing a disease selected from coronary artery disease ('CAD'), ischemic heart disease ('IHD' also known as coronary heart disease ('CHD')), acute coronary syndrome ('ACS') (for example myocardial infarction and/or angina), left ventricular thrombus ('LVT'), cerebral ischemia, critical limb ischemia, peripheral artery disease ('PAD'), deep vein thrombosis ('DVT') and pulmonary embolism.
  • a disease selected from coronary artery disease ('CAD'), ischemic heart disease ('IHD' also known as coronary heart disease ('CHD')), acute coronary syndrome ('ACS') (for example myocardial infarction and/or angina), left ventricular thrombus ('LVT'), cerebral ischemia, critical limb ischemia, peripheral artery disease ('PAD'), deep vein thrombosis ('DVT') and pulmonary embolism.
  • administration of the arginine methylation inhibitor as described herein has a reduced risk of bleeding.
  • administration of the arginine methylation inhibitor is for use as an antiplatelet agent and/or antithrombotic agent.
  • administration of the arginine methylation inhibitor as described herein maintains protective haemostatic platelet function whilst reducing the risk of bleeding.
  • the method is for treating and/or preventing thrombosis (for example preventing of thrombosis) in a subject undergoing a surgical procedure, for example an angioplasty procedure.
  • an arginine methylation inhibitor in the treatment and/or prevention of cardiovascular disease.
  • the use is for treating and/or preventing a thromboembolic condition.
  • the use is for treating and/or preventing an atherothrombotic disease.
  • the use is for treating and/or preventing a disease selected from coronary artery disease ('CAD'), ischemic heart disease ('IHD' also known as coronary heart disease ('CHD')), acute coronary syndrome ('ACS') (for example myocardial infarction and/or angina), left ventricular thrombus ('LVT'), cerebral ischemia, critical limb ischemia, peripheral artery disease ('PAD'), deep vein thrombosis ('DVT') and pulmonary embolism.
  • a disease selected from coronary artery disease ('CAD'), ischemic heart disease ('IHD' also known as coronary heart disease ('CHD')), acute coronary syndrome ('ACS') (for example myocardial infarction and/or angina), left ventricular thrombus ('LVT'), cerebral ischemia, critical limb ischemia, peripheral artery disease ('PAD'), deep vein thrombosis ('DVT') and pulmonary embolism.
  • arginine methylation inhibitor is as an antiplatelet agent and/or antithrombotic agent.
  • use of the arginine methylation inhibitor as described herein maintains protective haemostatic platelet function whilst reducing the risk of bleeding.
  • arginine methylation inhibitor is for use in the treatment and/or prevention of thrombosis (for example the prevention of thrombosis) in a subject undergoing a surgical procedure, for example an angioplasty procedure.
  • thrombosis for example the prevention of thrombosis
  • a surgical procedure for example an angioplasty procedure.
  • an arginine methylation inhibitor as a research tool for the discovery, development and/or commercialisation of treatments and/or preventative methods for cardiovascular disease such as thromboembolic conditions (for example atherothrombotic disease).
  • an arginine methylation inhibitor which inhibits arginine methylation of VASP for use in the treatment and/or prevention of cardiovascular disease as described herein such as thromboembolic conditions (for example atherothrombotic disease).
  • the arginine methylation inhibitor is a nucleic acid molecule e.g. an aptamer or an antibody or fragment thereof.
  • Washed platelets were incubated with PGI2 (100 nM) or thrombin (0.1 U/mL) for 1 min, then lysed.
  • PGI2 100 nM
  • thrombin 0.1 U/mL
  • the observed decrease in the signal for ArgMe (#8015 Cell Signalling Technologies) after thrombin activation suggests that proteins modified by ArgMe participate in the response of platelets to thrombin.
  • the blot was re-probed for actin as loading control.
  • VASP was immunoprecipitated from washed, untreated platelet lysates.
  • VASP EVH1 domains highlighting hydrogen bonds between R10 and D98 (human numbering), note the 2.4-3.2 A distance between donors and acceptors. These interactions by hydrogen bonding between R and D (or E) at equivalent positions are conserved in other proteins containing EVH1 domains including Mena (2IYB), Spred2 (2JP2) and Falafel (4WSF). R10 methylation blocks a possible hydrogen bond and could thus impair the close packing of the EVH1 C-terminal a-helix against the ⁇ -barrel. Genetic variants at VASP R10 have been reported (rs 190763638). Mutation of the acidic partner of R10 leads to reduced expression of EVH1 -containing proteins.
  • E. provides data demonstrating that ArgMe regulates cAMP signalling in platelets.
  • Washed human platelets were incubated with various concentrations of AMI-1 for4h and lysed. Lysates were probed with a phosphor-specific anti-VASP antibody that recognizes Ser 157 , a target for PKA.
  • As a positive control platelets were stimulated with 100 ⁇ prostaglandin (PGI2) for 5 min in the absence of AMI-1. The blot was re-probed for actin as a loading control.
  • PKI2 prostaglandin
  • Data from three independent experiments were quantitated and presented as average ⁇ SD fold increase relative to basal. All differences are statistically significant (ANOVA) relative to basal.
  • Figure 2E shows a Western blot showing 5-fold increase in VASP S157 phosphorylation, a hallmark for platelet quiescence, upon treatment with AMI-1 Figure 3.
  • Washed platelets were incubated with various concentrations of AMI-1 (A), GSK591 (B) or furamidine, effective in the ⁇ range (C) for 4 h prior to platelet aggregation experiments.
  • the bar diagrams show percentage aggregation (average ⁇ SEM, 3 biological replicates) after 4 min relative to basal (no inhibitor, assigned 100%). All ArgMe inhibitors and concentrations caused a significant reduction of aggregation response (ANOVA).
  • Figure 4 illustrates the amino acid sequence of human VASP protein.
  • the invention provides that in some embodiments, the arginine methylation inhibitor is a PRMT inhibitor.
  • the arginine methylation inhibitor is a PRMT1 inhibitor.
  • the arginine methylation inhibitor is a PRMT2 inhibitor.
  • the arginine methylation inhibitor is a PRMT3 inhibitor.
  • the arginine methylation inhibitor is a PRMT4 inhibitor (also known as a CARM1).
  • the arginine methylation inhibitor is a PRMT5 inhibitor.
  • the arginine methylation inhibitor is a PRMT6 inhibitor.
  • the arginine methylation inhibitor is a PRMT7 inhibitor.
  • the arginine methylation inhibitor is a PRMT 8 inhibitor.
  • the arginine methylation inhibitor is a PRMT9 inhibitor.
  • the arginine methylation inhibitor is an inhibitor of arginine methylation of VASP.
  • VASP vasodilator stimulated phosphoprotein
  • Ena- VASP family members contain an EHV1 N-terminal domain that binds proteins containing E/DFPPPPXD/E motifs and targets Ena-VASP proteins to focal adhesions.
  • the NCBI accession number of VASP (homo sapiens) is NP_003361 (version 1).
  • the amino acid sequence of human VASP is shown in Figure 4 (SEQ. ID. No. 1).
  • the arginine methylation inhibitor inhibits methylation of an arginine residue at a position selected from 10, 35, 52, 236 and 308 of the VASP protein. In certain embodiments, the arginine inhibitor inhibits methylation of an arginine residue at position 10 of the VASP protein. In certain embodiments, the arginine inhibitor inhibits methylation of an arginine residue at position 35 of the VASP protein. In certain embodiments, the arginine inhibitor inhibits methylation of an arginine residue at position 52 of the VASP protein. In certain embodiments, the arginine methylation inhibitor inhibits methylation of an arginine residue at position 236 of the VASP protein. In certain embodiments, the arginine methylation inhibitor inhibits methylation of an arginine residue at position 308 of the VASP protein.
  • the arginine methylation inhibitor is a compound selected from:
  • the arginine methylation inhibitor is a compound referred to herein as AMI-1, or a pharmaceutically acceptable salt or solvate thereof.
  • the arginine methylation inhibitor is a compound referred to herein as AIM-5, or a pharmaceutically acceptable salt or solvate thereof.
  • the arginine methylation inhibitor is a compound referred to herein as AIM-6, or a pharmaceutically acceptable salt or solvate thereof.
  • the arginine methylation inhibitor is a PRMT1 inhibitor selected from a compound referred to herein as AM 1-1 , AMI-5 and AMI-6, or a pharmaceutically acceptable salt or solvate thereof.
  • the arginine methylation inhibitor is a compound selected from stilbamide and allantodapsone or pharmaceutically acceptable salt or solvate thereof.
  • the arginine methylation inhibitor is a compound selected from:
  • the arginine methylation inhibitor is a compound selected from:
  • the arginine methylation inhibitor is a compound selected from:
  • the arginine methylation inhibitor is a compound selected from:
  • the arginine methylation inhibitor is a PRMT5 inhibitor selected from a compound referred to herein as GSK591 , or a pharmaceutically acceptable salt or solvate thereof.
  • the arginine methylation inhibitor is a compound selected from:
  • the arginine methylation inhibitor is a PRMT4 inhibitor selected from a compound referred to herein as 111, 112, EPZ025654, EZM2303 and 113, or a pharmaceutically acceptable salt or solvate thereof.
  • the arginine methylation inhibitor is a compound selected from:
  • the arginine methylation inhibitor is a PRMT5 inhibitor selected from a compound referred to herein as C1, C2, C3, C4, C5, C6, C7, C9 and C11, or a pharmaceutically acceptable salt or solvate thereof.
  • the arginine methylation inhibitor is a PRMT5 inhibitor selected from a compound referred to herein as C2, C5, C6, C9 and C11 , or a pharmaceutically acceptable salt or solvate thereof.
  • the arginine methylation inhibitor is a compound selected from:
  • the arginine methylation inhibitor is a PRMT5 inhibitor selected from a compound referred to herein as EPZ015666, CMP5, 3, 4, and 5(A9145C), or a pharmaceutically acceptable salt or solvate thereof.
  • the arginine methylation inhibitor is a compound selected from:
  • the arginine methylation inhibitor is a PRMT5 inhibitor selected from a compound referred to herein as 6 (DC_P04), 7 (DC_P05), 8 (DC_P63), 9 (DC_P88), DC_P65) and 11 (DC_P08), or a pharmaceutically acceptable salt or solvate thereof.
  • the arginine methylation inhibitor is a compound selected from:
  • the arginine methylation inhibitor is a PRMT5 inhibitor selected from a compound referred to herein as any compound selected from compounds 12 to 42, or a pharmaceutically acceptable salt or solvate thereof.
  • the arginine methylation inhibitor is methyl 2-(2-((5-methoxy-1H-benzo[d]imidazol-2- yl)thio)-acetamido)benzoate (17); or a pharmaceutically acceptable salt or solvate thereof.
  • the arginine methylation inhibitor is a compound selected from formula (I):
  • R is selected from 3-COOCH 3 , 4-COOCH 3 , 3-CF 3 , 4-CF 3 , 3-OCF 3 , 4-OCF 4 , 3-OCH 3 , 4-OCH 3 , 3-CN, and 4-CN;
  • R is selected from 3-COOCH 3 , 3-CF 3 , 4-CF 3 , 3-OCF 3 and 4-OCF 4 .
  • the arginine methylation inhibitor is a PRMT1 inhibitor selected from a compound wherein R is selected from 3-COOCH 3 , 3-CF 3 , 4-CF 3 , 3-OCF 3 and 4-OCF 4 , or a pharmaceutically acceptable salt or solvate thereof.
  • the arginine methylation inhibitor is a compound selected from formula (II):
  • R is selected from 3-COOCH 3 , 4-COOCH 3 , 3-CF 3 , 4-CF 3 , 3-OCF 3 , 4-OCF 4 , 3-OCH 3 , 4-OCH 3 , 3-CN, and 4-CN;
  • R is 3-OCF 3 .
  • the arginine methylation inhibitor is a PRMT1 inhibitor selected from a compound wherein R is 3-OCF3 or a pharmaceutically acceptable salt or solvate thereof.
  • the arginine methylation inhibitor is a compound selected from:
  • the arginine methylation inhibitor is a PRMT1 inhibitor selected from a compound referred to herein as DCLC069, DCLX078, DB75 or MS023; or a pharmaceutically acceptable salt or solvate thereof.
  • the arginine methylation inhibitor is a PRMT1 inhibitor selected from a compound referred to herein as DB75; or a pharmaceutically acceptable salt or solvate thereof.
  • DB75 is also known as furamidine.
  • the invention provides that in some embodiments, the arginine methylation inhibitor is a compound selected from:
  • the arginine methylation inhibitor is a PRMT1 inhibitor selected from a compound referred to herein as Ligand 1, Ligand 2 and Ligand 3 (hereinabove); or a pharmaceutically acceptable salt or solvate thereof.
  • the arginine methylation inhibitor is a compound selected from:
  • the arginine methylation inhibitor is a PRMT1 inhibitor selected from a compound referred to herein as methamidine, ethamidine, propamidine, butamidine, pentamidine or hexamidine; or a pharmaceutically acceptable salt or solvate thereof.
  • the arginine methylation inhibitor is a compound selected from:
  • the linker may be selected from -0(CH2) m O- wherein m is an integer selected 0, 1, 2, 3, 4, 5, 6, 7, 8, 9 and 10.
  • the arginine methylation inhibitor is a PRMT1 inhibitor selected from a compound referred to herein as compounds 2a to 2j (hereinabove); or a pharmaceutically acceptable salt or solvate thereof.
  • the arginine methylation inhibitor is a compound selected from:
  • DC_C04 2-(((2-((4-fluorobenzyl)oxy)naphthalen-1-yl)methyl)amino)-1-phenylethan-1-ol
  • DC_C05 2-(((2-((4-chlorobenzyl)oxy)naphthalen-1-yl)methyl)amino)-1-phenylethan-1-ol
  • DC_C06 4-(((1-(((2-hydroxy-2-phenylethyl)amino)methyl)naphthalen-2-yl)oxy)methyl) benzonitrile (DC_C07);
  • DC_C18 N-((2-(benzyloxy)naphthalen-1-yl)methyl)-N-methyl-2-phenylethan-1-amine
  • DC_C 19 2-((2-(benzyloxy) benzyl)am ino)- 1 -phenylethan- 1 -ol
  • the arginine methylation inhibitor is a PRMT5 inhibitor selected from a compound referred to herein a DC_C01 to DC_C19 (hereinabove); or a pharmaceutically acceptable salt or solvate thereof.
  • the arginine methylation inhibitor is a compound of formula (III):
  • the arginine methylation inhibitor is a PRMT5 inhibitor selected from a compound referred to herein as DC_P33 and DC_S01 to DC_S14 (hereinabove); or a pharmaceutically acceptable salt or solvate thereof.
  • the arginine methylation inhibitor is a nucleic acid molecule.
  • the nucleic acid molecule may be single-stranded or double-stranded.
  • the nucleic acid is an inhibitor of arginine methylation of VASP.
  • nucleic acid molecule refers to deoxyribonucleotide molecules, ribonucleotide molecules, or modified nucleotides, and polymers thereof.
  • the nucleic acid molecule may be in a single- or double-stranded form.
  • the term encompasses a nucleic acid molecule which contains known nucleotide analogs or modified backbone residues or linkages, which are synthetic, naturally occurring, and non-naturally occurring, which have similar binding properties as the reference nucleic acid molecule, and which are metabolized in a similar manner.
  • Examples of such analogs include, without limitation, phosphorothioates, phosphoramidates, methyl phosphonates, chiral-methyl phosphonates, 2-O-methyl ribonucleotides and peptide-nucleic acids (PNAs).
  • nucleic acid molecule comprises a plurality of nucleotides.
  • nucleotide refers to a ribonucleotide or a deoxyribonucleotide, or a modified form thereof.
  • Nucleotides include species that include purines (e.g., adenine, hypoxanthine, guanine, and the like) as well as pyrimidines (e.g., cytosine, uracil, thymine, and the like).
  • purines e.g., adenine, hypoxanthine, guanine, and the like
  • pyrimidines e.g., cytosine, uracil, thymine, and the like.
  • the nucleic acid molecule may be synthetic or naturally-occurring.
  • naturally occurring may refer to ssomething found in an organism without any intervention by a person; it could refer to a naturally-occurring wildtype or mutant molecule.
  • a synthetic nucleic acid molecule may be an analogue of a naturally-occurring nucleic acid molecule or may be different.
  • the nucleic acid molecule is selected from a miRNA, an RNA aptamer and a DNA aptamer.
  • the nucleic acid molecule may be a miRNA.
  • miRNA is used according to its ordinary and plain meaning and refers to a microRNA molecule found in eukaryotes that is involved in RNA-based gene regulation. See, e.g., Carrington et al., 2003, which is hereby incorporated by reference. As used herein, the term is used to refer to the single-stranded RNA molecule processed from a precursor.
  • the nucleic acid molecule is an aptamer. Aptly, the aptamer is an RNA aptamer or a DNA aptamer.
  • aptamer refers to a non-naturally occurring nucleic acid that has a desirable action on a target molecule. Desirable actions include, but are not limited to, binding of the target, inhibiting the activity of the target, enhancing the activity of the target, altering the binding properties of the target (such as, for example, increasing or decreasing affinity of the target for a ligand, receptor, cofactor, etc.), inhibiting processing of the target (such as inhibiting protease cleavage of a protein target), enhancing processing of the target (such as increasing the rate or extent of protease cleavage of a protein target), and inhibiting or facilitating the reaction between the target and another molecule.
  • An aptamer may also be referred to as a "nucleic acid ligand.”
  • an aptamer specifically binds a target molecule, wherein the target molecule is a three-dimensional chemical structure other than a polynucleotide that binds to the aptamer through a mechanism which is independent of Watson/Crick base pairing or triple helix formation, and wherein the aptamer is not a nucleic acid having the known physiological function of being bound by the target molecule.
  • aptamers to a given target include nucleic acids that are identified from a candidate mixture of nucleic acids, by a method comprising: (a) contacting the candidate mixture with the target, wherein nucleic acids having an increased affinity to the target relative to other nucleic acids in the candidate mixture can be partitioned from the remainder of the candidate mixture; (b) partitioning the increased affinity nucleic acids from the remainder of the candidate mixture; and (c) amplifying the increased affinity nucleic acids to yield a ligand- enriched mixture of nucleic acids, whereby aptamers to the target molecule are identified.
  • An aptamer can include any suitable number of nucleotides.
  • Aptamers may comprise DNA, RNA, both DNA and RNA, and modified versions of either or both, and may be single stranded, double stranded, or contain double stranded or triple stranded regions, or any other three- dimensional structures.
  • aptamers may be obtained by a technique called the systematic evolution of ligands by exponential enrichment (SELEX) process (Tuerk et al., Science 249:505-10 (1990), U.S. Patent Number 5,270,163, and U.S. Patent Number 5,637,459, each of which is incorporated herein by reference in their entirety).
  • the arginine methylation inhibitor is a double-stranded nucleic acid molecule.
  • the double-stranded nucleic acid molecule is selected from siRNA, pDNA, a gene, e.g. a synthetic gene (linear, 5' and 3' end-hairpin ligated expression cassette), mRNA e.g. synthetic messenger RNA (mRNA).
  • siRNA short interfering RNA
  • siRNA is a term used in the art and refers to a short double stranded RNA complex, typically 19-28 base pairs in length and which operates in the RNAi pathway where it interferes with the expression of specific genes with complementary nucleotide sequences by degrading mRNA after transcription.
  • siRNA is a is double- stranded nucleic acid molecule comprising two nucleotide strands, each strand having about 19 to about 28 nucleotides (i.e. about 19, 20, 21, 22, 23, 24, 25, 26, 27, or 28 nucleotides).
  • the complex often includes a 3'-overhang.
  • SiRNA can be made using techniques known to one skilled in the art and a wide variety of siRNA is commercially available.
  • the nucleic acid molecule is a PRMT inhibitor.
  • the nucleic acid molecule is a PRMT1 inhibitor.
  • the nucleic acid molecule is a PRMT2 inhibitor.
  • the nucleic acid molecule is a PRMT3 inhibitor.
  • the nucleic acid molecule is a PRMT4 inhibitor (also known as a CARM1).
  • the nucleic acid molecule is a PRMT5 inhibitor.
  • the nucleic acid molecule is a PRMT6 inhibitor.
  • the nucleic acid molecule is a PRMT7 inhibitor.
  • the nucleic acid molecule is a PRMT 8 inhibitor.
  • the nucleic acid molecule is a PRMT9 inhibitor.
  • the arginine methylation inhibitor may be an antibody or fragment thereof.
  • antibody herein is meant a protein consisting of one or more polypeptides substantially encoded by all or part of the recognized immunoglobulin genes.
  • the recognized immunoglobulin genes include the kappa ( ⁇ ), lambda ( ⁇ ), and heavy chain genetic loci, which together comprise the myriad variable region genes, and the constant region genes mu (u), delta ( ⁇ ), gamma ( ⁇ ), sigma (a), and alpha (a) which encode the IgM, IgD, IgG (lgG1 , lgG2, lgG3, and lgG4), IgE, and IgA (lgA1 and lgA2) isotypes respectively.
  • Antibody herein is meant to include full length antibodies and antibody fragments, and may refer to a natural antibody from any organism, an engineered antibody, or an antibody generated recombinantly for experimental, therapeutic, or other purposes.
  • the term “antibody” herein is used in the broadest sense and encompasses various antibody structures, including but not limited to monoclonal antibodies, polyclonal antibodies, multispecific antibodies (e.g., bispecific antibodies), and antibody fragments so long as they exhibit the desired antigen-binding activity.
  • An “antibody fragment” refers to a molecule other than an intact antibody that comprises a portion of an intact antibody that binds the antigen to which the intact antibody binds.
  • antibody fragments include but are not limited to Fv, Fab, Fab', Fab'-SH, F(ab')2; diabodies; linear antibodies; single-chain antibody molecules (e.g. scFv); and multispecific antibodies formed from antibody fragments.
  • the arginine methylation inhibitor may be an antibody or fragment thereof which inhibits the methylation of one or more than one arginine residue of a VASP protein.
  • Arginine methylation inhibitors are compounds and other molecules e.g. nucleic acid molecules and antibodies that inhibit the post translational methylation of arginine.
  • an arginine methylation inhibitor may be a compound or other molecule e.g. nucleic acid molecules and antibodies that partially or completely inhibits the activity of one or more protein arginine N-methyltransferase (PRMT).
  • PRMT protein arginine N-methyltransferase
  • nucleic acid molecules and antibodies is an arginine methylation inhibitor are known and, for example, can be found in Cheng et a/., J. Biol. Chem. 2004, 279, 23892-23899, the content of which are incorporated herein by reference.
  • the arginine methylation inhibitors and/or pharmaceutical compositions of the invention may be used for the treatment and/or prevention of cardiovascular disease as described herein as a sole therapy or may be used in combination with other conventional surgery or therapies.
  • the arginine methylation inhibitors and/or pharmaceutical compositions of the invention may be used in combination with other antiplatelet agents and/or anticoagulant agents and/or thrombolytic agents.
  • the arginine methylation inhibitors and/or pharmaceutical compositions of the invention may be used in combination with aspirin.
  • the arginine methylation inhibitors and/or pharmaceutical compositions of the invention may be used in combination with clopidogrel.
  • the arginine methylation inhibitors and/or pharmaceutical compositions of the invention may be used in combination with vorapaxar.
  • the arginine methylation inhibitors and/or pharmaceutical compositions of the invention may be used in combination with warfarin.
  • the arginine methylation inhibitors and/or pharmaceutical compositions of the invention may be used in combination with streptokinase.
  • Other antiplatelet and/or anticoagulant and/or thrombolytic agents for use in combination with the arginine methylation inhibitors and/or pharmaceutical compositions of the invention can be found in Antiplatelet Therapy - A Summary for the General Physicians (Thachil J. Clin Med, 2016, 152-160), the contents of which are incorporated herein by reference.
  • the arginine methylation inhibitors of the present disclosure may be formulated as pharmaceutical compositions prepared for storage or administration for use in the treatment and/or prevention of cardiovascular disease (for example thromboembolic conditions) as described herein.
  • Such a composition typically comprises a therapeutically effective amount of an arginine methylation inhibitor, in the appropriate form, and a pharmaceutically acceptable carrier.
  • an arginine methylation inhibitor as described herein will depend on the route of administration, the type of mammal being treated, and the physical characteristics of the specific mammal under consideration. These factors and their relationship to determining this amount are well known to skilled practitioners in the medical arts. This amount and the method of administration can be tailored to achieve optimal efficacy, and may depend on such factors as weight, diet, concurrent medication and other factors, well known to those skilled in the medical arts.
  • the arginine methylation inhibitors of the present disclosure may be particularly useful for treatment of humans.
  • An effective dosage and treatment protocol may be determined by conventional means, starting with a low dose in laboratory animals and then increasing the dosage while monitoring the effects, and systematically varying the dosage regimen as well. Numerous factors may be taken into consideration by a clinician when determining an optimal dosage for a given subject. Such considerations are known to the person skilled in the art.
  • pharmaceutically acceptable carrier includes any of the standard pharmaceutical carriers. Pharmaceutically acceptable carriers for therapeutic use are well known in the pharmaceutical art, and are described, for example, in Remington's Pharmaceutical Sciences, Mack Publishing Co. (A.R.Gennaro edit. 1985). For example, sterile saline and phosphate- buffered saline at slightly acidic or physiological pH may be used.
  • pH buffering agents may be phosphate, citrate, acetate, tris/hydroxymethyl)aminomethane (TRIS), N- Tris(hydroxymethyl)methyl-3-aminopropanesulphonic acid (TAPS), ammonium bicarbonate, diethanolamine, histidine, arginine, lysine, or acetate or mixtures thereof.
  • TIS tris/hydroxymethyl)aminomethane
  • TAPS N- Tris(hydroxymethyl)methyl-3-aminopropanesulphonic acid
  • ammonium bicarbonate diethanolamine
  • histidine histidine
  • arginine arginine
  • lysine or acetate or mixtures thereof.
  • salts include pharmaceutically acceptable salts such as acid addition salts and basic salts.
  • acid addition salts include hydrochloride salts, citrate salts and acetate salts.
  • basis salts include salts where the cation is selected from alkali metals, such as sodium and potassium, alkaline earth metals, such as calcium, and ammonium ions + N(R 3 )3(R 4 ), where R 3 and R 4 independently designates optionally substituted C -e-alkyl, optionally substituted C2-e-alkenyl, optionally substituted aryl, or optionally substituted heteroaryl.
  • solvate in the context of the present disclosure refers to a complex of defined stoichiometry formed between a solute (e.g., an arginine methylation inhibitor or pharmaceutically acceptable salt thereof according to the present disclosure) and a solvent.
  • the solvent in this connection may, for example, be water, ethanol or another pharmaceutically acceptable, typically small-molecular organic species, such as, but not limited to, acetic acid or lactic acid.
  • a solvate is normally referred to as a hydrate.
  • Treatment is an approach for obtaining beneficial or desired clinical results.
  • beneficial or desired clinical results include, but are not limited to, alleviation of symptoms, diminishment of extent of disease, stabilized (i.e., not worsening) state of disease, delay or slowing of disease progression, amelioration or palliation of the disease state, and remission (whether partial or total), whether detectable or undetectable.
  • Treatment can also mean prolonging survival as compared to expected survival if not receiving treatment.
  • Treatment' is an intervention performed with the intention of preventing the development or altering the pathology of a disorder. Accordingly, “treatment” refers to both therapeutic treatment and prophylactic or preventative measures in certain embodiments.
  • Those in need of treatment include those already with the disorder as well as those in which the disorder is to be prevented.
  • treatment is meant inhibiting or reducing an increase in pathology or symptoms when compared to the absence of treatment, and is not necessarily meant to imply complete cessation of the relevant condition.
  • compositions for use in the treatment of a cardiovascular disease can be in unit dosage form.
  • the composition is divided into unit doses containing appropriate quantities of the active component
  • the unit dosage form can be a packaged preparation, the package containing discrete quantities of the preparations, for example, packeted tablets, capsules, and powders in vials or ampoules.
  • the unit dosage form can also be a capsule, cachet, or tablet itself, or it can be the appropriate number of any of these packaged forms. It may be provided in single dose injectable form, for example in the form of a pen.
  • packaged forms include a label or insert with instructions for use.
  • Compositions may be formulated for any suitable route and means of administration.
  • Pharmaceutically acceptable carriers or diluents include those used in formulations suitable for oral, rectal, nasal, topical (including buccal and sublingual), vaginal or parenteral (including subcutaneous, intramuscular, intravenous, intradermal, and transdermal) administration.
  • the formulations may conveniently be presented in unit dosage form and may be prepared by any of the methods well known in the art of pharmacy.
  • an "effective” amount or a “therapeutically effective amount” of an inhibitor or agent refers to a nontoxic but sufficient amount of the inhibitor or agent to provide the desired effect.
  • the amount that is “effective” will vary from subject to subject, depending on the age and general condition of the individual, mode of administration, and the like. An appropriate “effective” amount in any individual case may be determined by one of ordinary skill in the art using routine experimentation.
  • patient may be used interchangeably and refer to either a humans or non-human mammal. Aptly, the subject is a human.
  • the arginine methylation inhibitors described herein may be used to treat and/or prevent a cardiovascular disease (for example a thromboembolic condition) e.g. an atherothrombotic disease.
  • a cardiovascular disease for example a thromboembolic condition
  • the arginine methylation inhibitor is for use in the treatment and/or prevention of a disease selected from coronary artery disease ('CAD'), ischemic heart disease ('IHD' also known as coronary heart disease ('CHD')), acute coronary syndrome ('ACS') (for example myocardial infarction and/or angina), left ventricular thrombus ('LVT'), cerebral ischemia, critical limb ischemia, peripheral artery disease ('PAD'), deep vein thrombosis ('DVT') and pulmonary embolism.
  • CAD' coronary artery disease
  • 'IHD' also known as coronary heart disease ('CHD')
  • 'ACS' acute coronary syndrome
  • the arginine methylation inhibitor is for use in the treatment and/or prevention of coronary artery disease ('CAD').
  • the arginine methylation inhibitor is for use in the treatment and/or prevention of ischemic heart disease ('IHD').
  • the arginine methylation inhibitor is for use in the treatment and/or prevention of coronary heart disease ('CHD'). In one embodiment, the arginine methylation inhibitor is for use in the treatment and/or prevention of acute coronary syndrome ('ACS') (for example myocardial infarction and/or angina).
  • 'ACS' acute coronary syndrome
  • the arginine methylation inhibitor is for use in the treatment and/or prevention of left ventricular thrombus ('LVT').
  • the arginine methylation inhibitor is for use in the treatment and/or prevention of cerebral ischemia. In one embodiment, the arginine methylation inhibitor is for use in the treatment and/or prevention of critical limb ischemia.
  • the arginine methylation inhibitor is for use in the treatment and/or prevention of peripheral artery disease ('PAD').
  • the arginine methylation inhibitor is for use in the treatment and/or prevention of deep vein thrombosis ('DVT').
  • the arginine methylation inhibitor is for use in the treatment and/or prevention of pulmonary embolism. In one embodiment, the arginine methylation inhibitor is for use in the treatment and/or prevention of thrombosis (for example the prevention of thrombosis) in a subject undergoing a surgical procedure, for example an angioplasty procedure.
  • thromboembolic condition occurs when a blood clot (a thrombus) occurs in a blood vessel. The thrombus can break loose and be carried to by the blood stream to partially or fully block another vessel. The blockage is referred to as an embolism.
  • a thromboembolism which occurs in an artery is known as an arterial thromboembolism and results from atherothrombotic disease.
  • An arterial thromboembolism can in principle affect any organ in the body resulting in tissue damage and/or death.
  • Coronary artery disease results when one or more artery that supplies blood to heart muscle becomes hardened and narrowed, usually as a result of atherosclerosis. Such a blockage may result in an ischemia.
  • Ischemia is a restriction in blood supply to tissues, causing a shortage of oxygen and glucose needed for cellular metabolism (to keep tissue alive). Ischemia is generally caused by problems with blood vessels, with resultant damage to or dysfunction of tissue. It also means local anemia in a given part of a body sometimes resulting from congestion (such as vasoconstriction, thrombosis or embolism). Ischemia comprises not only insufficiency of oxygen, but also reduced availability of nutrients and inadequate removal of metabolites.
  • Ischemic heart disease is a disease characterized by reduced blood supply to the heart. Ischemic heart disease is also known as coronary heart disease ('CHD').
  • Acute coronary syndrome is a syndrome (set of signs and symptoms) resulting from decreased blood flow in the coronary arteries such that part of the heart muscle is unable to function properly or dies.
  • Examples of ACS include myocardial infarction and angina.
  • ACS is often associated with rupture of an atherosclerotic plaque and partial or complete thrombosis of the infarct-related artery.
  • the signs and symptoms of ACS which usually begin abruptly, may include one or more of:
  • LVT Left ventricular thrombus
  • Cerebral ischemia is an ischemia occurring in the brain and can be acute or chronic.
  • Acute ischemic stroke is a neurologic emergency that may be reversible if treated rapidly.
  • Chronic ischemia of the brain may result in a form of dementia called vascular dementia.
  • a brief episode of ischemia affecting the brain is called a transient ischemic attack (TIA).
  • Peripheral vascular disease refers to a narrowing of blood vessels other than those that supply the heart and brain.
  • the disease is known as peripheral artery disease ('PAD') although the terms are often used interchangeably.
  • PAD most commonly affects the legs but other arteries may also be involved.
  • PAD can be caused by arteriosclerosis (hardening and/or stiffening of the artery), which is most commonly due to the formation of atheromatous plaques.
  • arteriosclerosis hardening and/or stiffening of the artery
  • the classic symptom is leg pain when walking which resolves with rest, known as intermittent claudication. Other symptoms including skin ulcers, bluish skin, cold skin, or poor nail and hair growth may occur in the affected leg.
  • Complications may include an infection or tissue death which may require amputation; coronary artery disease, or stroke. Up to 50% of cases of PAD are without symptoms. Complications of PAD include critical limb ischaemia (CLI) where blood flow to the limbs is severely restricted.
  • CLI critical limb ischaemia
  • Critical limb ischemia is an ischemia occurring in the arteries of the lower extremities. It is a serious form of PAD. It is a chronic condition that results in severe pain in the feet or toes, even while resting. Complications of poor circulation can include sores and wounds that won't heal in the legs and feet. Left untreated, the complications of CLI will result in amputation of the affected limb.
  • the most prominent features of critical limb ischemia (CLI) are called ischemic rest pain— severe pain in the legs and feet while a person is not moving, or nonhealing sores on the feet or legs. Symptoms may include one or more of: • Pain or numbness in the feet
  • a pulmonary embolism refers to a blockage in an artery to the lungs. Signs and symptoms of a pulmonary embolism include chest pain, shortness of breath and coughing. In particular, symptoms can include:
  • coughing this is usually dry, but may include coughing up blood or mucus that contains blood;
  • DVT deep vein thrombosis
  • DVT Deep vein thrombosis
  • a blood clot within a deep vein in the body, usually in the leg.
  • DVT usually occurs in a deep leg vein, a larger vein that runs through the muscles of the calf and the thigh. It can cause pain and swelling in the leg and may lead to complications such as pulmonary embolism. This is a serious condition that occurs when a piece of blood clot breaks off into the bloodstream and blocks one of the blood vessels in the lungs (see below).
  • Angioplasty non-invasive or percutaneous is a surgical procedure by which a catheter with a balloon is inserted into a vessel and directed to a position where there is a stenosis (narrowing).
  • the balloon is inflated in order to widen a narrowed or obstructed artery or vein.
  • a stent may or may not be placed to maintain the vessel dilated.
  • the procedure is mainly (but not exclusively) used to treat stenosis caused by atherosclerosis. The most common procedure is coronary angioplasty.
  • Angioplasty is also used to treat carotid artery stenosis and some cases of venous stenosis.
  • arginine methylation inhibitors can be used to treat any one or more of the diseases listed herein by virtue of their ability to disrupt platelet aggregation and/or other platelet function.
  • Platelets and Megakaryocytes Volume 1: Functional Assays. Methods in Molecular Biology Vol. 272 Edited by J. M. Gibbins and M. P. Mahaut-Smith. Humana Press. 2004;
  • ArgMe occurs in platelets: analysis of available proteomic data suggests that hundreds of platelet proteins undergo arginine methylation in platelets, PRMTs are expressed (Fig. 1A) and ArgMe can be demonstrated immunologically and can be specifically inhibited (Fig. 1 B). Furthermore it has been shown that ArgMe inhibition by AMI-1 dramatically affects the dynamics of platelet activation by thrombin and collagen, to the extent that platelet aggregation is significantly delayed.
  • VASP plays a central role in platelet signalling pathways (Massberg et al. Blood. 2004; 103(1): 136-42). VASP phosphorylation correlates with platelet inhibition and is paralleled by the inhibition of fibrinogen receptor GPIIb-llla (Aszodi et al. EMBO J. 1999;18(1):37-48). It is demonstrated here, for the first time, that VASP is modified by ArgMe in platelets. VASP was immunoprecipitated using an anti-VASP antibody (from Abeam) from washed platelet lysates (isolated from healthy human donors, and subjected to no treatment).
  • VASP was recognised by an anti- ArgMe antibody (commercially available from Cell Signalling Technologies, product reference #8015, Figure 2A). VASP is predicted to be modified by ArgMe at sites including R10 ( Figure 2B). Immunoprecipitated VASP was isolated by SDS- PAGE and the coomassie stained band was digested with Lys-C protease post reduction with DTE and alkylation with iodoacetamide. The resulting peptides were analysed by LC-MS/MS using our Orbirap Fusion mass spectrometer with elution from an easy spray 50 cm PepMap column over a 35 min gradient.
  • Thermo .raw files were converted to .mgf using MSconvert before submitting to Mascot database searching against the human subset of the SwissProt database.
  • the Mascot .DAT result file was imported into Scaffold and a second search of the same database was performed using X!Tandem.
  • Peptide and protein identifications were filtered in Scaffold to require a global false discovery rate of ⁇ 1 % at both the protein and peptide level. Protein matches also required a minimum of two unique peptide identifications.
  • VASP EVH1 domain is in fact a critical regulator of actin dynamics and platelet aggregation.
  • platelet treatment with ArgMe inhibitors increased VASP phosphorylation at S157, a hallmark for platelet quiescence ( Figure 2E).

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Abstract

L'invention concerne le traitement et/ou la prévention d'une maladie cardiovasculaire, par exemple des affections thromboemboliques, telles qu'une maladie athérothrombotique. En particulier, l'invention concerne un inhibiteur de méthylation de l'arginine ('inhibiteur de 'MéArg') destiné à être utilisé dans le traitement et/ou la prévention d'une maladie cardiovasculaire telle que des affections thromboemboliques (par exemple, une maladie athérothrombotique).
PCT/GB2018/050080 2017-01-12 2018-01-12 Inhibiteur de méthylation de l'arginine pour le traitement ou la prévention d'une maladie cardiovasculaire Ceased WO2018130840A1 (fr)

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US20210402196A1 (en) * 2018-04-26 2021-12-30 West Affum Holdings Corp. Wearable medical (wm) system monitoring ecg signal of ambulatory patient for heart condition
CN117265097A (zh) * 2023-09-20 2023-12-22 复旦大学 一种bach1分子在胰岛素抵抗中的应用
CN117503740A (zh) * 2023-11-09 2024-02-06 复旦大学 一种bach1抑制剂及其应用

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